Department of Horticulture Faculty of Crop Production Sciences the University of Agriculture Peshawar-Pakistan

SPECIES DIVERSITY AND NUTRITIONAL ANALYSIS OF MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN

BY SHAH MASAUD KHAN

A thesis submitted to The University of Agriculture Peshawar, in partial fulfillment of the requirement for the degree of

DOCTOR OF PHILOSOPHY IN AGRICULTURE (HORTICULTURE)

DEPARTMENT OF HORTICULTURE FACULTY OF CROP PRODUCTION SCIENCES THE UNIVERSITY OF AGRICULTURE PESHAWAR-PAKISTAN

JANUARY, 2015 SPECIES DIVERSITY AND NUTRITIONAL ANALYSIS OF MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN

BY SHAH MASAUD KHAN

A thesis submitted to The University of Agriculture Peshawar, in partial fulfillment of the requirement for the degree of

DOCTOR OF PHILOSOPHY IN AGRICULTURE (HORTICULTURE)

APPROVED BY:

______Chairman Prof. Dr. Noor ul Amin Supervisory Committee

______Co-Supervisor Prof. Dr. Habib Ahmad (T I). Vice Chancellor Hazara University Mansehra.

______Member (Major Field) Dr. Muhammad Sajid

______Member (Minor Field) Prof. Dr. Ahmad-Ur-Rahman Saljoqi

______Chairman & Convener Board of Studies Prof. Dr. Noor ul Amin

______Dean Prof. Dr. Muhammad Afzal Faculty of Crop Production Sciences

______Director Prof. Dr. Farhatullah Advanced Studies and Research

DEPARTMENT OF HORTICULTURE FACULTY OF CROP PRODUCTION SCIENCES

THE UNIVERSITY OF AGRICULTURE PESHAWAR-PAKISTAN JANUARY, 2015

SPECIES DIVERSITY AND NUTRITIONAL ANALYSIS OF MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN

SHAH MASAUD KHAN

THESIS APPROVED BY:

EXTERNAL EXAMINER:

Professor. Dr. Hong Bo China Agricultural University Beijing, 100193, China.

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SPECIES DIVERSITY AND NUTRITIONAL ANALYSIS OF MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN

SHAH MASAUD KHAN

THESIS APPROVED BY:

EXTERNAL EXAMINER:

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Faculty of Agriculture University Putra Malaysia.

dedication

I dedicate my this modest endeavor to my parents, especially to my father, late aziz ur rahman khan, who loved education and always encouraged me for higher studies

and to my niece, malala yousafzai, the noble laureate, who is an icon of hope for millions and an ambassador for the right of women to education.

shah masaud khan yousafzai

TABLE OF CONTENTS

CHAPTER NO. TITLE PAGE NO.

GENERAL ABSTRACT ...... I ACKNOWLEDGEMENTS ...... III LIST OF TABLES ...... IV LIST OF FIGURES ...... IV LIST OF APPENDICES ...... IX

CHAPTER NO. 1

GENERAL INTRODUCTION ...... 1

CHAPTER NO. 2

REVIEW OF LITERATURE ...... 7

CHAPTER NO. 3

EXPERIMENT-1

SPECIES DIVERSITY AND ETHNO BOTANICAL STUDY OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

ABSTRACT ...... 26 INTRODUCTION ...... 27 MATERIALS AND METHODS ...... 30 RESULTS AND DISCUSSIONS ...... 34

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SUMMARY, CONCLUSIONS AND RECOMMENDATIONS .... 40

EXPERIMENT-2

ECOLOGICAL ANALYSIS OF THE VEGETATION OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

ABSTRACT ...... 43 INTRODUCTION ...... 44 MATERIALS AND METHODS ...... 46 RESULTS AND DISCUSSIONS ...... 49 SUMMARY, CONCLUSIONS AND RECOMMENDATIONS. ....79 EXPERIMENT-3

STUDY OF BIO-CHEMICAL SUBSTANCES OF SELECTED MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

ABSTRACT ...... 82 INTRODUCTION ...... 84 MATERIALS AND METHODS ...... 87 RESULTS AND DISCUSSIONS ...... 93 SUMMARY, CONCLUSIONS AND RECOMMENDATIONS .....164

EXPERIMENT-4

THE STUDY OF HORTICULTURAL ATTRIBUTES OF SELECTED MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

ABSTRACT ...... 168 INTRODUCTION ...... 169 MATERIALS AND METHODS ...... 171 RESULTS AND DISCUSSIONS ...... 174 SUMMARY,CONCLUSIONS AND RECOMMENDATIONS ...... 197

CHAPTER NO. 4 OVERALL CONCLUSIONS AND RECOMMENDATIONS ...... 200 CHAPTER NO. 5

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LITERATURE CITED ...... 202 CHAPTER NO. 6 APPENDICES ...... 236

SPECIES DIVERSITY AND NUTRITIONAL ANALYSIS OF MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

Shah Masaud Khan and Noor ul Amin. The University of Agriculture Peshawar, Khyber Pakhtunkhwa, Pakistan.

GENERAL ABSTRACT

The study on "Species diversity and nutritional analysis of medicinal plants of Khanpur Valley, in the sub Himalayan mountains of Pakistan" was conducted during 2010-2013. The valley was divided into four ecologically diverse sites and data was collected in both summer and winter seasons. A total of 202 plant species belonging to 48 families were recorded. Maximum species (19) were from family Asteraceae (Compositae), followed by Poaceae (18), Fabaceae (10), Solanaceae (10), Euphorbeaceae (9), Brassicaceae (9) and Moraceae (9). Similarly maximum species were herbs (141), followed by trees (31), while minimum species were shrubs (30). Moreover, out of total 202 plant species, 71 species (34%) belonging to 42 families were identified as medicinally important. The study concluded that whole plant in case of herbs and leaves of woody plants were preferred by the local people for use in various recipes against a variety of diseases most preferably associated with digestive system. Adhatoda vasica was the most preferred medicinal plant species while powder form was the most preferred form of utilization of the people of Khanpur valley. A total of 116 ecologically important species (Frequency≥1%) were found. Maximum number of plant species (102) were found at Dabola site in summer season while minimum species (73) were observed at Dam site during winter season. The maximum density was recorded for plant species associated with Dabola and Jabri sites. The results also established that majority of the species fall in the class of Rare (45.37%), followed by Occasional (19.72%), then Frequent (5.06%) and least fall in the class of Abundant (2.05%).

Nutritional analysis of the most preferred medicinal plant, Adhatoda vasica showed significantly higher values for ash (19.5%) at Jabri during summer, crude proteins (14.7%) at Dabola during summer, crude fibers (12.6%) at Jabri during winter, essential oil (2.5%) at Mang during summer, Nitrogen Free Extractable Substances (NFES) (55.8%) at Mang during summer, Net Free Energy Estimates (NFEE) (194.6%) at Dam during summer, potassium (179.4mg/100g) at Dam during summer, phosphorus (171mg/100g) at Dabola during summer, copper (0.58mg/100g) at Dabola during summer and zinc (8.07mg/100g) at Mang during summer. Similarly, Fumaria officinalis revealed higher significant results for ash (27.7%) at Jabri during summer, crude proteins (15.2%) at Mang during summer,

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crude fibers (13.1%) at Jabri during winter, crude fats (9.6%) at Mang during summer, essential oil (2.2%) at Mang during winter, NFES (47.04%) at Mang during winter, NFEE (186.13%) at Mang during summer, potassium (494.1mg/100g) at Dabola during summer, phosphorus (265.3mg/100g) at Mang during summer, magnesium (184.3mg/100g) at Jabri during summer and manganese (14.03mg/100g) at Dabola during summer. Third most important medicinal plant species, Euphorbia hirta gave higher significant contents for ash (17.95%) at Jabri during summer, crude proteins (15.42%) at Jabri during summer, crude fibers (12.78%) at Mang during winter, crude fats (10.64%) at Jabri during summer, essential oil (1.94%) at Dabola during summer, NFES (59.49%) at Dam during winter, NFEE (198.69%) at Jabri during summer, calcium (199.17mg/100g) at Dam during summer, phosphorus (185.37mg/100g) at Jabri during winter. All six medicinal plants under study gave maximum yield during summer (last week of July) except Fumaria officinalis which performed best during last week of May. Similarly Ajuga bracteosa gave higher yield at Dabola site; Euphorbia hirta at Dam site while the rest of medicinal plants under study showed maximum yield at Mang site.

It was concluded from the results of the current study that 202 total plant species, 116 ecologically important species and 71 medicinal plants were identified at Khanpur valley. Among the top six most preferred medicinal plants, Ajuga bracteosa and Euphorbia hirta gave higher values for bio-chemicals during last week of October,

Fumaria officinalis performed best during last week of May while Adhatoda vasica, Recinus communis and Calatropis procera showed maximum bio-chemicals and yield during the last week of July.

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ACKNOWLEDGEMENTS

It is a moment of great honor for me to submit with all obedience and humbleness and pay gratitude to Almighty Allah, the Most Merciful and the Beneficent, Who showered upon me His special blessings and enabled me to accomplish this much awaited job and contribute to the noble cause of knowledge creation. Cordial gratitude to the Prophet Muhammad (P.B.U.H), who is forever a torch of guidance and knowledge for humanity.

I pay my profound regards to Prof. Dr. Muhammad Zubair (late); Allah may rest his soul in paradise, who guided me to initiate this project.

I wish to express my deepest gratitude and regards to my honorable supervisor Prof. Dr. Noor ul Amin, Chairman, Department of Horticulture for his professional guidance and friendly supervision, which was a great stimulus for me to achieve this tremendous success. He dealt me like his fellow scientist and always encouraged and supported me in all these endeavors.

I am really grateful to my Co-Supervisor Prof. Dr. Habib Ahmad (TI) Dean faculty of Science, Hazara University, Mansehra, for his continuous advice and supervision of my field research and providing practical support of his professional team and herbarium. I feel pleasure to thank member of my supervisory committee, Prof. Dr. Ahmad ur Rahman Saljoqi, Department of Plant Protection, for his cooperative behavior, guidance and valuable suggestions. I am also thankful to Dr. Muhammad Sajid, Department of Horticulture, Prof. Dr. Farhatullah, Director Advance Studies and Research, Prof. Dr. Muhammad Afzal, Dean Faculty of Crop Production Sciences for their cooperation. I am highly indebted to Dr. Ishrat Naz, Department of Plant Pathology, for her tremendous support in data analysis and Dr. Muhammad Arif,

Department of Agronomy, for his help in use of statistical models. I am grateful to Dr Saleemullah, Department of Agriculture Chemistry for his support and supervision in bio-chemical analysis. I am also thankful to Hakeem Naseem ijaz for providing me full support and helping in the identification of local medicinal plants and their uses. I admire

the cooperation of my fellow faculty members of the Department of Agricultural Sciences, University of Haripur.

Finally, I would feel incomplete without thanking my parents, my wife, my children(especially Zikra Salar Dalokhela), brothers and sisters who tolerated me patiently during this critical period and their prayers enabled me to complete my Ph.D work successfully.

Shah Masaud Khan Yousafzai

LIST OF TABLES

Table No. Title Page No

Table 1.1. Summary of Enlistment of Total Plant Species and Medicinally Important Plants found at Khanpur Valley...... 35 Table-2.1. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the density of plant species found at Khanpur Valley ...... 50

Table-2.2. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the frequency of plant species found at Khanpur Valley...... 55

Table-2.3. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the coverage of plant species found at Khanpur Valley...... 59 Table-2.4. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Relative Frequency of plant species at Khanpur Valley...... 63

Table-2.5. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Relative Density of plant species found at Khanpur Valley...... 67

Table-2.6. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Relative Coverage of plant species found at Khanpur Valley...... 71

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Table-2.7. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Importance Value(IV) of plant species found at Khanpur Valley...... 74 Table-3.1. Effect of Different Seasons and Sites on Proximate analysis of Adhatoda vasica, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... i ...... 97

Effect of Different Seasons and Sites on Elemental analysis of Adhatoda vasica, indigenous to Khanpur Valley, in sub-Himalayan mountains of Table-3.2. Pakistan...... 103 Table-3.3. Effect of Different Seasons and Sites on proximate analysis of Calatropis procera, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 109

Table-3.4. Effect of Different Seasons and Sites on Elemental analysis of Calatropis procera, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 115 Table-3.5. Effect of Different Seasons and Sites on Proximate analysis of Recinus communis, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 121

Table-3.6. Effect of Different Seasons and Sites on Elemental analysis of Recinus communis, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 127

Table-3.7. Effect of Different Seasons and Sites on Proximate analysis of Ajuga bracteosa, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 132

Table-3.8. Effect of Different Seasons and Sites on Elemental analysis of Ajuga bractiousa, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 138

Table-3.9. Effect of Different Seasons and Sites on Proximate Analysis of Euphorbia hirta, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 145

Table-3.10. Effect of Different Seasons and Sites on Elemental analysis of Euphorbia hirta, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 151

Table-3.11. Effect of Different Seasons and Sites on Proximate analysis of Fumaria officinalis, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 156

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Table-3.12. Effect of Different Seasons and Sites on Elemental analysis of Fumaria officinalis, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 162 Table-4.1. ii Effect of Different Seasons a nd Sites on Horticultural Attributes of Adhatoda vasica indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 177 Table-4.2. Effect of Different Seasons and Sites on Horticultural Attributes of Calatropis procera indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 181 Table-4.3. Effect of Different Seasons and Sites on Horticultural Attributes of Recinus communisa indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 184

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Table-4.4. Different Seasons and Sites on Horticultural Attributes of Ajuga bracteousa indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 187 Table-4.5. Effect of Different Seasons and Sites on Horticultural Attributes of Euphorbia hirta indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 191 Table-4.6. Effect of Different Seasons and Sites on Horticultural Attributes of Fumaria officinalis indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan...... 195

LIST OF FIGURES

Figures No. Title Page No

Figure 1.1. Percentage of respondent`s preferences for species of medicinal plants...... 36 Figure 1.2. Percentage of respondent`s preferences for category of use, of medicinal plants...... 37 Figure 1.3. Percentage of respondent`s preference for plant part used of medicinal plants...... 38 Figure1.4. Percentage of respondent`s preference for traditional use (recipe) of medicinal plants ...... 39 Figure 2.1. pCCA Ordination biplot showing the effect of different seasons on the density of plant species found at Khanpur Valley...... 52 Figure 2.2. pCCA Ordination biplot showing the effect of different sites on the density of plant species found at Khanpur Valley...... 53 Figure 2.3. pCCA Ordination biplot showing the effect of different seasons on the frequency of plant species found at Khanpur Valley...... 56 Figure 2.4. pCCA Ordination biplot showing the effect of different sites on the frequency of plant species found at Khanpur Valley...... 57 Figure 2.5. pCCA Ordination biplot showing the effect of different seasons on the coverage of plant species found at Khanpur Valley...... 60 Figure 2.6. pCCA Ordination biplot showing the effect of different sites on the coverage of plant species found at Khanpur Valley...... 61 Figure 2.7. pCCA Ordination biplot showing the effect of different seasons on the relative density of plant species found at Khanpur Valley...... 64 Figure 2.8. pCCA Ordination biplot showing the effect of different sites on the relative density of plant species found at Khanpur Valley...... 65

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Figure 2.9. pCCA Ordination biplot showing the effect of different seasons on the relative frequency of plant species found at Khanpur Valley...... 68 Figure 2.10. pCCA Ordination biplot showing the effect of different sites on the relative frequency of plant species found at Khanpur Valley...... 69 Figure 2.11. pCCA Ordination biplot showing the effect of different seasons on the relative coverage of plant species found at Khanpur Valley...... 72 Figure 2.12. pCCA Ordination biplot showing the effect of different sites on the relative coverage of plant species found at Khanpur Valley...... 73 Figure 2.13. pCCA Ordination biplot showing the effect of different seasons on the importance value of plant species found at Khanpur Valley...... 76 Figure 2.14. CCA Ordination biplot showing the effect of different sites on the importance value of plant species found at Khanpur Valley...... 77 Figure-2.15. Summary showing the effect of different seasons and sites on the classification of Plant Species found at Khanpur Valley...... 78 Figure-3.1. Effect of different seasons and sites on Ash % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 97 Figure-3.2. Effect of different seasons and sites on Crude Proteins % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 98

Figure-3.3. seasons and sites on Crude fibers % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 98 Figure-3.4. Effect of different seasons and sites on Essential oil % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 98 Figure-3.5. Effect of different seasons and sites on NFES % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 99 Figure-3.6. Effect of different seasons and sites on NFEE in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 99 Figure-3.7. Effect of different seasons and sites on Potassium (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 103 Figure-3.8. Effect of different seasons and sites on Phosphorus (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 104 Figure-3.9. Effect of different seasons and sites on Copper (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 104

Figure-3.10. Effect of different seasons and sites on Zinc (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 104 Figure-3.11. Effect of different seasons and sites on Ash % in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 109 Figure-3.12. Effect of different seasons and sites on Crude Proteins % in leaves of Calatropis procera collected from Khanpur valley in the sub- Himalayan mountains of Pakistan...... 110 Figure-3.13. Effect of different seasons and sites on Crude Fats % in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 110 Figure-3.14. Effect of different seasons and sites on Crude Fibers % in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 110 Figure-3.15. Effect of different seasons and sites on Essential Oil % in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 111 Figure-3.16. Effect of different seasons and sites on NFES % in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 115 Figure-3.17. Effect of different seasons and sites on NFEE in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 116 Figure-3.18. Effect of different seasons and sites on Potassium (mg/100g) in leaves of Calatropis procera collected from Khanpur valley in the sub-Himalayan mountains of Pakistan...... 116

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Figure-3.19. Phosphorus (mg/100g) in leaves of Calatropis procera collected from Khanpur valley in the sub- Himalayan mountains of Pakistan...... 116 Figure-3.20. Effect of different seasons and sites on Magnesium (mg/100g) in leaves of Calatropis procera collected from Khanpur valley in the sub- Himalayan mountains of Pakistan...... 117 Figure-3.21. Effect of different seasons and sites on Manganese (mg/100g) in leaves of Calatropis procera collected from Khanpur valley in the sub- Himalayan mountains of Pakistan...... 117 Figure-3.22. Effect of different seasons and sites on Ash contents in Euphorbia hirta...... 117 Figure-3.23. Effect of different seasons and sites on Crude Protein contents in Recinus communis...... 122 Figure-3.24. Effect of different seasons and sites on Crude Fats contents in Recinus communis...... 122 Figure-3.25. Effect of different seasons and sites on Crude Fibers contents in Recinus communis...... 122 Figure-3.26. Effect of different seasons and sites on Essential Oil contents in Recinus communis...... 123 Figure-3.27. Effect of different seasons and sites on NFES contents in Recinus communis ...... 123 Figure-3.28. Effect of different seasons and sites on NFEE in Recinus communis. .127 Figure-3.29. Effect of different seasons and sites on Calcium contents in Recinus communis ...... 128 Figure-3.30. Effect of different seasons and sites on Phosphorus contents in Recinus communis...... 128 Figure-3.32. Effect of different seasons and sites on Ash contents of Ajuga bracteosa...... 132 Figure-3.33. Effect of different seasons and sites on Crude Protein contents of Ajuga bracteosa...... 133 Figure-3.34. Effect of different seasons and sites on Crude Fats contents of Ajuga bracteosa...... 133 Figure-3.35. Effect of different seasons and sites on Essential Oil contents of Ajuga bracteosa...... 133 Figure-3.36. Effect of different seasons and sites on NFES contents of Ajuga bracteosa...... 134 Figure-3.37. Effect of different seasons and sites on NFEE of Ajuga bracteosa. ....134 Figure-3.38. Effect of different seasons and sites on Sodium content of Ajuga bracteosa...... 134 Figure-3.39. Effect of different seasons and sites on Potassium (K) contents of Ajuga bracteosa...... 139 Figure-3.40. Effect of different seasons and sites on Calcium (Ca) contents of Ajuga bracteosa...... 139

Figure-3.41. Effect of different seasons and sites on Magnesium (Mg) contents of Ajuga bracteosa...... 139 Figure-3.42. Effect of different seasons and sites on Copper (Cu) contents of Ajuga bracteosa...... 140 Figure-3.43. Effect of different seasons and sites on Zinc (Zn) contents of Ajuga bracteosa...... 140 Figure-3.44. Crude Protein contents in leaves of Euphorbia hirta...... 140 Figure-3.45. Effect of different seasons and sites on Crude Fats contents in leaves of Euphorbia hirta...... 145 Figure-3.46. Effect of different seasons and sites on Crude Fibers contents in leaves of Euphorbia hirta ...... 146 Figure-3.47. Effect of different seasons and sites on Essential Oil contents in leaves of Euphorbia hirta...... 146 Figure-3.48. Effect of different seasons and sites on NFEE in leaves of Euphorbia hirta 146 Figure-3.49. Effect of different seasons and sites on Sodium content in leaves of Euphorbia hirta...... 147 Figure-3.50. Effect of different seasons and sites on Iron (Fe) contents in leaves of Euphorbia hirta ...... 147 Figure-3.51. Effect of different seasons and sites on Manganese (Mn) contents in leaves of Euphorbia hirta...... 147 Figure-3.52. Effect of different seasons and sites on Zinc (Zn) contents in leaves of Euphorbia hirta...... 151 Figure-3.53. Effect of different seasons and sites on Crude Protein contents in leaves of Fumaria officinalis...... 152 Figure-3.54. Effect of different seasons and sites on Crude Fats contents in leaves of Fumaria officinalis...... 156 Figure-3.55. Effect of different seasons and sites on Crude Fibres contents in leaves of Fumaria officinalis...... 157 Figure-3.56. Effect of different seasons and sites on NFES contents in leaves of Fumaria officinalis...... 157 Figure-3.57. Effect of different seasons and sites on NFEE in leaves of Fumaria officinalis...... 157 Figure-3.58. Effect of different seasons and sites on Potassium (K) contents in leaves of Fumaria officinalis ...... 158 Figure-3.59. Effect of different seasons and sites on Calcium (Ca) contents in leaves of Fumaria officinalis ...... 158 Figure-3.60. Effect of different seasons and sites on Magnesium (Mg) contents in leaves of Fumaria officinalis ...... 158 Figure-3.61. Effect of different seasons and sites on Copper (Cu) contents in leaves of Fumaria officinalis ...... 162 Figure-3.62. Effect of different seasons and sites on Iron (Fe) contents in leaves of Fumaria officinalis ...... 163

Figure- 4.5. Yield Meter-2 (gms) of Fumaria officinalis...... 191 Figure-4.6. Effect of different seasons and sites on yield Hectare-2 (Kg) of Fumaria officinalis...... 195 Figure-4.7. Effect of different seasons and sites on number of Plants Meter-2 of Euphorbia hirta...... 196 Figure-4.8. Effect of different seasons and sites on number of Leaves plant-1 of Ajuga bracteosa...... 196 Figure-3.63. Effect of different seasons and sites on Manganese (Mn) contents in leaves of Calatropis procera...... 163 Figure-3.64. Effect of different seasons and sites on Zinc (Zn) contents in leaves of Fumaria officinalis ...... 163 Figure-4.1. Effect of different seasons and sites on number of plants Meter-2 of Adhatoda vasica...... 177 Figure-4.2. Effect of different seasons and sites on yield meter-2 (g) of Adhatoda vasica...... 178 Figure-4.3. Effect of different seasons and sites on yield hectare-1 (kg) of Adhatoda vasica...... 178 Figure-4.4. Effect of different seasons and sites on number of leaves plant-1 of Fumaria officinalis...... 188

LIST OF APPENDICES

Appendices No. Title Page No

Appendix-1.1. List of total plant species identified at Khanpur Valley...... 236 Appendix-1.2. List of Medicinally Important Plant Species Identified at Khanpur Valley...... 243 Appendix-1.3. Questionnaire...... 248 Appendix-3.1. ANOVA for Proximate analysis of Adhatoda vasica ...... 249 Appendix-3.2. ANOVA for Elemental analysis of Adhatoda vasica ...... 249 Appendix-3.3. ANOVA for Proximate analysis of Calatropis procera ...... 249 Appendix-3.4. ANOVA for Elemental analysis of Calatropis procera...... 249 Appendix-3.5. ANOVA for Proximate analysis of Recinus communis...... 249 Appendix-3.6. ANOVA for Elemental analysis of Recinus communis ...... 250 Appendix-3.7. ANOVA for Proximate analysis of Ajuga bracteosa ...... 250 Appendix-3.8. ANOVA for Elemental analysis of Ajuga bracteosa...... 250 Appendix-3.9. ANOVA for Proximate analysis of Euphorbia hirta ...... 250 Appendix-3.10. ANOVA for Elemental analysis of Euphorbia hirta...... 250

Appendix-3.11. ANOVA for Proximate analysis of Fumaria officinalis ...... 251 Appendix-3.12. ANOVA for Elemental analysis of Fumaria officinalis ...... 251 Appendix-4.1. ANOVA for Horticultural attributes of Adhatoda vasica...... 251 Appendix-4.2. ANOVA for Horticultural attributes of Calatropis procera. ..251 Appendix-4.3. ANOVA for Horticultural attributes of Recinus communis. ....252 Appendix-4.4. ANOVA for Horticultural attributes of Ajuga bracteosa...... 252 Appendix-4.5. ANOVA for Horticultural attributes of Euphorbia hirta...... 252 Appendix-4.6. ANOVA for Horticultural attributes of Fumaria officinalis. ...252

GENERAL INTRODUCTION

Plant species that are indigenous to various countries and regions are one of the most important natural resources, on which the future of its agriculture, environment, forestry, wildlife, pharmaceutical and food industries relies and these landraces have always defined the course of research and development. Unfortunately species diversity is facing a cumbersome pressure from the exploding human population, as demand for earth‟s natural resources has increased manifold. Renewable and non- renewable resources play a vital role in human well being and hence it should be used in a sustainable manner adopting some simple techniques like differed gratification (Hunter, 1996).

Diversity of Plant species has a universal acceptance as a source of medicinally active ingredients (Pearce and Puroshothaman, 1992). The name Ethno Botany was first used by Harshberger in 1896 for the plants used by local population for traditional health care (Plotkin, 1991). Currently ethnobotany is considered as part of Economic Botany, which relates to the economic utilization of plants for human wellbeing (Heiser, 1993; Wickens, 2001).

Ethno-pharmacological studies were reported to be first carried out by the personal physician of the King of Spain who was sent to study medicinal plants used by the Mayans and Aztecs in the fifteenth century (Sheng-Ji, 2001). Similarly, the use of herbal medicine in Asia has a long history of human interaction with the plants as Himalayan herbs were being used for medicinal purposes since ancient times. The earliest use was recorded in the Vedas about 4500 to 600 B.C. This is known to be the oldest collection of human knowledge containing the medicinal use of 67 plant species (Sheng-Ji, 2001). In China, Sheng-Nongs Herbal book is considered to be the earliest source of indigenous knowledge on the utilization of herbs. It contains information of 365 plants, animals, and minerals useful as medication in the period (3000 B.C.) of Sheng-Nong (Sheng-Ji, 1987). Now a days the Traditional Chinese Medicine includes about 11,146 plant species, of which 492 species are cultivated and the remaining 10,654 species are regarded as natural or wild plants (Adnan et al., 2006).

The Islamic traditions of Prophet Muhammad (PBUH) collectively known as Tibb-i-Nabvi (Prophet‟s way of treating ailments and diseases) contains information regarding the use of a number of herbs as a remedy for curing common diseases. For example, the use of barley preparation for loss of appetite, Senna for constipation and olive, black cumin, chicory, endive fenugreek, ginger, marjorum, saffron, vinegar, water-Cress etc. for other various diseases. According to the Prophet, black cumin is a remedy for

every known disease except for death. The Prophet expressed similar views on the efficacy of senna and cress (Farooqi, 2006).

Unani Medicine was started by the Greco-Arabic society in Indo-Pak subcontinent. It is developed on many concepts and recipe introduced by Ayurveda, a tradition that started the use of medicinal plants as early as 2500 BC (Mahmood et al., 2004). It enlist the details for the medicinal use of as many as 290 herbal drugs (Manandhar, 1980). Now a days, this local medicine system is extensively used in India, Pakistan and other Asian countries.

Medicinal plants not only serve as an important source of raw materials for the manufacture of traditional medicines but also are used for the preparation of a number of modern allopathic medicines such as Senokot® and Fybogel® (Reckitt and Colman Pharmaceuticals, UK) are derived from Senna leaves and Plantago seeds respectively, and are actively used for constipation and diarrhea. Similarly some of the cough syrups are being prepared from certain alkaloid containing plants, such as Cough Nill® (T.J and P Laboratories, Pakistan) from Justicia adhatoda, Tinospora cordifolia and Ephedra vulgaris.

The major demand in medicinal plants results in a huge business from local to global level. In the 1990s, the reported annual world-wide demand of herbal plants was 400,000 t and valued at USD 1,224 million. The international trade is dominated by only few countries. About 80 % of the world-wide imports and exports are controlled by only 12 countries with the dominance of temperate Asian and European countries. Pakistan is one of these 12 countries with higher imports bill than exports. Whereas Japan and the Republic of Korea are the main consumers of pharmaceutical plants, and China and India are the leading producers at international level. It is very pertinent to mention here that , in terms of numbers, about 90 % of the 1,200-1,300 European medicinal species are

mostly collected from the nature (Adnan et al., 2006). Also in China, the 1,000 commonly used herbal plants, 80 % in terms of numbers originate from the wild (Adnan et al., 2006). Besides the use of wild plant species in traditional and modern medicines, this natural treasure can be used in agriculture as a cheap source of bio-pesticide. For example Azadirachta indica is used as systemic poison and insect repellent, Carica papaya L is used as larvicide, Nicotiana tobacum L is a good source of contact poison and insect repellent and Terminalia catappa L is used as insecticide.

Pakistan, currently exports plants and earn US$ 5.45 million per year. Over 60% of the total export originates from the Hindukush-Himalayan mountains of the country (Sher & Hussain, 2009). The destinations of such exports include India, Germany, USA, Middle East, Iran, etc. However, Pakistan also imports a huge quantity of medicinal plants, worth US$ 130 million, from the above countries (Sher & Hussain, 2009). Such imports have increased over the last ten years. Studies (Adnan et al., 2006) have revealed that about 70% of the herbs that are being imported to Pakistan actually grow wild in the Hindukush-Himalayan mountains of Pakistan but these have neither been explored fully nor their economic and medicinal importance is known to the local communities. The potential to increase the density and cover of most of medicinal plants does exist, if these plants are conserved in wild and promoted through cultivation and treated as a cash crop in the country. This will not only save the valuable foreign exchange but shall also provide opportunity for the income of the poor communities.

Pakistan has a unique geographical location stretching from the seashore to K-2 (8611 m above sea level), the second highest peak of the world. It occupies an area of 79.6 million hectares within the geographical limits between 24-37o N and 61-75o E. The country is mostly arid with 75% of its parts receiving an annual rain fall of less than 250 mm and 20% of it is receiving less than 125 mm. Only 10% of the area in the northern

mountain ranges receives rainfall from 500 mm to 1500 mm (Ahmad and Waseem, 2004).

The vegetation in different parts of Pakistan varies with elevation, soil type, and precipitation. Forests are mainly present on mountain ranges in the north west, where coniferous alpine and sub-alpine trees are found. The southern ranges of the Himalayas, which are of lower elevation, receive heavy rainfall and have dense forests of deodar, pine, poplar, and willow trees. The more arid Suleiman and Salt mountain ranges mostly contain Delbergia sisso. Dry-temperate vegetation, such as coarse grasses, scrub plants, and dwarf palm, predominates in the valleys of Khyber Pakhtunkhwa and Baluchistan Plateau (Shinwari et al., 2005). Pakistan has a vast diversity of landscapes. The high snow caped mountain ranges of the Hindu Kush, Himalayas and Karakuram, long glaciers between them, the high and long lying cold deserts in the north, the vast irrigated plains, the hot low lying Thar and Thal deserts, the long rocky Potohar in Punjab, Sindh and Baluchistan and the coastal shores provide all the possible habitats for the development of different plant communities (Ahmad and Waseem, 2004).

The paramount importance of medicinal plants/herbs can also be judged from the gigantic shares of medicinal plant species in plant species available/reported in Pakistan. It has been reported that in Pakistan out of 4950 total plant species 1500 are medicinally important which becomes 30.3% as compared to global share of medicinal plants which is 17.1% of total reported plant species (Govaerts, 2001 and Moerman, 1996). Some scientists state that nearly half of the flora of pakistan is enthno botanically important. The conventional medicine derived from almost 300 species of plants (Perveen and Hussain, 2007); whereas 16.8 % of the total documented flora is medicinally important which comprises of 1010 species (Shinwari et al., 2005). There are 86 recoginized institutions involved in making herbal/traditional medicine; producing 300-400

items. Majority (around 90%) of the required raw material is imported. These traditional medicines usually take care of 50% of the population. The herbalists of eastern medicines in Pakistan are about 50,000 (Zaidi 1998). proper attention is not been given to the local conventional herbals and spices although these have commendable economic benefits for the local community.

Pakistan is rich in the diversity of natural resources. To manipulate these resources and use them for the betterment of its people, it is necessary to identify the actual requirements and the source, from where it can be fulfilled. The two basic requirements of ordinary Pakistanis can be identified as food and health. To explore natural resources, for cheap medicines and additional food supplement, a scientific eye will concentrate on medicinal plants diversity in the potential valleys of northern Pakistan. The area comprising moist temperate to subtropical zones of Pakistan deserve specific attention for the exploration and conversation of species diversity. During the last decades, the zone has been subjected to major structural variations, resulting in decrease of the forest area. The decrease in forest cover, combined with major changes in community structure, has been responsible for the decline of indigenous medicinal plant resources and its utilization in the traditional healthcare. It is of utmost importance that research projects, be initiated in climatically diverse valleys of Pakistan and valuable medicinal flora needs to be documented and analyzed for its usefulness in human healthcare, in agriculture and as food supplement.

The Khanpur valley in the sub Himalayan mountains of Pakistan is located between longitudes 72° 35' to 73° 15' and latitudes 33° 44' to 34° 22', in the Haripur District of Khyber Pakhtunkhwa (Ejaz et al, 2012). The valley is rich in natural resources and has very important water reservoir, the Khanpur Dam. Geographically, it is a gateway between, on the one side, to the Khyber Pakhtunkhwa, and on the other side to the national

capital, Islamabad. This beautiful valley with a lake / dam on Haro River is 45 km from Islamabad on Taxila-Haripur Road. Khanpur Dam has been constructed in a narrow gorge on the Haro River. It is a multipurpose project which supplies drinking water to Islamabad and Rawalpindi and irrigation water to Khyber Pakhtunkhwa (110 cusecs) and Punjab (87 cusecs) (Ejaz et al, 2012). The Catchment Area of the Haro River comprises of four main streams i.e. Lora Haro: its source is in the Murree hills, Stora Haro: It starts from the base of Nathiagali hills, Neelan Stream: its base is in Nara hills and Kunhad Stream: Its base is in mountain of Siribang, mountain of Dubran and the surrounding areas.

The following categories of the Forests are found in the Khanpur Valley.

Reserved Forests: The Reserved Forests are located starting from New Khanpur Town, and scattered up to Nathiagali and Muree, along the sides of River Haro. These Forests covers a large area of Khanpur valley. All these Forests are the property of Government of Khyber Pakhtunkhwa province of Pakistan. These forests are called “Beer” in the local language.

Protected Forests or Guzaras: These forests are scattered all over the region, are owned by the people, but under the protection of Forest Department.

Cooperative Forests or Mehdoodas: The lands of people were taken by the Forest Department for Forestation/plantation about 50 years ago.

Private Forests & Farms: There are some private Farms and Forests in the area.

Graveyard Forests: Large and old Graveyards almost in every village have become thick forests.

All the above mentioned types of forests are full of medicinal plants indigenous to the valley. The species of medicinal plants, its frequency,

volume, and coverage is still unknown. The local Herbalists collect valuable medicinal flora and utilize it in herbal medicines preparation but the scientific exploration is of utmost importance. Furthermore, the valley is in close vicinity with Herbal industries of Hattar Industrial Area, Haripur and its exploration for its medicinal wealth can be a source of raw material supply for the Industries and source of income for the poor population of the valley. This research project is destined to open new venues of research for the domestication, cultivation and improvement of medicinal plants indigenous to other valleys of Pakistan. It will lead other research scholars to explore further aspects of the current research project. This research activity will definitely lead to the cultivation of medicinal plants indigenous to Khanpur valley and their commercial utilization by herbalists and pharmaceutical industries, located at the Industrial Estate Hattar, near Khanpur valley.

Having the above mentioned scenario in view, Department of Horticulture, the University of Agriculture Peshawar, initiated the current research studies, titled “Species diversity and agro chemical prospects of medicinal plants of Khanpur Valley in the sub-Himalayan Mountains of Pakistan” with the following objectives:

1. Enlistment of total plant species found in the valley.

2. Identification of the medicinally important species of the valley and its traditional utilization. 3. Determination of ecologically important species and its classification.

4. Selection of the best harvesting season and site for collection of medicinal plants. 5. Quantification of the nutritional components of selected medicinal plants for medicinal and nutritional utilizations.

6. Documentation of the growth geometry of the selected medicinal plants for their annual sustainable harvest potential.

II. REVIEW OF LITERATURE

Ethno Botany and Species Diversity

Indigenous knowledge of plants of different areas is as old as human civilization. However, the term “Ethno Botany” was first used by an American botanist Johan W. Harshberger in 1896, “to the study of plants used by primitive and aboriginal people”. In modern ecological terms ethno botany was described as “The study of direct interactions between human and plant populations (Plotkin, 1991; Heiser, 1993). Today ethno botany is widely accepted as a science of human interactions with plants and related ecosystems. Plants have been used as medicine since ancient times. Use of plants to improve economy is an old tradition of human history. In Pakistan, however, the field of ethno botany is relatively nascent and just a few research projects have been accomplished (Qureshi and Khan, 2001).

Leporatti and Lattanzi (1994) studied 27 medicinal plants ethno botanically in Makran (Southern Pakistan). Quershi and Khan (2001) recorded a total of 25 species of herbs belonging to 18 families and their medicinal uses by the people of Chakwal, Pakistan. Some of these species were used for the treatment of cholera, dyspepsia , fevers cure, herpes, eczema, jaundice and liver complaints. An ethno botanical use of plants is more common in remote valleys of Pakistan due to lack of medical facilities. Forty medicinally important plant species of twenty one

families were reported in Kala Chitta Hills (salt range) of District Attock (Mahmood et al., 2004) which were under common use of indigenous people. Due to increase in population, demands of people increases, causing greater pressure on the products of the area. To understand the indigenous knowledge of the local people, ethnomedicinal study is very important. This helps a lot for creating awareness among them regarding sustainable natural resource management. Local people, hakims and medicinal businessmen are very important in this regard (Mahmood et al., 2004).

In Africa, people use different plant extracts for treating trypanosomiasis (Atawodi et al., 2003 and Mulholland, 2005). Some of the indigenous plants are very important in the diets of postpartum women during which time it is considered that these spices and herbs help the contraction of the uterus. Spices and herbs are generally known to possess antibacterial and antioxidant properties (Iwu, 1989).

Murad et al., (2011) conducted regular field survey for two consecutive years in HazarNao forest of the tribal zone of Dargai, Malakand District, Pakistan. Information about medicinal plants were gathered through formal and informal interviews and questionnaires were filled from local people. The folk medicinal uses of 75 plant species were recorded for various human ailments.

Dar (2003) investigated the flora of Lawat in Muzaffarabad, Azad Jammu and Kashmir for ethno botanical utilization. He found 52 species of which 3 species were of 2 gymnosperm families while 49 species were belonging to 35 angiosperm families. Most of the plants were used medicinally. The investigation indicated that the medicinal plants were either used singly or with mixtures by local inhabitants. The area under investigation, due to unplanned utilization, had resulted in loss of medicinally important plant species.

Euphorbiaceae is an important plant family especially recognized for its anti-cancer and anti-hepatitis components. In the literature of ancient traditional Chinese medicine, 33 species of plants from 17 genera of Euphorbiaceae have been mentioned as medicines. Presently 111 species within 35 genera of medicinal Euphorbiaceous plants has been reported. Among them, 17 species were used to treat snakebites. It was observed that most of the species within the Euphorbiaceae family contained toxic components. Only a few species were utilized as widespread medicines. Most species were recognized only as associated with one tribe or another (Lai et al., 2005).

Sambucus nigra bush of family Caprifoliaceae is one of the plants which are most commonly used for medicinal and various other purposes by the inhabitants of Catalonia and in many Mediterranean regions. It is a most versatile plant, being used for food and medicine. In addition, almost every part of the plant, including the bark, roots, leaves, flowers, and fruit, has some uses (Vallès et al., 2004).

In Salt range and other areas, different plants are used for medicinal purposes. Leaves and roots of Justicia adhatoda L. (Acanthaceae) are used for coughs, bronchitis, asthma and rheumatism. Leaf buds are also used in diabetes and for joints and as antiseptic. Green leaves of Withania sominifera (L.) Dinal (Solonaceae) are used to relieve the joints ache and painful swelling. Roots are used as diuretic and tonic. Juice of the whole plant is useful in rheumatism. Seeds are used to coagulate milk. Whole plant of Buxus papillosa is used as diaphoretic, purgative and anti rheumatic. Different species of Diclepter shoots are used as tonic. Whole plant of Hermal (Peganum harmala) is used as an analgesic, aphrodisiac, hypnotic and antispasmodic. Salvia virgata leaves are applied to tumors and ulcers. Solanum indicum roots, leaves and fruits are used as expectorant, carminative analgesic and febrifuge. Solanum surattense

whole plant is used as vasodilator, astringent and expectorant. Sophora mollis Khumbi seeds are used as anthelmintic (Ahmad et al., 2002; Khan, 1951).

Khan et al., (2012) identified use of Medicinal Plants in Folk Recipes by the Inhabitants of Himalayan Region Poonch Valley Azad Kashmir (Pakistan). Total 68 species of plants belonging to 44 families were recorded as used medicinally for preparations of folk recipes of 68 ailments. The survey indicated that 72% source of indigenous knowledge related to the medicinal use of plants comes from people of age of 50 years and above, while 28% of it contributed by people between age 30 and 50 years. The survey also found that old men were more informative of folk knowledge of medicinal plants than women in the area. It was also indicated that about 60% of the homemade drugs were used by people above the age of 50 years, 30% by children below age of 15 years especially infants. While remaining 10% of the herbal-medicines were utilized by people between ages of 15-50 years.

Rasool et al., (2010) studied indigenous knowledge of folk medicine by the women of Kalat and Khuzdar regions of Baluchistan, Pakistan and concluded that women use medicinal plant resources of the area for their ailments mainly digestive complaints, stomach problems, fevers, liver complaints, diabetes, children diseases and birth related problems.

Abbasi et al., (2009) studied medicinal plants used for the treatment of jaundice and hepatitis based on socio-economic documentation. A total of 30 plant species belonging to 24 families were reported by local practitioners for the treatment of jaundice and hepatitis. Leporatti and Lattanzi, (1994) studied 27 medicinal plants ethno botanically in Makran (Southern Pakistan). They reported and discussed their traditional medicinal uses. The inhabitants of Kahuta use the medicinal plants for

various purposes and are dependent on surrounding plant resources for their food, shelter and health.

Jan et al., (2008) discovered 27 herbal recipes used for gastrointestinal disorders in Kaghan Valley, KP, Pakistan. Edeoga et al., (2005) analyzed ten medicinally important plants belonging to different families and compared for alkaloids, tannins, saponins, terpenoid, flavonoids and phenolics. The medicinal plants investigated were found very important in traditional medicine. Qureshi and Khan, (2001) recorded a total of 25 species of herbs belonging to 18 families and their medicinal uses by indigenous people form the area. Some of these species were used for the treatment of cholera, dyspepsia, fever, herpes, eczema, jaundice and liver complaints. Dar (2003) investigated the vegetation of Lawat in District of Muzaffarabad, Azad Jammu and Kashmir for ethnobotanical purposes. He recorded 52 species, out of which 3 species were of 2 gymnospermic families while 49 species were of 35 angiospermic families. Most of the plants were used medicinally. The area under investigation, due to unplanned utilization, had resulted in loss of medicinally important plant species.

Fazal et al., (2010) undertaken taxonomic studies about the species diversity of District Haripur. The flora of the area consists of 211 species of 170 genera and 66 families. Similarly Qureshi et al., (2009) investigated medico-ethnobotanical inventory of tehsil chakwal, Pakistan and recorded 29 species belonging to 25 genera and 18 families used by the local people for curing various human ailments.

Hussain et al., (2007) studied some plants of Mastuj, district chitral, Pakistan. They determiined that 111 species of 46 families. Family Asteraceae (11 spp.), Papilionaceae (10 spp.) and Rosaceae (9 spp.) were declared as having larger number of species. The traditional uses revealed that there were 90 fodder, 52 medicinal, 40 firewood, 19 vegeTable, 15

thatching/fencing, 13 timber and 9 fruit species. Ibrar et al., (2007) worked on ethno botanical studies on plant resources of Ranyal hills, district Shangla, Pakistan. Ethno botanical information were collected on 97 plant species from Ranyal Hills District Shangla, Pakistan.

Haq et al., (2010) found out species diversity of vascular plants of Nandiar valley western Himalaya, Pakistan. Field observations showed that vegetation of the area was threatened due to injudicious use by the local people. The trend of urbanization, deforestation, over grazing, habitat fragmentation, unscientific extraction of natural vegetation, introduction of the exotic taxa and habitat loss were the visible threats. Total of 402 species belonging to 110 families of vascular plants were judged. Among the 402 species reported, 237 species were herbs, 71 shrubs, 68 trees, 06 climbing shrubs, 18 climbers and 03 epiphytes. The plants were classified according to local, traditional and economic value. Based on local uses, there were 178 medicinal plants, 21 were poisonous, 258 were fodder species, 29 were hedge plants, 122 were fuel wood species, 37 were timber yielding plants, 41 were thatching and sheltering plants, 71 were wild ornamental, 100 were weeds, 47 species yield edible fruits and seeds, 43 were used as vegeTable and pot herb.

Adnan et al., (2006) studied the Ethno-Medicinal uses in Northern Pakistan and documented over 300 plant species with medicinal value. Similarly, Qamar et al., (2010) studied the ethnobotanical importance of medicinal plants of Neelam Valley, Azad Kashmir and revealed that leaves were the most important part of medicinal plants used in the traditional healthcare. While Alam et al., (2011) studied the indigenous knowledge of the residents of Chagharzai Valley, Buner and revealed that whole plant and leaves are the most preferred parts of medicinal plants collected by local people.

Ahmad and Waseem (2004) found that anthropogenic stresses are also very common, which has depleted vegetation in general and medicinal flora in particular. From 29 species investigated locally; the conservation status of critically endangered, endangered, vulnerable and low risk species was 6, 9, 4 and 10, respectively. Immediate and complete protection, efficient recovery systems and effective community involvement for long term conservation, are pertinent for sustainable utilization of plant resources.

Biochemical Analysis (proximate analysis)

Biochemical analysis described in terms of main classes of substances is called proximate analysis. In proximate analysis the groups are measured as such, instead of individual proteins or specific minerals (FAO, 2001). It mostly includes proteins, fats, minerals and carbohydrates. Almost all plants contain these substances and are initially analyzed proximately.

Green, (1992) and Bianco et al., (1998) stated that now interest has been developed in wild species for their possible medicinal values in diets. Wild plant species provide minerals, fibers, vitamins and essential fatty acids and enhance taste and colour in diets. In addition, they have antibacterial, hepato-protective and anti-carcinogenic properties, and therefore, have medicinal value.

Sofowara, (1993) found that leaves and stems of most of the plants were rich in alkaloids, flavonoids, tannins and phenolic compounds. They had already been examined to show medicinal activity as well as exhibiting physiological activity.

Morinda citrifolia is an important medical plant in Southeast Asian countries. It was analyzed proximately and biochemically to make a more modern drug from a traditional product. In order to obtain better understanding of the medicinal attributes of the M. citrifolia a fruit

cultivated in Cambodia, fatty acids, proteins, amino-acids and sugars of juices were studied (Chunhieng et al., 2005).

Kochhar et al., (2006) analyzed three traditional medicinal plants namely bitter gourd, fenugreek and jumbo seeds. It was found that these plants were a good source of protein, minerals, crude fiber, fat and energy. They recommended to diabetic patients to include these medicinal plants in their daily diet to maintain blood sugar level.

In Southern Nijeria, Okwu and Ekeke, (2003) analyzed ripe fruits of Dennettia tripetala for proximate composition. Dennettia tripetala contained crude protein (15.31%), total carbohydrate (62%), crude fibres (9.84%), crude lipids (3.47%) and moisture (8.0%). It had energy value of 480.24 cal·100 g-1 of fresh fruit which justified the use of Dennettia tripetala fruits as food and as drug. Another fruit plant, Cymbopogonjwa rancusa, was found useful in diseases of blood, skin, vomiting, abdominal tumors, unconsciousness and fever. This plant was proximately studied by Mahmud et al., (2002) and was found to contain moisture contents, 67.02%; ash contents, 9.52%; carbohydrates, 1.8%; reducing sugar, 1.07%; nitrogen, 0.67%; crude proteins, 5.02% and crude fiber, 9.50%.

In another study, four medicinal plants belonging to the family Lamiaceae were chemically analysed by Edeoga et al., (2006) for their chemical constituents and nutritional value. The medicinal plants contained crude protein (9.19 to 17.94%), crude fibre (4.88 to 9.04%), ash (5.68 to 6.88%), and carbohydrate (66.24 to 75.87%), crude lipid (3.48 to 4.90%) and food energy (357.68 to 373.26 mg/cal). These plants play an important role not only in nutrition but also in traditional medicine and in pharmaceutical industry.

Fagonia arabica is amongst the widely used medicinal plants in Pakistan. Generally the plant is located on dry calcareous rocks, distributed in most parts of the Mediterranean region to South Africa, Afghanistan, India, Pakistan especially Sindh, Punjab and Khyber Pakhtunkhwa (Rizvi et al., 1996). Proximate analysis of this plant revealed that leaves and seeds have maximum moisture content (58.51±0.50) followed by shoots and roots (43.29±0.42, 29.45±0.28 respectively). Ash and protein (1.85±0.12, 0.64±0.01 respectively) increased in different parts in descending order i.e. rootsshoots>leaves and seeds (Shad et al., 2002).

Interest has been developed in wild species for their possible medicinal values in diets. Wild plant species provide minerals, fibers, vitamins and essential fatty acids and enhance taste and color in diets. In addition, they have anti-bacterial, hepato-protective and anti-carcinogenic properties, and therefore have medicinal value (Green, 1992; Bianco et al., 1998 and Yildirim et al., 2001) analyzed eight plant species in Turkey for dry matter, ascorbic acid, nitrogen and protein which is important nutritionally as well as for medicinal value.

Piliostigma thonningii is a leguminous medicinal plant belonging to the family Caesalpiniacea, used for the treatment of dysentery, fever, infections, respiratory, ailments, snake bites, hookworm and skin diseases (Jimoh and Oladiji, 2005). Proximate composition of Piliostigma thonningii seeds showed that seeds contained moisture contents 6.71 %, ash 3.50 %, crude proteins 30.33 %, crude fibers 35.03 %, lipids 1.42 % and carbohydrates 23.00 %.

Two rural settled Fulani villages, northeastern Nigeria, were surveyed for the use of wild plants as food or medicine (Lockett et al., 2000). Edible wild species available during the wet season generally were inferior in

energy and mineral content as compared to dry season plants. Fruits commonly eaten by children were poor sources of protein and minerals but rich in carbohydrate and fibers. Shiwaka leaves (Veronia colorate) that were mostly consumed by pregnant women to increase breast milk production and to expel intestinal worms contained high fibers contents. In another study it was proved that plants can be used not only in traditional medicine but also in food and in pharmaceutical industries (Edeoga et al., 2006).

Carica papaya belonging to the family Caricaceae is an important and common medicinal plant in tropical Africa. Proximate analysis of the unripe pulp of Carica papaya was analyzed for the presence of different phytochemicals and minerals (Oloyede, 2005). It showed that the pulp contained starch (43.28%), sugars (15.15%), crude protein (13.63%), crude fat (1.29%), moisture (10.65%) and fiber contents up to 1.88%. These results indicated that the pulp of mature unripe Carica papaya contained nutrients and mineral elements useful in nutrition. The presence of some phytochemicals like saponins and cardenolides explained the astringent action of the plant encountered in the numerous therapeutic uses.

Arubi, (2003) analyzed papaya kernel flour for proximate composition and functional properties. The flour was high in protein (32.4%) but moderate in available carbohydrates (49.9 %) and low in moisture (7.5 %) contents. The total minerals and fiber contents were 5.3% and 4.2%, respectively. Oil and water absorption capacities of the flour sample were high. The flour had very good foaming and emulsifying properties. These results suggested that papaya kernel flour can be used in a number of food formulations. The underground codex of the cycad, Stangeria eriopus is used extensively by several communities in South Africa, mainly as an emetic. It was found that only in the month of July during 1992, 3410

plants were sold, which threatened the remaining plant populations. Proximate analysis of the caudex material gives high carbohydrate content with only small percentages of fat, protein, fiber and ash, (Osborne et al., 1994).

Hassan and Umar, (2006) studied the nutritive value of Momordica balsamina L. leaves and found high moisture contents (71.00±0.95% fresh weights). The concentration of estimated crude protein and available carbohydrates on dry weight (DW) basis were 11.29±0.07% and 39.05±2.01% respectively. The leaves also have high mineral (18.00±0.56% DW) and crude fiber (29.00±1.23% DW) contents; while crude lipid contents (2.66±0.13% DW) and energy value (191.16kcal/100g DW) were low. The results indicated that the Momordica balsamina leaves could be a good supplement for mineral, protein, carbohydrate and fiber contents.

Sundriyal and Sundriyal, (2004) studied wild edible plants in the Himalaya. Total 190 species were screened as edible species, out of which nearly 47 species came to the market. Twenty seven plant species were analyzed proximately for their nutritive values, 22 were edible for their fruits and 5 for leaves and shoots. Among different plant parts, generally higher nutrient concentration was recorded for leaves, followed by new shoots and fruits.

Chunhieng et al., (2005) revealed that Morinda citrifolia, a fruit cultivated in Cambodia, is an important medical plant in Southeast Asian countries. It was analyzed proximately and biochemically to make a more modern drug from a traditional product. In this regard the juice of M. citrifolia was analyzed for fatty acids, proteins, aminoacids and sugars.

Edeoga et al., (2006) screened four medicinal plants belonging to the family Lamiaceae for their chemical constituents and nutritional value.

The medicinal plants contained crude protein (9.19 to 17.94%), crude fibre (4.88 to 9.04%), ash (5.68 to 6.88%), and carbohydrate (66.24 to 75.87%) crude lipid (3.48 to 4.90%) and food energy (357.68 to 373.26 mg/cal). These plants play an important role not only in nutrition but also in traditional medicine and in pharmaceutical industry. Edeoga et al., (2006) concluded that the active particles differ from plant to plant due to their biodiversity and they produce a definite physiological action on the human body.

Biochemical Analysis (Elementology)

The importance of elemental contents of medicinal plants was first highlighted by Hakim Abdul Hamid, President Hamdard National Foundation, India, who is considered the founder of the discipline “Elementology”. Health depends upon the organized state of elements in the body and their imbalance causes diseases and restoration of balance by drugs can cure diseases (Golden, 1988).

Medicinal plants show therapeutic effects for the treatment of different diseases due to the presence of certain chemical compounds in these plants. These are mostly organic compounds which have biological activities, but none of these act independently and they perform the functions of the medicinal plant collectively (Mutaftchiev, 2003).

Analysis showed that medicinal plants are rich in many trace elements, and it was suggested that this was an important factor in the curative effect of these plants (Olabanji et al., 1997; Pereira and Felcman, 1998). Remington, (1995) observed that the trace elements can be found in free states or organically bound in a complex. It is well established fact that different states and forms can have different functions in its physiological activities such as biotoxicity and percent absorption in the body. Trace elements co-exist with various organic compounds in medicinal plants

and mostly they are bound to organic compounds. So the concentration of the free trace elements will be very low (Remington, 1995).

Sahito et al., (2001) investigated two varieties of medicinal herb Catharanthus roseus for elemental composition as Ca, Na, K, Mg, Zn, Fe, Cu, Co, Mn, Ni, Cd, Pb, Ba and Al. The level of essential elements such as Zn, Fe, Mn, and Cu was present in considerable amount. In decoction the level of essential elements was high as compared to toxic elements.

Most of the medicinal plants qualify as nonprescription drugs and some of them are taken in low doses as food drugs in these days (Obiajunwa et al., 2002) for example, Se, Zn, vitamin E and other antioxidants of plant origin are proving to be reliable factors in the effort against premature ageing and the delay of degenerative diseases. There are at least 50 elements which are vital for the well being of humans. Now the people are very much interested in trace elements in the area of medical science.

Obiajunwa et al., (2002) investigated different major and minor elements in different plants. Fourteen different elements, namely K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, Rb and Sr were detected in varied amounts in different plants. The concentrations of Ca and K were the highest where as Br and Se were the least. All the essential elements were present in required dose but some plants such as C. procera, A. indica and A. wilkensiana showed toxicity due to their high Cr level.

The high concentration of Ca is very important as Ca enhances the qualities of bones and teeth and also of neuromuscular systemic and cardiac functions. Iron is another important element present in all the specimens which plays a role in oxygen and electron transport in human beings. The high amount of Fe and Ca in C. alata showed that it could be especially useful in the treatment of constipation in nursing and pregnant mothers at 5 g./dose, as its main medicinal activity is as a mild laxative.

E. hirta, A. melegueta, M. indica, G. kola contained high concentration of Zn and could be used in cases of Zn deficiency which includes impairment in healing, taste and growth (Obiajunwa et al., 2002).

More than 25 naturally occurring elements perform essential functions in the human body. Some of them such as zinc, copper, selenium, cobalt, chromium, molybdenum, manganese and iodine, are required in small amounts and each consists less than 0.01% of the body weight and they are known as essential trace elements. They work in a similar way in the body. Most of them are at the active site of enzymes and other physiologically active substances of the body. Dietary deficiency of these elements causes various problems, which are consistent with the decreased activity of these active substances (Wada and Yanagisawa, 1996).

Ahmad and Chaudhary, (2009) found that Ajuga bractiousa can be utilized for diabetes and hypertension due to higher quantity of chromium and potassium respectively. Zinc is one of the most important trace elements in the body as it performs various biological activities. It is an essential catalytic component for more than 200 enzymes, and also acts as a structural component of many proteins, hormones, neuropeptides, hormone receptors, and most likely polynucleotide ( Fabris and Mocchegiani, 1995). Zinc play role in cell division, differentiation, programmed cell death, gene transcription, biomembrane functioning and obviously many enzymatic activities, and hence considered the most important element in the accurate fuctioning of human organisms (Fabris and Mocchegiani, 1995). The supplementation of Zinc is useful in most of the problems. It is said to be a therapeutic support instead of a simple dietary supplement. The relevance of zinc to many age-associated diseases and the aging itself of the major homeostatic mechanisms of the body, i.e., the nervous, neuroendocrine and immune systems, places zinc

in an essential position in the economy of the aging organism (Fabris and Mocchegiani, 1995).

Rajurkar and Damame, (1998) studied elemental composition of some Ayurvedic medicinal plants used for healing urinary tract disorders. Fourteen elements were estimated in different plants, among these Cu, Cr, Co and Cd were found to be present at the trace level; Mn, Pb, Zn, Ni, Na, Fe and Hg at minor level and K, Ca and Cl at major level. The differences in the concentration of the elements are attributed to soil composition and the climate in which the plant grows. It was found that these elements play an important role in treatment of urinary tract disorders.

Some inorganic trace elements such as V, Zn, Cr, Cu, Fe, K, Na, and Ni play an important role in maintaining normal glycemia by activating the beta-cells of the pancreas. Leaves of four traditional medicinal plants (Murraya koenigii, Mentha piperitae, Ocimum sanctum, and Aeglem armelos) widely used in the treatment of diabetes and related metabolic disorders, were analyzed for different inorganic elements. The levels of Cu, Ni, Zn, K, and Na were found to be in trace amounts and Fe, Cr and V levels were found in minor levels (Narendhirakannan et al., 2005).

According to Anke, (1986) seven quantitative elements and eighteen trace elements are of vital importance for the animals. Their metabolism is antagonistically or synergistically influenced by the inorganic and organic constituents of the food of different types. More than 30 elements (Cu, Zn, Mg, Mn, Cr, V etc) are useful in the treatment in the process of arteriosclerosis.

Singh and Garg, (1997) analyzed specific plant parts of several plants (fruits, leaves or roots) often used as medicines in the Indian Ayurvedic system for 20 elements (As, Ba, Br, Ca, Cl, Co, Cr, Cu, Fe, K, Mn, Mo,

Na, P, Rb, Sb, Sc, Se, Sr and Zn). Most of the medicinal herbs were found to be rich in one or more of the elements under study.

Dennettia tripetala or pepper fruit plant is a well-known Nigerian spicy medicinal plant which contains protein, carbohydrate, fibres, lipids, mineral contents like calcium (1.80%), phosphorus (0.33%), potassium (2.50%) and magnesium (0.42%). This justifies the use of Dennettia tripetala fruits as food and drug in herbal medicine in Southeastern Nigeria (Donatus and Morah, 2004).

The level of iron amongst all minerals analyzed was found to be the highest (782 ppm) .This might be of nutritional importance especially in the part of the world where anemia and iron deficiency is more common. (Jimoh and Oladiji, 2005) P. thonningii seeds are also good sources of calcium (43.11 ppm) while zinc (0.016 ppm), manganese (1.00ppm) and phosphorus (0.02 ppm) levels were quite low when compared with iron and calcium but comparable with values for some legumes (Elegbede, 1998). Iron, selenium, zinc and manganese are antioxidant micronutrients and their presence could thus boost the immune system.

Naz et al., (2013) found that Fumaria parviflora derived extracts and the phytochemicals particularly the cis- and trans- prtopinium carry antibiotic properties and these compounds could be utilized in the making of novel chemotheraputic agents.

Fagoina arabicais amongst the widely used medicinal plants in Pakistan. Its mineral composition showed that Zn and Na were maximum in roots and minimum in leaves and seeds. Concentrations of Fe, P, K, and Ca decreased in order of leaves, seeds, shoots and then roots. Similary, Fagonia Arabica contained Ca, Na, P, Cu, Fe, Mn and Zn. Zn plays an important role as an antioxidant in animals (Bray and Betterger, 1990) as well as in plant membranes (Cakmak and Marschner, 1988). Fagonia

arabica contains lower amounts of heavy metals. The possible reason for low concentrations of heavy metal could be due to the fact that this herb is found mostly in desert and dry calcareous rocks where industrial pollutions are not found, which might have resulted in least amount of heavy metal. However, the macro elements (P, K, Na, Ca) were found to be maximum. Phosphorus is mainly involved in RNA, DNA- sugar phosphate backbone, in the process of energy transfer and it is the inorganic phosphate that appears as an intermediate product during photosynthesis and respiration pathways of metabolism (Shad et al., 2002).

Fagonia arabica contains a fair amount of K and Na. Due to this reason it is mostly used in diseases like diarrhea, stomatitis and deobstruent (Dey et al., 1980) where mostly fluid losses take place (Whitney and Hamilton, 1984).Whereas Cu, Zn, Mn and Fe are considered as trace elements due to their relatively minute quantity that is essential to the body. Copper is important for red blood cell formation, mitochondria function and a component of ribonucleic acid, whereas Zn, Mn and Fe are necessary for the development of bones and connective tissues (Nielsen, 1987).

Unlike other compounds, living organisms cannot synthesize mineral elements. Only small fraction of the Ca, Mg, and P and most of the Na, K, and Cl are present as electrolytes in the body fluids and soft tissues. Electrolytes present in blood or cerebrospinal fluid maintain acid -base and water balance, and adjust osmotic pressure.

They regulate membrane permeability and exert characteristic effects on the excitability of muscles and nerves (Nielsen, 1987; Bukhari et al., 1987).

The uptake of mineral constituents depends on the amount of mineral elements present in soil, their availability, moisture contents of soil and

the plant factor. The variation in mineral composition was found in different varieties of the same species (Sahito et al., 2001). Narendhirakannan et al., (2005) found that some trace elements such as V, Zn, Cr, Cu, Fe, K, Na, and Ni play an important role in maintaining mormoglycemia by stimulating the beta-cells of the pancreas.

Spatio-temporal Variations (site-season variation)

The work on spatial and temporal variations and its impacts goes back to the days of Theophrastus during 371-286 BC (Thanos, 1994) and surprisingly even now plant population models do not give spatially realistic descriptions of key environmental factors which outline various plant development and recruitment processes. This can be explained by the great complexity of natural habitats, and the difficulty in identifying key factors which take part in these processes and measuring the variation over large spatial and long temporal scales. Survival, growth, and reproduction of plant species varied significantly through space and time (Horvitz and Schemske, 1995). Plant population dispersal processes take place with variation in temporal conditions. Temporal variations in climatic conditions take place mostly due to changes in rainfall and temperature which may cause significant change in the conditions for seed germination (Horvitz and Schemske, 1995).

Buried seeds, in soil, do not germinate without suiTable amount of rainfall and surface temperature even with improved environmental and biotic conditions. Matured plants produce seeds with considerable spatiotemporal variations among individuals growing at different sites. Seed production and release is followed by the seed dispersal which is the only stage during which individual plants move in space (Chambers and MacMahon, 1994).

Plant demography has been found to be concerned with the temporal sequence of events from seed dispersal to germination, seedling emergence, and subsequent survival, maturation, and reproduction. All these are affected by various environmental factors (Harper, 1977; Houle et al., 2001). Not only the number, but also the dispersion pattern of the seeds from the seed rain to different soil conditions, affects the pattern of seedling recruitment and subsequent demographic processes because the environment is spatially heterogeneous (Houle, 1995).

Harper, (1977) and Houle et al., (2001) found that plant demography is concerned with the temporal sequence of events form seed dispersal to germination, seedling emergence, and subsequent survival, maturation, and reproduction. All these are affected by various environmental factors.

Sen and Datta, (1986) found that determinations of appropriate harvesting time for a plant for synthesis of maximum quantity of secondary metabolites and changes in plants in different seasons are very important in standardization of herbal medicines. In E. coronaria and R. serpentine, the total alkaloids as well as some individual alkaloids were studied during different seasons using latest scientific techniques. Total and individual alkaloids showed significant temporal variations. The experimental results clearly suggested definite changes in different seasons, which suggest the necessity of selecting a proper season and favorable environment (e.g. day length, temperature, rainfall etc.) for harvesting. Amaral et al., (2004) investigated the association between the decrease of phenolics in the walnut leaves in June and the rapid development of the fruits at the time when most of the nutrients and photo assimilates are employed in fruit growth considering that the light plays an important role in phenol biosynthesis (Treutter, 2001). Solar et al., (2006) stated the concentration of flavonoids increased from May (during vigorous spring growth) to December or from spring to winter.

Horticultural Aspects:

The growth and yield related attributes are mostly studied in cultivated farms and fields, while its analysis in the natural vegetation or wild population is rarely conducted, though of immense importance.

Mir-Ashfaq et al., (2012) stated that M. longifolia locally called as Yen belonging to family Lamiaceae, is a tall perennial herb of length 1 to 1.5 m with hairy stout stem. Leaves 4.7 cm long ovate and lanceolate. Flowers 0.4 cm to 0.8 cm in diameter having colour light and dark green or grey. The leaves of this plant are of great medicinal value.

Krishnan et al., (2000) studied the medicinal herb Acalyphaindica Linn. (Family: Euphorbiaceae) in deciduous forest of Alagar hill, South India to assess the effect of season and altitudinal variations on growth performance. They found that growth performance of A. indicain terms of number of leaves per plant and shoot height was better during monsoon than the summer season. Similarly, its best growth was recorded on hill top (550 m) than at mid hill (350 m) and foot hill (275 m). One way ANOVA revealed significant effect of altitude on all growth parameters.

Sher et al., 2010. concluded that summer temperature boost growth and yield of plant species. He further revealed that plant growth and development was directly related to the environmental factors. Summer rains and high water availability to plants enhance yield and yield related parameters. Furthermore, the increased over exploitation of medicinal plants is so alarming that many reputable organizations are of the view that cultivation of these plants is the only way to preserve this national wealth (WHO, 1993; BAH, 2004; Lambert et al., 1997.).

Khan. et al., (2011) conducted an experiment on antimicrobial activity of selected medicinal plants of Margalla Hills, Islamabad, Pakistan. The present studies focus on the antimicrobial activities of the crude

methanolic extracts of different plant parts of thirteen selected medicinal plants including Adhatoda vasica and Euphorbia hirta. E. hirta was most affective against E. coli by inhibiting the growth showing MIC value of 1.0 mg/ml.

Italo et al., (2009) investigated physical and chemical characteristics and insecticidal worth of melia (Melia azedarach L.). The diameter of M. azedarach fruit was comparatively lower. The flour obtained from green fruit had an average dry weight lesser than that of mature fruit. The average dry leaf weights were equal in both juvenile and mature stages. The green fruits had 50% initial humidity, similar to juvenile (60%) and mature (57%) leaves, but greater than the mature fruits (44%). The chemical analysis of the fruit maturity stages determined a slight increase in crude fiber content as maturity increased. There was a decrease in the lipid content of leaves close to 60% at maturity. Finally, the aqueous fruit and leaf extracts of M. azedarach were claimed to be effective insecticides on D. melanogaster.

Carpinella et al., (2003) studied anti feedant and Insecticide Properties of a Limonoid from Melia azedarach (Meliaceae) and concluded that M. azedarach fruit extract and its active principle could be effectively used in pest control programs. Ohiokpeha, (2003) worked on promoting the nutritional goodness of traditional food products. Nutrition/food security is a complex issue, which is affected by a national food security status. Shafique et al., (2007) identified that a mealy bug species could be used as an agent of biological control of Achyranthes aspera and Xanthium strumarium in Pakistan.

Ecological analysis of Vegetation

Research scientist have worked on different ecological aspects of medicinal plants in different ecological zones which are reviewed as below.

The forests of Himalayan mountains are full of medicinal plants indigenous to the valleys (Khan et al., 2013). Many of the natural products are derived from these medicinal plants. Majority of indigenous medicinal plant species are threatened or have become rare or endangered by large- scale exploitation (Nalawade et al., 2003; Khan et al., 2011) quantified the importance values of all the wild plant species identified at the University of Agriculture Peshawar, using quadrate method and concluded that higher the importance value of a weed, the greater is its competitiveness. The importance value of plant species is directly related to its relative density. Similarly, Liu et al., 2014 stated that plant density can be predicted with the change in topography of sites.

Skarpe, (1990) stated that the density and cover of woody plants were higher in the over-grazed areas than in the less disturbed vegetation. It was found that the differences in total abundance of plant species depend on variations in the amount of soil water and minerals available for plant growth while the differences in species composition, height and distribution were controlled by the site and seasonal variation of water in the soil. Uniyal et al., (2006) stated that different species had different habitat requirements. Steep slopes had the highest species richness and diversity, while rocky areas had lesser density. Maximum similarity regarding species distribution was observed between steep slopes and undulating meadows. Similarly temperature variation also predict species diversity, like low temperature encourage the growth of grasses and other herbs and the elevated temperature enhances shrub production and reduces non-vascular plants (Chapin, 1995; Chapin and Gaius, 1996; Escosa et al., 2000). Plant coverage also depends on the biotic factors and

the severity of disturbance significantly affects the coverage and forest structural development with different responses among sites (Kane et al., 2014).

Adnan et al., (2006) revealed threats to the sustainability of Ethno- Medicinal uses in Northern Pakistan (A Case Study of Miandam Valley, Swat, Kyber Pakhtunkhwa, Pakistan). They reported about 300 plant species, majority of which were medicinal and concluded that deforestation was the main threat behind the fact that majority of medicinal plants found were endangered, rare and vulnerable. They further revealed that each year 8053 trees are felled at the high deforestation rate of 2% per year over the last 30 years and hence badly affecting the ethno-medicinal uses and the socioeconomic conditions of the associated people. Kane et al., 2014 observed that biotic factors and disturbance significantly affected the relative coverage and importance value of wild plants. Type of soil and its structure influence species diversity and growth, The availability of macronutrients and high field capacity affect the importance value of various land races (Ahmad et al., 2008).

Utilization in modern medicines

Natural products have been an important source of drugs for centuries and about half of the pharmaceutical market presently depends on natural products (Clark, 1996).

Kumar et al., (2005) investigated the potential hypoglycemic and hypo lipidemic effect of Morus indica and Asystasia gangetica in alloxan induced diabetes mellitus. They concluded that the ethanolic extracts of leaves of Morus Indica and Asystasia gangetica had good hypoglycemic and hypolipidemic effect.

Arfan et al., (2009) conducted research on antioxidant activity of phenolic compounds of Mallotus philippinensis extract of its bark and Asia et al., (2004) conducted an experiment to investigate the active constituents of Portulaca oleraceae (portulacaceae) growing in jordan. The analysis of the fresh aerial parts of Portulaca oleracea (Portulacaceae), growing in Jordan, using conventional chromatographic procedures resulted in the isolation of ß-sitosterol, ß-sitosterol-glucoside, N,N`- dicyclohexylurea,and allantoin.

Ahmad et al., (1995) researched on studies of Punica granatum-I isolation and identification of some constituents from the seeds of Punica granatum. Initial phytochemical screening of the seeds of Punica granatum has revealed the presence of Ursolic acid and fl-Sitosterol along with a long straight chain hydrocarbon - nonacosene. Presence of estrogens and glycosides have also been detected.

Sahreen et al., (2011) noticed phenolic compounds and antioxidant activities of Rumex hastatus D. Don. Leaves. In this study different solvent fractions (n-hexane, ethyl acetate, chloroform, butanol and aqueous) of the methanol extract of Rumex hastatus (RH) leaves were evaluated for antioxidant activities. They substantiated that RH leaves can be used as a good source of potential antioxidant or functional food material due to the presence of sufficient amount of phenolics such as luteolin and kaempherol.

Sajid et al., (1991) indicated inhibition of adrenaline-induced aggregation of human platelets by Pakistani medicinal plants. The effects of extracts of Pakistani plants claiming medicinal value were observed on adrenaline-induced aggregation of human platelets. The minds manifested significant inhibitory activity in doses of 0.625 mgml-1 to 2.5 mgml-1 in this invitro model. The potency of the plant extracts in inhibiting platelet aggregation induced by adrenaline is in the following

order: Sida pakistanica, IC 50 = 0.90 mgml -1 >Tribulus terrestris, IC >Solanum strattense, IC 50 =1.34 mgml -1 50 = 0.97 mgml >Tephrosia subtnflora, IC =1.40 mgml -1 >Glycyrrhiza glabra (butanolic extract), IC 50 =166 mgml>Urtica dioica IC 50=2.17 mgml-1.

EXPERIMENT No-1

SPECIES DIVERSITY AND ETHNO BOTANICAL STUDY OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

Shah Masaud Khan and Noor ul Amin Department of Horticulture, The University of Agriculture Peshawar, Pakistan

ABSTRACT

Khanpur Valley, in the sub Himalayan Mountains of Pakistan, is a rich repository of diverse flora of immense medicinal importance. A detailed survey was conducted during 2010-2011 for documentation of indigenous flora of the valley. On the basis of preliminary survey of the valley and discussion with the inhabitants, four ecologically diverse sites namely Dam, Dabola, Jabri and Mang were selected for current study. The study comprised of three parts; firstly, all plants were documented, secondly, the available medicinal plants were identified and thirdly, the preferences of local people regarding the use of plants, its parts and forms for treatment of various ailments were studied. The data were collected through a balanced questionnaire and percent preferences were worked out. A total of 202 plant species belonging to 48 families were recorded. Maximum species were from family Asteraceae (19), followed by Poaceae (18), Fabaceae (12), Solanaceae (12), Euphorbeaceae (9), Brassicaceae (9) and Moraceae (9). Similarly maximum species were herbs (141), followed by trees

(31), while minimum species were shrubs (30). Moreover, out of total 202 plant species, 71 species (34%) belonging to 42 families were identified as medicinally important. The results revealed that the top 12 medicinally important species in terms of percent preferences were Adhatoda vasica (17.32%), Fumaria officinalis (13.22%), Ajuga bracteosa (12.27%), Euphorbia hirta (12.02%), Calostapis procera (8.12%), Recinus communis (7.42%), Mintha royaleana (7.06%), Berberis lyceum (6.41%), Punica granatum (5.61%), Xanthoxylum armatum (4.22%), Artemisia bevifolia (3.25%),and Solanum nigrum (3.08%). The findings further established that the largest number of ailments cured with medicinal plants were associated with the digestive system (36.05%) followed by respiratory disorders (14.83%), blood purification (14.42%), reproductive diseases (9.7%), skin problems (6.64%), urinary diseases (5.62%), nutritional & tonic supplement (5.21), brains & nerves (4.73%) and bones and joints (2.8%). Furthermore, the survey concluded that maximum preferences (37.52%) were expressed for whole plant in case of herbs followed by leaves (23.32%) of woody plants, roots (10.32%), rhizomes (8.36%), fruits (7.61%), bark (5.14%), seed (4.62%), flowers (2.02%) and shoots (1.21%) of both herbs and woody plants. Finally, the study found that maximum utilization of medicinal plants was in the form of powder (39.14%) followed by decoction (21.22%), tea (8.41%), paste (7.13%), fresh or raw form (7.1%), juice (6.04%), cooked (4.31%), cream (3.84%) and tincture (2.81%). The study concluded that out of 202 plant species found in Khanpur valley, 71 species were medicinally important. Whole plant in case of herbs and leaves of woody plants were preferred by the local people for use in various recipes against a variety of diseases most preferably associated with digestive system. Adhatoda vasica was the most preferred medicinal plant species while powder form was the most preferred form of utilization by the people of Khanpur valley. INTRODUCTION

Species diversity contributes to the main ingredients of medicines in traditional systems of healing and has been the source of inspiration for a large number of major pharmaceutical drugs. There are more than 8,000 plants species in South Asia used in the alternative medicines and are an integral part of traditional health care systems (Hussain et al., 2007). Pakistan, though, not among the biodiversity hot spots of the world, still faces immense challenges of conservation and sustainable utilization of biological resources. Degradation in natural resources is visible, caused by increased human activities related to the growing population coupled with, human destruction of natural habitats and migration of human population (Haq et al. 2010). This resulted in the change of land use

pattern, spread of invasive species, the growing demand for natural resources and its inappropriate management. In addition, neither systematic work has been carried out on the status and threats to ecosystems, nor the effects of global climate change are grasped by ecosystem managers. The management of an appropriate combination of resources, in various locations and under diverse conditions would be one of the efficient ways to conserve ecosystem that offers the medicinal wealth.

In developing countries it is estimated that birth attendants (dhais) assist in up to 95 % of rural and 70 % of urban births (Shinwari et al., 2005) and mainly relies on herbal medicines as pre and post maternity care. The herbal medicine have been used by locals since time immemorial. Research suggested that a large number of medicinces have been derived from the folk-use by the traditional civilizations (Shinwari et al., 2005). About 50 drugs have been discovered from ethno-botanical technologies through translating folk knowledge into new pharmaceuticals. Moreover, very few of the wide medicinal species have been domesticated globally and most of these species are still collected from their wild habitats (Gupta and Chadha 1995). Very little work has been undertaken on their selection and improvement, for developing suitable varieties.

Plant species have contributed significantly to the development of modern drugs. The use of medicinal plants is increasing worldwide due to expansion of traditional medicine and a growing interest in herbal treatments. Traditional Greek (Unani) medicine is quite a popular practice. It originated in Greece and was developed and documented by Muslims during the glorious period of Islamic civilization. This trade of medicine was introduced to the subcontinent by Muslim scholars and practiced successfully for centuries. In subcontinent, it gets benefited from the Ayurvedic system of medicine, which was an important component of Hindu civilization and all these systems of healing greatly

depend on wild plant collections. It has been estimated that around 95 % of the medicinal plants are harvested and collected in wild (Lucy and DaSilva 1999). Such excessive exploitation of natural resources is threatening to some plant species. The alarming levels of deforestation and ecosystem degradation have severely reduced the availability of medicinal plants and the overall environmental sustainability of the subcontinent. Due to high market and community demand many medicinal plants today, face either extinction or loss of genetic diversity (Lucy and DaSilva 1999).

Most of the wild plants have remarkable medicinal and economic value, but often only known to indigenous communities. Unfortunately, very little attention has been paid to the ethnobotanical aspect of plants as hakims (local healers) are only concerned with the floral and vegetative parts of medicinal plants without any regard to their botanical characteristics, or distribution in the various ecological zones of Pakistan. Herbs are not only used in the eastern system of treatment but also in the preparation of many allopathic and homeopathic drugs: despite the fact that these herbs are now being commercially exploited for the extraction of various active ingredients.

Pakistan is among the reasonably diverse countries in plant resources, where people's reliance on biological sources for the survival and wellbeing is very strong (Ahmad et al., 2008). Additionally, the country has rich and unbroken traditions of the use of medicinal plants and its natural products for healthcare needs (Zaman et al., 1997). A focus on medicinal plants raises some major questions of conservation and endangered species. Conservation and livelihoods are closely linked with medicinal plants. If conserved, medicinal plants will continue to be available to provide benefits for healthcare, income and support of cultural heritage. The overexploitation of habitats in terms of farm conversions and human interference is a major threat to biodiversity (Khan et al., 2013). The only

way to conserve the biological diversity is the involvement of all stakeholders and professionals to take it on war footings. like humans, animals do have been treated by herbal medicines, since long. It has been observed that livestock raisers and healers everywhere in the world have traditional ways of classifying, diagnosing, preventing and treating common animal diseases. Many of these "ethnoveterinary" practices offer viable alternatives or complement to conventional, western style veterinary medicines-especially where the latter is hardly available and costly or inappropriate. The use of medicinal plants constitutes major part of ethnoveterinary medicine (EVM) in Pakistan. Use of medicinal plants as an anthelmintic (de-wormer) is an example (Iqbal et al., 2005).

The sub Himalayan mountainous valley of Khanpur has a unique ecosystem which provides all eco-physiological support to its inhabitants. It has reserve forests, cultivated lands, range lands, water reservoirs, uplands, diverse plants and wildlife, as well as climatic extremes. The valley is at the gate way of great Himalayan Mountains and easily approached from different population centers like the capital city Islamabad and Taxila on its one side and Haripur and Abbotabad on the other side. So there is a huge pressure on the natural resources especially on medicinal flora of the valley, which otherwise magnify its immense scope for conservation.

Keeping in view the importance of species diversity, its traditional uses and conservation of the medicinal wealth, the current study was initiated by the Department of Horticulture University of Agriculture, Peshawar. The Khanpur Valley was selected for this study because the valley is very rich in indigenous medicinal flora and is located adjacent to medicinal and food industries at Hattar Industrial Estate. The Khanpur Valley can become a source of raw material supply to Hattar industries, if scientific methods of collection and utilization are adopted.

The current study was designed to achieve the following objectives:

1. To enlist the available plant species in the valley

2. To identify the medicinally important species of the valley

3. To determine the locally preferred medicinal plant species and its parts used in the preparation of different recipes for curing various ailments.

MATERIALS AND METHODS

The study titled “Species Diversity and Ethno Botanical Study of Khanpur Valley in the Sub Himalayan Mountains of Pakistan” was conducted during 2010-2011. An expert team of Hazara University, Mansehra accompanied the scholar in the initial visits for technical assistance to undertake this important study. At first instance during early 2010, the whole valley was extensively visited and agricultural, industrial & forest experts, local elders, herbalists and leaders were interviewed. These interviews were randomly face to face meetings and group discussions and the information collected was utilized in the designing and planning of the research project. The valley was divided into four ecologically diverse sites to cover all ecological diversities of the study area.

Selection of Sites:

On the basis of preliminary survey of the valley and discussion with stakeholders, four ecologically diverse sites namely Mang, Dam, Dabola and Jabri were identified and selected. These sites were different from one another in their environmental attributes especially variation in altitude,

slope, topography, habitat, vegetation type and plant community. A brief description of the selected sites is given below:

Dam Site: It is situated in the center of the valley with a beautiful lake (Khanpur Dam), Sarra reserved forests and downstream Khanpur village and fruit orchards, especially of citrus. Some prominent villages of this site are Darra, Khanpur old, Khanpur new, Tarnawa, Bhera, Julia, Pepla and March abad. Its central point the Dam is located at longitude 72o 55' 52.38 E and latitude 33o 48' 45.87 N with an altitude of 1940 feet.

Dabola Site: This site of Khanpur valley comprises area upstream from Tarnawa to Kohala villages. The Choi, Kamalpur, Najafpur, Denya, Dabola, Ranja, Chaskala, Shara, Besman, Desara, Bagla, Hally and Kohala are important villages of Dabola site. Dabola is located at longitude 73o 04' 33.36 E and latitude 33o 49' 50.73 N with an altitude of 3940 feet.

Jabri Site: It is situated at the north-east of the Khanpur valley. It comprises of area located between Stora and Jabri villages upto lora town. The main villages in this site are Stora, Jabri, Nulla, Banda, Ropra, Palla, Gambher, and Lora. Jabri is located at longitude 73o 10' 08.98 E and latitude 33o 54' 11.69 N with an altitude of 3120 feet.

Mang Site: It is situated at north-west of the khanpur valley. It comprises of both plain and hilly areas. Most of the area is dry scrub rangeland and rain fed fields surrounded by villages. The prominent village in Mang site are Mang, Pindmuneen, Gujar Nulla, Khui Nara, Masoom abad, Pind Gujran, Wajjian, Suraj Gali and Awan Rah on one side while Pind Kamal Khan, Kahil Pain, Thala, Noordi, Dooria, Kamal Poor, Rani Wah, Koka and Butri on the other side. Mang is located at longitude 72o 54' 49.24 E and latitude 33o 54' 21.12 N with an altitude of 1885 feet.

Selection of Seasons:

Medicinal plants of Khanpur valley were studied in two major seasons: winter and summer. Plants specimens for identification and data on various parameters were collected during October to March for winter and during April to September for summer season.

The study was divided into three parts.

1. Enlistment of the total plant species available at Khanpur Valley:

The research area was visited on weekly basis in both summer & winter and specimens of all the available species were collected from all of the four sites and brought to the herbarium of Hazara University, Mansehra, for identification by the experts and with the help of flora of Pakistan. The local names, common names, technical names, family names and type of plant or growth habit was properly documented.

2. Identification of Medicinally Important Plant Species.

Local herbalists, agricultural and forest experts, local elders, and leaders were interviewed and the plant species enlisted in the first part of the study were discussed with them. Moreover, the available literature on the subject was thoroughly studied and relevant information was used to help document the plant species which were medicinally important and available in the valley.

3. Determination of Community Preferences The preferences of local people for the treatment of various ailments in terms of medicinal plant used, part used, the category of diseases treated and the form of therapeutic use (recipe) were analyzed with the help of a questionnaire survey conducted in the whole valley. The questionnaire (as annexed in appendix 1.3) covered the following four major themes:

I. Most Preferred Medicinal Plant Species. The respondents were asked to mention the name of a medicinal plant species which was mostly preferred by the local people for traditional healthcare. The percent preferences for each mentioned species was calculated with the help of the following formula:

Percent Preferences of a Species = Number of respondents who termed it as most preferred Species*100 Total number of respondents

II. Most preferred Type/category of ailment, cured with medicinal plants. The respondents were asked to mention the type of an ailment which was mostly preferred by the local people for traditional healthcare. The percent preferences for each mentioned species was calculated with the help of the following formula:

Percent Preferences of an ailment = Number of respondents who termed it as most preferred ailment*100 Total number of respondents

III. Most preferred type of plant part, used. The respondents were asked to mention the name of a plant part which was mostly preferred by the local people for traditional healthcare. The percent preferences for each mentioned plant part was calculated with the help of the following formula:

Percent Preferences of a Plant part = Number of respondents who termed it as most preferred Part*100 Total number of respondents

IV. Most preferred form of utilization (recipe). The respondents were asked to mention the form of utilization or recipe which was mostly preferred by the local people for traditional healthcare. The percent preferences for each mentioned form or recipe was calculated with the help of the following formula:

Percent Preferences of a Recipe = Number of respondents who termed it as most preferred Recipe*100 Total number of respondents

A Total of one hundred (100) respondents, 25 from each of the four sites, were interviewed. In each site 5 most populous villages were selected and from each village 5 available eldest respondents were interviewed (Khan et al., 2012) and the data on all the parameters was recorded.

Statistical Analysis:

The data recorded was tabulated theme wise and Microsoft Excel program was used in the calculation of percent preferences and its presentation in graphic form.

RESULTS AND DISCUSSIONS

1. Enlistment of the total plant species available at Khanpur Valley:

The detailed list of total plant species enlisted at Khanpur valley is given in Appendix 1.1, while its summary of classification is given in Table1.1. Total species identified were 202 belonging to 48 families. Maximum species 19 were from family Asteraceae (Compositae), followed by 18 species belonging to Poaceae family while Fabaceae and Solanaceae families were found with 12 species each and Euphorbeaceae, Brassicaceae and Moraceae were revealed with 9 species each. Similarly, maximum species were of growth habit herbs (141), followed by shrubs (31), while minimum plant species were trees (29) in nature.

It is an obvious fact that the people of rural Pakistan extensively depends on herbal medicines for their traditional healthcare. The inhabitants of Khanpur valley are lucky to have hundreds of wild species available for use, as part of their ecosystem. The results show that Khanpur valley is very rich in terms of species diversity, as there were 202 total plant species belonging to 48 families, with 71 medicinally important species. Maximum species were from family Asteraceae (Compositae), followed by Poaceae family. Asteraceae and poaceae were two larger families of plant species indigenous to the valleys of Pakistan (Fazal et al., 2010). Similarly maximum species were of growth habit herbs, followed by trees, while minimum plant species were shrubs in nature. Similar results were obtained by Murad et al., (2011) for Malakand Agency, KP, Pakistan; Qureshi et al., (2009) for Chakwal district of Punjab, Alam et

al., (2011) for Chagharzai area Buner Pakistan, and Fazal et al., (2010) who documented 211 species of wild and cultivated plants of Haripur, Pakistan, with all mention of plant type and part used.

2. Identification of Medicinally Important Plant Species.

As a second part of the experiment, 71 species (35.15%) out of 202 total plant species were found to be medicinally important (Appendix 1.2). These medicinally important species were belonging to 42 families (Table 1.1). Maximum species were from family Solanaceae (8) followed by Asteraceae (7) followed by Euphorbeaceae (4) while Fabaceae and Moraceae were found with 3 species each. Similarly maximum species were of growth habit herbs (42) followed by shrubs (15) while minimum plant species were of growth habit tree (14).

The efforts on the ethnobotany and documentation of valuable flora have been carried out since long. Abbasi et al., (2009) reported 30 plant species belonging to 24 families used by local practitioners for the treatment of jaundice and hepatitis. Alam et al., (2011) concluded that 141 plant species of medicinal importance are found at Chagharzai Area of Buner district, Pakistan. Khan (1985) conducted another survey and reported that 95 species were used by Hakims and the annual consumption of medicinal plants was more than 5.65 million kg which valued approximately up to Rs. 36 million. Khan et al., (2012) identified use of Medicinal Plants in Folk Recipes by the Inhabitants of Himalayan Region Poonch Valley Azad Kashmir (Pakistan). Total 68 species of plants belonging to 44 families were recorded as used medicinally for preparations of folk recipes of 68 ailments. Leporatti and Lattanzi, (1994) studied 27 medicinal plants ethnobotanically in Makran (Southern Pakistan). They reported and discussed their traditional medicinal uses.

Table 1.1 Summary of Enlistment of Total Plant Species and Medicinally Important Plants found at Khanpur Valley.

Plant Species Families Total Species Asteraceae Poaceae Solanaceae Fabaceae Euphorbiaceae Moraceae Brassicaceae /Families

Total 19 18 12 12 9 9 9 202/ 48 Enlisted Medicinally 7 0 8 3 4 3 0 71 / 42 Important Growth Habit

Herbs %age Shrubs %age Trees %age Total enlisted 141 69.8 31 15.35 30 14.85 202/ 48

Medicinally 42 59.15 14 19.72 15 21.13 71 / 42 Important 3. Determination of Community Preferences

Community preferences for medicinal plant species, part used, ailment addressed and mode of utilization or recipe, was judged with a scientific survey and the results are given below:

I. Most preferred medicinal plant:

In terms of species preferences survey revealed that the top 12 medicinally important species were Adhatoda vasica (17.32%), Fumaria officinale (13.22%), Ajuga bracteosa (12.27%), Euphorbia hirta (12.02%), Calostapis procera (8.12%), Recinus communis (7.42%), Mintha royaleana (7.06%), Berberis lyceum (6.41%), Punica granatum (5.61%), Xanthoxylum armatum (4.22%), Artemisia bevifolia (3.26%),and Solanum nigrum (3.08%).

The people of various valleys and regions prefer specific medicinal plants, locally available. Lai, et al., 2005 studied medicinal plants and concluded that Euphorbiaceae is an important plant family especially recognized for its anti-cancer components, antihepatitis B components and carcinogenic factors. They further stated that only a few species were preferred as widespread medicines while most of its species were recognized only as preferred by one tribe or another. Similarly, Vallès et al., 2004. identified Sambucus nigra bush of family Caprifoliaceae as one of the most commonly used medicinal plant by the inhabitants of Catalonia and in many Mediterranean regions. The most preferred plant species of

Khanpur valley is Adhatoda vasica, which is among the dominant vegetation of hilly areas and frequently utilized in the treatment of respiratory disorders like cough, chronic bronchitis, asthma and other chest diseases (Amin et al., 1959).

Figure 1.1 Percentage of respondent`s preferences for species of medicinal plants.

II. Most preferred Type/category of ailment, cured with medicinal plants:

The results obtained on this parameter are given in Figure 1.2. The results revealed that the largest number of ailments cured with medicinal plants were associated with the digestive system (36.05%) followed by those associated with respiratory, blood purification, reproductive, skin, urinary, nutritional and tonic, brain and nerves and bones and tonic (14.83%, 14.42%, 9.7%, 6.64%, 5.21%, 5.62, 4.73 and 2.8% respectively).

Further finding were regarding ailments cured. The largest number of ailments cured with medicinal plants were associated with the digestive system followed by those associated with respiratory, blood purification, reproductive, skin, urinary, nutritional and tonic, brain and nerves and

bones and joints. The logic behind this finding could be the most commonplace diseases are associated with the digestive and respiratory disorders (Rasool et al., 2010; Jan et al., 2008) and hence the people of the research area use medicinal plants for these ailments. Similar results were obtained by Khan et al., (2012). who identified the use of Medicinal Plants in Folk Recipes by the Inhabitants of Himalayan Region, Poonch Valley of Azad Kashmir (Pakistan).

Figure 1.2 Percentage of respondent`s preferences for category of use, of medicinal plants. III. Most preferred type of plant part, used:

The questionnaire survey indicated that in maximum treatments whole plants (37.52%), in case of herbs and leaves (23.2%), in case of woody plants, followed by roots (10.32%), rhizomes (8.36%), fruits (7.61%), bark (5.14%), seed (4.62%), flowers (2.02%) and shoots (1.21%) were used by the local community.

Similarly the questionnaire survey showed that in maximum treatments whole plants in case of herbs and leaves in case of woody plants were used by the local community. This could be based on centuries old experiences for the efficacy of these parts.

Contrary to these results, Khan et al., (2013) concluded that whole plant and rhizomes are the most preferred parts utilized by the people of Naran valley. The possible reason for rhizome being the second most preferred part can be the temperate ecology of Naran valley contrary to the sub tropical climate of Khanpur valley. While Qamar et al., (2010) found that leaves and root are the preferred parts of medicinal importance in the Neelam Valley of Azad Kashmir, Pakistan.

Figure 1.3 Percentage of respondent`s preference for plant part used of medicinal plants.

IV. Most preferred form of utilization (recipe):

The third outcome of the survey was regarding the type of use (recipe). Which revealed that maximum utilization of medicinal plants was in the form of powder (39.14%) followed by decoction (21.22%), tea (8.41%), paste (7.13%), fresh (7.1%), juice (6.04%), cooked (4.31%), cream (3.84%) and tincture (2.81%).

The survey resulted in the finding that powder form followed by decoction were the most preferred forms of utilization by the local community of Khanpur valley. This may be due to ease of utilization in the preferred forms and is in connivance with the findings of other ethno botanists (Rasool et al., 2010; Jan et al., 2008; Khan et al, 2012). In line

with the findings of this experiment, Khan et al. (2013) has also reported the preferences of respondents for part used, ailments cured and form of recipe, in Naran Valley of Pakistan.

Figure 1.4 Percentage of respondent`s preference for traditional use (recipe) of medicinal plants

SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

This is an important aspect of ecosystem and socio-economic interaction which reveals ethno botanical trends and unveils the dynamics of rural Pakistan. Species diversity study which results in the enlisting and documentation of the indigenous plants always provides base line information for any scientific undertaking. Khanpur Valley, in the sub Himalayan Mountains of Pakistan, is a rich repository of diverse flora of great medicinal importance. A detailed survey was conducted during 2010-2011 for documentation of indigenous wild plant species of the valley. On the basis of preliminary survey of the valley and discussion with the inhabitants, four ecologically diverse sites namely Dam, Dabola, Jabri and Mang were selected for this important study. The study comprised of three parts; firstly, all plants were documented, secondly, the available medicinal plants were identified and thirdly, the preferences of local people regarding the use of plants, its parts and forms for treatment of various ailments were studied. The data were collected through a balanced questionnaire and percent preferences were determined.

A total of 202 plant species belonging to 48 families were recorded. Maximum species were from family Asteraceae (19), followed by Poaceae (18), Fabaceae (10), Solanaceae (10), Euphorbeaceae (9), Brassicaceae (9) and Moraceae (9). Similarly maximum species were herbs (141), followed by trees (31), while minimum species were shrubs (30). Moreover, out of total 202 plant species, 71 species (34%) belonging to 42 families were identified as medicinally important. The results revealed that the top 12 medicinally important species in terms of percent preferences were Adhatoda vasica (17.32%), Fumaria officinalis (13.22%), Ajuga bracteosa (12.27%), Euphorbia hirta (12.02%),

Calostapis procera (8.12%), Recinus communis (7.42%), Mintha royaleana (7.06%), Berberis lyceum (6.41%), Punica granatum (5.61%), Xanthoxylum armatum (4.22%), Artemisia bevifolia (3.25%),and Solanum nigrum (3.08%). The findings further established that the largest number of ailments cured with medicinal plants were associated with the digestive system (36.05%) followed by respiratory disorders (14.83%), blood purification (14.42%), reproductive diseases (9.7%), skin problems (6.64%), urinary diseases (5.62%), nutritional and tonic supplement (5.21), brain and nerves (4.73%) and bones and joints (2.8%). Furthermore, the survey concluded that maximum preferences (37.52%) were expressed for whole plant in case of herbs followed by leaves (23.32%) of woody plants, roots (10.32%), rhizomes (8.36%), fruits (7.61%), bark (5.14%), seed (4.62%), flowers (2.02%) and shoots (1.21%) of both herbs and woody plants. Finally, the study found that maximum utilization of medicinal plants was in the form of powder (39.14%) followed by decoction (21.22%), tea (8.41%), paste (7.13%), fresh or raw form (7.1%), juice (6.04%), cooked (4.31%), cream (3.84%) and ticture form (2.81%).

During this experiment three important spheres of the research area were addressed. At the beginning the whole valley was searched for the available plant species and 202 plant species, which is reasonably high number of plant species were recorded for the first time in the valley. After enlisting the indigenous species, the medicinally important plants were investigated and 71 plants were identified to be of medicinal worth. And finally the local preferences for species utilization as medicinal plant (Adhatoda vasica), part of plant preferred (leaves and whole plant), ailment thought to be preferably treated (digestive disorders) and trends of recipe of utilization (powder form) were quantified on the bases of community responses to the questionnaire survey.

Conclusions: The following conclusions can be derived from the current study.

• The 202 species, belonging to 48 different families, recorded at Khanpur valley shows that the valley is rich in species diversity. • The 71 medicinally important species were identified at the research area which means that 35.15% of the flora of khanpur is pharmaceutically important. • The local preference for species revealed that Adhatoda vasica, Fumaria officinallis and Ajuga bracteosa are the most preferred medicinal plants of the valley. • It was concluded that the most preferred part of plant used is whole plant in case of herbs and leaves in case of woody plants. • The survey found that the people of Khanpur prefer medicinal plants used in the cure of ailments related to digestive and respiratory systems and they prefer powder and decoction form of its utilization.

Recommendations: Following are some of the recommendations synthesized from the study and based on the research findings and conclusions.

• Being a repository of 202 plant species with 71 medicinal plants, it will be worthwhile if a herbal garden of indigenous medicinal flora is established in the valley to preserve this national wealth. • The people of Khanpur valley prefer medicinal plants (Adhatoda vasica, Fumaria officinallis and Ajuga bracteosa) used in the cure of ailments related to digestive and respiratory systems. These plant species are recommended to be brought into cultivation for promotion and conservation.

• As khanpur valley is in close vicinity of Hattar Industrial Estate a linkage between the local community and industry at Hattar, may be established to supply raw material from the wild for products preparation. Adhatoda vasica which is used in Joshanda can be the one to start with. • Local community need trainings and awareness about the usefulness and sustainable utilization of the herbal plants to make this invaluable source of living available to the generations to come.

EXPERIMENT No-2

ECOLOGICAL ANALYSIS OF THE VEGETATION OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

Shah Masaud Khan and Noor ul Amin

Department of Horticulture, The University of Agriculture Peshawar, Pakistan

ABSTRACT

An ecological analysis of the vegetation of Khanpur valley in the sub Himalayan Mountains of Pakistan was undertaken during the years 2011- 2012. The valley was divided into four ecologically diverse sites, namely Dam, Dabola, Jabri and Mang. Data was collected both in summer and winter seasons separately. A total of thirty quadrates, fifteen each for herbs and woody plants, were taken to study frequency, density, coverage, relative frequency(RF), relative density(RD), relative coverage(RC) and importance value(IV) for all recorded plant species. The data was analyzed using Canoco Computer Package for windows (version 4.5). Findings revealed that a total of 116 ecologically important species (Frequency≥1%) were found in all the four sites in both seasons. Maximum number of plant species (102) were found at Dabola site in summer season while minimum species (73) were observed at Dam site during winter season. The maximum density was recorded for plant species associated with Dabola and Jabri sites. The ordination plots revealed that Dam site is ecologically correlated with Mang site while Dabola site is correlated with Jabri site for all the parameters. The results further showed that species ecologically associated with summer were Adiantum venusetum, Cyperus retendus, Euphorbia granulate, Euphorbia hirta, Hedera hilex, Matricaria chamomile, Medicago polymorpha, Mintha ryaliana, Nasturtium officinale, Plantago major, Pteridium equilinium, Skimmi lanreola and Xanthium strumarium, while those associated with winter were Foeniculum vulgarus and Plantago ovata. Species like Ajuga bractiosa, Fragaria vulgaris, Mallotus phillipensis, Melia azadaracta, Punica granatum and Urtica dioica were found associated with Dabola site, species Acacia modesta, Medicago denticulate, Rumex dentatus and Themeda triandra with Dam site, species Olea ferrugineae Mentha arvensis, Nasturtium officinale and Stellaria media at Jabri site while species Adhatoda vasica, Fumaria officinalus, Rumax longifolius and Urtica dubia were found more frequent at Mang site. The results also established that majority of the species fall in the class of Rare (45.37%), followed by Occasional (19.72%), then Frequent (5.06%) and least fall in the class of Abundant (2.05%). While there was not a single species found as Very Abundant during any season and at any site of the valley. The overall finding of the experiment was the fact that species were ecologically associated with summer and Dabola site to be collected therein, while majority of the species were rare and hence need conservation.

INTRODUCTION

An effective plan for conservation and sustainable utilization of plant species needs the in-depth study of species dynamics at all growing

seasons, various habitats and sites as well as analysis of prevailing ecological and ethno botanical factors. The Khanpur valley is located at the door step of Himalayan Mountains and its ecosystem presents an introduction to the ecology of the Mountains. The occurrence of various species and its association with various ecological zones and temporal variations reflects the strengths of biodiversity and ethno-ecological value of the indigenous flora (Alam et al., 2011). The study of ecological importance of local flora and its value can help the industries utilizing the raw medicinal plant parts to determine the magnitude and time of collection and thus the judicious use of indigenous plant species can be ensured.

The temperate and subtropical zones of Pakistan deserve specific focus for the exploration and conversation of species diversity. During the last hundred years, the zone has been subjected to major structural variations, resulting in the decrease of about fifty percent of the forest area (Iqbal, 2012). The decrease in forest cover as well as major shifts in community structure, are the causes of decline of indigenous plant resources and its utilization in the traditional healthcare (Kane et al., 2014). It is of utmost importance that research projects, be initiated in climatically diverse valleys of Pakistan and valuable medicinal flora needs to be documented and analyzed for its usefulness in human healthcare, in agriculture and as food supplement. The current study at khanpur valley was taken as a test case in this regard.

The Khanpur valley in the sub Himalayan mountain range of Pakistan is located between longitudes 72° 35' to 73° 15' and latitudes 33° 44' to 34° 22', in the Haripur District of Khyber Pakhtunkhwa (Fazal et al., 2010). The valley is rich in natural resources and has very important water reservoir, the Khanpur Dam. This beautiful valley with a lake / dam on Haro River is 45 km from Islamabad on Taxila-Haripur Road. Khanpur

Dam is a multipurpose project which supplies drinking water to the twin cities of Islamabad & Rawalpindi and irrigation water to Khyber Pukhtoonkhwa with 110 cusecs and Punjab with 87 cusecs (Wassi-ur- Rehamn. 2012). The Catchment area of the Haro River comprises of four main streams i.e. Lora Haro, Stora Haro, Neelan Stream and Kunhad Stream.

There are five types of forests at Khanpur Valley (Wassi-ur-Rehamn. 2012). Reserved Forests or Beer: The area of Reserved Forests, starts from New Khanpur Town, and scattered up to Nathiagali and Muree, along the sides of River Haro. All these forests are under the control of Khyber Pakhtukhwa Forest Department. Protected Forests or Guzaras: These forests are owned by the local people and protected by Forest Department. Cooperative Forests or Mehdoodas: The land of people was taken by the Forest Department for plantation about 50 years ago and is under management of Forest Department. Private Forests & Farms: There are some private Farms and Forests in the area which are exclusively owned and managed by the local people as their personal enterprises. Graveyard Forests: Large and old Graveyards almost in every village have become thick forests. These are owned by community as combined property and are often less interfered.

The forests of Himalayan mountains are full of medicinal plants indigenous to the valleys (Khan et al. 2013). Many of these natural resources are derived from species that are threatened or have become rare or endangered by large-scale exploitation (Nalawade et al. 2003; Cole et al. 2007). The species of medicinal plants, its frequency, volume, and coverage is still unknown. The local Herbalists collect valuable medicinal flora and utilize it in herbal medicines preparation. Furthermore, the valley is in close vicinity with Herbal industries of Hattar Industrial Area, Haripur and its exploration for its medicinal wealth can be a source of raw

material supply for the Industries and a source of income for the poor population of the valley. This research project is destined to open new venues of research for the domestication, cultivation and improvement of medicinal plants indigenous to other valleys of Pakistan. It may lead to the cultivation of medicinal plants indigenous to Khanpur valley and their commercial utilization by herbalists and pharmaceutical industries, located at the Industrial Estate Hattar, near Khanpur valley.

Having the above mentioned scenario in view, the current study was carried out with the aims to:

1. Find out the distribution pattern of the vegetation at Khanpur Valley. 2. Identify the ecologically important species (EIS) indigenous to Khanpur Valley. 3. Classify the existing flora on the basis of its frequency. 4. Highlight the most suiTable season and site for plant collection.

MATERIALS AND METHODS

The study on ecological analysis of vegetation of Khanpur valley in the sub Himalayan mountains of Pakistan was conducted during 2011 and 2012. The whole valley was divided into four different sites namely: Dam, Dabola, Jabri and Mang (details of the sites are documented in the materials and methods of experiment-1 page 6-7). The study aimed to assess the site-season variation of plant species indigenous to the region. Data was recorded in both summer (April-September) and winter (October-March) seasons separately. The ecological data was recorded using quadrate sampling method. Fixed quadrates of 1m2 for herbs and 25m2 for shrubs and trees were used. All individual plants in each quadrate were taken into consideration. Randomly 15 quadrates for herbs and 15 quadrates for shrubs and trees were taken at each site in both the

seasons. The data was recorded on properly designed data sheets as per methods suggested by Cox (1996). The recorded data was used for the calculation of density, frequency, cover/dominance, relative density, relative frequency, relative cover/dominance, importance value and classification of species, with the help of standard formulas (Bonham, 1989).

Parameters Studied: The ecological data on each species found was recorded with the help of the following parameters:

1. Density: Total number of individual plants of all species were counted in each quadrate and recorded in the data sheet followed by species wise number of plants per quadrate. The percent density of each species found was calculated with the help of the following formula:

Total Number of individuals of a species in a quadrate x 100 Density % = Total Number of individuals of all species in a quadrate

2. Frequency: The occurrence (1 for present and 0 for absent) of plant species in all the quadrates was noted down on the data sheet. The frequency of individual species for all species found in all sites and seasons were calculated with the help of the following formulas:

Number of quadrates in which a species occurred x 100 Frequency % = Total Number of quadrates taken Species found in at least one of any quadrat taken in the whole valley (frequency%≥1) was considered as ecologically important.

3. Coverage/Dominance: Percent Coverage of a species was recorded with the help of 1 meter2 quadrate with 100 squares, specially designed for herbs, 100 meter long plastic rope, measuring tap and iron poles. Line intercept method was used to measure the coverage

in case of shrubs and trees. With the help of these tools area covered per quadrate by all species and by individual species was recorded and percent coverage per species was calculated with the help of the following formula:

Area covered by a species in a quadrate x 100 Coverage/Dominance % = Total area covered by all species in a quadrate

4. Relative Density: Relative density of a species was calculated with the help of the following formula:

Density of aparticular species in a site x 100 Relative Density % = Total density of all species in a site

5. Relative Frequency (%): Relative Frequency of a species was calculated with the help of the following formula:

Frequency value of a species in a site x 100 Relative Frequency % = Total frequency values of all the species in a site

6. Relative Coverage (%): Relative Coverage of a species was calculated with the help of the following formula:

Coverage of a species in site x 100 Relative Coverage % = Total coverage for all the species in a site

7. Importance value (IV): Importance value of a species was calculated with the help of the following formula:

Importance value = Relative Frequency +Relative Density +Relative Coverage

8. Classification of Species:

Ecologically Species were classified on the basis of its frequency at various sites and seasons with the help of the key (System Kult) developed by Kult (1947) based on the following criteria: Frequency Range (%) Category Symbol

F = ˂1 Threatened T

1------20 Rare R

21------40 Occasional O

41------60 Frequent F

61------80 Abundant A

81------100 Very Abundant VA Statistial Analysis:

The data for various ecological attributes in all the four sites and two seasons was analyzed using Canoco Computer Package for Windows (version 4.5).

RESULTS AND DISCUSSIONS

Total 116 species were identified at all the quadrates taken in both summer and winter at all the four sites of Khanpur valley. The detailed list of these ecologically important species is given in legends below each Table. Furthermore, the serial number given in legends has been used as species number and the species number represent the species in Ordination Biplots (figures) and in results & discussions.

1. Density

The Partial Canonical Correspondence Analysis (pCCA) of density data regarding seasons as environmental variables and sites as co-variables revealed highly significant response of density of various plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.034) along axis 1 (Table 2.1). The pCCA Ordination biplot of species density and seasons as environmental variables showed that most of the species were centroid (Figure 2.1). The density of a number of species was closely associated with summer and few (with higher density values) were associated with winter. Most of the species had equal density values for summer and winter. Species no 64, 29 and 76 were densely populated, their density was significantly affected by seasons and were closely associated with summer while species no 3, 6, 30, 38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were found with low density values but observed only at summer. similarly species no 84, 77, 94, 39 and 19 were densely populated, their density was significantly affected by seasons and were closely associated with winter while species no 60, 71 and 82 were found with low density values but associated with both seasons.

The pCCA Ordination biplot of density data with respect to sites as environmental variables and seasons as co-variable also showed highly

significant variation (P=0.0020 and eigenvalue=0.043) along axis1 (Table 2.1). pCCA ordination bi plot of species density and different sites showed that majority of the species were associated with Jabri site (Figure 2.2). Most of species were dense at Jabri and Dabola sites. Species no 68, 72 and 102 were associated with Jabri site while species number 8 and 50 were densely found in association with Dabola site. While species number 106, 92 and 64 were associated with Dam site and species number 51, 94 and 110 were found dense at Mang site. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang and jabri sites while Dabola is correlated with Jabri and Mang sites. Maximum plant species (102) were found at Dabola site in summer season while minimum species (73) were observed at Dam site during winter season. From the results it can be concluded that among medicinal plants, Adhatoda vasica and Fumaria officinalis were the most abundant species in the Khanpur Valley.

The spatial variations might be due to the soil type and its composition, elevation of sites, moisture contents of soil, nature of disturbance like grazing pressure, human interference and isolation of study site populated regions (Hussain, 2007). The seasonal variations in plant density were related to rain fall, temperature changes and availability of essential nutrients during different seasons. The medicinal plants Adhatoda vasica and Fumaria officinalis were the most abundant species in the Khanpur Valley that might be due to its deep root system and adaptations to various types of environments (sites and seasons) (Ahmed, 2002; Hussain, 2007; Skarpe, 1990; Nanette et al., 2007).

The association of density of a number of species with summer might be due to favorable environmental conditions and association with winter was due the growth habit of such herbs to sprout in winter (Zaman, 1997). The results of this experiment were in accordance with sharma et al.,

(1983); Cole et al., (1987); Dalsted, (1988); Austin and Heylgens, (1989); Smitheman and Perry, (1990) who supported the principles mentioned above and discussed the plant communities of different sites of the world. Liu et al., (2014) stated that plant density can be predicted with the change in topography of sites.

Table-2.1. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the density of plant species found at Khanpur Valley. Parameters and data Axes Total F ratio P value 1 2 3 4 Inertia Density (Seasons/Sites) Eigenvalues 0.034 0.031 0.000 0.000 0.065 103.662 0.0020 Sum of all canonical 0.034 51.831 0.0020 Eigenvalues Density (Sites/Seasons) Eigenvalues 0.043 0.019 0.006 0.037 0.141 44.856 0.0020 Sum of all canonical 0.068 24.039 0.0020 Eigenvalues Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.

1.Acacia modesta 2.Acacia muricata 3.Acacia victoriae 4.Adhatoda vasica 5.Adiantum pedatum 6. Adiantum venusetum 7.Ailanthus altissima 8.Ajuga bracteosa 9.Arandu domax 10.Artemisia absenthium 11.Artimisia bevifolia 12.Avena fetua 13.Barleria priontis 14.Berberis vulgarus/lyceum 15.Boerhavia deffusa 16.Bohina variegata 17.Bombax malabaricum 18.Calatropis procera 19.Canabus sativa 20.Capsella bursa 21.Carthamus oxycantha 22.Cassia fistula 23.Cassia sophera 24.Celtis australis 25.Chemopodium album 26.Clitaria annua 27.Convulvolus arvensus 28.Conyza candensis 29.Cynodan Dactylon 30. Cyperus retendus 31. Dalburgia sisso 32. Datura alba 33. Datura stramonium 34. Desmodium elegans 35. Dodonia viscose 36. Echinochloa colona 37.Echinops echinatus 38.Euphorbia granulate 39.Euphorbia helioscopia 40.Euphorbia heterophylla 41.Euphorbia hirta 42.Euphorbia humifusa 43.Ferula assafoetida 44.Ficus hispida 45.Ficus jahaninsis 46.Ficus palmate 47.Ficus varigata 48.Foeniculum vulgarus 49.Fragaria orientalis 50.Fragaria vulgaris 51.Fumeria officinalis 52.Grewia optivia 53.Grewia tenax 54.Hedera hilex 55.Hedera nepalensis 56.Imperata cylindrical 57.Indigofera spicata 58.Iris ensata 59.legerstroemia speciosa 60.lyceum barbarum 61.Mallotus phillipensis 62.Malvestrum caromandilianum 63.Matricaria chamomile 64.Medicago denticulate 65.Medicago polymorpha 66.Melia azadaracta 67.Melilotus parviflora 68.Mentha arvensis 69.Mintha royaliana 70.Morus indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium

oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta 104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

Figure 2.1. pCCA Ordination biplot showing the effect of different seasons on the density of plant species found at Khanpur Valley.

Figure 2.2. pCCA Ordination biplot showing the effect of different sites on the density of plant species found at Khanpur Valley.

2. Frequency

The partial canonical corresponding analysis (pCCA) of frequency data regarding seasons as environmental variables and sites as co-variables showed highly significant variation in frequency of different plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.014) along axis 1(Table 2.2). The pCCA Ordination biplot of species frequency and seasons as environmental variables showed that most of the species were centroid (Figure 2.3). The frequency of a large number of species was closely associated with summer and few were associated with winter. 85

Most of the species had equal frequency values for summer and winter. Species no 82, 23 and 48 were closely associated with winter while species no 3, 6, 30, 38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were associated with summer.

The pCCA Ordination biplot of frequency data with respect to sites as environmental variables and seasons as co-variable also showed highly significant variation (P=0.0020 and eigenvalue=0.090) along axis1 (Table 2.2). pCCA ordination bi plot of frequency of species and different sites showed that majority of the species were associated with Dabola site (Figure 2.4). Most of species were frequent at Dabola and Dam sites. species no 87, 61, 109 and 66 were frequent at Dabola site. At Dam species no 1 at Mang species no 4 and at Jabri site species no 74, were found frequently associated. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site.

Khanpur Valley is very rich in species diversity, entailing a treasure of valuable plant population. Seasons and sites both had a significant effect on the frequency of plant species, though most of the species were centroid . Plant demography is concerned with the temporal sequence of events form seed dispersal to germination, seedling emergence, and subsequent survival, maturation, and reproduction. All these are affected by various environmental factors (Harper, 1977; Houle et al., 2001). In this study, the frequency of a large number of species were closely associated with summer which most probably was due to the presence of suiTable temperature, enough moisture and nutrients during summer (Skarpe, 1990; Zaman, 1997; Nanette et al., 2007). Another reason can be that the elevated temperature enhances shrub production and reduces non- vascular plants (Chapin, 1995 and Chapin & Gaius, 1996).

Similarly the reason behind the association of a large number species with Dabola site, can be the steep hilly topography of the site. In a similar experiment Uniyal et al. 2006 had concluded that steep slopes had the highest species richness and diversity than plains. Moisture loving and moderately moisture requiring species were associated with Dam Site which bear moist soil and humid environment due to the presence of water reservoir and low altitude (Misbahuzzaman & Alam, 2006; Malik et al., 2007; Ahmad et al., 2008). At Dam Acacia modesta, at Mang site Adhatoda vasica and at Jabri site Olea ferrugineae were found frequently associated. The correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site. The reason for this correlation is the temporal, topographic and altitudinal similarities between the two correlated sites (Chapin, 1995; Chapin & Gaius, 1996 and Liu et al., 2014).

Table-2.2. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the frequency of plant species found at Khanpur Valley. Parameters and data Axes Total F ratio P value 1 2 3 4 Inertia Frequency (Seasons/Sites) Eigenvalues 0.014 0.019 0.000 0.000 0.033 84.657 0.0020 Sum of all canonical 0.014 42.329 0.0020 Eigenvalues Frequency (Sites/Seasons) Eigenvalues 0.090 0.011 0.008 0.051 0.180 111.271 0.0020 Sum of all canonical 0.109 42.802 0.0020 Eigenvalues Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.

18.Calotropis procera 19.Canabus sativa 20.Capsella bursa 21.Carthamus oxycantha 22.Cassia fistula 23.Cassia sophera 24.Celtis australis 25.Chemopodium album 26.Clitaria annua 27.Convulvolus arvensus 28.Conyza candensis 29.Cynodan Dactylon 30. Cyperus retendus 31. Dalburgia sisso 32. Datura alba 33. Datura stramonium 34. Desmodium elegans 35. Dodonia viscose 36. Echinochloa colona 37.Echinops echinatus 38.Euphorbia granulate 39.Euphorbia helioscopia 40.Euphorbia heterophylla 41.Euphorbia hirta 42.Euphorbia humifusa 43.Ferula assafoetida 44.Ficus hispida 45.Ficus jahaninsis 46.Ficus palmate 47.Ficus varigata 48.Foeniculum vulgarus 49.Fragaria orientalis 50.Fragaria vulgaris 51.Fumeria officinale 52.Grewia optivia 53.Grewia tenax 54.Hedera hilex 55.Hedera nepalensis 56.Imperata cylindrical 57.Indigofera spicata 58.Iris ensata 59.legerstroemia speciosa 60.lyceum barbarum 61.Mallotus phillipensis 62.Malvestrum caromandilianum 63.Matricaria chamomile 64.Medicago denticulate 65.Medicago polymorpha 66.Melia azadaracta 67.Melilotus parviflora 68.Mentha arvensis 69.Mintha ryaliana 70.Morus indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta 104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

Figure 2.3. pCCA Ordination biplot showing the effect of different seasons on the frequency of plant species found at Khanpur Valley.

Figure 2.4. pCCA Ordination biplot showing the effect of different sites on the frequency of plant species found at Khanpur Valley.

3. Plant Cover

The Partial Canonical Correspondence Analysis (pCCA) of coverage data regarding seasons as environmental variables and sites as co-variables showed highly significant variation in coverage of different plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.018) along axis 1(Table 2.3 ). pCCA Ordination biplot of species coverage and seasons as environmental variables showed that most of the species were centroid (Figure 2.5). The coverage of a number of species was closely associated with summer and few were associated with winter. Most of the species had equal coverage values for summer and winter. Species no 82, 23 and 48 were closely associated with winter while species no 3, 6, 30,

38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were observed only at summer.

The pCCA Ordination biplot of coverage data with respect to sites as environmental variables and seasons as co-variable also showed highly significant variation (P=0.0020 and eigenvalue=0.075) along axis1 (Table 2.3). pCCA ordination bi plot of coverage of species and different sites showed that majority of the species were associated with Dabola site (Figure 2.6). Most of species were having maximum coverage at Dabola.site. Species no 87, 61and 109 were of high coverage at Dabola site. At Dam site, species no 1, at Mang site species number 4 and at Jabri site species no 74, were found associated. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site.

The obvious reason for more coverage at summer than winter might be the sufficient availability of moisture, nutrients and sunlight at summer and their deficiency during winter as concluded by Skarpe, (1990) and Nanette et al., (2007). Similar results were obtained by Ahmad et al., (2008) who pointed out another reason for low coverage at winter as the excessive use of the aerial parts of trees and shrubs by the residents as fuel and fodder during winter.

Most of species were having maximum coverage at Dabola.site followed by Jabri site depending upon the availability of enough nutrients, conducive temperatures and sufficient water as stated by Dupont and Plummer, (1997) and Nanette et al., (2007). Here it is worth mentioning that at Dabola and Jabri which are at high altitude and comparatively cool climate sites were having herbs or annuals prevailing everywhere. On other hand the Dam and Mang sites which are comparatively at low altitude and hot were found with more vascular plants (shrubs and trees) than herbs. Chapin, (1995) and Chapin & Gaius, (1996) concluded that

elevated temperature enhances shrub production and reduces non- vascular plants. Similarly, Liu et al., (2014) found that plant coverage depends on the biotic factors and the severity of disturbance.

Table-2.3. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the coverage of plant species found at Khanpur Valley.

Parameters and Axes Total F ratio P value data Inertia 1 2 3 4 Plant Cover (Seasons/Sites) Eigenvalues 0.018 0.023 0.000 0.000 0.041 90.528 0.0020 Sum of all canonical 0.0018 45.256 0.0020 Eigenvalues Plant Cover (Sites/Seasons) Eigenvalues 0.075 0.016 0.011 0.065 0.204 65.033 0.0020 Sum of all canonical 0.102 28.097 0.0020 Eigenvalues Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.

indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta 104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

Figure 2.5. pCCA Ordination biplot showing the effect of different seasons on the coverage of plant species found at Khanpur Valley.

Figure 2.6. pCCA Ordination biplot showing the effect of different sites on the coverage of plant species found at Khanpur Valley.

4. Relative Density

The Partial Canonical Correspondence Analysis (pCCA) of relative density data regarding seasons as environmental variables and sites as co- variables showed highly significant variation in relative density of different plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.016) along axis 1(Table 2.4). pCCA Ordination biplot of

species relative density and seasons as environmental variables showed that most of the species were centroid (Figure 2.7). The relative density of a large number of species was closely associated with summer and few were associated with winter. Most of the species had equal relative density values for summer and winter. Species no 82, 23, 20 and 48 were closely associated with winter while species no 3, 6, 30, 38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were having relative density only at summer.

The pCCA Ordination biplot of relative density data with respect to sites as environmental variables and seasons as co-variable also showed highly significant variation (P=0.0020 and eigenvalue=0.055) along axis1 (Table 2.4). pCCA ordination bi plot of relative density of species and different sites showed that the species were associated equally with all the four sites (Figure 2.8). The species number 73, 61 and 109 were associated with Dabola site, species no 8, 85 and 74 were found associated with Jabri site, 89, 110, and 59 were associated with Mang site and species number 7 and 47 were found associated with Dam site. however species number 114 and 87 were equally relatively frequent between Dabola and Jabri sites. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site.

Relative Density of a species is directly proportional to its density and it is an indicator of importance value of a species. The relative density of plant species was of the same trend as density and hence the variations at sites might be due to the soil type and its composition, elevation of sites, moisture contents of soil, nature of disturbance like grazing pressure, human interference and isolation of study site populated regions (Hussain, 2007). The seasonal variations in the relative density were related to rain

fall, temperature changes and availability of essential nutrients during different seasons.

The results of this experiment were in accordance with many scientests (sharma et al., 1983; Cole et al., 1987; Dalsted, 1988; Austin and Heylgens, 1989; Smitheman and Perry, 1990) who supported the principles mentioned above and discussed the plant communities of different sites of the world. Liu et al., (2014) stated that the topography of different sites determine the relative density.

Table-2.4. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Relative Density of plant species found at Khanpur Valley. Parameters and data Axes Total F ratio P value 1 2 3 4 Inertia Relative Density (Seasons/Sites) Eigenvalues 0.016 0.025 0.000 0.000 0.041 69.842 0.0020 Sum of all canonical 0.016 34.921 0.0020 Eigenvalues Relative Density (Sites/Seasons) Eigenvalues 0.055 0.012 0.007 0.073 0.178 49.654 0.0020 Sum of all canonical 0.075 20.030 0.0020 Eigenvalues Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.

Cyperus retendus 31. Dalburgia sisso 32. Datura alba 33. Datura stramonium 34. Desmodium elegans 35. Dodonia viscose 36. Echinochloa colona 37.Echinops echinatus 38.Euphorbia granulate 39.Euphorbia helioscopia 40.Euphorbia heterophylla 41.Euphorbia hirta 42.Euphorbia humifusa 43.Ferula assafoetida 44.Ficus hispida 45.Ficus jahaninsis 46.Ficus palmate 47.Ficus varigata 48.Foeniculum vulgarus 49.Fragaria orientalis 50.Fragaria vulgaris 51.Fumeria officinale 52.Grewia optivia 53.Grewia tenax 54.Hedera hilex 55.Hedera nepalensis 56.Imperata cylindrical 57.Indigofera spicata 58.Iris ensata 59.legerstroemia speciosa 60.lyceum barbarum 61.Mallotus phillipensis 62.Malvestrum caromandilianum 63.Matricaria chamomile 64.Medicago denticulate 65.Medicago polymorpha 66.Melia azadaracta 67.Melilotus parviflora 68.Mentha arvensis 69.Mintha ryaliana 70.Morus indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta 104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

Figure 2.7. pCCA Ordination biplot showing the effect of different seasons on the relative density of plant species found at Khanpur Valley.

Figure 2.8. pCCA Ordination biplot showing the effect of different sites on the relative density of plant species found at Khanpur Valley.

5. Relative Frequency (RF)

The Partial Canonical Correspondence Analysis (pCCA) of relative frequency data regarding seasons as environmental variables and sites as co-variables revealed highly significant variation in relative frequency of different plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.018) along axis 1(Table 2.5). pCCA Ordination biplot of

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species relative frequency and seasons as environmental variables showed that most of the species were centroid (Figure 2.9). The relative frequency of a large number of species was closely associated with summer and few were associated with winter. Most of the species had equal relative frequency values for summer and winter. Species no 82, 23 and 48 were closely associated with winter while species no 3, 6, 30, 38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were having relative frequency only at summer.

The pCCA Ordination biplot of relative frequency data while taking sites as environmental variables and seasons as co-variable also showed highly significant variation (P=0.0020 and eigenvalue=0.093) along axis1 (Table 2.5). pCCA ordination bi plot of relative frequency of species and different sites showed that the species were associated equally with all the four sites (Figure 2.10). The species number 73, 61 and 109 were associated with Dabola.site, species no 8, 85 and 74 were found associated with Jabri site, 89, 110, and 59 were associated with Mang site and species number 7 and 47 were found associated with Dam site. however species number 114 and 87 were equally relatively frequent between Dabola and Jabri sites. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site.

Seasons and sites both had a significant effect on the relative frequency of plant species in line with the frequency. Sites-seasons variations affect seed dispersal, seed germination, seedling growth and subsequent survival, maturation, and reproduction (Harper, (1977); Houle et al., (2001). In this study, most of the species had equal RF values for summer and winter, still it was observed that the RF of a large number of shrubs was closely associated with summer which might be due to the presence of suiTable temperature, enough moisture and macronutrients at summer

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(Skarpe, 1990; Zaman, 1997; Nanette et al., 2007). Topography of the sites was a determining factor for variation in RF of plant species. In a similar experiment Uniyal et al. 2006 had concluded that steep slopes had the highest species richness and diversity than plains. Moisture loving and moderately moisture requiring species were associated with Dam (Misbahuzzaman & Alam, 2006; Malik et al., 2007; Ahmad et al., 2008). At Dam Acacia modesta, at Mang site Adhatoda vasica and at Jabri site Olea ferrugineae were found frequently associated. The reason for the correlation of sites is the temporal, topographic and altitude similarities between the two correlated sites (Chapin, 1995; Chapin & Gaius, 1996 and Liu et al., 2014).

Table-2.5. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Relative Frequency of plant species at Khanpur Valley. Parameters and data Axes Total F ratio P value Inertia 1 2 3 4 Relative Frequency (Seasons/Sites) Eigenvalues 0.018 0.019 0.000 0.000 0.036 106.452 0.0020 Sum of all canonical 0.018 53.226 0.0020 Eigenvalues Relative Frequency (Sites/Seasons) Eigenvalues 0.093 0.013 0.011 0.065 0.203 94.925 0.0020 Sum of all canonical 0.118 38.645 0.0020 Eigenvalues Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.1.Acacia modesta 2.Acacia muricata 3.Acacia victoriae 4.Adhatoda vasica 5.Adiantum pedatum 6. Adiantum venusetum 7.Ailanthus altissima 8.Ajuga bractiosa 9.Arandu domax 10.Artemisia absenthium 11.Artimisia bevifolia 12.Avena fetua 13.Barleria priontis 14.Berberis vulgarus/lyceum 15.Boerhavia deffusa 16.Bohina variegate 17.Bombax malabaricum 18.Calotropis procera 19.Canabus sativa 20.Capsella bursa 21.Carthamus oxycantha 22.Cassia fistula 23.Cassia sophera 24.Celtis australis 25.Chemopodium album 26.Clitaria annua 27.Convulvolus arvensus 28.Conyza candensis 29.Cynodan Dactylon 30. Cyperus retendus 31. Dalburgia sisso 32.

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Datura alba 33. Datura stramonium 34. Desmodium elegans 35. Dodonia viscose 36. Echinochloa colona 37.Echinops echinatus 38.Euphorbia granulate 39.Euphorbia helioscopia 40.Euphorbia heterophylla 41.Euphorbia hirta 42.Euphorbia humifusa 43.Ferula assafoetida 44.Ficus hispida 45.Ficus jahaninsis 46.Ficus palmate 47.Ficus varigata 48.Foeniculum vulgarus 49.Fragaria orientalis 50.Fragaria vulgaris 51.Fumeria officinale 52.Grewia optivia 53.Grewia tenax 54.Hedera hilex 55.Hedera nepalensis 56.Imperata cylindrical 57.Indigofera spicata 58.Iris ensata 59.legerstroemia speciosa 60.lyceum barbarum 61.Mallotus phillipensis 62.Malvestrum caromandilianum 63.Matricaria chamomile 64.Medicago denticulate 65.Medicago polymorpha 66.Melia azadaracta 67.Melilotus parviflora 68.Mentha arvensis 69.Mintha ryaliana 70.Morus indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta 104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

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Figure 2.9. pCCA Ordination biplot showing the effect of different seasons on the relative frequency of plant species found at Khanpur Valley.

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Figure 2.10. pCCA Ordination biplot showing the effect of different sites on the relative density of plant species found at Khanpur Valley.

6. Relative Cover

The Partial Canonical Correspondence Analysis (pCCA) of relative coverage data regarding seasons as environmental variables and sites as co-variables showed highly significant variation in relative coverage of different plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.018) along axis 1(Table 2.6). pCCA Ordination biplot of species relative coverage and seasons as environmental variables showed

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that most of the species were centroid (Figure 2.11). The relative coverage of a large number of species was closely associated with summer and few were associated with winter. Most of the species had equal relative coverage values for summer and winter. Species no 82, 23, 67 and 48 were closely associated with winter while species no 3, 6, 30, 38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were having relative coverage only at summer.

The pCCA Ordination biplot of relative coverage data with respect to sites as environmental variables and seasons as co-variable also showed highly significant variation (P=0.0020 and eigenvalue=0.080) along axis1 (Table 2.6). pCCA ordination bi plot of relative coverage of species and different sites showed that the species were associated equally with all the four sites (Figure 2.12). The species number 1 was associated with Dam site, species no 61 was found associated with Dabola site, species number 74 was associated with Jabri site and species number 4, 59, 89 and 43 were found associated with Mang site. However species number 60 and 87 were having equal relative coverage between Dabola and Jabri sites. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site.

The higher relative coverage at summer than winter might be due to availability of sufficient moisture, nutrients and sunlight at summer and their deficiency during winter (Skarpe, 1990; Nanette et al., 2007). Another reason for low relative coverage at winter is the excessive use of the aerial parts of trees and shrubs by the residents as fuel and fodder during winter (Ahmad et al., 2008). Most of species were having maximum coverage at Dabola.site followed by Jabri site depending upon the availability of enough nutrients, conducive temperatures and sufficient water (Dupont and Plummer, 1997; Nanette et al., 2007). Kane

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et al., (2014) observed the same trend and concluded that biotic factors and disturbance significantly affected the relative coverage and forest structural development with different responses among sites.

Table-2.6. Summary of the partial CCA and Monte Carlo Test (MCT) showing the effect of different seasons and sites on the Relative Coverage of plant species found at Khanpur Valley. Parameters and Axes Total F ratio P value data 1 2 3 4 Inertia Relative Cover (Seasons/Sites) Eigenvalues 0.018 0.024 0.000 0.000 0.042 84.129 0.0020 Sum of all canonical 0.018 42.064 0.0020 Eigenvalues elative Cover (Sites/Seasons) Eigenvalues 0.080 0.015 0.012 0.069 0.209 69.232 0.0020 Sum of all canonical 0.107 28.899 0.0020 Eigenvalues Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.

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47.Ficus varigata 48.Foeniculum vulgarus 49.Fragaria orientalis 50.Fragaria vulgaris 51.Fumeria officinale 52.Grewia optivia 53.Grewia tenax 54.Hedera hilex 55.Hedera nepalensis 56.Imperata cylindrical 57.Indigofera spicata 58.Iris ensata 59.legerstroemia speciosa 60.lyceum barbarum 61.Mallotus phillipensis 62.Malvestrum caromandilianum 63.Matricaria chamomile 64.Medicago denticulate 65.Medicago polymorpha 66.Melia azadaracta 67.Melilotus parviflora 68.Mentha arvensis 69.Mintha ryaliana 70.Morus indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta 104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

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Figure 2.11. pCCA Ordination biplot showing the effect of different seasons on the relative coverage of plant species found at Khanpur Valley.

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Figure 2.12. pCCA Ordination biplot showing the effect of different sites on the relative coverage of plant species found at Khanpur Valley.

7. Importance Value

The Partial Canonical Correspondence Analysis (pCCA) of importance value data regarding seasons as environmental variables and sites as co- variables showed highly significant variation in importance value of different plant species indigenous to Khanpur Valley (P=0.0020 and eigenvalue=0.022) along axis 1(Table 2.7). pCCA Ordination biplot of species importance value and seasons as environmental variables showed that most of the species were centroid (Figure 2.13). The importance value of a large number of species was closely associated with summer and few were associated with winter. Most of the species had equal importance values for summer and winter. Species no 82, 84, 20, 23 and 48 were 110

closely associated with winter while species no 3, 6, 30, 38, 41, 46, 54, 63, 65, 69, 72, 79, 81, 86, 97 and 113 were having importance value only at summer.

The pCCA Ordination biplot of importance value data with respect to sites as environmental variables and seasons as co-variable also showed highly significant variation (P=0.0020 and eigenvalue=0.074) along axis1 (Table 2.7). pCCA ordination bi plot of importance value of species and different sites showed that the species were associated equally with all the four sites (Figure 2.14). The species number 1 was associated with Dam site, species no 39, 66, 109, 87, 19 and 61 were found associated with Dabola site, species number 74 was associated with Jabri site and species number 4, 89, 77 and 64 were found associated with Mang site. The ordination plots reveals the correlation among different sites, which show that Dam is correlated with Mang site and Dabola is correlated with Jabri site.

The computation of importance value is a good verdict for deciding the standing of a given plant species in a community. The higher the importance value of a plant, the greater is its competitiveness (Adrees et al., 2011). The higher IV of species at different sites and seasons was due to their association with the availability of macronutrients and high field capacity (Ahmad et al., 2008). Salt and drought tolerant species were recorded around Mang site as it provides the same environment. Moisture loving and moderately moisture requiring species were associated with Jabri and Dabola sites due to availability of the requisite climate at the two sites (Misbahuzzaman and Alam, 2006; Malik et al., 2007a; Malik et al., 2007b; Ahmad et al., 2008).

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Parameters and 1 2 3 4 Total data Inertia Importance value (Seasons/Sites) Eigenvalues 0.014 0.022 0.000 0.000 0.035 71.938 0.0020 Sum of all canonical 0.014 35.929 0.0020 Eigenvalues Importance Value (Sites/Seasons) Eigenvalues 0.074 0.010 0.007 0.069 0.188 72.685 0.0020 Sum of all canonical 0.092 26.549 0.0020 Eigenvalues

Legends: List of plant species with species number, identified during ecological survey of vegetation of Khanpur Valley.

1.Acacia modesta 2.Acacia muricata 3.Acacia victoriae 4.Adhatoda vasica 5.Adiantum pedatum 6. Adiantum venusetum 7.Ailanthus altissima 8.Ajuga bractiosa 9.Arandu domax 10.Artemisia absenthium 11.Artimisia bevifolia 12.Avena fetua 13.Barleria priontis 14.Berberis vulgarus/lyceum 15.Boerhavia deffusa 16.Bohina variegate 17.Bombax malabaricum 18.Calotropis procera 19.Canabus sativa 20.Capsella bursa 21.Carthamus oxycantha 22.Cassia fistula 23.Cassia sophera 24.Celtis australis 25.Chemopodium album 26.Clitaria annua 27.Convulvolus arvensus 28.Conyza candensis 29.Cynodan Dactylon 30. Cyperus retendus 31. Dalburgia sisso 32. Datura alba 33. Datura stramonium 34. Desmodium elegans 35. Dodonia viscose 36. Echinochloa colona 37.Echinops echinatus 38.Euphorbia granulate 39.Euphorbia helioscopia 40.Euphorbia heterophylla 41.Euphorbia hirta 42.Euphorbia humifusa 43.Ferula assafoetida 44.Ficus hispida 45.Ficus jahaninsis 46.Ficus palmate 47.Ficus varigata 48.Foeniculum vulgarus 49.Fragaria orientalis 50.Fragaria vulgaris 51.Fumeria officinale 52.Grewia optivia 53.Grewia tenax 54.Hedera hilex 55.Hedera nepalensis 56.Imperata cylindrical 57.Indigofera spicata 58.Iris ensata 59.legerstroemia speciosa 60.lyceum barbarum 61.Mallotus phillipensis 62.Malvestrum caromandilianum 63.Matricaria chamomile 64.Medicago denticulate 65.Medicago polymorpha 66.Melia azadaracta 67.Melilotus parviflora 68.Mentha arvensis 69.Mintha ryaliana 70.Morus indica/nigrum 71.Myrsine Africana 72.Nasturtium officinale 73.Nirium oleander 74.Olea ferrugineae 75.Otostegia limbata 76.Oxalis corniculata 77.Parthenium hystrophorus 78.Phalaris aquatic 79.Pinus roxbergy 80.Plantago lanceolata 81.Plantago major 82.Plantago ovate 83.Poa anna 84.Portulaca oleraceae 85.Prunus serotina 86.Pteridium equilinium 87.Punica granatum 88.Ranunculus repens 89.Recinus cammunis 90.Rubus fruticosus 91.Rumax vasicarius 92. Rumex dentatus 93.Rumex histatus 94.Rumex longifolius 95.Saccharum spontanum 96.Silybum marianum 97.Skimmi laureola 98.Solanum nigrum 99.Solanum pseudocapsicum 100.Solanum surratense 101.Solanum xanthocarpum 102.Stellaria media 103.Tagetes minuta

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104.Taraxacum officinale 105.Tecomela undulate 106.Themeda triandra 107.Tinosporia cordifolia 108.Trianthema portuclacastrum 109.Urtica dioica 110.Urtica dubia 111.Verbascum thapsum 112.Vitex nigudo 113.Xanthium strumarium 114.Xanthoxylum armatum 115.Ziyphus nemularia 116.Zizyphus vulgarus

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Figure 2.13. pCCA Ordination biplot showing the effect of different seasons on the importance value of plant species found at Khanpur Valley.

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Figure 2.14. CCA Ordination biplot showing the effect of different sites on the importance value of plant species found at Khanpur Valley.

8. Classification of Species:

These results show that out of 116 ecologically important species found in the valley, majority fall in the class of Rare (R) (45.37%), followed by Occasional (O) (19.72%), then Frequent (F) (5.06%) and least fall in the class of Abundant (A) (2.05%). While there was not a single species found as very abundant. Similarly out of 202 total species documented in Khanpur Valley (in experiment-1 page no9) 116 species (57.43%) were ecologically important while the rest of 86 species (42.57%) were threatened.

The criteria of classification set in the present study were strongly supported by Dyksterhuis (1949) while Hussain (2007) classified various

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plant species based on density and frequency percentage. The reason behind low occurrence and threat to natural plant population of most of the plant species might be the increased pressure on plant recourses and deforestation (Adnan et al., 2006), arid land salinity (Keighery, 2000) injudicious use of plants (IUCN, 2004) and urbanization (Rouget et al., 2003). Many of the herbal products are derived from species that are threatened or have become rare or endangered by large-scale exploitation (Nalawade et al. 2003). Similar study was conducted by Adnan et al., (2006) who revealed threats to the sustainability of Ethno-Medicinal uses in Miandam Valley, Swat, Kyber Pakhtunkhwa, Pakistan and concluded that the valley has over 300 plant species, mostly medicinal and rare or endangered. They further revealed that decrease in medicinal plants has been observed in the last 30 years due to various threats and issues, like high deforestation rate of 2% per year under which 8,053 trees are felled every year.

Figure-2.15. Summary showing the effect of different seasons and sites on the classification of Plant Species found at Khanpur Valley.

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SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

Ecological analysis of vegetation is the basic technique to present a true picture of species diversity and distribution. The study on ecological analysis of vegetation of Khanpur valley in the sub Himalayan mountains of Pakistan was conducted in 20112012. The whole valley was divided into four different sites namely: Dam, Dabola, Jabri and Mang. Data was recorded in both summer (April-September) and winter (October-March) separately. The ecological data was recorded using quadrate sampling method. Fixed quadrates of 1m2 for herbs and 25m2 for shrubs and trees were used and all individual plants in the quadrate were taken into consideration. Randomly 15 quadrates for herbs and 15 quadrates for shrubs and trees were taken at each site in both the seasons. The data was recorded on properly designed data sheets (Cox, G.W. 1996. Laboratory Manual of General Ecology, 7th Edition, Wm.C. Brown Publishers, Boston). The recorded data was used for the calculation of density, frequency, cover/dominance, relative density, relative frequency, relative cover/dominance, importance value and classification of species. The data was analyzed using Canoco Computer Package for windows (version 4.5).

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Findings based on quadrate analysis, revealed that a total of 116 ecologically important species (Frequency≥1%) were found in all the four sites in both seasons. Maximum number of plant species (102) were found at Dabola site in summer season, while minimum species (73) were observed at Dam site during winter season. The maximum density was recorded for plant species associated with Dabola and Jabri sites. The ordination plots, figured through the statistical tool, revealed that Dam site is ecologically correlated with Mang site while Dabola site is correlated with Jabri site for all the parameters. The results further showed that species ecologically associated with summer were Adiantum venusetum, Cyperus retendus, Euphorbia granulate, Euphorbia hirta, Hedera hilex, Matricaria chamomile, Medicago polymorpha, Mintha royaleana, Nasturtium officinale, Plantago major, Pteridium equilinium, Skimmi lanreola and Xanthium strumarium, while those associated with winter were Foeniculum vulgarus and Plantago ovata. Species like Ajuga bractiosa, Fragaria vulgaris, Mallotus phillipensis, Melia azadaracta, Punica granatum and Urtica dioica were found associated with Dabola site, species Acacia modesta, Medicago denticulate, Rumex dentatus and Themeda triandra with Dam site, species Olea ferrugineae Mentha arvensis, Nasturtium officinale and Stellaria media at Jabri site while species Adhatoda vasica, Fumaria officinalus, Rumax longifolius and Urtica dubia were found more frequent at Mang site. The results also revealed that majority of the species fall in the class of Rare (45.37%), followed by Occasional (19.72%), then Frequent (5.06%) and least fall in the class of Abundant (2.05%). While there was not a single species found as Very Abundant during any season and at any site of the valley. Many of the phytopharmaceuticals are prepared from species that are threatened or have become rare due to large-scale exploitation . The ordination plots revealed that Dam is correlated with Mang site and Dabola is correlated with Jabri site for all the parameters.

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The following conclusions were drawn from the current study:

1. Khanpur Valley was documented with 116 ecologically important plant species (57.43%) and 86 threatened or endangered plant species (42.57%), out of total 202 wild indigenous plant species (experiment-1 page number 9). 2. Out of total of 116 ecologically important species maximum plant species (102) were found at Dabola site in summer season while minimum species (73) were observed at Dam site during winter season. 3. The results revealed that number of species associated with summer were higher than those associated with winter. Similarly maximum species in terms of frequency, density and coverage were associated with Dabola site followed by Jabri site. 4. It was concluded that the six most preferred medicinally important species of the valley, namely Adhatoda vasica, Ajuga bracteosa, Calatropis procera, Euphorbia hirta, fumaria officinalis and Recinus communis (experimint-1 page 24-25) were found to be ecologically important as well in all the four sites and two seasons except Euphorbia hirta which was found only in summer and was not available in winter in any quadrate studied at all the sites. 5. The results revealed that out of 116 ecologically important species found in the valley, majority fall in the class of Rare (R) (45.37%), followed by Occasional (O) (19.72%), then Frequent (F) (5.06%) and least fall in the class of Abundant (A) (2.05%). While there was not a single species found as very abundant. 6. The ordination plots revealed that Dam is correlated with Mang site and Dabola is correlated with Jabri site for all the

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parameters. Which means the soil and environmental conditions of the two correlated sites were in connivance with each other.

Following are some of the recommendations synthesized from the study and based on the research findings;

• Being an ecosystem of 57.43% ecologically important species and 42.57% threatened species, the flora require a sustainable harvesting and collection mechanism to maintain biodiversity and ensure a sTable supply of raw material for herbal medicines. • As majority of the ecologically important species fall in the class rare (45.37%) and an alarming number of total plant species as threatened (42.57%), the flora require a conservation strategy on urgent basis, both in nature and in nursery. • Maximum plant species were ecologically associated with summer, so plants or parts collection may be restricted to summer season and felling of plants in winter for firewood may also be controlled. • Awareness campaigns must be launched to involve the local community in the conservation and management process and acquaint them about the usefulness and sustainable utilization of the herbal plants to make this invaluable source of food, medicines and biodiversity intact and growing. • Detailed ecological study on threatened plant species is recommended to be conducted in future to find out the magnitude and causes of loss to the precious plant resources.

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EXPERIMENT-3

THE STUDY OF BIO-CHEMICAL SUBSTANCES OF SELECTED MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

Shah Masaud Khan and Noor ul Amin

Department of Horticulture, The University of Agriculture Peshawar, Pakistan

ABSTRACT

The experiment on the bio-chemical substances of selected medicinal plants of Khanpur Valley in the sub Himalayan mountains of Pakistan was conducted during 2012-2013. The samples were collected from Adhatoda vasica, Calatropis procera, Recinus communis, Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis and chemical analysis was carried out at the University of agriculture Peshawar. Whole plants (in case of herbs) and leaves (in case of woody plants) were collected from their natural habitats from all the four sites and two seasons. Significant effects were observed at different seasons and sites upon various bio-chemical substances of all six medicinal plants. Adhatoda vasica showed significantly higher values for ash (19.5%) at Jabri during summer, crude proteins (14.7%) at Dabola during summer, crude fibers (12.6%) at Jabri during winter, essential oil (2.5%) at Mang during summer, NFES (55.8%) at Mang during summer, NFEE (194.6%) at Dam during summer, potassium (179.4mg/100g) at Dam during summer, phosphorus (171mg/100g) at Dabola during summer, copper (0.58mg/100g) at Dabola during summer and zinc (8.07mg/100g) at Mang during summer. Fumaria officinalis revealed higher significant results for ash (27.7%) at Jabri during summer, crude proteins (15.2%) at Mang during summer, crude fibers (13.1%) at Jabri during winter, crude fats (9.6%) at Mang during summer, essential oil (2.2%) at Mang during winter, NFES (47.04%) at Mang during winter, NFEE (186.13%) at Mang during summer, potassium (494.1mg/100g) at Dabola during summer, phosphorus (265.3mg/100g) at Mang during summer, magnesium (184.3mg/100g) at Jabri during summer and manganese (14.03mg/100g) at Dabola during summer. Euphorbia hirta 121

gave higher significant contents for ash (17.95%) at Jabri during summer, crude proteins (15.42%) at Jabri during summer, crude fibers (12.78%) at Mang during winter, crude fats (10.64%) at Jabri during summer, essential oil (1.94%) at Dabola during summer, NFES (59.49%) at Dam during winter, NFEE (198.69%) at Jabri during summer, calcium (199.17mg/100g) at Dam during summer, phosphorus (185.37mg/100g) at Jabri during winter. Ajuga bracteosa revealed higher significant values for ash (34.13%) at Jabri during summer, crude proteins (15.35%) at Dam during summer, crude fats (10.30%) at Jabri during summer, essential oil (2.48%) at Mang during summer, NFES (46.17%) at Dabola during winter, NFEE (186.28%) at Dam during summer, sodium (6.96mg/100g) at Mang during winter, potassium (559.67mg/100g) at Jabri during winter, calcium (254.20mg/100g) at Jabri during summer, magnesium (264.31mg/100g) at Mang during winter, copper (1.017mg/100g) at Dabola during winter and zinc (6.53mg/100g) at Dam during winter. Recinus communis revealed higher significant values for crude proteins (15.26%) at Mang during summer, crude fibers (13.85%) at Dabola during winter, crude fats (10.24%) at Jabri during summer, essential oil (3.48%) at Mang during winter, NFEE (195.29%) at Jabri during summer, sodium (4.91mg/100g) at Dam during summer, iron (300.37mg/100g) at Jabri during summer, manganese (13.43mg/100g) at Dabola during summer and zinc (8.07mg/100g) at Mang during summer. Calatropis procera revealed higher significant values for crude proteins (14.58%) at Mang during summer, crude fibers (13.39%) at Dabola during winter, crude fats (11.98%) at Jabri during summer, NFES (50.80%) at Dam during winter, NFEE (203.1%) at Jabri during summer, potassium (500.33mg/100g) at Dam during summer, calcium (274.07mg/100g) at Dabola during winter, magnesium (291.57mg/100g) at Dabola during winter, copper (0.72mg/100g) at Jabri during summer, iron (291.6mg/100g) at Dam during summer manganese (19.27mg/100g) at Dam during summer and zinc (6.03mg/100g) at Mang during winter. It was concluded from the results of the current experiment that medicinal plants under study were rich in bio-chemical substances. Ajuga bracteosa and Euphorbia hirta gave higher values for bio- chemicals during last week of October, Fumaria officinalis performed best during last week of May while Adhatoda vasica, Recinus communis and Calatropis procera showed maximum bio-chemicals in the samples collected during last week of July.

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INTRODUCTION

Medicinal plants are staging a comeback and herbal „renaissance‟ is happening all over the world (Ramirez et al., 2006). The herbal medicines today symbolize safety in contrast to the synthetics that are considered as unsafe to human and environment. Plants are the natural resources which contribute major portion to human nourishment and medication. Historically, plants are harvested or collected from nature by men and women since the birth of life, for their wellbeing. There are plant species which are used in the preparation of functional food, which serves both the purposes of nutritional as well as medicinal requirements. Such dual purpose species are of immense importance and need comprehensive investigation in the natural flora of ecologically diverse habitats of its existence.

Phyto-medicines are also being used on large scale in Western Europe. Recently the US

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Government has established the “Office of Alternative Medicine” at the National Institute of Health at Bethesda. Ayurveda, a system of herbal medicine in India, Sri Lanka and South-East Asia has more than 8000 traditional uses while 35,000-70,000 plant species are medicinal (Farnsworth and Soejarto 1991). China has demonstrated the best use of traditional medicine in providing the health care. China has pharmacologically validated and improved many phyto pharmaceuticals and eventually integrated them in modern medicines (Liu et al., 2011).

Green plants synthesize and preserve a variety of biochemical products, many of which are extracTable and used as chemical feed stocks or as raw material for various scientific investigations (Edeoga et al., 2005). Many secondary metabolites of plant are economically very important and find use in a number of medicinal compounds. However, a constant supply of the source material often becomes difficult due to a number of factors like environmental changes, cultural practices, diverse geographical distribution, labour cost, selection of the superior plant stock and over exploitation by pharmaceutical industry.

Human body needs essential nutrients, bio-chemical compounds (protiens, fats, carbohydrate etc), major minerals (Ca, P, K, Na and Mg) and trace elements (Fe, Zn, Cu, Mn and others), which are essential for general health, growth and reproduction. We must bear in mind that the consequences of essential trace mineral deficiency may be just as severe as those of a deficiency of a major essential mineral. Many elements are associated with one another in maintaining our normal growth and health. The medicinal plants may prove to be a useful remedy for many common and complicated ailments and part of essential nutrients required for human nourishment.

A major shortfall in plant medicine is the lack of drug standardization, information and quality control. Most of the plant medicines are in the

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form of crude extracts which are a mixture of several ingredients and the active principles when isolated individually fail to give desired activity (Ahmad et al., 2011). This implies that the activity of the extract is the synergistic effect of its various components. In the absence of pharmacopoeia data on the various plant extracts, it is not possible to isolate or standardize the active contents having the desired effects. Plant pharmaceuticals compiled on modern lines and updated periodically is an urgent requirement.

An integrated technology combining herbal and allopathic ways of treatments, whereby the side effects and adverse reactions could be controlled can be very useful for the ailing persons. Studies can show that the toxic effects of radiations and chemotherapy in cancer treatment could be reduced by herbal medications and similarly surgical wound healing could be accelerated by the use of herbal medicines (Liu et al., 2011). Modern science and technology have an essential role to play in the process. An holistic approach for the cultivation, conservation and preservation of important plant species used by the local communities of various regions is the need of the hour (Sharma, 1997). Pakistan is very rich in species diversity and its various ecological zones are unique in its ethno botanical and traditional healthcare systems. Khanpur valley in the sub-Himalayan mountains of Pakistan was selected by the Department of Horticulture, the University of Agriculture Peshawar to study the bio- chemical attributes of selected medicinal plants with the aim to achieve the following objectives:

1. To find out the comparative suitability of seasons and sites for the best harvest of bio-chemical attributes of medicinal plants of Khanpur valley. 2. To elaborate the prospects of medicinal plants of bio-chemical importance, for possible use as source of food supplement.

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3. To recommend potential medicinal plants to the pharmaceutical industries and for further promotion through cultivation and research.

MATERIAL AND METHODS

Impact of different seasons and sites on nutritional/bio-chemical Attributes of 6 most important medicinal plants of khanpur valley was conducted during 2012-2013. Medicinal plants were evaluated on the basis of medicinal importance and three most important herbs and three most important shrubs were selected for biochemical study. Medicinal importance was calculated through a questionnaire survey as presented in experiment 1 page# 24-25. Medicinal plants, whole plant in case of herbs and leaves in case of woody plants (as these were the most preferred materials used by the local people, results obtained in experiment 1 page # 27), were collected from their natural habitat at all the four sites and two seasons (details of sites and seasons are mentioned in experiment 1 page # 6-7). Plant material were collected in summer and winter at different specified periods as below: 1. Adhatoda vasica Last week of July and last week of December.

2. Calatropis procera Last week of July and last week of December.

3. Recinus communis Last week of July and last week of December.

4. Ajuga bracteosa Last week of July and last week of October.

5. Euphorbia hirta Last week of July and last week of October.

6. Fumaria officinalis Last week of May and last week of February. Through quadrate transact method, three transacts (replications) were taken and in every transact, the material were collected from different available plants and fresh weights were recorded. The samples were brought to the laboratory of the Agricultural Chemistry, The University of Agriculture Peshawar. The sample was thoroughly cleaned manually and then made into powder by laboratory grinder. The samples in the

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grinded form were then analyzed for the following bio-chemical attributes, using standard procedures:

Adhatoda vasica

Fumaria officinalis

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Euphorbia hirta

Ajuga bracteosa

Recinus communis

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Calastopis procera

Moisture and Dry Matter content:

Percent moisture and dry matter of the samples were measured while following AOAC,

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(2000) and the standard procedure of weight loss during heating in the oven at about 105 °C until weight remains, was followed.

Ash content:

Ignition method was followed, where organic materials of the samples under consideration were burnt at 550-600 °C in muffle furnace (AOAC, 2000).

Ash content was measured from the remaining residues with the following formula.

(W2 W3) x 100 % Ash = Wt. of sample

Crude Fat content: Crude fat was measured by solvent extraction method using soxhtec apparatus (AOAC, 2000). Crude fat was calculated as follow:

Wt. of ether extract x 100 % Crude Fat = Wt. of sample

Crude Protein content: 1 gram samples were introduced into micro

Kjeldahl flask. 1.5 g of catalytic mixture (7K2SO4:1CuSO4) and concentrated H2SO4 (7 ml) were added to the sample and then mixed thoroughly. The flask containing samples were heated first at 100 oC and then gradually raised to 300 oC. The heating continued till whole of the materials were converted to clear inorganic solution. That acid digest was then diluted to 100mL in volumetric flask with distilled water.

Then 10mL of acid digest plus 10 mL of 40% NaOH was added in micro distillation apparatus and was heated with steam and the NH3 thus released was collected in 20 ml of 4% boric acid containing modified

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methyl red indicator in 100mL conical flask. The content in the flask was then titrated with 0.05% HCl using phenolphthalein as indicator. The crude protein content was then calculated as below:

(S- B) x N of acid x 0.014 x dilution x 100 % Nitrogen = Wt. of sample x aliquot of digest taken for distillation

Where S = Sample titration reading

B = Blank titration reading

0.14 = Miliequivalent of Nitrogen Crude Protein was determined by multiplying the nitrogen content with the standard factor.

Crude Fiber contents

First two g sample was put in a beaker containing 200ml HCl and boiled for 30 minutes. The residue was filtered and was transferred to a beaker containing 20ml NaOH and was boiled for 30 minutes on a water bath. The residue was transferred to crucible and was put in the furnace for 4 hrs at 550 and was reweighed after cooling.

% Crude Fiber = (weight of oven dried residue- weight after ignition) x 100 Weight of sample

Nitrogen Free Extractable Substances (NFES): Nitrogen Free Extract was calclated as below:

NFES % = 100-(Ash%+Crude Proteins%+Crude Fats%+Crude Fibers%)

Net Free Energy Estimation(NFEE)= Crude Proteins%*4+Crude fats%*8.7+NFES%

Elementology (Mineral Analysis) (Na, K, Ca, P, Mg, Cu, Fe, Mn and Zn)

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The samples (dried powdered) were digested with nitric acid and perchloric acid following standard method of AOAC (2000). Samples (0.5g) were weighed with an electronic balance and were taken in a digestion flask. Nitric acid (10mL) was added and were kept overnight for complete reaction. Then 10mL Percloric acid was added into the flasks and heated on a control heater, initially at 100 and then temperature was raised up to 250 oC to completely hydrolyze the samples into clear inorganic solution. The acid digests were then used for mineral determination.

Statistical Analysis:

The data on bio-chemical parameters were analyzed by using the statistical software " Statistix-8.1", using linear model and general ANOVA with specific model statement. The mean data, for factors and their interaction, were compared by using LSD at 0.05.

RESULTS AND DISCUSSIONS

The results and discussions on bio-chemical/nutritional analysis of all six selected medicinal plants are presented under two major headings: I. Proximate Analysis: The bio-chemical compounds which include Moisture Contents, Dry matter, Ash/Minerals, Crude Fiber, Crude Protein, Fat Contents, Essential oil, NFES & NFEE and II. Elementology: The bio-chemical elements which include Na, K, Ca, P, Mg, Cu, Fe, Mn and Zn.

The first three of the six selected medicinal plants were shrubs (Adhatoda vasica, Calatropis procera and Recinus communis) and the subsequent three were herbs (Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis).

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3.1. ADHATODA VASICA

I. Proximate Analysis:

1. Moisture%:

The mean data of moisture % for various seasons and sites is shown in Table-3.1. The results revealed that the effect of seasons and sites on moisture % was significant while the effect of their interactions was non- significant.

Maximum moisture % was observed at summer (71.46%) while minimum at winter (66.86%). Similarly maximum moisture % was recorded at Dam site (71.365%) while minimum at Mang site (67.18%).

2. Dry Matter %:

The mean data of dry matter % for various seasons and sites is shown in Table-3.1. The results revealed that the effect of seasons and sites on dry matter % was significant while the effect of their interactions was non- significant.

Maximum dry matter % was observed at winter (33.14%) while minimum at summer (28.54%). Similarly maximum moisture % was recorded at Mang site (32.83%) while minimum at Dam site (28.64%).

3. Ash / Minerals %:

The mean data of ash / minerals % for various seasons and sites is shown in Table-3.1, while their interaction is shown in figure-3.1. The results revealed that the effect of seasons, sites and their interactions on ash / minerals % was significant.

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Maximum ash / minerals % was observed at summer (17.54%) while minimum at winter (14.82%). Similarly maximum ash / minerals % was recorded at Jabri site (17.76%) while minimum at Mang site (14.84%). In case of interactions maximum ash % was recorded at Jabri site (19.53%) while minimum of it was observed at Dabola site during winter (13.30%).

4. Crude Proteins %:

The mean data of crude proteins % for various seasons and sites is shown in Table-3.1, while their interaction is shown in figure-3.2. The results revealed that the effect of seasons, sites and their interactions on crude proteins % was significant.

Maximum crude proteins % was observed at summer (13.21%) while minimum at winter (11.87%). Similarly maximum crude proteins % was recorded at Dabola site (13.70%) while minimum at mang site (11.43%). In case of interactions maximum crude proteins % was recorded at Dabola site during summer (14.72%) followed by Dam site during summer (14.12%) while minimum of it was observed at Mang site during summer (11.22%).

5. Crude Fibers %:

The mean data of crude fibers % for various seasons and sites is shown in Table-3.1, while their interaction is shown in Figure-3.3. The results revealed that the effect of seasons, sites and their interactions on crude fibers % was significant.

Maximum crude fibers % was observed at winter (11.28%) while minimum at summer (9.97%). Similarly maximum crude fibers % was recorded at jabri site (12.31%) while minimum at Dam site (9.54%). In case of interactions maximum crude fibers % was recorded at Jabri site

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during winter (12.61%) followed by Jabri site during summer (12%) while minimum of it was observed at Mang site during summer (8.71%).

6. Crude Fats %:

The mean data of crude fats % for various seasons and sites is shown in Table-3.1. The results revealed that the effect of seasons and sites on crude fats % was significant and the effect of their interaction was non- significant.

Maximum crude fats % was observed at summer (9.50%) while minimum at winter (7.99%). Similarly maximum crude fats % was recorded at Dam site (9.25%) while minimum at Mang site (8.25%).

7. Essential Oils%:

The mean data of essential oil % for various seasons and sites is shown in Table-3.1, while their interaction is shown in figure-3.4. The results revealed that the effect of seasons on essential oil % was non-significant while the effect of sites and interactions was significant.

Maximum essential oil % was recorded at Mang site (2.51%) while minimum at Dabola site (1.35%). In case of interactions maximum essential oil % was recorded at Mang site during summer (2.58%) followed by Jabri site during winter (2.43%) while minimum of it was observed at Dabola site during summer (0.93%).

8. Nitrogen Free ExtracTable Substances (NFES) %:

The mean data of NFES % for various seasons and sites is shown in Table- 3.1, while their interaction is shown in figure-3.5. The results revealed that the effect of seasons, sites and their interactions on NFES % was significant.

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Maximum NFES % was observed at winter (54.04%) while minimum at summer (49.79%). Similarly maximum NFES % was recorded at Mang site (55.56%) while minimum at Jabri site (48.82%). In case of interactions maximum NFES % was recorded at Mang site during summer (55.78%) followed by Mang site during winter (55.34%) while minimum of it was observed at Jabri site during summer (45.69%).

9. Net Free Energy Estimation (NFEE):

The mean data of NFEE for various seasons and sites is shown in Table- 3.1, while their interaction is shown in Figure-3.6. The results revealed that the effect of seasons, sites and their interactions on NFEE was significant.

Maximum NFEE was observed at summer (185.26) while minimum at winter (171.01). Similarly maximum NFEE was recorded at Dam site (183.29) while minimum at mang site (173.07). In case of interactions maximum NFEE was recorded at Dam site during summer (194.59) followed by Dabola site during summer (189.47) while minimum of it was observed at Jabri site during winter (166.53).

It is evident from the results that summer and winter seasons have significantly affected the proximate analysis. The bio-chemical compounds like crude proteins, crude fats and ashes or total minerals were higher in summer season than winter season in all the four sites of the study area. In adhatoda vasica, which is an evergreen species, the plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and to keep nutrients for winter stresses. Summer rains and high water availability to plants facilitated the accumulation of these compounds in leaves during summer. These results were confirmed by

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many reports (McDowell, 2003; Ramirez et al., 2006). Amaral et al., (2004) attributed the increase in bio-chemicals in July to the highest value of solar radiation during this month.High protein and total minerals found in summer season may be due to availability of high moisture due to heavy rainfall in the valley.

On the other hand dry matter, crude fibers, essential oils and nitrogen free extracts were found significantly higher during winter than summer in all of the sites. The accumulation of these compounds may be due to low temperature and deficiency of water and stresses (Ashraf et al., 1994; Harborne and Williams, 2000; Ali and Abbas, 2003). Furthermore, plants grown in cold areas accumulate oils and sugar during early winter to cope with severe cold and provide support to plant survival and protect it from frost injury (Rig and Cain, 1992).

The sites comparison revealed that Dam site produced maximum value for moisture and crude fats in the leaves of adhatoda. It was observed that the soil of dam site was having higher water holding capacity, as a result of which high moisture were recorded in leaves of adhatoda (McDowell, 2003). Dabola site gave maximum value for crude proteins and jabri site showed maximum results for crude fibers and ash which might be associated with environmental stresses as both the sites are at higher altitude (Harborne and Williams, 2000; Ali and Abbas, 2003). While Mang site produced maximum values for dry matter, essential oil and NFES which are closely associated with the mineral composition of the soil of the site (Demeyer and Dejaegere, 1996).

Table-3.1. Effect of Different Seasons and Sites on Proximate analysis of Adhatoda vasica, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan.

Dry Ash/mi Crude Moistur Crude Crude Essentia Matter nerals Proteins e% Fibers% Fats% l Oils% NFES Seasons % % % NFEE %

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Summer 71.46a 28.54b 17.54a 13.21a 9.97b 9.50a 1.83 49.79b 185.26a

Winter 66.86b 33.14a 14.82b 11.87b 11.28a 7.99b 2.06 54.04a 171.01b LSD at α 0.05 0.876 0.876 0.500 0.580 0.224 0.533 n.s 1.373 5.144 Sites

Dam 71.36a 28.64c 16.88b 12.83b 9.54d 9.25a 1.61b 51.50b 183.29a

Dabola 68.39bc 31.61ab 15.22c 13.70a 10.75b 8.55bc 1.35c 51.78b 181.00ab

Jabri 69.70ab 30.30bc 17.76a 12.21bc 12.31a 8.91ab 2.32a 48.82c 175.17bc

Mang 67.18c 32.83a 14.84c 11.43c 9.91c 8.25c 2.51a 55.56a 173.07c LSD at α 0.05 2.466 2.466 0.443 1.104 0.430 0.557 0.205 1.756 5.854 Interactions

Seasons*Sites Ns ns * * * ns * * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability. 25

15 Summer 10 Winter 5

0 Dam Dabola Jabri Mang

Figure-3.1: Effect of different seasons and sites on Ash % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

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10 Summer Winter 5

0 Dam Dabola Jabri Mang

Figure-3.2: Effect of different seasons and sites on Crude Proteins % in leaves of Adhatoda vasica collected from Khanpur valley in the sub- Himalayan mountains of Pakistan. 14 12 10 8 Summer 6 4 Winter 2 0 Dam Dabola Jabri Mang

Figure-3.3: Effect of different seasons and sites on Crude fibers % in leaves of Adhatoda vasica collected from Khanpur valley in the sub- Himalayan mountains of Pakistan. 3.5 3 2.5 2 Summer 1.5 1 Winter 0.5 0 Dam Dabola Jabri Mang

Figure-3.4: Effect of different seasons and sites on Essential oil % in leaves of Adhatoda vasica collected from Khanpur valley in the sub- Himalayan mountains of Pakistan.

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70 60 50 40 Summer 30 Winter 20 10 0 Dam Dabola Jabri Mang

Figure-3.5: Effect of different seasons and sites on NFES % in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 250

200

150 Summer 100 Winter 50

0 Dam dabola Jabri Mang

Figure-3.6: Effect of different seasons and sites on NFEE in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

II. Elementology:

1. Sodium (Na) mg/100g:

The mean data of Na for various seasons and sites is shown in Table-3.2. The results revealed that the effect of seasons on Na was significant while the effect of sites and of their interaction was non-significant.

Maximum Na was observed at summer (1.24 mg/100g) while minimum at winter (0.88 mg/100g).

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2. Potassium (K) mg/100g: The mean data of K for various seasons and sites is shown in Table-3.2, while their interaction is shown in Figure-3.7. The results revealed that the effect of seasons and sites on K was non-significant while the effect of their interaction was significant.

Maximum K was observed at Dam site during summer (179.4 mg/100g) followed by dabola site during summer (179 mg/100g) followed by Jabri site during winter (177 mg/100g). While minimum K was observed at Mang site during summer (102.77 mg/100g)

3. Calcium (Ca) mg/100g:

The mean data of Ca for various seasons and sites is shown in Table-3.2. The results revealed that the effect of seasons and sites on Ca was significant while the effect of their interactions was non-significant.

Maximum Ca was observed at summer (230.72 mg/100g) while minimum at winter (187.77 mg/100g). Similarly maximum Ca was recorded at Dam site (244.60 mg/100g) while minimum at Mang site (183.97 mg/100g).

4. Phosphorus (P) mg/100g:

The mean data of Phosphorus for various seasons and sites is shown in Table-3.2, while their interaction is shown in figure-3.8. T The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum P was observed at summer (128.52 mg/100g) while minimum at winter (113.52 mg/100g). Similarly maximum P was recorded at Dam site (137.67 mg/100g) while minimum at jabri site (100.88 mg/100g). In case of interactions maximum P was observed at Dabola site during summer (171 mg/100g) followed by Dam site during Winter (151.97

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mg/100g). While minimum P was observed at Dabola site during winter (101.23 mg/100g)

5. Magnesium (Mg) mg/100g:

The mean data of Mg for various seasons and sites is shown in Table-3.2. The results revealed that the effect of seasons on Mg was significant while the effect of sites and of their interaction was non-significant.

Maximum Mg was observed at summer (235.52 mg/100g) while minimum at winter (157.09 mg/100g).

6. Copper (Cu) mg/100g:

The mean data of Cu for various seasons and sites is shown in Table-3.2, while their interaction is shown in Figure-3.9. T The results revealed that the effect of seasons, sites and their interaction on Cu was significant.

Maximum Cu was observed at summer (0.48 mg/100g) while minimum at winter (0.16 mg/100g). Similarly maximum Cu was recorded at Jabri site (0.37 mg/100g) while minimum at Dam site (0.23 mg/100g). In case of interactions maximum Cu was observed at Dabola site during summer (0.58 mg/100g) followed by Jabri site during summer (0.49 mg/100g). While minimum Cu was observed at Dam site during winter (0.08 mg/100g)

7. Iron (Fe) mg/100g:

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The mean data of Fe for various seasons and sites is shown in Table-3.2. The results revealed that the effect of seasons and sites on Fe was significant while the effect of their interactions was non-significant.

Maximum Fe was observed at summer (322.03 mg/100g) while minimum at winter (250.52 mg/100g). Similarly maximum Fe was recorded at Dabola site (301.42 mg/100g) while minimum at Dam site (270.37 mg/100g).

8. Manganese (Mn) mg/100g:

The mean data of Mn for various seasons and sites is shown in Table-3.2. The results revealed that the effect of seasons and sites on Mn was significant while the effect of their interactions was non-significant.

Maximum Mn was observed at summer (17.85 mg/100g) while minimum at winter (10.14 mg/100g). Similarly maximum Mn was recorded at Mang site (16.15 mg/100g) while minimum at Dam site (11.70 mg/100g).

9. Zinc (Zn) mg/100g:

The mean data of Zn for various seasons and sites is shown in Table-3.2, while their interaction is shown in Figure-3.10. T The results revealed that the effect of seasons, sites and their interaction on Zn was significant.

Maximum Zn was observed at summer (7.13 mg/100g) while minimum at winter (5.25 mg/100g). Similarly maximum Zn was recorded at Mang site (7.53 mg/100g) while minimum at Jabri site (5.03 mg/100g). In case of interactions maximum Zn was observed at Mang site during summer (8.07 mg/100g) followed by Dabola site during summer (7.43 mg/100g). While minimum Zn was observed at Jabri site during winter (4.1 mg/100g)

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The above mentioned results revealed that bio-chemical elements, Na, Ca, P, Mg, Cu, Fe, Mn and Zn, were found significantly higher in summer season than in winter season while Potassium concentration in the leaves of Adhatoda vasica was non-significantly higher in summer than in winter. This might be due the finding that during summer, the mineral concentration remain high in most of the species (Ramireze et al., 2006) because mineral absorption is associated with high rate of transpiration due to high temperature during summer (Vidyarthi, 2002, Romney et al., 1960). Another reason for higher concentration of minerals during summer might be that summer is more active period bio-chemically and photo synthetically (Vidyarthi, 2002).

The sites comparison revealed that dam site produced maximum value for K, Ca and Mg in the leaves of Adhatoda. It was observed that the soil of Dam site was rich in K salts and having higher water holding capacity, as a result of which high K, Ca and Mg were recorded in leaves of Adhatoda (McDowell, 2003). Dabola site gave maximum value for P, Cu and Fe and Jabri site showed maximum results for Mn, which might be associated with environmental stresses as both the sites are at higher altitude (Harborne and Williams, 2000; Ali and Abbas, 2003). While Mang site produced maximum values for Na and Zn, which is closely associated with the mineral composition of the soil of the site (Demeyer and Dejaegere, 1996). This site to site variation shows that the soil texture and environmental interactions at sites were different from each other and that many reports confirm the present findings (Hu and Wang, 2004; Zhang et al., 2004).

Table-3.2. Effect of Different Seasons and Sites on Elemental analysis of Adhatoda vasica, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan.

Sodium Potassiu Calciu Phosph Magnesi Copper Mangan Zinc Iron (Fe) (Na) m (K) m (Ca) orus (P) um(Mg) (Cu) ese(Mn) (Zn) Seasons

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Summer 1.24a 148.87 128.52a 235.52a 0.48a 322.03a 17.85a 7.13a 230.72a 187.77b Winter 0.88b 144.49 113.52b 157.09b 0.16b 250.52b 10.14b 5.25b LSD at α 0.05 0.135 Ns 12.61 7.566 12.162 0.027 16.526 0.773 0.404 Sites

Dam 0.94 159.58 137.67a 201.32 0.23b 11.70d 5.57c 244.6a 270.37b 216.1b 301.42a Dabola 0.97 145.92 136.12a 197.25 0.35a 13.15c 6.63b 192.33c 298.78a Jabri 1.15 155.67 100.88c 201.28 0.37a 14.98b 5.03c 183.97c 274.53b Mang 1.17 125.57 109.40b 185.37 0.33a 16.15a 7.53a LSD at α 0.05 Ns Ns 22.30 5.458 ns 0.095 29.927 1.294 0.813 Interactions

Seasons*Sites Ns * Ns * ns * Ns ns * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

250

200

150 Summer 100 Winter 50

0 Dam dabola Jabri Mang

Figure-3.7: Effect of different seasons and sites on Potassium (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 200

150

100 Summer Winter 50

0 Dam dabola Jabri Mang

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Figure-3.8: Effect of different seasons and sites on Phosphorus (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 0.7 0.6 0.5 0.4 Summer 0.3 Winter 0.2 0.1 0 Dam dabola Jabri Mang

Figure-3.9: Effect of different seasons and sites on Copper (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub- Himalayan mountains of Pakistan. 10

6 Summer 4 Winter 2

0 Dam dabola Jabri Mang

Figure-3.10: Effect of different seasons and sites on Zinc (mg/100g) in leaves of Adhatoda vasica collected from Khanpur valley in the sub- Himalayan mountains of Pakistan.

2. CALATROPIS PROCERA.

I. Proximate Analysis:

1. Moisture%:

The mean data of moisture % for various seasons and sites is shown in Table-3.3. The results revealed that the effect of seasons on moisture %

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was significant while the effect of sites and their interactions was non- significant.

Maximum moisture % was observed at summer (92.15%) while minimum at winter (88.87%).

2. Dry Matter %:

The mean data of dry matter % for various seasons and sites is shown in Table-3.3. The results revealed that the effect of seasons on dry matter % was significant while the effect of sites and their interactions was non- significant.

Maximum dry matter % was observed at winter (11.13%) while minimum at summer (7.85%).

3. Ash / Minerals %:

The mean data of ash / minerals % for various seasons and sites is shown in Table-3.3. The results revealed that the effect of seasons and sites on Ash% was significant while that of their interactions on % was non- significant.

Maximum ash / minerals % was observed at summer (21.25%) while minimum at winter (17.84%). Similarly maximum ash / minerals % was recorded at Jabri site (20.98%) while minimum at Dam site (18.33%).

4. Crude Proteins %:

The mean data of crude proteins % for various seasons and sites is shown in Table-3.3, while their interaction is shown in Figure-3.11. The results revealed that the effect of seasons, sites and their interactions on crude proteins % was significant.

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Maximum crude proteins % was observed at summer (14.32%) while minimum at winter (12.47%). Similarly maximum crude proteins % was recorded at Jabri site (13.810%) while minimum at Dam site (12.72%). In case of interactions maximum crude proteins % was recorded at Mang site during summer (14.58%) followed by Jabri site during summer (14.54%) while minimum of it was observed at Dam site during winter (11.19%).

5. Crude Fibers %:

The mean data of crude fibers % for various seasons and sites is shown in Table-3.3, while their interaction is shown in Figure-3.12. The results revealed that the effect of seasons and their interactions on crude fibers % was significant while that of sites was non-significant..

Maximum crude fibers % was observed at winter (11.71%) while minimum at summer (9.06%). In case of interactions maximum crude fibers % was recorded at Dabola site during winter (13.39%) followed by Mang site during winter (11.64%) while minimum of it was observed at Dabola site during summer (8.64%).

6. Crude Fats %:

The mean data of crude fats % for various seasons and sites is shown in Table-3.3, while their interaction is shown in Figure-3.13. The results revealed that the effect of seasons and interactions on crude fats % was significant while the effect of sites was non-significant.

Maximum crude fats % was observed at summer (10.99%) while minimum at winter (9.56%). In case of interactions maximum crude fats % was recorded at Jabri site during summer (11.98%) followed by Dam site during summer (11.12%) while minimum of it was observed at Jabri site during winter (7.13%).

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7. Essential Oils%:

The mean data of essential oil % for various seasons and sites is shown in Table-3.3. The results revealed that the effect of sites on essential oil % was significant while the effect of seasons and interactions was non- significant.

Maximum essential oil % was recorded at Jabri site (2.03%) while minimum at Dam site (1.36%).

8. Nitrogen Free ExtracTable Substances (NFES) %:

The mean data of NFES % for various seasons and sites is shown in Table- 3.3, while their interaction is shown in Figure-3.14. The results revealed that the effect of seasons, sites and their interactions on NFES % was significant.

Maximum NFES % was observed at winter (48.42%) while minimum at summer (44.28%). Similarly maximum NFES % was recorded at Dam site (48.44%) while minimum at Dabola site (44.78%). In case of interactions maximum NFES % was recorded at Dam site during winter (50.80%) followed by Jabri site during winter (50.27%) while minimum of it was observed at Jabri site during summer (40.72%).

9. Net Free Energy Estimation (NFEE):

The mean data of NFEE for various seasons and sites is shown in Table- 3.3, while their interaction is shown in Figure-3.15. The results revealed that the effect of seasons and their interactions on NFEE was significant while that of sites was non-significant.

Maximum NFEE was observed at summer (197.18) while minimum at winter (181.48).

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In case of interactions maximum NFEE was recorded at Jabri site during summer (203.10) followed by Dam site during summer (199.86) while minimum of it was observed at Jabri site during winter (164.59).

It is evident from the results that summer and winter seasons have significantly affected the bio-chemical attributes except essential oils. The bio-chemical compounds like moisture, crude proteins, crude fats, ashes or total minerals and NFEE were higher in summer season than winter season in all the four sites of the study area. In Calatropis procera, which is an evergreen species, the plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and to keep nutrients for winter stresses. Summer rains and high water availability to plants facilitated the accumulation of these compounds in leaves during summer. These results were confirmed by many reports (McDowell, 2003; Ramirez et al., 2006). High protein and total minerals found in summer season may be due to availability of high moisture due to heavy rainfall in the valley.

On the other hand dry matter, crude fibers and nitrogen free extracts were found significantly higher during winter than summer in all of the sites. The accumulation of these compounds may be due to low temperature and deficiency of water and stresses (Ashraf et al., 1994; Harborne and Williams, 2000; Ali and Abbas, 2003). Furthermore, plants grown in cold areas accumulate oils and sugar during early winter to cope with severe cold and provide support to plant survival and protect it from frost injury (Rig and Cain, 1992). During winter plants faced the low temperature stress as a result of which high accumulation of dry matter and nitrogen free extracts were recorded. High soil EC was also observed during winter which also caused stressed conditions increasing the accumulation of these compounds (Smirnoff, 1993; Miranda- Ham et al., 2007). High NFES might be due to deficiency of nitrogen during winter. Low

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temperature also reduced absorption of N which further increased accumulation of NFES.

The sites comparison revealed that Dam site produced maximum value for NFES in the leaves of adhatoda. It was observed that the soil of dam site was having higher water holding capacity, as a result of which high moisture were recorded in leaves of Calatropis procera (McDowell, 2003). Dabola site gave maximum value for crude fibers and Jabri site showed maximum results for crude proteins, essential oils and ash which might be associated with environmental stresses as both the sites are at higher altitude (Harborne and Williams, 2000; Ali and Abbas, 2003). While Mang site produced maximum values for dry matter, crude fats and NFEE which are closely associated with the mineral composition of the soil of the site (Demeyer and Dejaegere, 1996; Augustus and Rutgers, 2006) and NFE at KG was higher due to high fat and protein contents in plants growing at this site which contain maximum energy. Increase in crude proteins, essential oils and ash might also be related with some environmental stress or anthropogenic stresses as clear cutting and grazing (Whittaker, 1970; Buchanan et al., 2000; Barroso et al., 2001) which are common at the site.

Table-3.3. Effect of Different Seasons and Sites on Proximate analysis of Calatropis procera, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan. Dry Ash/mi Crude Moistur Crude Essentia Matter nerals Proteins Fats% e% % % % Fibers% l Oils% Seasons NFES% NFEE Summer 92.15a 7.85b 21.25a 14.32a 9.06b 10.99a 1.73 44.28b 197.18a

Winter 88.87b 11.13a 17.84b 12.47b 11.71a 9.56b 1.64 48.42a 181.48b LSD at α 0.05 1.312 1.312 0.847 0.297 0.795 0.527 Ns 1.525 4.300 Sites

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Dam 92.00 8.00 18.33b 12.72b 9.75 10.75 1.36b 48.44a 192.90

Dabola 90.53 9.47 20.71a 13.66a 11.02 9.83 1.47b 44.78b 184.98

Jabri 90.78 9.22 20.98a 13.81a 10.17 9.55 2.03a 45.49b 183.85

Mang 88.72 11.27 18.37b 13.38ab 10.60 10.96 1.87a 46.69ab 195.59 LSD at α 0.05 Ns ns 1.294 0.692 ns ns 0.354 2.227 ns Interactions

Seasons*Sites Ns ns Ns * * * Ns * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

20 15 10 Summer 5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.11: Effect of different seasons and sites on Crude Protein contents in leaves of Calatropis procera.

14 12 10 8 6 Summer 4 2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.12: Effect of different seasons and sites on Crude Fats contents in leaves of Calatropis procera.

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16 14 12 10 8 6 Summer 4 Winter 2 0 Dam Dabola Jabri Mang Sites

Figure-3.13: Effect of different seasons and sites on Crude Fibres contents in leaves of Calatropis procera. 60 50 40 30 20 Summer 10 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.14: Effect of different seasons and sites on NFES contents in leaves of Calatropis procera.

250 200 150 100 Summer Winter 50 0 Dam Dabola Jabri Mang Sites

Figure-3.15: Effect of different seasons and sites on NFEE in leaves of Calatropis procera.

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II. Elementology:

1. Sodium (Na) mg/100g:

The mean data of Na for various seasons and sites is shown in Table-3.4. The results revealed that the effect of seasons and sites on Na was significant while the effect of their interaction was non-significant.

Maximum Na was observed at summer (7.16 mg/100g) while minimum at winter (5.41 mg/100g). Similarly maximum Na was recorded at Mang site (8.09 mg/100g) while minimum at Dam site (5.12 mg/100g).

2. Potassium (K) mg/100g:

The mean data of K for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.16. The results revealed that the effect of seasons, sites and their interaction on K was significant.

Maximum K was observed at summer (390.79 mg/100g) while minimum at winter (346.92 mg/100g). Similarly maximum K was recorded at Dabola site (447.50 mg/100g) while minimum at Jabri site (276.25 mg/100g). In case of interactions maximum K was observed at Dam site during summer (500.33 mg/100g) followed by dabola site during summer (454.00 mg/100g) and then followed by Mang site during summer (439.67 mg/100g). While minimum K was observed at Jabri site during summer (169.17 mg/100g)

3. Calcium (Ca) mg/100g:

The mean data of Ca for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.17. The results revealed that the effect of seasons, sites and their interaction on Ca was significant.

Maximum Ca was observed at winter (187.00 mg/100g) while minimum at summer (167.67 mg/100g). Similarly maximum Ca was recorded at 154

Dabola site (196.13 mg/100g) while minimum at Dam site (157.38 mg/100g). In case of interactions maximum Ca was observed at Dabola site during winter (274.07 mg/100g) followed by Dam site during summer (198.90 mg/100g). While minimum Ca was observed at Dabola site during summer (118.20 mg/100g)

4. Phosphorus (P) mg/100g:

The mean data of Phosphorus for various seasons and sites is shown in Table-3.4. The results revealed that the effect of seasons and sites on P was significant while that of their interactions was non-significant.

Maximum P was observed at winter (99.49 mg/100g) while minimum at summer (84.75 mg/100g). Similarly maximum P was recorded at Dabola site (100.25 mg/100g) while minimum at Mang site (81.87 mg/100g).

5. Magnesium (Mg) mg/100g:

The mean data of Mg for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.18. The results revealed that the effect of seasons on Mg was non-significant while the effect of sites and of their interaction was significant.

Maximum Mg was recorded at Dabola site (244.68 mg/100g) while minimum at Dam site (198.53 mg/100g). In case of interactions maximum Mg was observed at Dabola site during winter (291.57 mg/100g) followed by Mang site during winter (228.67 mg/100g). While minimum Mg was observed at Dam site during winter (193.90 mg/100g)

6. Copper (Cu) mg/100g:

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The mean data of Cu for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.19. The results revealed that the effect of seasons, sites and their interaction on Cu was significant.

Maximum Cu was observed at winter (0.65 mg/100g) while minimum at summer (0.59 mg/100g). Similarly maximum Cu was recorded at Jabri site (0.67 mg/100g) while minimum at Dabola and Mang sites (0.0.58 mg/100g). In case of interactions maximum Cu was observed at Jabri site during summer (0,72 mg/100g) followed by Dam site during winter (0.68 mg/100g). While minimum Cu was observed at Mang site during summer (0,50 mg/100g)

7. Iron (Fe) mg/100g:

The mean data of Fe for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.20. The results revealed that the effect of seasons, sites and their interaction on Cu was significant.

Maximum Fe was observed at summer (252.35 mg/100g) while minimum at winter (202.54 mg/100g). Similarly maximum Fe was recorded at Dam site (243.57 mg/100g) while minimum at Mang site (207.13 mg/100g). In case of interactions maximum Fe was observed at Dam site during summer (291.60 mg/100g) followed by Jabri site during summer (249.63 mg/100g). While minimum Fe was observed at Mang site during winter (179.30 mg/100g)

8. Manganese (Mn) mg/100g:

The mean data of Mn for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.21. The results revealed that the effect of seasons and interactions on Mn was significant while the effect of sites was non-significant.

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Maximum Mn was observed at summer (16.38 mg/100g) while minimum at winter (12.28 mg/100g). In case of interactions maximum Mn was observed at Dam site during summer (19.27 mg/100g) followed by Dabola site during summer (18.97 mg/100g). While minimum Mn was observed at Dam site during winter (10.53 mg/100g)

9. Zinc (Zn) mg/100g:

The mean data of Zn for various seasons and sites is shown in Table-3.4, while their interaction is shown in Figure-3.22. The results revealed that the effect of sites and interactions on Mn was significant while the effect of seasons was non-significant.

Maximum Zn was recorded at Dabola site (5.55 mg/100g) while minimum at Dam site (4.27 mg/100g). In case of interactions maximum Zn was observed at Mang site during winter (6.03 mg/100g) followed by Dam site during summer (5.97 mg/100g). While minimum Zn was observed at Dam site during winter (2.57 mg/100g).

The statistical analysis of the data recorded on bio-chemical attributes revealed that biochemical elements, Na, K, Fe and Mn were found significantly higher in summer season than in winter season while Mg and Zn concentration in the leaves of Calatropis procera was non- significantly higher in summer than in winter. This might be due the finding that during summer, the mineral concentration remain high in most of the species (Ramireze et al., 2006) because mineral absorption is associated with high rate of transpiration due to high temperature during summer (Vidyarthi, 2002, Romney et al., 1960). Another reason for higher concentration of these minerals during summer might be due to the reason that summer is more active period bio-chemically and photo synthetically (Zhang et al., 2004). Similarly Ca, P and Cu were significantly higher in winter than summer

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The sites comparison revealed that Dabola site produced maximum value for K, Ca, P, Mg, Fe, Mn and Zn, while Na at Mang and Cu at Jabri was in maximum concentration in the leaves of Calatropis procera. It was observed that the soil of Dabola site was rich in K salts and having higher water holding capacity, which might have caused the accumulation of higher concentrations of K, Ca, P, Mg, Fe, Mn and Zn in leaves of Calatropis procera (McDowell, 2003). Jabri site showed maximum results for Cu which might be associated with environmental stresses as Jabri is at higher altitude (Harborne and Williams, 2000; Ali and Abbas, 2003). While Mang site produced maximum values for Na which is closely associated with the mineral composition of the soil of the site (Demeyer and Dejaegere, 1996). This site to site variation shows that the soil and environmental conditions at sites were different from each other which resulted in the accumulation of nutrients in the plants with different levels of concentrations and that many reports confirm the present findings (Hu and Wang, 2004; Zhang et al., 2004). The availability of more bio-chemical elements in the leaves of Calatropis procera at Dabola site might be due to stressed conditions like low temperature in winter and soil conditions (Harborne and Williams, 2000; Ali and Abbas, 2003; Eskin, 1989; Essafi et al., 2006, Ahmad et al. 2014 ).

Table-3.4 Effect of Different Seasons and Sites on Elemental analysis of Calatropis procera, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan. Sodium Potassi Calciu Phospho Magnesi Copper Iron (Fe) Mangan Zinc Seasons (Na) um (K) m (Ca) rus (P) um(Mg) (Cu) ese(Mn) (Zn) Summer 7.16a 390.79a 167.67b 84.75b 215.93a 0.59b 252.35a 16.38a 5.27a

Winter 5.41b 346.92b 187.00a 99.49a 234.25a 0.65a 202.54b 12.28b 4.71a LSD at α 0.05 0.403 18.237 12.989 4.124 ns 0.044 18.958 0.976 ns Sites

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Dam 5.12c 432.33a 157.38c 91.50b 198.53b 0.65a 243.57a 14.90 4.27b

Dabola 5.19c 447.50a 196.13a 100.25a 244.68a 0.58b 236.52ab 15.08 5.55a

Jabri 6.75b 276.25c 186.28a 94.87ab 234.73a 0.67a 222.57bc 13.80 4.82ab

Mang 8.09a 319.33b 169.55b 81.87c 222.40ab 0.58b 207.13c 13.53 5.33a LSD at α 0.05 0.977 32.016 10.457 7.033 30.058 0.047 19.721 ns 0.762 Interactions

Seasons*Sites Ns * * ns * * * * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

700 600 500 400 300 Summer 200 100 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.16: Effect of different seasons and sites on Potassium (K) contents in leaves of Calatropis procera.

350 300 250 200 150 Summer 100 Winter 50 0 Dam Dabola Jabri Mang Sites

Figure-3.17: Effect of different seasons and sites on Calcium (Ca) contents in leaves of Calatropis procera.

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350 300 250 200 150 Summer 100 50 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.18: Effect of different seasons and sites on Magnesium (Mg) contents in leaves of Calatropis procera. 1 0.8 0.6

0.4 Summer 0.2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.19: Effect of different seasons and sites on Copper (Cu) contents in leaves of Calatropis procera.

350 300 250 200 150 Summer 100 50 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.20: Effect of different seasons and sites on Iron (Fe) contents in leaves of Calatropis procera.

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25 20 15

10 Summer 5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.21: Effect of different seasons and sites on Manganese (Mn) contents in leaves of Calatropis procera. 8 7 6 5 4 3 Summer 2 Winter 1 0 Dam Dabola Jabri Mang Sites

Figure-3.22: Effect of different seasons and sites on Zinc (Zn) contents in leaves of Calatropis procera.

3.3. RECINUS COMMUNIS.

I. Proximate Analysis:

1. Moisture%:

The mean data of moisture % for various seasons and sites is shown in Table-3.5. The results revealed that the effect of seasons on moisture % was significant while the effect of sites and their interactions was non- significant. Maximum moisture % was observed at summer (74.52%) while minimum at winter (69.23%). 161

2. Dry Matter %:

The mean data of dry matter % for various seasons and sites is shown in Table-3.5. The results revealed that the effect of seasons on dry matter % was significant while the effect of sites and their interactions was non- significant.

Maximum dry matter % was observed at winter (30.82%) while minimum at summer (25.47%).

3. Ash / Minerals %:

The mean data of ash / minerals % for various seasons and sites is shown in Table-3.5. The results revealed that the effect of seasons and sites on ash / minerals % was significant while a non-significant effect was observed for interactions on ash/minerals %..

Maximum ash / minerals % was observed at summer (16.97%) while minimum at winter (14.47%). Similarly maximum ash / minerals % was recorded at Jabri site (17.11%) while minimum at Mang site (14.24%).

4. Crude Proteins %:

The mean data of crude proteins % for various seasons and sites is shown in Table-3.5, while their interaction is shown in Figure-3.23. The results revealed that the effect of seasons and their interactions on crude proteins % was significant while that of sites was non-significant.

Maximum crude proteins % was observed at summer (13.21%) while minimum at winter (11.87%). In case of interactions maximum crude proteins % was recorded at Mang site during summer (15.26%) followed by Jabri site during summer (14.78%) while minimum of it was observed at Dabola site during winter (11.77%).

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5. Crude Fibers %:

The mean data of crude fibers % for various seasons and sites is shown in Table-3.5, while their interaction is shown in Figure-3.24. The results revealed that the effect of seasons, sites and their interactions on crude fibers % was significant.

Maximum crude fibers % was observed at winter (12.26%) while minimum at summer (9.82%). Similarly maximum crude fibers % was recorded at Dabola site (12.53%) while minimum at Dam site (9.36%). In case of interactions maximum crude fibers % was recorded at Dabola site during winter (13.85%) followed by Mang site during winter (13.57%) while minimum of it was observed at Mang site during summer (9.26%).

6. Crude Fats %:

The mean data of crude fats % for various seasons and sites is shown in Table-3.5, while their interaction is shown in Figure-3.25. The results revealed that the effect of seasons and their interactions on crude fats % was significant while that of sites was non-significant.

Maximum crude fats % was observed at summer (9.63%) while minimum at winter (7.82%). In case of interactions maximum crude fats % was recorded at Jabri site during summer (10.24%) followed by Dam site during summer (9.79%) while minimum of it was observed at Mang site during winter (7.35%).

7. Essential Oils%:

The mean data of essential oil % for various seasons and sites is shown in Table-3.5, while their interaction is shown in Figure-3.26. The results

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revealed that the effect of seasons, sites and their interactions on essential oil % was significant.

Maximum essential oil % was observed at winter (2.76%) while minimum at summer (2.17%). Similarly maximum essential oil % was recorded at Mang site (3.47%) while minimum at Dam site (1.58%). In case of interactions maximum essential oil % was recorded at Mang site during winter (3.48%) followed by Mang site during winter (3.47%) while minimum of it was observed at Dam site during summer (1.52%).

8. Nitrogen Free ExtracTable Substances (NFES) %:

The mean data of NFES % for various seasons and sites is shown in Table- 3.5. The results revealed that the effect of seasons and sites on NFES was significant while that of their interactions was non-significant.

Maximum NFES % was observed at winter (53.25%) while minimum at summer (49.45%). Similarly maximum NFES % was recorded at Dam site (52.97%) while minimum at Jabri site (49.27%).

9. Net Free Energy Estimation (NFEE):

The mean data of NFEE for various seasons and sites is shown in Table- 3.5, while their interaction is shown in Figure-3.27. The results revealed that the effect of seasons and their interactions on NFEE % was significant while that of sites was non-significant.

Maximum NFEE was observed at summer (189.74) while minimum at winter (170.09). In case of interactions maximum NFEE was recorded at Jabri site during summer (195.29) followed by Dabola site during summer (189.04) while minimum of it was observed at Mang site during winter (166.31).

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It is evident from the results that summer and winter seasons have significantly affected the bio-chemical attributes. The bio-chemical compounds like moisture, ashes, crude proteins, crude fats and NFEE were higher in summer season than winter season in the leaves of wild Jatropa. In Recinus communis, which is an evergreen shrub, the plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and to keep nutrients for winter stresses. Summer rains and high water availability to plants facilitated the accumulation of these compounds in leaves during summer. These results were confirmed by many reports (McDowell, 2003; Ramirez et al., 2006).

The sites comparison revealed that majority of the parameters were not significantly affected by sites and only ashes, crude fibers, essential oils and NFES were significantly varied among sites. Jabri site showed maximum results for essential oils and ashes while Dabola site gave maximum crude fiber which might be associated with environmental stresses as both the sites are at higher altitude and the plant prefer low plains (Harborne and Williams, 2000; Ali and Abbas, 2003). highest value of NFES at Dam site

Table-3.5. Effect of Different Seasons and Sites on Proximate Analysis of Recinus communis, indigenous to Khanpur Valley, in subHimalayan mountains of Pakistan.

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Dry Ash/mi Crude Moistur Crude Essentia Matter nerals Proteins Fats% e% % % % Fibers% l Oils% Seasons NFES% NFEE Summer 74.52 25.47 16.97a 14.13a 9.82b 9.63 2.17 49.45 189.74

Winter 69.23 30.82 14.47b 12.20b 12.26a 7.82 2.76 53.25 170.09 LSD at α 0.05 1.958 1.958 1.117 0.385 0.548 0.265 0.206 1.160 1.892 Sites

Dam 73.90 26.19 16.39a 12.64 9.36c 8.63 1.58d 52.97a 178.63

Dabola 71.46 28.54 15.16b 12.77 12.53a 9.13 2.12c 50.41b 180.92

Jabri 71.67 28.33 17.11a 13.68 10.84b 9.10 2.69b 49.27b 183.19

Mang 70.48 29.52 14.24c 13.58 11.41b 8.03 3.47a 52.74a 176.95 LSD at α 0.05 Ns ns 0.750 ns 0.947 ns 0.499 1.399 ns Interactions

Seasons*Sites Ns ns ns * * * * ns * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

20 15 10 Summer 5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.23: Effect of different seasons and sites on Crude Protein contents in leaves of Recinus communis.

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12 10 8 6 Summer 4 2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.24: Effect of different seasons and sites on Crude Fats contents in leaves of Recinus communis. 16 14 12 10 8 6 Summer 4 Winter 2 0 Dam Dabola Jabri Mang Sites

Figure-3.25: Effect of different seasons and sites on Crude Fibers contents in leaves of Recinus communis.

5 4 3

2 Summer 1 Winter 0 Dam Dabola Jabri Mang Sites

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Figure-3.26: Effect of different seasons and sites on Essential Oil contents in leaves of Recinus communis.

200 190 180 170 160 Summer 150 Winter 140 Dam Dabola Jabri Mang Sites

Figure-3.27: Effect of different seasons and sites on NFEE in leaves of Recinus communis.

II. Elementology:

1. Sodium (Na) mg/100g:

The mean data of Na for various seasons and sites is shown in Table-3.6, while their interaction is shown in Figure-3.28. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum Na was observed at summer (1.55 mg/100g) while minimum at winter (1.37 mg/100g). Similarly maximum Na was recorded at Dam site (3.07 mg/100g) while minimum at Mang site (1.41 mg/100g). In case of interactions maximum Na was observed at Dam site during summer (4.91 mg/100g) followed by Jabri site during summer (2.22 mg/100g). While minimum Na was observed at Dam site during winter (1.22 mg/100g)

2. Potassium (K) mg/100g:

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The mean data of K for various seasons and sites is shown in Table-3.6. The results revealed that the effect of seasons and sites on K was significant while the effect of their interaction was non-significant.

Maximum K was observed at summer (83.67 mg/100g) while minimum at winter (70.17 mg/100g). Similarly maximum K was recorded at Dam site (96.08 mg/100g) while minimum at Jabri site (57.25 mg/100g).

3. Calcium (Ca) mg/100g:

The mean data of Ca for various seasons and sites is shown in Table-3.6. The results revealed that the effect of seasons and sites on Ca was significant while the effect of their interactions was non-significant.

Maximum Ca was observed at summer (105.79 mg/100g) while minimum at winter (84.17 mg/100g). Similarly maximum Ca was recorded at Dabola site (152.57 mg/100g) while minimum at Dam site (71.42 mg/100g).

4. Phosphorus (P) mg/100g:

The mean data of Phosphorus for various seasons and sites is shown in Table-3.6. The results revealed that the effect of seasons on P % was significant while the impact of sites and their interaction on P was non- significant.

Maximum P was observed at winter (171.72 mg/100g) while minimum at summer (135.72 mg/100g).

5. Magnesium (Mg) mg/100g:

The mean data of Mg for various seasons and sites is shown in Table-3.6. The results revealed that the effect of sites on Mg was significant while the effect of seasons and of their interaction was non-significant.

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Maximum Mg was observed at Dam site (294.17 mg/100g) while minimum at Mang site (236.68 mg/100g).

6. Copper (Cu) mg/100g:

The mean data of Cu for various seasons and sites is shown in Table-3.6. The results revealed that the effect of seasons and sites on Cu was significant while the effect of their interactions was non-significant.

Maximum Cu was observed at summer (0.62 mg/100g) while minimum at winter (0.37 mg/100g). Similarly maximum Cu was recorded at Dabola and Jabri sites (0.57 mg/100g) while minimum at Dam site (0.39 mg/100g).

7. Iron (Fe) mg/100g:

The mean data of Fe for various seasons and sites is shown in Table-3.6, while their interaction is shown in Figure-3.29. The results revealed that the effect of seasons, sites and their interaction on Fe was significant.

Maximum Fe was observed at summer (260.12 mg/100g) while minimum at winter (208.25 mg/100g). Similarly maximum Fe was recorded at Jabri site (279.08 mg/100g) while minimum at Mang site (197.58 mg/100g). In case of interactions maximum Fe was observed at Jabri site during summer (300.37 mg/100g) followed by Dabola site during summer (294.50 mg/100g). While minimum Fe was observed at Dabola site during winter (186.17 mg/100g)

8. Manganese (Mn) mg/100g:

The mean data of Mn for various seasons and sites is shown in Table-3.6, while their interaction is shown in Figure-3.30. The results revealed that the effect of seasons, sites and their interaction on Mn was significant.

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Maximum Mn was observed at summer (12.41 mg/100g) while minimum at winter (8.70 mg/100g). Similarly maximum Mn was recorded at Dam site (12.33 mg/100g) while minimum at Mang site (9.75 mg/100g). In case of interactions maximum Mna was observed at Dabola site during summer (13.43 mg/100g) followed by Dam site during summer (13.20 mg/100g). While minimum Mn was observed at Dabola site during winter (7.27 mg/100g)

9. Zinc (Zn) mg/100g:

The mean data of Zn for various seasons and sites is shown in Table-3.6, while their interaction is shown in Figure-3.31. The results revealed that the effect of seasons, sites and their interaction on Zn was significant.

Maximum Zn was observed at summer (6.50 mg/100g) while minimum at winter (3.67 mg/100g). Similarly maximum Zn was recorded at Mang site (7.15 mg/100g) while minimum at Jabri site (3.89 mg/100g). In case of interactions maximum Zn was observed at Mang site during summer (8.07 mg/100g) followed by Dam site during summer (7.13 mg/100g). While minimum Zn was observed at Dam site during winter (2.23 mg/100g).

The statistical analysis of the data recorded on bio-chemical attributes revealed that biochemical elements, Na, Ca, P, K, Cu, Fe, Mn and Zn, were found significantly higher in summer season than in winter season while Magnisium concentration in the leaves of Recinus communis was non-significantly higher in summer than in winter. This might be due the finding that during summer, the mineral concentration remain high in most of the species (Ramireze et al., 2006) because mineral absorption is associated with high rate of transpiration due to high temperature during summer (Vidyarthi, 2002, Romney et al., 1960). Another reason for higher concentration of minerals during summer might be that summer is

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more active period bio-chemically and photo synthetically as stated by Zhang et al., 2004.

The sites comparison revealed that Dam site produced maximum value for Na, K, Mg and Mn in the leaves of Recinus communis. It was observed that the soil of Dam site was rich in K salts and having higher water holding capacity, as a result of which high K, Na, Mn and Mg were recorded in leaves of Recinus communis (McDowell, 2003). Dabola site gave maximum value for Ca and Cu and Jabri site showed maximum results for Fe, P and Cu, which might be associated with environmental stresses as both the sites are at higher altitude (Harborne and Williams, 2000; Ali and Abbas, 2003). While Mang site produced maximum values for Zn, which is closely associated with the mineral composition of the soil of the site (Demeyer and Dejaegere, 1996). This site to site variation show that the soil texture and environmental interactions at sites were different from each other and that many reports confirm the present findings (Hu and Wang, 2004; Zhang et al., 2004).

Table-3.6. Effect of Different Seasons and Sites on Elemental analysis of Recinus communis, indigenous to Khanpur Valley, in subHimalayan mountains of Pakistan.

Sodium Potassi Calciu Phospho Magnesi Copper Mangan Zinc Iron (Fe) (Na) um (K) m (Ca) rus (P) um(Mg) (Cu) ese(Mn) (Zn) Seasons Summer 2.55a 83.67a 105.79a 135.72b 264.20b 0.62a 260.12a 12.41a 6.50a

Winter 1.37b 70.17b 84.17b 171.72a 284.25a 0.37b 208.25b 8.70b 3.67b LSD at α 0.05 0.215 13.457 20.594 15.957 ns 0.061 15.129 0.813 0.555 Sites

Dam 3.07a 96.08a 71.42c 147.62 294.17a 0.39b 219.73bc 12.33a 4.68b

Dabola 1.61b 60.25b 122.57a 170.33 282.98a 0.57a 240.33b 10.35b 4.65c

Jabri 1.74b 57.25b 82.87bc 159.03 283.07a 0.57a 279.08a 9.78b 3.89c

Mang 1.41b 94.08a 103.08ab 137.35 236.68b 0.45b 197.58c 9.75b 7.15a LSD at α 0.05 0.775 16.963 30.282 Ns 28.111 0.067 30.735 1.130 0.791

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Interactions

Seasons*Sites * ns Ns ns ns ns * * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

7 6 5 4 3 Summer 2 1 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.28: Effect of different seasons and sites on Sodium content in leaves of Recinus communis.

350 300 250 200 150 Summer 100 50 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.29: Effect of different seasons and sites on Iron (Fe) contents in leaves of Recinus communis.

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16 14 12 10 8 6 Summer 4 Winter 2 0 Dam Dabola Jabri Mang Sites

Figure-3.30: Effect of different seasons and sites on Manganese (Mn) contents in leaves of Recinus communis. 10 8 6

4 Summer 2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.31: Effect of different seasons and sites on Zinc (Zn) contents in leaves of Recinus communis.

3.4. AJUGA BRACTIOSA.

I. Proximate Analysis:

1. Moisture%:

The mean data of moisture % for various seasons and sites is shown in Table-3.7. The results revealed that the effect of seasons on moisture % was significant while the effect of sites and their interactions was non- significant. Maximum moisture % was observed at summer (89.27%) while minimum at winter (86.80%). 174

2. Dry Matter %:

The mean data of dry matter % for various seasons and sites is shown in Table-3.7. The results revealed that the effect of seasons on dry matter % was significant while the effect of sites and their interactions was non- significant. Maximum dry matter % was observed at winter (13.21%) while minimum at summer (10.73%).

3. Ash / Minerals %:

The mean data of ash / minerals % for various seasons and sites is shown in Table-3.7, while their interaction is shown in Figure-3.32. The results revealed that the effect of seasons, sites and their interactions on ash / minerals % was significant.

Maximum ash / minerals % was observed at summer (30.00%) while minimum at winter (22.84%). Similarly maximum ash / minerals % was recorded at Jabri site (29.75%) while minimum at Mang site (23.97%). In case of interactions maximum ash % was recorded at Jabri site during summer (34.13%) while minimum of it was observed at Dabola site during winter (21.69%).

4. Crude Proteins %:

The mean data of crude proteins % for various seasons and sites is shown in Table-3.7, while their interaction is shown in Figure-3.33. The results revealed that the effect of seasons and their interactions on crude proteins % was significant while the effect of sites is non-significant.

Maximum crude proteins % was observed at summer (14.20%) while minimum at winter (11.83%). In case of interactions maximum crude proteins % was recorded at Dam site during summer (15.35%) followed

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by Dabola site during summer (14.73%) while minimum of it was observed at Dam site during winter (11.25%).

5. Crude Fibers %:

The mean data of crude fibers % for various seasons and sites is shown in Table-3.7, while their interaction is shown in Figure-3.34. The results revealed that the effect of seasons and sites on crude fibers % was significant while that of interactions is nonsignificant.

Maximum crude fibers % was observed at winter (12.64%) while minimum at summer (9.44%). Similarly maximum crude fibers % was recorded at Jabri site (11.65%) while minimum at Mang site (10.48%).

6. Crude Fats %:

The mean data of crude fats % for various seasons and sites is shown in Table-3.7, while their interaction is shown in Figure-3.35. The results revealed that the effect of seasons and their interactions on crude fats % was significant while the effect of sites is non-significant.

Maximum crude fats % was observed at summer (9.65%) while minimum at winter (7.83%). In case of interactions maximum crude fats % was recorded at Jabri site during summer (10.30%) followed by Dam site during summer (10.11%) while minimum of it was observed at Jabri site during winter (6.93%).

7. Essential Oils%:

The mean data of essential oil % for various seasons and sites is shown in Table-3.7, while their interaction is shown in Figure-3.36. The results revealed that the effect of seasons on essential oil % was non-significant while the effect of sites and interactions was significant.

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Maximum essential oil % was recorded at Mang site (2.33%) while minimum at Jabri site (0.91%). In case of interactions maximum essential oil % was recorded at Mang site during summer (2.48%) followed by Mang site during winter (2.18%) while minimum of it was observed at Jabri site during summer (0.40%).

8. Nitrogen Free ExtracTable Substances (NFES) %:

The mean data of NFES % for various seasons and sites is shown in Table- 3.7, while their interaction is shown in Figure-3.37. The results revealed that the effect of seasons, sites and their interactions on NFES % was significant.

Maximum NFES % was observed at winter (44.87%) while minimum at summer (36.70%). Similarly maximum NFES % was recorded at Mang site (44.35%) while minimum at Jabri site (37.22%). In case of interactions maximum NFES % was recorded at Dabola site during winter (46.17%) followed by Dam site during winter (45.87%) while minimum of it was observed at Jabri site during summer (31.53%).

9. Net Free Energy Estimation (NFEE):

The mean data of NFEE for various seasons and sites is shown in Table- 3.7, while their interaction is shown in Figure-3.38. The results revealed that the effect of seasons, sites and their interactions on NFEE was significant.

Maximum NFEE was observed at summer (177.49) while minimum at winter (160.29). Similarly maximum NFEE was recorded at Dam site (173.95) while minimum at Jabri site (163.24). In case of interactions maximum NFEE was recorded at Dam site during summer (186.28) followed by Jabri site during summer (176.54) while minimum of it was observed at Jabri site during winter (149.94).

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The results revealed that the effect of seasons on all parameters were highly significant. In summer, maximum values were found for moisture, crude protein, crude fats, minerals/ashes, essential oils and NFEE while in winter maximum values were found for dry matter, crude fibers and NFES. This seasonal effect was reasonably due to the plant growth habit and environmental conduciveness. Ajuga sprouts during summer and during last week of July, which is the metabolically most active period of its growth (Zavodnik, 1998); most of the compounds were in high concentration. Another reason of high moisture, essential nutrients and mineral during summer was high soil moisture contents (McDowell, 2003) due to more rain fall and high rate of transpiration due to high temperature (Vidyarthi, 2002). So when more growth took place, more dry biomass was also produced. Ajuga plants sprout naturally in May-July and plants matures in September-December. The rapid growth stage of the plant and availability of more moisture at summer in most of the sites was the possible reason for higher percentages of moisture, CP, CFts and NFEE during summer at these sites (McDowell, 2003; Ramirez et al., 2006). Higher values for dry matter, crude fibers and NFES at winter is definitely an indirect proportion to the low moisture and other nutrients accumulation in the plant tissues due to environmental and physiological stresses (Miranda- Ham et al., 2007 and Ahmad et al. 2014)

The effect of sites on moisture, dry matter, crude proteins and crude fats was nonsignificant. Maximum values were found for NFEE at Dam site while maximum values for essential oil and NFES were found at Mang site. At Jabri, maximum values were recorded for minerals, and crude fibers. High values of ashes and crude fibers at Jabri might be due to favorable conditions of the soil, as most of the plant species contain these compounds higher at maturity (Sher et al. 2010).

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Table-3.7. Effect of Different Seasons and Sites on Elemental analysis of Ajuga bracteosa, indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan.

Dry Ash/mi Crude Moistur Crude Essentia Matter nerals Proteins Fats% e% Fibers% l Oils% NFES Seasons % % % NFEE % Summer 89.27a 10.73b 30.00a 14.20a 9.44b 9.65a 1.38 36.70b 177.49a

Winter 86.80b 13.21a 22.84b 11.83b 12.64a 7.83b 1.55 44.87a 160.29b LSD at α 0.05 0.826 0.826 0.586 0.338 0.386 0.521 ns 0.791 4.805

Sites

Dam 86.99 13.01 24.84c 13.29 11.32ab 9.12 1.46b 41.42b 173.95a

Dabola 88.75 11.25 27.10b 13.28 10.70bc 8.75 1.15bc 40.16b 169.42ab

Jabri 87.84 12.16 29.75a 12.76 11.65a 8.62 0.91c 37.22c 163.24b

Mang 88.55 11.45 23.97c 12.72 10.48c 8.67 2.33a 44.35a 168.96ab LSD at α 0.05 Ns Ns 1.712 Ns 0.863 ns 0.539 2.351 6.386 Interactions

Seasons*Sites ns Ns * * ns * * * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant.and * = Significant at 5 % level of probability.

40 35 30 25 20 15 Summer 10 Winter 5 0 Dam Dabola Jabri Mang Sites

Figure-3.33: Effect of different seasons and sites on Ash contents of Ajuga bracteosa.

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20 15 10 Summer 5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.34: Effect of different seasons and sites on Crude Protein contents of Ajuga bracteosa. 12 10 8 6 Summer 4 2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.35: Effect of different seasons and sites on Crude Fats contents of Ajuga bracteosa.

3.5 3 2.5 2 1.5 Summer 1 Winter 0.5 0 - 0.5 Dam Dabola Jabri Mang Sites

Figure-3.36. Effect of different seasons and sites on Essential Oil contents of Ajuga bracteosa. 180

250 200 150 100 Summer 50 Winter 0 - 50 Dam Dabola Jabri Mang - 100 Sites

Figure-3.37. Effect of different seasons and sites on NFES contents of Ajuga bracteosa. 50

20 Summer 10 Winter

0 Dam Dabola Jabri Mang Sites

Figure-3.38: Effect of different seasons and sites on NFEE of Ajuga bracteosa.

II. Elementology:

1. Sodium (Na) mg/100g:

The mean data of Na for various seasons and sites is shown in Table-3.8, while their interaction is shown in Figure-3.39. The results revealed that the effect of seasons, sites and their interaction on Na was significant.

Maximum Na was observed at summer (5.85 mg/100g) while minimum at winter (4.24 mg/100g). Similarly maximum Na was recorded at Mang site (6.60 mg/100g) while minimum at Dabola site (4.18 mg/100g). In

181

case of interactions maximum Na was observed at Mang site during winter (6.96 mg/100g) followed by Jabri site during summer (6.45 mg/100g). While minimum Na was observed at Dam site during winter (2.74 mg/100g)

2. Potassium (K) mg/100g:

The mean data of K for various seasons and sites is shown in Table-3.8, while their interaction is shown in Figure-3.40. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum K was observed at winter (475.25 mg/100g) while minimum at summer (162.79 mg/100g). Similarly maximum K was recorded at Jabri site (377.33 mg/100g) while minimum at Dabola site (244.50 mg/100g). In case of interactions maximum K was observed at Jabri site during winter (559.67 mg/100g) followed by Dam site during Winter (485.00 mg/100g). While minimum K was observed at Dam site during summer (83.67 mg/100g)

3. Calcium (Ca) mg/100g:

The mean data of Ca for various seasons and sites is shown in Table-3.8, while their interaction is shown in Figure-3.41. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum Ca was observed at summer (194.64 mg/100g) while minimum at winter (127.66 mg/100g). Similarly maximum Ca was recorded at Jabri site (197.95 mg/100g) while minimum at Mang site (120.98 mg/100g). In case of interactions maximum Ca was observed at Jabri site during summer (254.20 mg/100g) followed by Dam site during summer (199.00 mg/100g). While minimum Ca was observed at Mang site during winter (95.10 mg/100g)

4. Phosphorus (P) mg/100g: 182

The mean data of Phosphorus for various seasons and sites is shown in Table-3.8. The results revealed that the effect of seasons and sites on Fe was significant while the effect of their interactions was non-significant.

Maximum P was observed at winter (196.73 mg/100g) while minimum at summer (138.13 mg/100g). Similarly maximum P was recorded at Jabri site (212.78 mg/100g) while minimum at Mang site (98.87 mg/100g).

5. Magnesium (Mg) mg/100g:

The mean data of Mg for various seasons and sites is shown in Table-3.8, while their interaction is shown in Figure-3.42. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum Mg was observed at winter (264.31 mg/100g) while minimum at summer (174.94 mg/100g). Similarly maximum Mg was recorded at Mang site (233.48 mg/100g) while minimum at Jabri site (204.42 mg/100g). In case of interactions maximum Mg was observed at Mang site during winter (288.57 mg/100g) followed by Dabola site during Winter (263.40 mg/100g). While minimum Mg was observed at Jabri site during summer (152.50 mg/100g)

6. Copper (Cu) mg/100g:

The mean data of Cu for various seasons and sites is shown in Table-3.8, while their interaction is shown in Figure-3.43. The results revealed that the effect of seasons, sites and their interaction on Cu was significant.

Maximum Cu was observed at winter (0.92 mg/100g) while minimum at summer (0.56 mg/100g). Similarly maximum Cu was recorded at Dabola site (0.86 mg/100g) while minimum at Dam and Jabri sites (0.69 mg/100g). In case of interactions maximum Cu was observed at Dabola site during winter (1.017 mg/100g) followed by Mang site during winter

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(0.097 mg/100g). While minimum Cu was observed at Mang site during summer (0.47 mg/100g)

7. Iron (Fe) mg/100g:

The mean data of Fe for various seasons and sites is shown in Table-3.8. The results revealed that the effect of seasons on Fe was significant while the effect of sites and their interactions was non-significant.

Maximum Fe was observed at winter (238.88 mg/100g) while minimum at summer (178.13 mg/100g).

8. Manganese (Mn) mg/100g:

The mean data of Mn for various seasons and sites is shown in Table-3.8. The results revealed that the effect of seasons and sites on Mn was significant while the effect of their interactions was non-significant.

Maximum Mn was observed at winter (6.03 mg/100g) while minimum at summer (3.58 mg/100g). Similarly maximum Mn was recorded at Jabri site (8.03 mg/100g) while minimum at Dabola site (2.78 mg/100g).

9. Zinc (Zn) mg/100g:

The mean data of Zn for various seasons and sites is shown in Table-3.8, while their interaction is shown in Figure-3.44. The results revealed that the effect of seasons, sites and their interaction on Zn was significant.

Maximum Zn was observed at inter (5.23 mg/100g) while minimum at summer (3.03 mg/100g). Similarly maximum Zn was recorded at Dam site (4.65 mg/100g) while minimum at Mang site (3.37 mg/100g). In case of interactions maximum Zn was observed at Dam site during winter (6.53 mg/100g) followed by Jabri site during winter (5.73 mg/100g). While minimum Zn was observed at Mang site during summer (2.70 mg/100g).

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Ajuga starts sprouting or its seed germinate naturally in May-July when conditions become favorable for germination. It initiate flowering in August to October and set seed in October to December. After first week of January no plants are seen in the natural vegetation.

The results can be summarized as the effect of seasons on these bio- chemical elements was highly significant. Maximum values were obtained at winter (October-November) and minimum at summer for all of the tested elements except Sodium and Calcium. The simplest explanation for this result can be that Ajuga matures at early winter season and hence the minerals accumulate in the plant parts like seed and roots. Similarly at early winter high moisture contents remain available in the soil which causes higher nutrients uptake (Ramirez et al., 2006). The effect on all elements except Iron (Fe) was significant which demonstrate the diversity of soil and climatic condition among various sites of Khanpur Valley.

It is evident from the results that early winter (last week of October) season is the best season for maximum medicinal and nutritional components in Ajuga at the cooler points of all the four sites of Khanpur valley. Jabri site showed maximum results for some parameters and so it is pertinent that hilly and moist areas with humid climate are the natural habitats for Ajuga (Abbas, 2003).

Table-3.8. Effect of Different Seasons and Sites on Elemental analysis of Ajuga bractiousa, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan.

Sodium Potassiu Calciu Phospho Magnesi Copper Mangan Iron (Fe) Zinc (Zn) (Na) m (K) m (Ca) rus (P) um(Mg) (Cu) ese(Mn) Seasons Summer 5.85a 162.79b 138.13b 174.94b 0.56b 178.13b 3.58b 3.03b 194.64a 127.66b Winter 4.24b 475.25a 196.73a 264.31a 0.92a 238.88a 6.03a 5.23a LSD at α 0.05 0.319 40.153 14.847 7.312 10.535 0.049 12.592 0.524 0.366

185

Sites

Dam 4.20c 165.87a 171.63c 229.63a 0.69b 206.07 2.90c 4.65a 284.33b Dabola 4.18c 159.80ab 186.43b 210.97b 0.86a 221.82 2.78c 4.15a 244.50b 377.33a Jabri 5.19b 197.95a 212.78a 204.42b 0.69b 208.10 8.03a 4.37a 369.92a Mang 6.60a 120.98b 98.87d 233.48a 0.72b 198.03 5.52b 3.37b LSD at α 0.05 0.579 79.277 40.288 12.165 14.843 0.102 ns 0.413 0.690 Interactions

Seasons*Sites * * * Ns * * ns ns * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

10 8 6

4 Summer 2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.39: Effect of different seasons and sites on Sodium content of Ajuga bracteosa. 700 600 500 400 300 Summer 200 100 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.40: Effect of different seasons and sites on Potassium (K) contents of Ajuga bracteosa. 186

300 250 200 150 Summer 100 Winter 50 0 Dam Dabola Jabri Mang Sites

Figure-3.41: Effect of different seasons and sites on Calcium (Ca) contents of Ajuga bracteosa.

350 300 250 200 150 Summer 100 Winter 50 0 Dam Dabola Jabri Mang Sites

Figure-3.42: Effect of different seasons and sites on Magnesium (Mg) contents of Ajuga bracteosa. 1.2 1 0.8 0.6 0.4 Summer 0.2 Winter 0 Dam Dabola Jabri Mang Sites

187

Figure-3.43: Effect of different seasons and sites on Copper (Cu) contents of Ajuga bracteosa. 8 7 6 5 4 3 Summer 2 Winter 1 0 Dam Dabola Jabri Mang Sites

Figure-3.44: Effect of different seasons and sites on Zinc (Zn) contents of Ajuga bracteosa.

3.5. EUPHORBIA HIRTA

I. Proximate Analysis:

1. Moisture%:

The mean data of moisture % for various seasons and sites is shown in Table-3.9. The results revealed that the effect of seasons and sites on moisture % was significant while the effect of their interactions was non- significant.

Maximum moisture % was observed at summer (87.31%) while minimum at winter (85.40%). Similarly maximum moisture % was recorded at Dam site (87.97%) while minimum at Mang site (83.94%).

2. Dry Matter %:

The mean data of dry matter % for various seasons and sites is shown in Table-3.9. The results revealed that the effect of seasons and sites on dry matter % was significant while the effect of their interactions was non- significant.

188

Maximum dry matter % was observed at winter (14.60%) while minimum at summer (12.69%). Similarly maximum moisture % was recorded at Mang site (16.06%) while minimum at Dam site (12.02%).

3. Ash / Minerals %:

The mean data of ash / minerals % for various seasons and sites is shown in Table-3.9, while their interaction is shown in Figure-3.45. The results revealed that the effect of seasons, sites and their interactions on ash / minerals % was significant.

Maximum ash / minerals % was observed at summer (15.99%) while minimum at winter (14.55%). Similarly maximum ash / minerals % was recorded at Jabri site (16.62%) while minimum at Dam site (13.84%). In case of interactions maximum ash % was recorded at Jabri site during summer (17.95%) while minimum of it was observed at Dam site during winter (13.16%).

4. Crude Proteins %:

The mean data of crude proteins % for various seasons and sites is shown in Table-3.9, while their interaction is shown in Figure-3.46. The results revealed that the effect of seasons, sites and their interactions on crude proteins % was significant.

Maximum crude proteins % was observed at summer (13.11%) while minimum at winter (11.18%). Similarly maximum crude proteins % was recorded at Jabri site (13.89%) while minimum at Mang site (10.40%). In case of interactions maximum crude proteins % was recorded at Jabri site during summer (15.42%) followed by Dam site during summer (15.12%) while minimum of it was observed at Mang site during winter (10.037%).

189

5. Crude Fibers %:

The mean data of crude fibers % for various seasons and sites is shown in Table-3.9, while their interaction is shown in Figure-3.47. The results revealed that the effect of seasons, sites and their interactions on crude fibers % was significant.

Maximum crude fibers % was observed at summer (10.90%) while minimum at winter (8.61%). Similarly maximum crude fibers % was recorded at Jabri site (10.92%) while minimum at Dam site (8.44%). In case of interactions maximum crude fibers % was recorded at Mang site during summer (12.78%) followed by Jabri site during summer (11.55%) while minimum of it was observed at Dam site during winter (6.51%).

6. Crude Fats %:

The mean data of crude fats % for various seasons and sites is shown in Table-3.9, while their interaction is shown in Figure-3.48. The results revealed that the effect of seasons, sites and their interactions on crude fats% was significant.

Maximum crude fats % was observed at summer (9.14%) while minimum at winter (7.98%). Similarly maximum crude fats % was recorded at Jabri site (9.71%) while minimum at Dabola site (7.45%). In case of interactions maximum crude fats % was recorded at Jabri site during summer (10.64%) followed by Dam site during summer (10.12%) while minimum of it was observed at Mang site during winter (7.15%).

7. Essential Oils%:

The mean data of essential oil % for various seasons and sites is shown in Table-3.9, while their interaction is shown in Figure-3.49. The results 190

revealed that the effect of sites on essential oil % was non-significant while the effect of seasons and interactions was significant.

Maximum essential oil % was observed at summer (1.80%) while minimum at winter (1.55%). In case of interactions maximum essential oil % was recorded at Dabola site during summer (1.94%) followed by Dam site during summer (1.89%) while minimum of it was observed at Mang site during winter (1.48%).

8. Nitrogen Free ExtracTable Substances (NFES) %:

The mean data of NFES % for various seasons and sites is shown in Table- 3.9, while their interaction is shown in Figure-3.50. The results revealed that the effect of seasons, sites and their interactions on NFES % was significant.

Maximum NFES % was observed at winter (57.68%) while minimum at summer (50.86%). Similarly maximum NFES % was recorded at Dabola site (58.55%) while minimum at Jabri site (49.15%). In case of interactions maximum NFES % was recorded at Dam site during winter (59.49%) followed by Dabola site during winter (59.30%) while minimum of it was observed at Jabri site during summer (44.44%).

9. Net Free Energy Estimation (NFEE):

The mean data of NFEE for various seasons and sites is shown in Table- 3.9, while their interaction is shown in Figure-3.51. The results revealed that the effect of seasons, sites and their interactions on NFEE was significant.

Maximum NFEE was observed at summer (182.85) while minimum at winter (171.79). Similarly maximum NFEE was recorded at Dam site (190.83) while minimum at Dabola site (168.09). In case of interactions

191

maximum NFEE was recorded at Jabri site during summer (198.69) followed by Dam site during summer (198.38) while minimum of it was observed at Mang site during winter (160.44).

The results revealed that the effect of seasons on all parameters were highly significant. In summer, maximum values have been found for moisture, crude protein, crude fats, crude fibers, minerals/ashes, essential oils and NFEE while in winter maximum values were found for dry matter and NFES. This seasonal effect was reasonably due to the plant growth habit and environmental conduciveness. E.hirta sprouts during late spring and summer which is the metabolically most active period of its growth (Zavodnik, 1998); so most of the minerals are absorbed during summer. Another reason of high moisture, essential nutrients and mineral during summer was high soil moisture contents (McDowell, 2003) due to more rain fall and high rate of transpiration due to high temperature (Vidyarthi, 2002). So when more growth took place, more dry biomass was also produced. E. hirta plants sprout naturally in May-July and plants matures in September-December. The rapid growth stage of the plant and availability of more moisture at summer in most of the sites was the possible reason for higher percentages of moisture, CP, CF and NFEE during summer at these sites (McDowell, 2003; Ramirez et al., 2006). Higher values for dry matter and NFES at winter is definitely an indirect proportion to the low moisture and other nutrients accumulation in the plant tissues due to environmental and physiological stresses (Miranda- Ham et al., 2007)

The effect of sites on all the parameters was significant except on essential oil. Maximum values were found for moisture and NFEE at Dam site while maximum values for NFES was found at Dabola site. This might be correlated with availability of high moisture contents in the soil and its water holding capacity at Dam site (McDowell, 2003). At Jabri,

192

maximum values were recorded for minerals, crude proteins and crude fats while at Mang, maximum values were found for dry matter and crude fibers. High proteins might be due to high nitrogen contents in the soil (Kayhan et al., 1997) available during summer at the site. High dry matter, ashes and crude fibers might be due to saline conditions of the soil, as most of the plant species contain these compounds higher at maturity when present at a saline environment (Akingbade, 2001). At Dabola site during summer, the analysis showed maximum value for essential oils.

Table-3.9. Effect of Different Seasons and Sites on Proximate analysis of Euphorbia hirta, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan.

Dry Ash/mi Crude Moistur Crude Essentia Matter nerals Proteins Fats% e% % % % Fibers% l Oils% Seasons NFES% NFEE Summer 87.31a 12.69b 15.99a 13.11a 10.90a 9.14a 1.80a 50.86b 182.85a

Winter 85.40b 14.60a 14.55b 11.18b 8.61b 7.98b 1.55b 57.68a 171.79b LSD at α 0.05 0.992 0.992 0.459 0.456 0.576 0.329 0.097 1.018 3.368 Sites

Dam 87.97a 12.02c 13.84b 13.68a 8.44b 9.36a 1.71 54.68b 190.83a

Dabola 85.88b 14.12b 14.08b 10.61b 9.04b 7.71b 1.73 58.55a 168.09b

Jabri 87.62ab 12.38bc 16.62a 13.89a 10.62a 9.71a 1.67 49.15c 189.26a

Mang 83.94c 16.06a 16.55a 10.40b 10.92a 7.45b 1.59 54.69b 161.10c LSD at α 0.05 1.760 1.760 1.197 0.793 1.320 0.670 ns 2.710 5.301 Interactions

Seasons*Sites ns Ns * * * * * * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

193

10 Summer 5 Winter

0 Dam Dabola Jabri Mang Sites

Figure-3.45: Effect of different seasons and sites on Ash contents in Euphorbia hirta.

10 Summer 5 Winter

0 Dam Dabola Jabri Mang Sites

Figure-3.46: Effect of different seasons and sites on Crude Protein contents in Euphorbia hirta. 12 10 8 6 4 Summer 2 Winter 0 Dam Dabola Jabri Mang Sites

194

Figure-3.47: Effect of different seasons and sites on Crude Fats contents in Euphorbia hirta. 16 14 12 10 8 6 Summer 4 Winter 2 0 Dam Dabola Jabri Mang Axis Title

Figure-3.48: Effect of different seasons and sites on Crude Fibers contents in Euphorbia hirta.

2.5 2 1.5

1 Summer 0.5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.49: Effect of different seasons and sites on Essential Oil contents in Euphorbia hirta. 70 60 50 40 30 Summer 20 Winter 10 0 Dam Dabola Jabri Mang Sites

195

Figure-3.50: Effect of different seasons and sites on NFES contents in Euphorbia hirta. 250

200 150

100 Summer 50 Winter

0 Dam dabola Jabri Mang Sites

Figure-3.51: Effect of different seasons and sites on NFEE in Euphorbia hirta.

II. Elementology:

1. Sodium (Na) mg/100g:

The mean data of Na for various seasons and sites is shown in Table-3.10. The results revealed that the effect of seasons and sites on Na was significant while the effect of their interaction was non-significant.

Maximum Na was observed at winter (2.87 mg/100g) while minimum at summer (2.61 mg/100g). Similarly maximum Na was recorded at Dam site (3.34 mg/100g) while minimum at Dabola site (2.21 mg/100g).

2. Potassium (K) mg/100g:

The mean data of K for various seasons and sites is shown in Table-3.10. The results revealed that the effect of sites on K was significant while the effect of seasons and interactions was non-significant.

Maximum K was observed at Dam site (81.74 mg/100g) while minimum K was at Mang site (68.96 mg/100g).

196

3. Calcium (Ca) mg/100g:

The mean data of Ca for various seasons and sites is shown in Table-3.10, while their interaction is shown in Figure-3.52. The results revealed that the effect of seasons, sites and their interaction on Ca was significant.

Maximum Ca was observed at winter (152.46 mg/100g) while minimum at summer (126.74 mg/100g). Similarly maximum Ca was recorded at Dam site (192.51 mg/100g) while minimum at Jabri site (117.63 mg/100g). In case of interactions maximum Ca was observed at Dam site during summer (199.17 mg/100g) followed by Dam site during Winter (185.85 mg/100g). While minimum Ca was observed at Dabola site during summer (79.17 mg/100g)

4. Phosphorus (P) mg/100g:

The mean data of Phosphorus for various seasons and sites is shown in Table-3.10, while their interaction is shown in Figure-3.53. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum P was observed at winter (143.74 mg/100g) while minimum at summer (89.27 mg/100g). Similarly maximum P was recorded at Jabri site (153.92 mg/100g) while minimum at Mang site (59.75 mg/100g). In case of interactions maximum P was observed at Jabri site during winter (185.37 mg/100g) followed by Dam site during Winter (176.69 mg/100g). While minimum P was observed at Mang site during summer (44.07 mg/100g)

5. Magnesium (Mg) mg/100g:

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The mean data of Mg for various seasons and sites is shown in Table- 3.10. The results revealed that the effect of seasons and sites on Fe was significant while the effect of their interactions was non-significant.

Maximum Mg was observed at summer (225.65 mg/100g) while minimum at winter (188.50 mg/100g). Similarly maximum Mg was recorded at Jabri site (282.47 mg/100g) while minimum at Dam site (225.48 mg/100g).

6. Copper (Cu) mg/100g:

The mean data of Cu for various seasons and sites is shown in Table-3.10. The results revealed that the effect of seasons and sites on Fe was significant while the effect of their interactions was non-significant.

Maximum Cu was observed at winter (2.37 mg/100g) while minimum at summer (1.45 mg/100g). Similarly maximum Cu was recorded at Mang site (2.34 mg/100g) while minimum at Dam site (1.49 mg/100g).

7. Iron (Fe) mg/100g:

The mean data of Fe for various seasons and sites is shown in Table-3.10. The results revealed that the effect of seasons and sites on Fe was significant while the effect of their interactions was non-significant.

Maximum Fe was observed at winter (267.98 mg/100g) while minimum at summer (206.57 mg/100g). Similarly maximum Fe was recorded at Mang site (289.00 mg/100g) while minimum at Dabola site (190.64 mg/100g).

8. Manganese (Mn) mg/100g:

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The mean data of Mn for various seasons and sites is shown in Table- 3.10. The results revealed that the effect of seasons and sites on Mn was significant while the effect of their interactions was non-significant.

Maximum Mn was observed at winter (29.43 mg/100g) while minimum at summer (20.40 mg/100g). Similarly maximum Mn was recorded at Jabri site (27.88 mg/100g) while minimum at Dam site (21.32 mg/100g).

9. Zinc (Zn) mg/100g:

The mean data of Zn for various seasons and sites is shown in Table-3.10. T The results revealed that the effect of seasons on Zn was significant while that of sites and their interactions was non-significant.

Maximum Zn was observed at winter (9.07 mg/100g) while minimum at summer (5.99 mg/100g).

The results can be summarized as the effect of seasons on these bio- chemical elements was highly significant. Maximum values were obtained at winter (October-November) and minimum at summer for all of the tested elements except Magnesium. The simplest explanation for this result can be that E.hirta matures at early winter season and hence the minerals accumulate in the plant parts like seed and roots. Ramirez et al., 2006 derived the same findings that at early winter high moisture contents remain available in the soil which causes higher nutrients uptake.

It is evident from the results that early winter (last week of October) season is the best season for maximum medicinal and nutritional components in E. hirta at Dam and Mang sites, lower Khanpur valley. Plain, shady areas with humid climate are the natural habitats for E.hirta (Stuart, 1979) which proved true for Mang and Dam sites.

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Table-3.10. Effect of Different Seasons and Sites on Elemental analysis of Euphorbia hirta, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan.

Sodium Potassi Calciu Phospho Magnesi Copper Mangan Zinc Iron (Fe) (Na) um (K) m (Ca) rus (P) um(Mg) (Cu) ese(Mn) (Zn) Seasons Summer 2.61b 71.23 126.74b 89.27b 225.65a 1.45b 206.57b 20.40b 5.99b

Winter 2.87a 75.19 152.46a 143.74a 188.50b 2.37a 267.98a 29.43a 9.07a LSD at α 0.05 0.251 Ns 5.019 8.186 22.092 0.214 17.043 1.744 0.568 Sites

Dam 3.34a 81.70a 192.51a 146.06a 225.48b 1.49c 232.33b 21.32b 7.61

Dabola 2.21c 70.35b 126.94b 106.31b 255.78ab 1.90b 190.64c 22.74b 8.12

Jabri 3.04b 71.83b 117.63b 153.92a 282.47a 1.91b 237.12b 27.88a 7.44

Mang 2.36c 68.96b 121.33b 59.75c 264.57a 2.34a 289.00a 27.73a 6.95 LSD at α 0.05 0.206 8.075 9.673 9.056 35.291 0.401 27.710 2.519 ns Interactions

Seasons*Sites ns Ns * * ns ns ns Ns ns Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

250

200

150

100 Summer

50 Winter

0 Dam dabola Jabri Mang Sites

Figure-3.52: Effect of different seasons and sites on Calcium contents in Euphorbia hirta.

200

250 200 150 100 Summer 50 0 Winter Dam dabola Jabri Mang Sites

Figure-3.53: Effect of different seasons and sites on Phosphorus contents in Euphorbia hirta.

3.6. FUMARIA OFFICINALIS

I. Proximate Analysis:

1. Moisture%:

The mean data of moisture % for various seasons and sites is shown in Table-3.11. The results revealed that the effect of seasons and sites on moisture % was significant while the effect of their interactions was non- significant.

Maximum moisture % was observed at summer (91.19%) while minimum at winter (88.43%). Similarly maximum moisture % was recorded at Dam site (91.62%) while minimum at Mang site (87.50%).

2. Dry Matter %:

The mean data of dry matter % for various seasons and sites is shown in Table-3.11. The results revealed that the effect of seasons and sites on dry matter % was significant while the effect of their interactions was non- significant.

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Maximum dry matter % was observed at winter (11.57%) while minimum at summer (8.81%). Similarly maximum moisture % was recorded at Mang site (12.50%) while minimum at Dam site (8.38%).

3. Ash / Minerals %:

The mean data of ash / minerals % for various seasons and sites is shown in Table-3.11, while their interaction is shown in Figure-3.54. The results revealed that the effect of seasons, sites and their interactions on ash / minerals % was significant.

Maximum ash / minerals % was observed at summer (26.07%) while minimum at winter (22.83%). Similarly maximum ash / minerals % was recorded at Dabola site (26.03%) while minimum at Mang site (22.17%). In case of interactions maximum ash % was recorded at Jabri site during summer (27.71) while minimum of it was observed at Mang site during winter (20.29%).

4. Crude Proteins %:

The mean data of crude proteins % for various seasons and sites is shown in Table3.11, while their interaction is shown in Figure-3.55. The results revealed that the effect of sites was non-significant while that of seasons and their interactions on crude proteins % was significant.

Maximum crude proteins % was observed at summer (14.22%) while minimum at winter (12.66%). In case of interactions maximum crude proteins % was recorded at Mang site during summer (15.22%) followed by Dabola site during summer (14.30%) while minimum of it was observed at Jabri site during winter (12.13%).

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5. Crude Fibers %:

The mean data of crude fibers % for various seasons and sites is shown in Table-3.11, while their interaction is shown in Figure-3.56. The results revealed that the effect of seasons, sites and their interactions on crude fibers % was significant.

Maximum crude fibers % was observed at winter (12.30%) while minimum at summer (10.30%). Similarly maximum crude fibers % was recorded at jabri site (12.04%) while minimum at Dam site (10.20%). In case of interactions maximum crude fibers % was recorded at Jabri site during winter (13.130%) followed by Dabola site during winter (12.887%) while minimum of it was observed at Mang site during summer (9.457%).

6. Crude Fats %:

The mean data of crude fats % for various seasons and sites is shown in Table-3.11, while their interaction is shown in Figure-3.57. The results revealed that the effect of seasons, sites and their interactions on crude fibers % was significant.

Maximum crude fats % was observed at summer (8.68%) while minimum at winter (7.38%). Similarly maximum crude fats % was recorded at Dam site (8.81%) while minimum at Dabola site (6.90%). In case of interactions maximum crude fats % was recorded at Mang site during summer (9.60%) followed by Jabri site during summer (9.21%) while minimum of it was observed at Dabola site during winter (6.61%).

7. Essential Oils%:

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The mean data of essential oil % for various seasons and sites is shown in Table-3.11, while their interaction is shown in Figure-3.58. The results revealed that the effect of seasons, sites and their interactions on crude fibers % was significant.

Maximum essential oil % was observed at winter (1.69%) while minimum at summer (1.17%). Similarly maximum essential oil % was recorded at Mang site (1.82%) while minimum at Jabri site (1.27%). In case of interactions maximum essential oil % was recorded at Mang site during winter (2.18%) followed by Jabri site during winter (1.70%) while minimum of it was observed at Jabri site during summer (0.84%).

8. Nitrogen Free ExtracTable Substances (NFES) %:

The mean data of NFES % for various seasons and sites is shown in Table- 3.11, while their interaction is shown in Figure-3.59. The results revealed that the effect of seasons, sites and their interactions on NFES % was significant.

Maximum NFES % was observed at winter (44.70%) while minimum at summer (40.67%). Similarly maximum NFES % was recorded at Mang site (44.34%) while minimum at Jabri site (41.29%). In case of interactions maximum NFES % was recorded at Mang site during winter (47.04%) followed by Dam site during winter (45.20%) while minimum of it was observed at Jabri site during summer (38.39%).

9. Net Free Energy Estimation (NFEE):

The mean data of NFEE for various seasons and sites is shown in Table- 3.11, while their interaction is shown in Figure-3.60. The results revealed that the effect of seasons, sites and their interactions on NFEE was significant.

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Maximum NFEE was observed at summer (173.31) while minimum at winter (159.76). Similarly maximum NFEE was recorded at Mang site (173.40) while minimum at Dabola site (156.28). In case of interactions maximum NFEE was recorded at Mang site during summer (186.13) followed by Jabri site during summer (173.50) while minimum of it was observed at Dabola site during winter (152.22).

The results revealed that the effect of seasons on all parameters was highly significant. In summer, maximum values have been found for moisture (91.186%) crude protein (14.277%) crude fats (8.6808%) NFEE (173.31 kcal/g) and minerals (26.070%), while in winter maximum values were found for dry matter (11.573%) crude fibers (12.302%) essential oil (1.6958%) and NFES (44.807%). This seasonal effect was reasonably due to the plant growth habit and environmental conduciveness (Zavodnik et al., 1998). Fumaria plants sprout naturally in October-February and matures in March-July. The availability of more moisture at summer in most of the sites was the possible reason for higher percentages of moisture, CP, CF and NFEE during summer (McDowell, 2003; Ramirez et al., 2006). During winter, low temperature reduces energy use and increases sugar storage which resulted in observation of high values of NFES

The effect of sites on all the parameters was significant except on crude fiber. Maximum values were found for moisture (91.628%) at Dam site while maximum values for minerals (26.037%) were found at Dabola site. At Jabri, maximum values were recorded for crude fiber (12.037%) and crude fats (8.5717%) while at Mang, maximum values were found for dry matter (12.498%), crude protein (13.838%), essential oil (1.8217%), NFES (44.345%) and NFEE (173.40 kcal/g). High proteins might be due to high nitrogen contents in the soil (Kayhan et al., 1997) available during summer. High essential oils, NFEE and NFES might be due to saline

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conditions of the soil of Mang site, as most of the plant species contain these compounds higher at maturity when present at a saline environment (Akingbade, 2001).

Table-3.11. Effect of Different Seasons and Sites on proximate analysis of Fumaria officinalis, indigenous to Khanpur Valley, in subHimalayan mountains of Pakistan.

Dry Ash/mi Crude Moistur Crude Crude Essentia Matter nerals Proteins e% Fibers% Fats% l Oils% NFES Seasons % % % NFEE % Summer 91.19a 8.81b 26.07a 14.22a 10.30b 8.68a 1.17b 40.67b 173.31a 44.80a Winter 88.43b 11.57a 22.83b 12.66b 12.30a 7.38b 1.69a 159.76b LSD at α 0.05 0.913 0.913 0.682 0.329 0.224 0.398 0.184 0.700 3.743 Sites

Dam 91.62a 8.37c 24.43b 13.48 10.20c 8.81a 1.32b 43.70a 168.87ab

Dabola 89.30b 10.70b 26.03a 13.63 11.79a 6.90b 1.32b 41.62b 156.28c

Jabri 90.79a 9.21c 25.17ab 12.93 12.04a 8.57a 1.27b 41.29b 167.59c

Mang 87.50c 12.50a 22.17c 13.83 11.17b 8.47a 1.82a 44.34a 173.40a LSD at α 0.05 1.323 1.323 1.00 Ns 0.531 0.686 0.210 0.986 5.303 Interactions

Seasons*Sites ns ns * * * * * * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

35 30 25 20 15 Summer 10 5 Winter 0 Dam Dabola Jabri Mang sites

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Figure-3.54: Effect of different seasons and sites on Ash % of Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

10 Summer 5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.55: Effect of different seasons and sites on Crude Proteins % of Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 12 10 8 6 4 Summer 2 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.56: Effect of different seasons and sites on Crude Fats % of Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

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16 14 12 10 8 6 Summer 4 Winter 2 0 Dam Dabola Jabri Mang Sites

Figure-3.57: Effect of different seasons and sites on Crude Fibers % of Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

2.5 2 1.5

1 Summer 0.5 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.58: Effect of different seasons and sites on Essential Oil % of Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 60 50 40 30 20 Summer 10 Winter 0 Dam Dabola Jabri Mang Sites

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Figure-3.59: Effect of different seasons and sites on NFES % of Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 250 200 150

100 Summer 50 Winter 0 Dam dabola Jabri Mang Sites

Figure-3.60: Effect of different seasons and sites on NFEE Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

II. Elementology:

1. Sodium (Na) mg/100g:

The mean data of Na for various seasons and sites is shown in Table-3.12. The results revealed that the effect of seasons and sites on Na was significant while the effect of their interactions was non-significant.

Maximum Na was observed at summer (2.65 mg/100g) while minimum at winter (1.84 mg/100g). Similarly maximum Na was recorded at Jabri site (2.91 mg/100g) while minimum at Mang site (1.72 mg/100g).

2. Potassium (K) mg/100g:

The mean data of K for various seasons and sites is shown in Table-3.12, while their interaction is shown in Figure-3.61. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum K was observed at summer (475.37 mg/100g) while minimum at winter (134.35 mg/100g). Similarly maximum K was recorded at Mang 209

site (336.33 mg/100g) while minimum at Dam site (287.40 mg/100g). In case of interactions maximum K was observed at Dabola site during summer (494.10 mg/100g) followed by Dam site during summer (489.03 mg/100g) followed by Jabri site during summer (467.17 mg/100g). While minimum K was observed at Dam site during winter (85.77 mg/100g)

3. Calcium (Ca) mg/100g:

The mean data of Ca for various seasons and sites is shown in Table-3.12. The results revealed that the effect of seasons and sites on Ca was significant while the effect of their interactions was non-significant.

Maximum Ca was observed at summer (199.13 mg/100g) while minimum at winter (125.58 mg/100g). Similarly maximum Ca was recorded at Jabri site (220.27 mg/100g) while minimum at Mang site (108.63 mg/100g).

4. Phosphorus (P) mg/100g:

The mean data of Phosphorus for various seasons and sites is shown in Table-3.12, while their interaction is shown in Figure-3.62. The results revealed that the effect of seasons, sites and their interaction on P was significant.

Maximum P was observed at summer (181.14 mg/100g) while minimum at winter (115.11 mg/100g). Similarly maximum P was recorded at Mang site (213.10 mg/100g) while minimum at Dam site (100.23 mg/100g). In case of interactions maximum P was observed at Mang site during summer (265.27 mg/100g) followed by Dabola site during summer (215.80 mg/100g). While minimum P was observed at Dam site during winter (85.60 mg/100g)

5. Magnesium (Mg) mg/100g:

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The mean data of Mg for various seasons and sites is shown in Table- 3.12, while their interaction is shown in Figure-3.63. The results revealed that the effect of seasons, sites and their interaction on Mg was significant.

Maximum Mg was observed at summer (156.48 mg/100g) while minimum at winter (116.79 mg/100g). Similarly maximum Mg was recorded at Jabri site (172.98 mg/100g) while minimum at Mang site (102.85 mg/100g). In case of interactions maximum Mg was observed at Jabri site during summer (184.30 mg/100g) followed by Dabola site during summer (182.53 mg/100g). While minimum Mg was observed at Dam site during winter (80.63 mg/100g)

6. Copper (Cu) mg/100g:

The mean data of Cu for various seasons and sites is shown in Table-3.12. The results revealed that the effect of seasons and sites on Cu was significant while the effect of their interactions was non-significant.

Maximum Cu was observed at summer (1.71 mg/100g) while minimum at winter (1.27 mg/100g). Similarly maximum Cu was recorded at Jabri site (0.53 mg/100g) while minimum at Dam site (0.42 mg/100g).

7. Iron (Fe) mg/100g:

The mean data of Fe for various seasons and sites is shown in Table-3.12. The results revealed that the effect of seasons and sites on Fe was significant while the effect of their interactions was non-significant.

Maximum Fe was observed at summer (242.42 mg/100g) while minimum at winter (144.60 mg/100g). Similarly maximum Fe was recorded at Mang site (235.25 mg/100g) while minimum at Dam site (152.00 mg/100g).

8. Manganese (Mn) mg/100g: 211

The mean data of Mn for various seasons and sites is shown in Table- 3.12, while their interaction is shown in Figure-3.64. The results revealed that the effect of seasons, sites and their interaction on Mn was significant.

Maximum Mn was observed at summer (13.10 mg/100g) while minimum at winter (8.49 mg/100g). Similarly maximum Mn was recorded at Mang site (12.26 mg/100g) while minimum at Dam site (8.55 mg/100g). In case of interactions maximum Mn was observed at Dabola site during summer (14.03 mg/100g) followed by Mang site during summer (13.90 mg/100g). While minimum Mn was observed at Dam site during winter (5.46 mg/100g).

9. Zinc (Zn) mg/100g:

The mean data of Zn for various seasons and sites is shown in Table-3.12. The results revealed that the effect of seasons and sites on Zn was significant while the effect of their interactions was non-significant.

Maximum Zn was observed at summer (3.75 mg/100g) while minimum at winter (2.18 mg/100g). Similarly maximum Zn was recorded at Dam site (3.55 mg/100g) while minimum at Dabola site (2.63 mg/100g).

The results showed that the effect of seasons on these bio-chemical elements was highly significant. Maximum values were obtained at summer and minimum at winter for all the tested elements. The simplest explanation for this result can be that Fumaria officinalis matures at summer season and hence the minerals accumulate in the plant parts. Similarly at summer high moisture contents remain available in the soil which causes higher nutrients uptake (Ramirez et al., 2006). Similarly the macro and micro nutrients in the plants of F. officinalis were significantly varied among various sites. This variation was due to the nutrients status in the soil of the site and its interaction with water, temperature and light

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availability (Kayhan et al., 1997). The most favorable season and site for the collection of this plant is summer and Mang site.

Table-3.12. Effect of Different Seasons and Sites on Elemental analysis of Fumaria officinalis, indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan.

Sodium Potassiu Calciu Phosph Magnesi Copper Mangan Zinc Iron (Fe) (Na) m (K) m (Ca) orus (P) um(Mg) (Cu) ese(Mn) (Zn) Seasons Summer 2.65a 275.37a 199.13a 181.14a 156.48a 0.71a 242.42a 13.10a 3.75a

Winter 1.84b 134.35b 125.58b 115.11b 116.79b 0.27b 144.60b 8.49b 2.18bb LSD at α 0.05 0.564 38.580 14.577 10.481 8.209 0.088 25.460 0.676 0.439 Sites

Dam 2.30ab 287.40 130.72c 100.23d 103.95b 0.42b 152.0c 8.55c 3.55a

Dabola 2.06b 300.02 189.80b 161.75b 166.77a 0.51ab 185.05b 10.98b 2.63b

Jabri 2.91a 295.68 220.27a 117.42c 172.98a 0.53a 201.78b 11.38b 2.95b

Mang 1.72b 336.33 108.63d 213.10a 102.85b 0.50b 235.25a 12.26a 2.73b LSD at α 0.05 0.610 Ns 16.113 13.317 15.99 0.071 21.301 0.762 0.472 Interactions

Seasons*Sites ns * ns * * ns ns * Ns Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

600 500 400 300 Summer 200 100 Winter 0 Dam dabola Jabri Mang Sites

Figure-3.61: Effect of different seasons and sites on Potassium (mg/100g) in Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan.

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350 300 250 200 150 Summer 100 Winter 50 0 Dam dabola Jabri Mang Sites

Figure-3.62: Effect of different seasons and sites on Phosphorus (mg/100g) in Fumaria officinalis collected from Khanpur valley in the subHimalayan mountains of Pakistan. 250 200 150

100 Summer 50 Winter 0 Dam Dabola Jabri Mang Sites

Figure-3.63: Effect of different seasons and sites on Magnesium (mg/100g) in Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 20

10 Summer 5 Winter 0 Dam dabola Jabri Mang Sites

Figure-3.64: Effect of different seasons and sites on Manganese (mg/100g) in Fumaria officinalis collected from Khanpur valley in the sub-Himalayan mountains of Pakistan. 214

SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

Human body needs essential nutrients, major minerals (Ca, P, K, Na and Mg), and trace elements (Fe, Zn, Cu, Mn and others), which are essential for general health, growth and reproduction. We must bear in mind that the consequences of essential trace mineral deficiency may be just as severe as those of a deficiency of a major essential mineral. Many elements are associated with one another in maintaining our normal growth and health, the herbs may prove to be a useful remedy for many common and complicated ailments and part of essential nutrients required for human nourishment.

The experiment on the bio-chemical attributes of selected medicinal plants of Khanpur Valley in the sub Himalayan mountains of Pakistan was conducted during 2012-2013. The samples were collected from the valley and chemical analysis was carried out at University of agriculture Peshawar. Out of 71 medicinal plant species identified in the research area, six most preferred medicinally important species of the valley, were selected for bio- chemical study. These species were Adhatoda vasica, Calatropis procera, Recinus communis, Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis. Medicinal plants, whole plant in case of herbs and leaves in case of woody plants were collected from their natural habitat at all the four sites and two seasons. The analysis revealed that the bio-chemical compounds moisture, crude proteins, crude fats, ashes or total minerals and NFEE were significantly higher in summer season while dry matter, crude fibers, essential oil and NFES were higher during winter season, in most of the plants under study, at all the four sites of the study area. Similarly maximum biochemical elements (Na, K, Ca, P, Mg, Cu, Fe, Mn and Zn) were observed during summer for all plants under study except

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E. hirta in which maximum bio-chemical elements were recorded in samples collected during winter.

The effect of sites on bio-chemical attributes varied in response to various species. The chemical analysis of leaves of shrubs (Adhatoda vasica, Calatropis procera and Recinus communis) depicted a unique trend (with little deviation) in bio-chemical composition at different sites. At Dam and Mang sites higher values for Na, K, P, Ca and Mg were recorded while at Dabola and Jabri sites high values for Fe, Mn, Cu and Zn were found. Similarly, these species gave maximum values for moisture, crude proteins, crude fats and NFEE at Dam and Dabola sites, while higher values of dry matter, ash/minerals, essential oil, crude fibers and NFES were found in the leaves of these shrubs at Jabri and Mang sites.

Significant effects were observed at different seasons and sites upon various biochemical attributes of all six medicinal plants. Adhatoda vasica showed significantly higher values for ash (19.5%) at Jabri during summer, crude proteins (14.7%) at Dabola during summer, crude fibers (12.6%) at Jabri during winter, essential oil (2.5%) at Mang during summer, NFES (55.8%) at Mang during summer, NFEE (194.6%) at Dam during summer, potassium (179.4mg/100g) at Dam during summer, phosphorus (171mg/100g) at Dabola during summer, copper (0.58mg/100g) at Dabola during summer and zinc (8.07mg/100g) at Mang during summer. Fumaria officinalis revealed higher significant results for ash (27.7%) at Jabri during summer, crude proteins (15.2%) at Mang during summer, crude fibers (13.1%) at Jabri during winter, crude fats (9.6%) at Mang during summer, essential oil (2.2%) at Mang during winter, NFES (47.04%) at Mang during winter, NFEE (186.13%) at Mang during summer, potassium (494.1mg/100g) at Dabola during summer, phosphorus (265.3mg/100g) at Mang during summer, magnesium (184.3mg/100g) at Jabri during summer and manganese (14.03mg/100g)

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at Dabola during summer. Euphorbia hirta gave higher significant contents for ash (17.95%) at Jabri during summer, crude proteins (15.42%) at Jabri during summer, crude fibers (12.78%) at Mang during winter, crude fats (10.64%) at Jabri during summer, essential oil (1.94%) at Dabola during summer, NFES (59.49%) at Dam during winter, NFEE (198.69%) at Jabri during summer, calcium (199.17mg/100g) at Dam during summer, phosphorus (185.37mg/100g) at Jabri during winter. Ajuga bracteosa revealed higher significant values for ash (34.13%) at Jabri during summer, crude proteins (15.35%) at Dam during summer, crude fats (10.30%) at Jabri during summer, essential oil (2.48%) at Mang during summer, NFES (46.17%) at Dabola during winter, NFEE (186.28%) at Dam during summer, sodium (6.96mg/100g) at Mang during winter, potassium (559.67mg/100g) at Jabri during winter, calcium (254.20mg/100g) at Jabri during summer, magnesium (264.31mg/100g) at Mang during winter, copper (1.017mg/100g) at Dabola during winter and zinc (6.53mg/100g) at Dam during winter. Recinus communis revealed higher significant values for crude proteins (15.26%) at Mang during summer, crude fibers (13.85%) at Dabola during winter, crude fats (10.24%) at Jabri during summer, essential oil (3.48%) at Mang during winter, NFEE (195.29%) at Jabri during summer, sodium (4.91mg/100g) at Dam during summer, iron (300.37mg/100g) at Jabri during summer, manganese (13.43mg/100g) at Dabola during summer and zinc (8.07mg/100g) at Mang during summer. Calatropis procera revealed higher significant values for crude proteins (14.58%) at Mang during summer, crude fibers (13.39%) at Dabola during winter, crude fats (11.98%) at Jabri during summer, NFES (50.80%) at Dam during winter, NFEE (203.1%) at Jabri during summer, potassium (500.33mg/100g) at Dam during summer, calcium (274.07mg/100g) at Dabola during winter, magnesium (291.57mg/100g) at Dabola during winter, copper (0.72mg/100g) at Jabri during summer, iron (291.6mg/100g) at Dam

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during summer manganese (19.27mg/100g) at Dam during summer and zinc (6.03mg/100g) at Mang during winter.

The chemical analysis of whole plants of herbs (Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis) revealed that there was a constant variation in the values of both compounds and elements from site to site and a set of parameters cannot be associated with one or the other site. It was concluded from the comparison of six species under bio- chemical study that dry matter, Ca and Fe were found maximum in Adhatoda vasica during summer; K was highest in Fumaria during summer; NFES, Cu, Zn and Mg were maximum in E. hirta during winter; ash/minerals, crude fibers and P were highest in Ajuga during winter; essential oil, Mg was maximum in R. communis during winter; moisture, crude proteins, crude fats, NFEE and Na were highest in C. procera during summer.

The following conclusions are derived from this whole exercise:

1. Medicinal plants or its leaves may be harvested in summer when higher quantity of proteins and fats is needed while medicinal plants or its leaves may be harvested in winter when higher quantity of sugar and essential oil is needed. 2. Medicinal Plants Adhatoda vasica, Fumaria officinalis and Recinus communis may be harvested in summer for maximum bio-chemical elements (Na, K, Ca, P, Mg, Cu, Fe, Mn and Zn) while Euphorbia hirta, Ajuga bracteosa and Calatropis procera, may be harvested in winter for highest bio-chemical elements. 3. Shrubs may be collected with maximum values for moisture, crude proteins, crude fats and NFEE at Dam and Dabola sites and with higher values of dry matter, ash/minerals, essential oil, crude fibers and NFES at Jabri and Mang sites. These can be

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collected or harvested at Dam and Mang sites with higher values for Na, K, P, Ca and Mg and at Dabola and Jabri sites with high values for Fe, Mn, Cu and Zn. 4. Herbs like Fumaria officinalis, which germinate in winter and mature in summer, can be collected in summer (last week of May) with maximum biochemical attributes. Similarly herbs which germinate in summer and mature before the onset of winter can be best collected in early winter (last week of October) for highest bio-chemical harvest.

Following are some of the recommendations based on the research findings of this research experiment:

• Human body needs essential nutrients, major minerals and trace elements which are essential for general health, growth and reproduction. Many elements are associated with one another in maintaining our normal growth and health, the herbs may prove to be a useful remedy for many common and complicated ailments and part of essential nutrients required for human nourishment. So a modern concept of functional food may be brought into practice. • Based on the findings of current research, production technologies may be developed for the promising medicinal plant species of Khanpur Valley and as such of other resourceful valleys. This will help bring the wild species under cultivation and preserved ex situ. • As these six most preferred medicinal plant species of khanpur Valley have proven worth of being used for treatment of various ailments and have tremendous bio-chemical importance, these indigenous landraces may be registered with the Federal

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Registration Agency and conserved in the gene bank for future research and biodiversity. • These plants can play an important role not only in nutrition but also in traditional medicine and in pharmaceutical industry. So these plant species are hereby recommended to herbal industries to be included in their medicinal and food products. These can be utilised in fuctional food to serve both purposes.

EXPERIMENT-4

THE STUDY OF HORTICULTURAL ATTRIBUTES OF SELECTED MEDICINAL PLANTS OF KHANPUR VALLEY IN THE SUB HIMALAYAN MOUNTAINS OF PAKISTAN.

Shah Masaud Khan and Noor ul Amin

Department of Horticulture, The University of Agriculture Peshawar, Pakistan

ABSTRACT

The experiment titled “The Study of Horticultural attributes of selected Medicinal Plants of Khanpur Valley in the sub Himalayan Mountains of Pakistan” was conducted during 2013. Data was recorded on plant height, leaves plant-1, stem diameter, leaf dry weight, plant meter-2, yield plant-1, yield meter-2 and yield hectare-1 for six selected medicinal plants of Khanpur valley (Adhatoda vasica, Calatropis procera, Recinus communis, Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis) at their natural habitats at all the four sites and two seasons. Significant effects were observed at different seasons and sites upon various horticultural attributes of all six medicinal plants. Adhatoda vasica showed significantly higher values for plants meter-2 (0.18) at Mang during summer, yield meter-2 (10.28g) at Mang during summer and yield hectare (102.76kg) at Mang during summer. Calatropis procera revealed higher values for yield hectare (530.02kg) at Mang during summer followed by Dabola site during summer (367.67kg). Recinus communis revealed higher values for yield hectare (248.82kg) at Mang during summer followed by Dam site during summer (220.55kg). Ajuga 220

bracteosa revealed higher significant values for leaves plant-1 (22.87) at Jabri during summer followed by Dabola during summer (22.08). Euphorbia hirta gave higher significant contents for plants meter-2 (0.69) at Dam during summer. Fumaria officinalis revealed higher significant results for leaves plant-1 (488.63) at Mang during summer, yield meter-2 (20.07g) at Mang during summer and yield hectare (200.72kg) at Mang during summer. It was concluded that medicinal plants under study gave maximum yield during summer (last week of July) except Fumaria officinalis which performed best during last week of May. Similarly Ajuga bracteosa gave higher yield at Dabola site; Euphorbia hirta at Dam site while the rest of medicinal plants under study showed maximum yield at Mang site. So medicinal plants may be collected during last week of May and July for best yield.

INTRODUCTION

Since pre-historic age, people have gathered plant and animal resources for their subsistence. Prime examples are edible nuts, mushrooms, fruits, herbs, spices, gums, game, fodder, fibers used in the making of shelters, houses, cloths, utensils, medicines, food, feed, cosmetics and cultural uses. Even today, hundreds of millions of people, mostly in developing countries, derive a significant part of their daily needs and economic benefits from plant and animal products (Walter 2001). Collection from indigenous flora of high-value products such as mushrooms (morels, matsutake, truffles) and medicinal plants (ginseng, gingko biloba, black cohosh, goldenseal) is also practiced in developed countries for social and financial reasons.

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Among these uses, medicinal plants play a central role, not only as traditional medicines used in many cultures but also as trade commodities that meet the demand of often distant markets overlapping fields of condiments, food and cosmetics. Demand for a wide variety of wild species is increasing with growth in human population, food & health needs and national & international trade. With the increased realization that some wild species are being over-exploited, a number of agencies are recommending that wild species be brought into cultivation systems (BAH 2004; Lambert et al. 1997; WHO 1993). Cultivation can also have conservation impacts, however, these need to be better understood. Medicinal plant production through cultivation, can reduce the extent to which wild populations are harvested, but it also may lead to environmental degradation and damage to genetic diversity as well as loss of incentives to conserve wild populations.

The relationship between in situ and ex situ conservation of species is an interesting topic with implications for local communities, public & private land owners, managers, entire industries and of course, wild species. Identifying the conservation benefits and costs of the different production systems for medicinal plants should help guide policies as to whether species conservation should take place in nature or the nursery, or both (Bodeker et al., 1997; Schippmann et al. 2005).

The most important component required to achieve a truly sustainable form of resource utilization is information. In reality, resource managers are always confronted with the lack of adequate information about the plants utilized, their distribution, the genetic diversity of wild plants & land races and, above all, the annual sustainable yield that can be harvested without damaging the plant populations. Research on the conservation and sustainable use of medicinal plants and their habitats has fallen far behind the demand for this globally important resource. Each

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species has unique ecological, socioeconomic, health and cultural associations that must be understood. Model research approaches are feasible, „one size fits all‟ solutions are not lasting solutions and these have to be tailored to local circumstances.

With the renewed interest in biodiversity, the knowledge pertaining to medicinal flora has assumed economic importance. Investigations on growth performance of medicinal plants have gained adequate attention in the worldwide. Quantitative information on the growth performance of medicinal plants under experimental field conditions (Bargali 1997; Karikanthimath et al. 1997; Pandey et al. 1998; Sangai 1995) as well as natural ecosystem conditions (Chauhan et al. 1997 and Jamwel & Kaul 1997) are enormous in literature. However, when considering the richness of Himalayan flora, the available information on the medicinal plants is rare (Shylaja et al. 1996; Singh & Singh 1998).

Quantitative information on the growth performance of medicinal flora of Pakistan have gained very little attention. This quantitative analysis in terms of horticultural attributes is a tremendous contribution towards understanding of the actual yield of the medicinally important part of plant and its sustainable collection without damaging the invaluable flora, indigenous to the valleys of Pakistan. The current study was undertaken to gauge the growth performance of six most preferred medicinal plants of Khanpur valley, by the Department of Horticulture, the University of Agriculture , Peshawar, with the following objectives:

1. To determine the annual yield for sustainable harvest of the selected medicinal plant species.

2. To find out the comparative suitability of seasons and sites for the growth and yield of six selected medicinal plants.

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MATERIAL AND METHODS

The experiment titled “Study of Horticultural Attributes of Selected Medicinal Plants of Khanpur Valley, in the sub Himalayan Mountains of Pakistan” was conducted during 2013. Medicinal plants were studied at their natural habitat. Six most preferred medicinal plants were selected for horticultural studies, three of which were shrubs (Adhatoda vasica, Calatropis procera and Recinus communis) and the three were herbs (Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis). Medicinal importance was calculated through a questionnaire survey as presented in experiment 1 page 24-25. Through quadrate transact method, three transacts (replications) were taken and in every transact data on horticultural attributes was recorded on the plants at their natural habitat at all the four sites and two seasons (details of sites and seasons are mentioned in experiment 1 page 6-7). For six selected plants the data was collected during different periods of summer and winter. The horticultural parameters were measured and data were collected during summer (April- September) and winter (October-March) at different specified periods as below:

1. Adhatoda vasica Last week of July and last week of December.

2. Calitropes procera Last week of July and last week of December. 3. Recenus communis Last week of July and last week of December.

4. Ajuga bracteosa Last week of July and last week of October.

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5. Euphorbia hirta Last week of July and last week of October.

6. Fumaria officinalis Last week of May and last week of February. Study Parameters: As Leaves of shrubs & trees while whole plant of herbs was determined to be the most preferred part of medicinal importance, so dry weight of leaves per plant for shrubs and whole plant for herbs was taken as yield per plant. The horticultural attributes were studied with the help of the following parameters.

1. Plant height (cm)

Plant height of the six selected medicinal species was measured with a standard ruler in cm. The average height of ten randomly selected plants was recorded, from each site in both seasons.

2. Dry Leaf Weight (g)

Leaves were excised with the help of cutter, dried with electric drier and weighed on electronic balance in grams. Leaf weight of the selected medicinal plants was calculated by taking the average of ten randomly selected leaves in the available plants, for all sites in both seasons.

3. Number of leaves plant-1

Number of leaves per plant was calculated by taking the average of number of leaves of ten randomly selected available plants, for all sites in both seasons.

4. Stem diameter (cm)

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Stem diameter was measured with the help of vernier caliper in case of herbs and with a standard ruler, in case of woody plants. Average data of ten randomly selected plants was recorded for all sites in both seasons.

5. Number of plants meter-2

Total number of individuals of a species in a quadrate were counted in all the quadrates (details are available in experiment-1 page 4) and its average was taken as number of plants per quadrate. Plants per quadrat for herbs and shrubs were then converted to plants meter2.

6. Yield plant-1 (Whole plant dry weight for herbs or total leaves dry weight for shrubs) (g).

In case of herbs, ten randomly selected whole plants of each medicinal species were taken and were dried. The dry weight of each plant was noted in grams and its average was taken as yield per plant. In case of shrubs the single leaf dry weight of a plant was multiplied with the total number of leaves per plant and was taken as yield per plant.

7. Yield meter2 (g).

This was calculated with the help of the following formula:

Yield per meter-2 = number of plants per meter2 × yield per plant

8. Yield hectare-1 (kg)

This was calculated with the help of the following formula:

Yield hectare-1 = yield per meter2 (g) × 10000/1000.

Statistical Analysis:

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The data on horticultural parameters were analyzed by using the statistical software "Statistics-8.1", using linear model and general ANOVA with specific AOV model statement. The mean data, for factors and their interactions, were compared by using LSD at 0.05.

RESULTS AND DISCUSSIONS

The study was conducted on six selected medicinal plants and the results & discussions on various parameters of these plants are given below, plant wise. The first three of the six selected medicinal plants were woody plants/shrubs (Adhatoda vasica, Calatropis procera and Recinus communis) and the subsequent three were herbs (Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis).

4.1. ADHATODA VASICA.

1. Plant Height (cm):

The mean data of plant height for various seasons and sites is shown in Table-4.1. The results revealed that the effect of seasons on plant height was significant while the effect of sites and that of their interactions was non-significant.

Maximum plant height was observed at summer (130.82cm) while minimum at winter (92.96cm).

2. Number of leaves plant-1:

The findings revealed that the impact of seasons on number of leaves plant-1 was significant while the effect of sites and that of their interactions was non-significant as shown in Table 4.1.

Maximum number of leaves plant-1 was observed at summer (238.75) while minimum at winter (170.15).

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3. Stem Diameter (cm):

The mean data on stem diameter for various seasons and sites is revealed in Table-4.1. The results discovered that the effect of seasons, sites and their interactions on stem diameter of Adhatoda vasica was non- significant.

4. Dry Leaf Weight (gm):

The results, shown in Table 4.1 on leaf dry weight for various seasons and sites, revealed that the effect of seasons, sites and their interactions on dry leaf weight of Adhatoda vasica was non-significant.

5. Plants Meter-2:

The mean data on plants meter-2 for various seasons and sites is shown in Table-4.1 while their interaction is shown in Figure-4.1. The results revealed that the effect of sites and their interactions on plants meter-2 was significant while the effect of seasons was non-significant.

Maximum number of plants meter-2 was observed at Mang site (0.17) followed by Dam site (0.14) while minimum of it was found at Jabri site (0.07). In case of interaction maximum plants meter-2 was observed at Mang site during summer (0.1833) followed by Mang site during winter (0.1633) while minimum plants meter-2 was observed at Jabri site during summer (0.0700).

6. Yield Plant-1 (dry leaves) (g):

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The results summarized in Table 4.1 revealed that the effect of seasons on yield plant-1 was significant while the effect sites and that of their interactions was non-significant.

Maximum yield plant-1 was observed at summer (51.90g) while minimum at winter (34.80g).

7. Yield Meter-2 (g):

The mean data on yield metert-2 for various seasons and sites is shown in Table-4.1 while their interaction is shown in Figure-4.2. The results discovered that the effect of seasons, sites and their interactions on yield meter-2 was significant.

Highest yield meter-2 was found at summer (6.86g) while lowest at winter (4.19g). Similarly maximum number of yield meter-2 was observed at Mang site (8.10gms) followed by Dam site (6.52g) while minimum of it was found at Jabri site (2.84g). In case of interaction maximum yield meter-2 was observed at Mang site during summer (10.277g) followed by Dam site during summer (8.570g) while minimum plants meter-2 was observed at Jabri site during winter (2.327g).

8. Yield Hectare-1 (kg):

Table 4.1 shows the mean data on number of yield hectare-1 for various seasons and sites, while figure-4.3 depicts their interaction. The results revealed that the impact of seasons, sites and their interactions on yield hectare-1 was significant.

Maximum yield hectare-1 was observed at summer (68.58kg) while minimum at winter (41.92kg). Similarly maximum yield hectare-1 was observed at Mang site (80.98kg) followed by Dam site (65.22kg) while minimum of it was found at Jabri site (28.41kg). In case of interaction

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maximum yield hectare-1 was observed at Mang site during summer (102.76kg) followed by site during (85.71kg) while minimum yield hectare-1 was observed at Jabri site during winter (23.30kg).

It is evident from the results that summer and winter seasons have significantly affected the horticultural attributes like plant height, leaves plant-1, yield plant-1, yield meter-2 and yield hectare-1 while stem diameter, leaf dry weight and plant quadrate-1 were affected with no significance. The plant performed best during summer. The obvious reason for these results can be the availability of required light, temperature, water and nutrients during summer. In Adhatoda vasica, which is an evergreen species, the plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and accumulate biomass at an increased rate. Summer rains and high water availability to plants were conducive for enhanced yield and yield related parameters (Krishnan et al., 2000). Similar results were obtained by Sher, et al. 2010, who concluded that summer temperature boost growth and yield of plant species.

Similarly the parameter like plant quadrate-1, yield meter-2 and yield hectare-1 were significantly affected by various sites while effect of sites on the rest of parameters was non-significant. The reason behind these results might be the soil and climatic conditions of sites, which are comparatively different from each other. Krishnan et al. (2000), contended that it was the altitude which affected the growth and performance of plants. Mang site with low altitude and dry hot climate and rough terrain was suiTable for the natural growth and abundance of Adhatoda vasica. The lowest performance at Jabri site might be due its high altitude and low temperature during winter. In connivance with these findings Liu et al., 2014 discovered that plant density and abundance depend on the climate and topography of sites.

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Table-4.1 Effect of Different Seasons and Sites on Horticultural attributes of Adhatoda vasica indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan. Plant Leaves Stem Leaf Plants Yield Yield Yield -2 Plant-1 -2 Height Plant-1 Diameter Dry Meter Meter- Ha-1 (cm) Weight (g) (g) (kg) Seasons (cm) (g) Summer 130.82a 238.75a 3.81 0.2183 0.13 51.90a 6.86a 68.58a

Winter 92.96b 170.15b 3.84 0.2058 0.12 34.80b 4.19b 41.92b LSD at α 0.05 16.40 17.69 ns ns Ns 3.8175 0.752 7.5151 Sites

Dam 116.75 205.00 3.87 0.2233 0.14b 45.50 6.52b 65.22b

Dabola 106.45 197.00 3.75 0.2150 0.11c 42.13 4.64c 46.38c

Jabri 103.10 188.43 3.83 0.2067 0.07d 39.62 2.84d 28.41d

Mang 121.25 227.63 3.85 0.2033 0.17a 46.12 8.10a 80.98a LSD at α 0.05 ns ns ns ns 0.0202 Ns 1.3815 13.822 Interactions

Seasons*Sites ns ns ns ns * Ns * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

Figure-4.1 Effect of different seasons and sites on number of plants Meter-2 of Adhatoda vasica.

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Figure-4.2 Effect of different seasons and sites on yield meter-2 (g) of Adhatoda vasica.

Figure-4.3 Effect of different seasons and sites on yield hectare-1 (kg) of Adhatoda vasica.

4.2. CALITROPES PROCERA.

1. Plant Height (cm):

The mean data of plant height for various seasons and sites is shown in Table-4.2. The results revealed that the effect of seasons and sites on plant height was significant while the effect of their interactions was non- significant.

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Maximum plant height was observed at summer (98.27cm) while minimum at winter (57.73cm). Similarly maximum plant height was observed at Mang site (88.59cm) followed by Dam site (87.00cm) while minimum of it was found at Jabri site (65.40cm).

2. Number of leaves plant-1:

The results on number of leaves plant-1 for various seasons and sites are shown in Table-4.2. The results showed that the effect of seasons and sites on number of leaves plant-1 was significant while the effect of their interactions was non-significant.

Higher number of leaves plant-1 was observed at summer (102.17) while lower at winter (69.92). Similarly maximum number of leaves plant-1 was observed at Dam site (99.00) followed by Mang site (95.83) while minimum of it was found at Jabri site (68.00).

3. Stem Diameter (cm):

The mean values on stem diameter for various seasons and sites are shown in Table4.6. The results revealed that the effect of seasons on stem diameter was significant while the effect of sites and their interactions was non-significant.

Maximum stem diameter was observed at winter (2.39cm) while minimum at summer (1.93cm).

4. Leaf Dry Weight (g):

The mean data on leaf dry weight for various seasons and sites is shown in Table-4.2. According to the results the effect of seasons and sites on leaf dry weight was significant while the effect of their interactions was non-significant.

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Maximum leaf dry weight was observed at winter (0.211g) while minimum at summer (0.177g). Similarly maximum leaf dry weight was observed at Jabri site (0.221g) followed by Mang site (0.195g) while minimum of it was found at Dam site (0.173g).

5. Plants Meter-2:

The Table-4.2 show the mean data on plants meter-2 for various seasons and sites. The results revealed that the effect of seasons and sites on plants meter-2 was significant while the effect of their interactions was non- significant.

Highest number of plants meter-2 was observed at summer (0.90) while lowest at winter (0.63). Similarly maximum number of plants meter-2 was observed at Mang site (0.91) followed by Dabola site (0.77) while minimum of it was found at Dam site (0.67).

6. Yield Plant-1 (g):

The mean data on yield plant-1 for various seasons and sites is shown in Table-4.2. The results revealed that the effect of seasons and sites on yield plant-1 was significant while the effect of their interactions was non- significant.

According to the results maximum yield plant-1 was observed at summer (42.73g) while minimum at winter (34.42g). Similarly maximum number of yield plant-1 was observed at Mang site (43.72g) followed by Dam site (39.85g) while minimum of it was found at Jabri site (34.62g).

7. Yield Meter-2 (g):

The results on yield metert-2 for various seasons and sites is shown in Table-4.2. The data revealed that the effect of seasons and sites on yield

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meter-2 was significant while the effect of their interactions was non- significant.

Maximum yield meter-2 was observed at summer (38.77g) while minimum at winter (21.77g). Similarly maximum yield meter-2 was observed at Mang site (40.82g) followed by Dabola site (27.99g) while minimum of it was found at Jabri site (25.52g).

8. Yield Hectare-1 (kg):

The mean data on number of yield hectare-1 for various seasons and sites is shown in Table-4.2. The results revealed that the effect of seasons and sites on yield hectare-1 was significant while the effect of their interactions was non-significant.

The results clearly showed that maximum yield hectare-1 was observed at summer (387.65kg) while minimum at winter (217.71kg). Similarly maximum number of yield plant-1 was observed at Mang site (408.19kg) followed by Dabola site (279.93kg) while minimum of it was found at Jabri site (255.16kg).

It is clear from the findings that summer and winter seasons have significantly affected all of the horticultural attributes. Maximum values were recorded during summer for all the parameters except stem diameter for which maximum data was recorded during winter. The obvious reason for these results can be the availability of required light, temperature, water and nutrients during summer. In C.procera, which is an evergreen species, the plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and accumulate biomass at an increased rate. Summer rains and high water availability to plants were conducive for enhanced yield and yield related parameters. Scientests have recommended the cultivation of wild medicinal pants as the only 235

successful startegy of conservation (BAH 2004; Lambert et al. 1997; WHO 1993).

The Precipitation intensity and water availability at sites might have triggered photosynthesis and growth performance (Samuelson et al., 2014). The most convincing explanation for these results can be the soil and climatic characteristics which were more feasible at Mang than other sites for the growth and development of C.procera. This performance is correlated with the soil and climatic conditions of habitat which directly and indirectly affect the growth performance of plant species (Laamrani et al. 2014 and Kane et al. 2014)

Table-4.2 Effect of Different Seasons and Sites on Horticultural attributes of Calatropis procera indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan.

Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

4.3. RECIMUS COMMUNIS.

1. Plant Height (cm):

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The mean data of plant height for various seasons and sites is shown in Table-4.3. The results revealed that the effect of seasons and sites on plant height was significant while the effect of their interactions was non- significant.

Maximum plant height was observed at summer (271.67cm) while minimum at winter (240.93cm). Similarly maximum plant height was observed at Dam site (284.67cm) followed by Dabola site (274.33cm) while minimum of it was found at Jabri site (193.67cm).

2. Number of leaves plant-1:

The results on number of leaves plant-1 for various seasons and sites are shown in Table-4.3. The results showed that the effect of seasons on number of leaves plant-1 was significant while that of sites and their interactions was non-significant.

Maximum number of leaves plant-1 was observed at summer (308.00) while minimum at winter (202.17).

3. Stem Diameter (cm):

The mean data on stem diameter for various seasons and sites is shown in Table-4.3. According to the results the effect of seasons, sites and their interactions on stem diameter was non-significant.

4. Leaf Dry Weight (g):

The results on leaf dry weight for various seasons and sites is shown in Table-4.3. The data revealed that the effect of seasons on leaf dry weight was significant while the effect of sites and their interactions was non- significant. Higher values for leaf dry weight were observed at winter (2.93g) while lower at summer (2.12g).

5. Plants Meter-2:

The mean data on plants meter-2 for various seasons and sites is shown in Table-4.3. According to the results the effect of sites on plants meter-2 was 237

significant while the effect of seasons and their interactions was non- significant.

Maximum number of plants meter-2 was observed at Dam and Mang sites (0.04) followed by Jabri site (0.03) while minimum of it was found at Dabola site (0.02).

6. Yield Plant-1 (g):

The results on yield plant-1 for various seasons and sites are shown in Table-4.3. The results revealed that the effect of seasons and sites on yield plant-1 was significant while the effect of their interactions was non- significant.

Maximum yield plant-1 was observed at summer (616.93g) while minimum at winter (477.70g). Similarly maximum number of yield plant- 1 was observed at Dam site (572.70g) followed by Mang site (571.37g) while minimum of it was found at Jabri site (513.88g).

7. Yield Meter-2 (g):

The Table-4.3 show the mean data on yield metert-2 for various seasons and sites. The results proved that the effect of seasons and sites on yield meter-2 was significant while the effect of their interactions was non- significant.

Highest yield meter-2 was observed at summer (18.35g) while lowest at winter (13.20g). Similarly maximum yield meter-2 was observed at Mang site (20.32g) followed by Dam site (20.07g) while minimum of it was found at Dabola site (9.42g).

8. Yield Hectare-1 (kg):

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The mean data on number of yield hectare-1 for various seasons and sites is shown in Table-4.3. According to the results the effect of seasons and sites on yield hectare-1 was significant while the effect of their interactions was non-significant.

Maximum yield hectare-1 was observed at summer (183.55kg) while minimum at winter (132.04kg). Similarly maximum yield plant-1 was observed at Mang site (203.25kg) followed by Dam site (200.72kg) while minimum of it was found at Dabola site (94.26kg).

It is evident from the results that summer and winter seasons have significantly affected the horticultural attributes like plant height, leaf dry weight, leaves plant-1, yield plant-1, yield meter-2 and yield hectare-1 while stem diameter and plants meter-2 were effected with no significance. Maximum values were recorded during summer for all the parameters except leaf dry weight whuch was observed higher during winter. The obvious reason for these results can be the availability of required light, temperature, water and nutrients during summer. In R.communis, which is an evergreen woody plant species, the plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and accumulate biomass at an increased rate (Liu et al 2014). Summer rains and high water availability to plants were conducive for enhanced yield and yield related parameters. The high values of leaf dry weight during winter might be due to the accumulation of minerals in leaves due to stress.

The sites comparison revealed that maximum plant height, plants meter-2 , yield plant, yield meter-2 and yield hectare-1 were recorded at Mang and Dam sites while its minimum values were observed at Jabri and Dabola sites. One possible reason for this variation can be affiliated with the topography of the site which affect the productivity directly (Lamrani et al., 2014). The most convincing reason for these results might be the soil

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and climatic conditions of the two sites, which are comparatively different from each other. Mang and Dam sites with dry hot climate and hilly rough terrain is suiTable for Recinus communis which grow best at tropical and subtropical climates while its growth was retarded by the cool climate and high altitude of Jabri and Dabola. It is due to the fact that growth and growth related parameters are directly related to the soil and climatic conditions of the site (Kane et al., 2014)

Table-4.3 Effect of Different Seasons and Sites on Horticultural attributes of Recinus communis indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan. - Plant Leaves Stem Leaf Dry Plants Yield Yield Yield Ha -2 -1 -2 Height (cm) Plant-1 Diameter Weight Meter Plant (g) Meter- 1 (kg) Seasons (cm) (g) (g) Summer 271.67a 308.00a 3.89 2.12b 0.03 616.93a 18.35a 183.55a Winter 240.93b 202.17b 3.88 2.39a 0.03 477.70b 13.20b 132.04b LSD at α 0.05 21.32 9.8250 Ns 0.1565 ns 26.117 1.54 15.403 Sites

Dam 284.67a 272.83 4.03 2.11 0.04a 572.70a 20.07a 200.72a Dabola 274.33a 259.50 3.97 2.12 0.02c 533.32b 9.42c 94.26c Jabri 193.67b 241.17 3.58 2.20 0.03b 513.88b 13.29b 132.95b Mang 272.53a 246.83 3.97 2.37 0.04a 571.37a 20.32a 203.25a LSD at α 0.05 35.56 Ns Ns ns 0.0066 34.120 3.1586 31.587 Interactions

Seasons*Sites Ns Ns Ns ns ns ns ns ns Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. and * = Significant at 5 % level of probability.

4.4. AJUGA BRACTEOSA

1. Plant Height (cm):

The results revealed that the effect of seasons on plant height was significant while the effect of sites and their interactions was non- significant as given in Table-4.4.

Maximum plant height was observed at summer (17.17cm) while minimum at winter (7.83cm).

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2. Number of leaves plant-1:

The mean data on number of leaves plant-1 for various seasons and sites is shown in Table-4.4 while their interaction is shown in Figure-4.4. The results revealed that the effect of seasons, and their interactions on number of leaves plant-1 was significant while that of sites was non-significant.

Maximum number of leaves plant-1 was observed at summer (20.70) while minimum at winter (15.77). In case of interaction maximum number of leaves plant-1 was observed at Jabri site during summer (22.867) followed by Dabola during summer (22.067) while minimum number of leaves plant-1 was observed at Dabola site during winter (14.833).

3. Stem Diameter (cm):

The results shown in Table-4.4, revealed that the effect of seasons on stem diameter was significant while the effect of sites and their interactions was non-significant.

Maximum stem diameter was observed at winter (1.32cm) while minimum at summer (0.87cm).

4. Leaf Dry Weight (g):

The results on leaf dry weight for various seasons and sites are shown in Table-4.4. The findings revealed that the effect of seasons on leaf dry weight was significant while the effect of sites and their interactions was non-significant.

Highest leaf dry weight was observed at winter (0.2533g) while minimum at summer (0.2233g).

5. Plants Meter-2:

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The mean data on plants meter-2 for various seasons and sites is shown in Table-4.4. The results revealed that the effect of seasons and sites on plants meter-2 was significant while the effect of their interactions was non-significant.

Maximum plants meter-2 was observed at summer (0.67) while minimum at winter (0.49). Similarly maximum number of plants meter-2 was observed at Dabola site (0.84) followed by Jabri site (0.73) while minimum of it was found at Mang site (0.36).

6. Yield Plant-1 (g):

The results given in Table-4.4, revealed that the effect of seasons and sites on yield plant-1 was significant while the effect of their interactions was non-significant.

Maximum yield plant-1 was observed at summer (10.57g) while minimum at winter (9.42g). Similarly maximum number of yield plant-1 was observed at Mang site (10.93g) followed by Jabri site (10.72g) while minimum of it was found at Dam site (8.92g).

7. Yield Meter-2 (g):

The mean data on yield metert-2 for various seasons and sites is shown in Table-4.4. The results revealed that the effect of seasons and sites on yield meter-2 was significant while the effect of their interactions was non- significant.

Maximum yield meter-2 was observed at summer (7.25g) while minimum at winter (4.64g). Similarly maximum yield meter-2 was observed at Dabola site (8.27g) followed by Jabri site (7.84g) while minimum of it was found at Dam site (3.69g).

8. Yield Hectare-1 (kg):

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The results on number of yield hectare-1 for various seasons and sites are given in Table-4.4. The findings revealed that the effect of seasons and sites on yield hectare-1 was significant while the effect of their interactions was non-significant.

Highest yield hectare-1 was observed at summer (72.52kg) while lowest at winter (46.37kg). Similarly maximum number of yield plant-1 was observed at Dabola site (82.70kg) followed by Jabri site (78.41kg) while minimum of it was found at Dam site (36.90kg).

The possible reason for these results can be the availability of required light, temperature, water and nutrients during summer (Liu et al., 2014). In Ajuga which is an annual herbaceous species, the plant acquired maximum bio-chemical compounds during summer to grow at higher rate and accumulate biomass at higher rate. Summer rains and high water availability to plants were conducive for enhanced yield and yield related parameters. As the plant gets matured at winter so the stem diameter and leaf dry weight were recorded higher in winter than summer.

The sites comparison revealed that maximum plants meter-2 (8.27) and yield hectare-1 (82.70) were recorded at Dabola site followed by those of Jabri site while its minimum values were observed at Mang and Dam sites. The most scientific reason for these results might be the soil and climatic conditions of the two sites, which are comparatively different from each other. Ajuga is a temperate to sub-tropical to temperate climate loving plant species which is apparently available at Dabola and Jabri sites. The low winter temperature and mild summer of these sites triggered higher growth and yield performance as found by Sher et al. (2010) and Laamrani et al. (2014). Similar results were obtained by Chauhan et al. (1997) and Jamwel & Kaul (1997) who studied growth performance of medicinal plants under natural ecosystem conditions.

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Table-4.4 Effect of Different Seasons and Sites on Horticultural attributes of Ajuga bracteosa indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan. Plant Leaves Stem Leaf Plants Yield Yield Yield -2 Plant-1 - Height Plant-1 Diameter Dry Meter Meter- Ha-1 2 Seasons (cm) (cm) Weight (g) (g) (kg) (g) Summer 17.17a 20.70a 0.87b 0.2233b 0.67a 10.57a 7.22a 72.52a Winter 7.83b 15.77b 1.32a 0.2533a 0.49b 9.42b 4.64b 46.37b LSD at α 0.05 1.6078 1.4946 0.1245 0.0222 0.0908 0.3438 0.9764 9.7799 Sites

Dam 11.75 18.27 1.08 0.2083 0.40b 8.92b 3.69b 36.90b Dabola 13.50 12.92 18.45 1.17 1.17 0.2300 0.84a 9.77b 8.27a 82.70a Jabri 11.83 19.08 0.97 0.2433 0.73a 10.72a 7.84a 78.41a Mang 17.13 0.2717 0.36b 10.93a 3.98b 39.77b LSD at α 0.05 Ns Ns ns ns 0.1325 1.2885 1.4442 14.441 Interactions

Seasons*Sites Ns * ns ns Ns Ns ns Ns

Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

Figure-4.4 Effect of different seasons and sites on number of Leaves plant-1 of Ajuga bracteosa.

4.5. EUPHORBIA HIRTA.

1. Plant Height (cm):

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The mean data of plant height for various seasons and sites is shown in Table-4.5. The results revealed that the effect of seasons and sites on plant height was significant while the effect of their interactions was non- significant.

Maximum plant height was observed at winter (54,08cm) while minimum at summer (28.67cm). Similarly maximum plant height was observed at Mang site (845.77cm) followed by Dabola site (43.30cm) while minimum of it was found at Jabri site (36.34cm).

2. Number of leaves plant-1:

The results on number of leaves plant-1 for various seasons and sites are shown in Table-4.5. The data revealed that the effect of seasons and sites on number of leaves plant-1 was significant while the effect of their interactions was non-significant.

Highest number of leaves plant-1 was observed at summer (80.77) while lowest at winter (67.17). Similarly maximum number of leaves plant-1 was observed at Mang site (82.60cm) followed by Dam site (479.36) while minimum of it was found at Dabola site (65.32).

3. Stem Diameter (cm):

The results revealed that the effect of seasons on stem diameter was significant while that of sites and their interactions was non-significant as shown in Table-4.5.

Maximum stem diameter was observed at summer (0.48cm) while minimum at winter (0.37cm).

4. Leaf Dry Weight (gm):

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The mean data on leaf dry weight for various seasons and sites is shown in Table-4.5. The findings revealed that the effect of seasons on leaf dry weight was significant while that of sites and their interactions was non- significant. Maximum leaf dry weight was observed at winter (0.0082g) while minimum at summer (0.0047g).

5. Plants Meter-2:

The results on plants meter-2 for various seasons and sites is shown in Table-4.5 while their interaction is depicted in Figure-4.5. The results revealed that the effect of seasons, sites and their interactions on plants meter-2 was significant.

Plants meter-2 were observed highest at summer (0.49) while lowest at winter (0.17). Similarly maximum number of plants meter-2 was found at Dam site (0.45) followed by Mang site (0.41) while minimum of it was found at Jabri site (0.13). In case of interaction maximum plants meter-2 were recorded at Dam site during summer (0.693) followed by Mang site during summer (0.550) while minimum plants meter-2 were observed at Jabri site during winter (0.067).

6. Yield Plant-1 (g):

The mean data on yield plant-1 for various seasons and sites is shown in Table-4.5. The results revealed that the effect of seasons on yield plant-1 was significant while the effect of sites and their interactions was non- significant.

Maximum yield plant-1 was observed at winter (1.56g) while minimum at summer (30.74g).

7. Yield Meter-2 (g):

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The results revealed that the effect of seasons and sites on yield meter-2 was significant while the effect of their interactions was non-significant as given in Table-4.5.

Maximum yield meter-2 was observed at summer (0.37g) while minimum at winter (0.26g). Similarly maximum number of yield meter-2 was observed at Mang site (0.45g) followed by Dam site (0.41g) while minimum of it was found at Jabri site (0.12g).

8. Yield Hectare-1 (kg):

The mean data on number of yield hectare-1 for various seasons and sites is shown in Table-4.5. The results revealed that the effect of seasons and sites on yield hectare-1 was significant while the effect of their interactions was non-significant.

Highest yield hectare-1 was observed at summer (3.68kg) while lowest at winter (2.64kg). Similarly maximum yield hectare-1 was observed at Mang site (4.41kg) followed by Dam site (4.13kg) while minimum of it was found at Jabri site (1.24kg).

The horticultural attributes like plant height, leaves plant-1, stem diameter, leaf dry weight, plants meter-2, yield plant-1, yield meter-2 and yield hectare-1 were significantly affected by seasons. Maximum values were recorded during summer for all the parameters except plant height, leaf dry weight and yield per plant which were higher during winter. The obvious reason for these results can be the availability of required light, temperature, water and nutrients during summer (Sher et al. 2010). In E.hirta which is an annual herbaceous plant species, the plant accumulated maximum biochemical compounds in summer to grow at higher rate and accumulate biomass at an increased rate. Summer rains and high water availability to plants were conducive for enhanced yield and yield related

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parameters. As the plant gets matured at winter so the plant height, leaf dry weight and yield per plant were recorded higher in winter than summer. Similar trend was reported by Ogbolie et al., (2007).

The sites comparison revealed that maximum values for all parameters except plants per meter-2 were recorded at Mang site while its minimum values were observed at Jabri and Dabola sites. The most scientific reason for these results might be the soil and climatic conditions of the two sites, which are comparatively much different from each other. Mang site with dry hot climate and shaddy sides were suiTable for E.hirta natural growth and vegetative abundance while the high altitude and cool climate of Jabri and Dabola sites retarded the growth of hirta. Similar findings were given by Ahmad et al., 2008 who found that plant performance depends on the climatic suitability of locations.

Table-4.5. Effect of Different Seasons and Sites on Horticultural attributes of Euphorbia hirta indigenous to Khanpur Valley, in sub- Himalayan mountains of Pakistan. Plant Leaves Stem Leaf Plants Yield Yield Yield -2 Plant-1 -2 Height Plant-1 Diameter Dry Meter Meter- Ha-1 Seasons (cm) (cm) Weight (g) (g) (kg) (g) Summer 28.67b 67.17b 0.37b 0.0047b 0.49a 0.74b 0.37a 3.68a Winter 54.08a 80.77a 0.48ba 0.0082a 0.17b 1.56a 0.26b 2.64b LSD at α 0.05 3.0274 3.7675 0.0428 0.0007 0.0338 0.1085 0.0641 0.6412 Sites

Dam 40.08b 79.36a 0.44 0.0057 0.45a 1.11 0.41a 4.13a Dabola 43.30a 65.32b 0.37 0.0070 0.33b 1.11 0.28b 2.76b Jabri 36.34c 68.62b 0.40 0.0070 0.13c 1.16 0.12c 1.24c Mang 45.77a 82.60a 0.48 0.0067 0.41ab 1.24 0.45a 4.41a LSD at α 0.05 2.9818 8.6698 Ns ns 0.1099 ns 0.1258 1.2565 Interactions

Seasons*Sites ns Ns Ns ns * ns ns Ns Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

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Figure-4.5 Effect of different seasons and sites on number of Plants Meter-2 of Euphorbia hirta.

4.6. FUMARIA OFFICINALIS.

1. Plant Height (cm):

The results revealed that the effect of seasons and sites on plant height was significant while the effect of their interactions was non-significant as shown in Table-4.6.

Maximum plant height was found at summer (48.03cm) while minimum at winter (21.11cm). Similarly maximum number of plant height was observed at Mang site (38.88cm) followed by Dam site (38.56cm) while minimum of it was found at Jabri site (29.73cm).

2. Number of leaves plant-1:

The mean data on number of leaves plant-1 for various seasons and sites is shown in Table-4.6 while their interaction is shown in Figure-4.6. The results revealed that the effect of seasons, sites and their interactions on number of leaves plant-1 was significant.

Highest number of leaves plant-1 was observed at summer (426.82) while minimum at winter (177.82). Similarly maximum number of leaves plant- 1 was recorded at Mang site (331.12) followed by Dam site (317.18) while minimum of it was found at Jabri site (264.27). In case of interaction maximum number of leaves plant-1 was observed at Mang site during summer (488.63) followed by Dam site during summer (441.13) while

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minimum number of leaves plant-1 was observed at Jabri site during winter (152.47).

3. Stem Diameter (cm):

The results on stem diameter for various seasons and sites are shown in Table-4.6. The data revealed that the effect of seasons on stem diameter was significant while the effect of sites and their interactions was non- significant.

Maximum stem diameter was observed at summer (0.82cm) while minimum at winter (0.35cm).

4. Leaf Dry Weight (g):

The mean data on leaf dry weight for various seasons and sites is shown in Table-4.6. The results revealed that the effect of seasons on leaf dry weight was significant while the effect of sites and their interactions was non-significant.

Maximum leaf dry weight was observed at summer (0.0124g) while minimum at winter (0.0031g).

5. Plants Meter-2:

The results revealed that the effect of sites on plants meter-2 was significant while the effect of seasons and their interactions was non- significant as shown in Table-4.6.

Highest number of plants meter-2 was observed at Mang site (1.54) followed by Dabola site (1.12) while minimum of it was found at Jabri site (1.04).

6. Yield Plant-1 (g):

The mean data on yield plant-1 for various seasons and sites is shown in Table-4.6. The results revealed that the effect of seasons on yield plant-1 was significant while the effect sites and that of their interactions was non- significant.

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Maximum yield plant-1 was observed at summer (12.63g) while minimum at winter (1.31g).

7. Yield Meter-2 (g):

The results on yield metert-2 for various seasons and sites are shown in Table-4.6 while their interaction is depicted in Figure-4.7. The data revealed that the effect of seasons, sites and their interactions on yield meter-2 was significant.

Maximum yield meter-2 was observed at summer (14.48g) while minimum at winter (1.68g). Similarly maximum yield meter-2 was observed at Mang site (11.12g) followed by Dam site (7.91g) while minimum of it was found at Jabri site (6.06g). In case of interaction maximum yield meter-2 was observed at Mang site during summer (20.07g) followed by Dam site during summer (14.12g) while minimum yield meter-2 was observed at Jabri site during winter (1.31g).

8. Yield Hectare-1 (kg):

The mean data on number of yield hectare-1 for various seasons and sites is shown in Table-4.6 while their interaction is shown in Figure-4.8. The results revealed that the effect of seasons, sites and their interactions on yield hectare-1 was significant. highest yield hectare-1 was observed at summer (144.83kg) while minimum at winter (16.76kg). Similarly maximum yield hectare-1 was observed at Mang site (11.15kg) followed by Dam site (79.13kg) while minimum of it was found at Jabri site (60.64kg). In case of interaction maximum yield hectare-1 was observed at Mang site during summer (200.72kg) followed by Dam site during summer (141.23kg) while minimum yield hectare-1 was observed at Mang site during winter (13.13kg).

Seasons have significantly affected the horticultural attributes like plant height, stem diameter, leaf dry weight, leaves plant-1, yield plant-1, yield meter-2 and yield hectare-1 while plants meter-2 were effected with no significance. Maximum values were recorded during summer for all the

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parameters except plants meter-2 which were nonsignificantly higher in winter. The obvious reason for these results can be the availability of required light, temperature and water during summer as stated by Liu et al., 2014. furthermore, excellent performance during summer was most probably due to the presence of suiTable temperature, enough moisture and nutrients during summer as mentioned by Skarpe, (1990); Zaman, (1997) and Nanette et al., (2007). In Fumeria officinalis which is a seasonal herb, which sprout during winter and bloom in spring. The plants gets maximum plant biomass and set seed in May-June. The plant accumulated maximum bio-chemical compounds in summer to grow at higher rate and accumulate biomass at an increased rate (Krishnan et al., 2000). Summer rains and high water availability to plants enhanced yield and yield related parameters except plants meter-2 which was higher in winter due to the fact that the seed of Fumaria, germinate in winter and hence maximum number of plants were available during winter which later on decreased due to climatic and anthropological pressure. The mild winter and low summer temperatures promote the growth and yield of many plant species (Sher et al. 2010)

Similarly, plant height, leaves plant-1, plant quadrate-1, yield meter-2 and yield hectare-1 were significantly different at various sites while the impact of sites on the rest of parameters was non-significant. The sites comparison revealed that maximum values for all parameters were recorded at Mang site while its minimum values were observed at Jabri site. The reason behind these findings might be the soil and climatic conditions of the two sites, which are comparatively much different from each other. Mang site with dry hot climate and farm sides were suitable for fumaria natural growth and vegetative abundance while the cool wet climate of Jabri site discouraged the growth of fumaria. Similar results were obtained by Laamrani et al., 2014, who concluded that spatial variation caused changes in the growth performance of plant species.

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Table-4.6 Effect of Different Seasons and Sites on Horticultural Parameters of Fumaria officinalis indigenous to Khanpur Valley, in sub-Himalayan mountains of Pakistan. Plant Leaves Stem Leaf Plants Yield Yield Yield -2 Plant-1 -2 Height Plant-1 Diameter Dry Meter Meter- Ha-1 Seasons (cm) (cm) Weight (g) (g) (kg) (g) 426.82a 0.82a 0.0124a 12.63a 14.48a 144.83a Summer 48.03a 1.14 177.82b 0.35b 0.0031b 1.31b 1.68b 16.76b Winter 21.11b 1.26 LSD at α 0.05 2.4464 18.587 0.1153 0.00103 Ns 0.5589 1.3807 13.807 Sites

Dam 38.56a 317.18ab 0.61 0.0080 1.11b 7.50 7.91b 79.13b Dabola 31.11b 296.70b 0.61 0.0078 1.12b 6.72 7.23bc 72.25bc Jabri 29.73b 264.27c 0.58 0.0080 1.04b 6.63 6.06c 60.64c Mang 38.88a 331.12a 0.55 0.0071 1.54a 7.04 11.12a 111.15a LSD at α 0.05 6.4318 25.193 Ns Ns 0.2230 ns 1.6098 16.097 Interactions

Seasons*Sites Ns * Ns ns Ns ns * * Means followed by similar letter(s) in column do not differ significantly. ns = Non Significant. * = Significant at 5 % level of probability.

Figure-4.6. Effect of different seasons and sites on number of leaves plant-1 of Fumaria officinalis.

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Figure 4.7. Effect of different seasons and sites on Yield Meter-2 (gms) of Fumaria officinalis.

Figure-4.8. Effect of different seasons and sites on yield Hectare-2 (Kg) of Fumaria officinalis.

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SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

This is an important and novel aspect of medicinal flora at their natural habitat studied for the first time ever in the history of research on plant sciences in the Himalayan region of Pakistan. The relationship between in situ and ex situ conservation of species is an interesting topic. sustainable resource utilization and conservation is a policy guideline as to whether species conservation should take place in nature or in the nursery, or both (Bodeker et al., 1997; Schippmann et al., 2005). BAH 2004; Lambert et al., 1997 and WHO 1993 have emphasized that, with the increased realization that some wild species are being over-exploited, the species shall be brought into cultivation systems.

In reality, resource managers are always confronted with the lack of adequate information about the plants used, their distribution and density, the local preferences, yield per plant, the annual sustained yield that can be harvested without damaging the populations (Bodeker et al., 1997). Research on the conservation and sustainable use of medicinal plants and their habitats has fallen far behind the demand for this globally important resource. To focus on the objective to measure the yield and yield related components of medicinally important plants in nature, a research titled “Effect of Different Seasons and Sites on the Horticultural attributes of the Most Preferred Medicinal Plants of Khanpur Valley” was conducted at Khanpur valley, in the sub Himalayan mountains of Khyber Pakhtunkhwa, Pakistan during 2013. Data was

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recorded on plant height, leaves plant-1, stem diameter, leaf dry weight, plants quadrate(488.63) at Mang during summer, yield meter-2 (20.07g) at Mang during summer and yield hectare (200.72kg) at Mang during summer. Euphorbia hirta gave higher significant contents for plants meter-2 (0.69) at Dam during summer. Ajuga bracteosa revealed higher significant values for leaves plant-1 (22.87) at Jabri during summer followed by Dabola during summer (22.08). Recinus communis revealed higher values for yield hectare (248.82kg) at Mang during summer followed by Dam site during summer (220.55kg). Calatropis procera revealed higher values for yield hectare (530.02kg) at Mang during summer followed by Dabola site during summer (367.67kg).

It was concluded that medicinal plants under study gave maximum yield during summer (last week of July) except Fumaria officinalis which performed best during last week of May. Similarly Ajuga bracteosa gave higher yield at Dabola site; Euphorbia hirta at Dam site while the rest of

1 , yield plant-1, yield meter-2 and yield hectare-1 for six most preferred medicinal plants of Khanpur valley (Adhatoda vasica, Calatropis procera, Recinus communis, Ajuga bracteosa, Euphorbia hirta and Fumaria officinalis) at their natural habitat at all the four sites and two seasons. The results obtained from the experiment were highly interesting.

Significant effects were observed at different seasons and sites upon various horticultural attributes of all six medicinal plants. Adhatoda vasica showed significantly higher values for plants meter-2 (0.18) at Mang during summer, yield meter-2 (10.28g) at Mang during summer and yield hectare (102.76kg) at Mang during summer. Fumaria officinalis revealed higher significant results for leaves plant-1

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medicinal plants under study showed maximum yield at Mang site. So medicinal plants may be collected during last week of May and July for best yield.

It was proved that summer season was the best period for collecting the plants or its parts to get maximum yield. Similarly Jabri and Dabola sites are best for collecting Ajuga and its relatives while Fumaria and Hirta can be best collected from the plains and farm bordering areas at Mang and Dam sites. The three woody medicinal plant species A.vasica, C.procera and R.communis were found with higher yield per hectare at Mang and Dam sites during the last week of July.

The following conclusions are drawn from the whole results and discussions:

• The last week of July is the most appropriate time than October for the collection of E. hirta and A. bracteosa while A. vasica, C. procera and R. communis may be better harvested during the last week of July than December. The most suitable time for collection of F. officinalis was the last week of May as compared to February. • It was proved that summer season was the best period for collecting the plants or its parts to get maximum yield.

• Similarly Jabri and Dabola sites are best for collecting Ajuga bracteosa and its relatives while Fumaria and Hirta can be best collected from the plains and farm bordering areas at Mang and Dam sites. • The three woody medicinal plant species A. vasica, C. procera and R. communis were found with higher yield per hectare at Mang and Dam sites.

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Following are some of the recommendations derived, after thorough consideration, from the study and based on the research findings;

• It is strongly recommended that while collecting the medicinal raw material from the indigenous flora, guidelines regarding appropriate season and site must be followed. Like last week of May for F. officinalis and last week of July for E. hirta, A. bracteosa, A. vasica, C. procera and R. communis are the most appropriate time for harvesting. • Where conservation of medicinal plant species in nature is not possible these may be brought into cultivation at nursery and be conserved. • Sustainable harvesting is the best policy to make the requirements fulfilled without any damage to the flora of these valleys. The farmers or anyone who is interested in the collection of medicinal plants must have the knowledge of the spatio-temporal existence of these plants.

OVERALL CONCLUSIONS • The 202 wild plant species, belonging to 48 different families, recorded at Khanpur valley shows that the valley is rich in species diversity. Similarly 71 medicinally important plant species were identified at the research area which means that 35.15% of the flora of khanpur is pharmaceutically important. • The local people prefer Adhatoda vasica, Fumaria officinalis and Ajuga bracteosa as medicinal plants and use whole plant in case of herbs and leaves in case of woody plants. The people of Khanpur prefer powder and decoction form of utilization and the cure of ailments related to digestive and respiratory systems. • Khanpur Valley was documented with 116 ecologically important plant species (57.43%) and 86 threatened plant species (42.57%), out of total 202 wild indigenous plant species. Out of 116 ecologically important species found in the valley, majority fall in the class of Rare

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(R) (45.37%), followed by Occasional (O) (19.72%), then Frequent (F) (5.06%) and least fall in the class of Abundant (A) (2.05%). • Medicinal plants demonstrated higher quantity of proteins and fats in summer and higher quantity of sugar and essential oil in winter. Medicinal Plants Adhatoda vasica, Fumaria officinalis and Recinus communis may be harvested in summer for maximum bio-chemical elements (Na, K, Ca, P, Mg, Cu, Fe, Mn and Zn) while Euphorbia hirta, Ajuga bracteosa and Calatropis procera, may be harvested in winter for highest biochemical elements. • It was proved that summer season was the best period for collecting the plants or its parts to get maximum yield. The last week of July was the most appropriate time than October for the collection of E. hirta and A. bracteosa while A. vasica, C. procera and R. communis may be better harvested during the last week of July than December. The most suiTable time for collection of F. officinalis was the last week of May as compared to February. • Similarly Jabri and Dabola sites are best for collecting Ajuga and its relatives while Fumaria and Hirta can be best collected from the plains and farm bordering areas at Mang and Dam sites. The three woody medicinal plant species A.vasica, C.procera and R.communis were found with higher yield per hectare at Mang and Dam sites.

OVER ALL RECOMMENDATIONS • Being a repository of 202 plant species with 71 medicinal plants, it will be worthwhile if a herbal garden of indigenous medicinal flora is established in the valley to preserve this national wealth. The people of Khanpur valley prefer medicinal plants (Adhatoda vasica, Fumaria officinalis and Ajuga bracteosa) used in the cure of ailments related to digestive and respiratory systems. These plant species are recommended to be brought into cultivation for promotion and conservation. • As khanpur valley is in close vicinity of Hattar Industrial Estate a linkage between the local community and industry at Hattar, may be established to supply raw material from the wild for products

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preparation. Adhatoda vasica which is used by Local Indutries in Joshanda can be the one to start with. Local community need trainings and awareness about the usefulness and sustainable utilization of the herbal plants to make this invaluable source of living available to the generations to come. • Being an ecosystem of 57.43% ecologically important species (116) and 42.57% threatened species (86) and yet majority of the ecologically important species fall in the class rare (45.37%), the flora require a conservation plan on urgent basis, both in nature and in nursery. • Maximum plant species were ecologically, biochemically and horticulturally associated with summer, so plants or it parts collection may be restricted to summer season and harvesting/felling of plants in winter may also be controlled. • Human body needs essential nutrients, major minerals and trace elements which are essential for general health, growth and reproduction. Many elements are associated with one another in maintaining our normal growth and health. The six medicinal plants studied may prove to be a useful remedy for many common and complicated ailments and part of essential nutrients required for human nourishment. So these plant species may be used in the modern concept of functional food. • Detailed ecological study on threatened plant species is recommended to be conducted in future to find out the magnitude and causes of loss to the precious plant resources. • Based on the findings of current research, production technologies may be developed for the promising medicinal plant species of Khanpur Valley. This will help bring the wild species under cultivation and the natural wealth shall be preserved. As the six most preferred medicinal plant species of khanpur Valley have proven worth of being used for treatment of various ailments and have tremendous bio- chemical importance, these indigenous landraces may be registered with the Federal Plant Variety Registration Agency and conserved in the national gene bank for future research and biodiversity.

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APPENDICES

Appendix 1.1: List of total plant species identified at Khanpur Valley. Serial Species Botanical Name Family Name local Name (English Growth habit No name) 1 Acacia modesta Memosaceae Palosa, Phulai (Acacia) Tree 2 Acacia muricata Fabaceae (spineless wattle) Tree 3 Acacia nilotica Memosaceae Kikar (Acacia) Tree 4 Acacia victoriae Fabaceae Khardar kikar (thorny Tree wattle)

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5 Achyranthes aspera L. Amaranthaceae Puthkanda Herb 6 Adhatoda vasika Acanthaceae Baikar, waikar (Malaber Shrub nut tree) 7 Adiantum pedatum Pteridaceae five fingered fern Herb (maidenhair fern) 8 Adiantum venustum Pteridaceae Round leaved fern Herb (Himalayan fern)

9 Ailanthus altissima Simaroubaceae Jangali darach, barabro Tree (Tree of heaven) 10 Ajuga bractiosa Labiatae Karvi Buti (Ajuga) Herb 11 Allium jaquemontii Alliaceae Jangli piaz (Wild onion) Herb 12 Alternanthera pungens Amaranthaceae Kanchari (Joy weed) Herb prostrate Kunth. 13 Amaranthus graecizans L. Amaranthaceae chulai (Pig weed) Herb 14 Amaranthus hybridus Amaranthaceae Shalkhay, chulai (wild Herb spinach) 15 Amaranthus spinosus L. Amaranthaceae Khardar Chulai Herb 16 Amaranthus viridis L. Amaranthaceae Chulai, shalkhay (wild Herb spinach) 17 Anagallis arvensis L. Primulaceae Olalai, dhabbar Herb prostrate (Jonkmari) 18 Androsace rotundifolia Primulaceae Arrow head alpine Herb 19 Anthemis cotula L. Asteraceae Tharkha babona Herb (Stinking chamomile) 20 Arenaria leptoclados Caryophyllaceae Thyme leaf (sand wart) Herb 21 Artemisia brevifolia Asteraceae(compositae) Tharkha (artimisia) Herb

22 Artemisia scoparia Asteraceae Jhao, lasaj, chaw, marua Herb woody (artimisia) type 23 Arundo donax Poaceae Kalam bote (Cane grass) Shrub 24 Asparagus gracilis Liliaceae Shahghandal (Halyun) Herb-woody 25 Avena sativa L. Poaceae Jangali jao (Wild oat) Herb 26 Barleria prionitis Acanthaceae (Porcupine flower, Shrub yellow picanier) 27 Bauhinia variegata Fabaceae Kachnar Tree

Serial Species Botanical Name Family Name local Name (English Growth habit No name) 28 Berberis lycium Berberidaceae Kwaray, simlo, kashmal, Shrub ziar largay

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29 Bidens biternata Asteraceae Spanish needle, gold cup Herb silver plate, black jack 30 Boerhavia diffusa Nyctaginaceae Tukhme ispast, Chalaray Herb (red hog weed) 31 Boerhavia procumbens Nyctangiaceae Sentori, itsit, wasao Herb 32 Bombax malabaricum Malvaceae Silk cotton tree 33 Brassica campestris L. Brasicaceae Sarson (Mustard) Herb 34 Broussonetia papyrifera Moraceae Gul toot (Paper Tree mulberry) 35 Calendula arvensis L. Asteraceae Ziar gula (field marigold) Herb 36 Calotropis procera Asclepidaceae Spalwaka, Mundar (Akk) Shrub 37 Canna indica L. Cannaceae Hakik (Indian shot) Harb 38 Cannabis sativa L. Cannabaceae Bhang (Canabis) Herb 39 Capsilla bursa-pastoris L. Brasicaceae Chambraka, (Shepherd Herb purse, mothers heart) 40 Carthamus oxycantha Asteraceae Jeweled distaff thistle Herb spiny leaved 41 Cassia absus L. Caesalpiniaceae Chaksu, jasmeejaz Herb shrubby type 42 Cassia fistula Fabaceae Amaltas ( golden shower Tree tree)

43 Cassia sophera Fabaceae senna, kasaundi 44 Celtis australis Canabbaceae Hack berry, wild falsa, Tree european nettle tree 45 Centaurea iberica Asteraceae Iberian knap weed, star Herb thistle,

46 Centaurium pulchellum Gentianaceae Lesser centaury, Herb branched centaury 47 Cerastium glomeratum Caryophyllaceae Sticky chick weed, sticky Herb mouse ear 48 Chenopodium album L. Chenopodiaceae Bathu (Wild spinach, Herb lambs quarter) 49 Chenopodium Chenopodiaceae Katto (Maxican tea, worm Herb ambrosioides L. seed) 50 Chenopodium murale L. Chenopodiaceae Bathu, kurund (Goose Heb foot) 51 Cleome viscose L. Capparidaceae Asian spider flower, tick Herb woody weed type

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52 Clitoria annua Fabaceae Desi banafsha (bombay Herb bean, butterfly pea) 53 Convolvulus arvensis L. Convolvulaceae leli, prewathai (Bind Herb prostrate weed)

Serial Species Botanical Name Family Name local Name (English Growth habit No name) 54 Conyza bonariensis Asteraceae Asthma weed Herb woody type 55 Coronopus didymus Brasicaceae Swiss cress Herb 56 Cousinia prolifera Asteraceae Cousinia Herb 57 Cuscuta reflexa Cuscutaceae Dodder, akash bail,amar Herb prostrate bail 58 Cynodon dactylon L Poaceae dub, kabbal (Bermuda Herbperennial grass) 59 Cyperus rotundus L. Cyperaceae Nut grass, Deela, Herbperennial

60 Dactyloctinium annulatum Poaceae Madhana (Egyptian Herb-annual L. finger grass) 61 Dalbergia sissoo Fabaceae Shisham Tree 62 Datura alba Solanaceae Mangaz butay, (devils Herb trumpit and metel) 64 Datura stramonium L. Solanaceae Datura siah (jimson Herb weed) 65 Desmodium elegans Fabaceae Tick clover, biggar lice soft wood Shrub 66 Dichanthium annulatum Poaceae Palwan (Marvel grass) Herbperennial

67 Dicliptera bubleuroides Acanthaceae Foldwing, kirch, somni. Herb Nees.

68 Digera muricata L. Amaranthaceae Sur gulay Herb 69 Dodonea viscosa Sapindaceae Sanata, ghwaraskay Shrub 70 Echinochloa colona Poaceae Jungle rice grass Herb 71 Echinops echinatus Asteraceae Ont katara (indian globe Herb thistle) 72 Eclipta prostrata L. Asteraceae Bhangra (False daisy) Herb aquatic 73 Ellusine indica L. Poaceae Chimber (Crab or crow Herb-annual foot grass) 74 Eragrostice minor Host, Poaceae Pungent meadow grass, Herb annual Gram.Auster love grass

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75 Eryngium coerolium Apiaceae Pimpiti (Sea holly) Herb MBieb. 76 Euphorbia falcata L. Euphorbiaceae Sickle spurge Herb 77 Euphorbia granulate Euohorbiaceae Prostrate spurge, lawn Herb weed 78 Euphorbia helioscopia L. Euphorbiaceae Chatri dodak, gandi buti, Herb zahar buti (Sun spurge) 79 Euphorbia heterophylla L. Euphorbiaceae Lechosa (Wild Herb Poinsettia, Cruel Plant) 80 Euphorbia hirta L. Euphorbiaceae Dhudi (Asthma weed, Herb spurge)

81 Euphorbia humifusa Euohorbiaceae Light spreading Herb

Serial Species Botanical Name Family Name local Name (English Growth habit No name) 82 Euphorbia prostrate Euphorbiaceae Prostrate sandman Herb 83 Fagonia indica Zygophyllaceae Dhamasa, Dhaman, Herb (woody Thand (Fagonia) stem) 84 Ferula assafoetida Apiaceae Landes, Hing, (Devils Tree dung) 85 Ficus benghalensis L. Moraceae Banyan, bar, bargad Tree 86 Ficus carica Moraceae Injir, kimri (wild fig) Tree 87 Ficus hispida Moraceae Injire dashti (wild fig) Tree 88 Ficus johannis Moraceae Lobed fig Tree 89 Ficus palmata Moraceae Phagwara, patguleri Tree 90 Ficus variegata Moraceae Wild fig, green fruited Tree fig 91 Foeniculum vulgare Apiaceae Kaga Herb 92 Fragaria orientalis Rosaceae Zmake totan (Wild Herb strawberry) 93 Fumaria indica Fumariaceae Pit-papra, berg-e-shahtra Herb 94 Fumaria officinalis Fumariaceae Papra, shahtra Herb 95 Gallium aparine L. Rubiaceae Sticky villy, goose grass, Herb cleevers climbing 96 Grewia optiva Malvaceae Bihul, bhimal (wild Tree falsa) 97 Hedera nepalensis Araliaceae Perennial iv plant, Herb himalayan iv 98 Imperata cylindrical L. Poaceae Blady grass, sword grass, Herb jap grass perennial

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99 Indigofera spicata Fabaceae Chelo, Herb 100 Ipomoea carnea Convolvulaceae Pink bush Shrub 101 Iris ensata Iridaceae Sword leaved, purple Herb flowered 102 Justica peploides Acanthaceae Dianthera (Water Herb willow) 103 Koeleria makrantha Poaceae Hair grass crested Herb (Ledeb) coelaria perennial 104 Lagerstroemia speciosa Lythraceae Banaba Tree 105 Lamium amplexicaule L. Lamiaceae Henbit (Dead nettle) Herb 106 Lantana camara L. Verbenaceae Panchphuli Shrub 107 Lathyrus aphaca L. Papilionaceae Sweet pea Herb trailing 108 Lepidium pinnatifidum L. Brasicaceae Lipidium Herb 109 Lycium barbarum Solanaceae Thornless berberes, Shrub hamalayan goji. 110 Mallotus philippensis Euphorbiaceae Kambela ( red kamala) Tree 111 Malva parviflora Malvaceae Sonchal (Least mallow) Herb 112 Malvastrum Malvaceae Sticky mallow Herb coromendilianum

Serial Species Botanical Name Family Name local Name (English Growth habit No name) 113 Malvestrum Malvaceae False mallow, broom Herb coromandelianum weed perennial 114 Matricaria auria Asteraceae Golden may weed Herb 115 Matricaria chamomile Asteraceae Babona Herb 116 Medicago denticulata Fabaceae Shpeshtaray (Wild Herb clover) 117 Medicago polymorpha L. Papilionaceae Sijee, pishtaray (wild Herb clover) 118 Melia azaderachta L. Meliaceae Bakayen, bikana, darech Tree (Persian lilac)

119 Melilotus indica Papilionaceae Ran methi, sinjee Herb 120 Menthe arvensis L. Lamiaceae podina (Pepper mint) Herb 121 Menthe longifolia Lamiaceae Chitta podina (Lamb Herb mint) 122 Menthe royleana Lamiaceae Velanay (wild mint) Herb 123 Mirabilis jalapa L. Nyctangiaceae Gule abbasi (4 o clock Herb flower)

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124 Monotheca buxifolia Sapotaceae But berries, Gurgura, Shrub thorny gargol 125 Morus alba L. Moraceae Chita toot (White Tree mulberry) 126 Morus nigra L. Moraceae Shahtoot, kala toot Tree (Black mulberry) 127 Myrsine Africana Myrsinaceae Purple beryy plant, short shrub thakisa 128 Narcissus tazitta L. Amaryrillidaceae Gul e nargas Herb 129 Nasturtium officinale Brasicaceae Tarmira (Water cress) Herb aquatic 130 Nerium oleander Apocynaceae Kaner (oleander) Shrub 131 Neslia apiculata Brasicaceae Neslia Herb 132 Olea ferruginea Oleaceae Khona, kao, jangali Tree zaitoon (Wild Olive) 133 Otostegia limbata Lamiaceae Spiny plant, Shrub 134 Oxalis corniculata L. Oxilidaceae Khatti biti, tharoka Herb creeping 135 Oxalis pescaprae L. Oxalidaceae Khatmit (Yellow sorrel) Herb 136 Papaver hybridum L. Papaveraceae Prickly headed poppy Herb 137 Papaver rhoeas L. Papaveraceae Gule lala Herb 138 Parthenium hysterophorus Asteraceae False rag weed, bitter Herb woody L. weed, carrot grass type 139 Phalaris aquatica Poaceae Ghuand wagay Herb 140 Pinus roxbergi Pewach, Peech Tree 141 Pistia stratiotus L. Araceae Water cabbage Herb-aquatic 142 Plantago lanceolata L. Plantaginaceae Ispaghol (Ribwort Herb plantain)

Serial Species Botanical Name Family Name local Name (English Growth habit No name) 143 Plantago major Plantaginaceae Gule ispagol (Greater Herb plantain)

144 Plantago ovata Plantaginaceae Isbaghol Herb 145 poa annua L. Poaceae Meadow grass, goose Herb annual grass, six weeks grass 146 Polygomnum plebejum Polygonaceae Macheche (Knot weed) Herb prostrate

147 Polypogan fugax Poaceae Rabit foot grass Herb annual 148 Portulaca oleracea Portulacaceae Warkharay Herb

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149 Prunus serotina Solanaceae Wild cherry, mountain Tree black cherry 150 Pteridium equilinium Dennstaedtiaceae Kuanjay Herb

151 Punica granatum Lythraceae Ananguray (wild Shrub pomegranate) 152 Randia Formosa Rubiaceae Blackberry jam fruit, wild Shrub pomegranate 153 Ranunculus arvensis L. Ranunculaceae Chambul (Devils claw) Herb 154 Ranunculus muricatus Ranunculaceae Spiny fruited butter cup, Herb cow foot 155 Ranunculus repens Renunculaceae Creeping butter cup, Herb Like parthinium 156 Recinus communis L. Euphorbiaceae Arand (Wild Jatropa, Shrub Castor Bean)

157 Robinia pseudo-acacia Papilionaceae Vilayati kikar (Black Tree locust) 158 Rubus fructicosus Rosaceae Gurguray, (Blackberry) Shrub 159 Rumax dentatus Polygonaceae Shalkhay, Ombavati Herb (Dock sorrel)

160 Rumax histatus Polygonaceae Gato Tharokay, Shrub 161 Rumax longifolius Polygonaceae Shalkhay Herb 162 Rumax vasicaricus Polygonaceae Shalkhay Herb 163 Saccharum spontaneum Poaceae Qalam butay Shrub 164 Scandix pectin-veneris L. Apiaceae Billi Puncha (Venus Herb Comb) 165 Silene conoidea L. Caryophyllaceae Takla, Qardi (Sand Herb catchfly) 166 Silybum marianum Asteraceae Kkandiari, Aunt Katara Herb spiny (Milk Thistle) 167 Sinapis arvensis Brasicaceae Jangali sarson, charlock Herb (Wild Mustard) 168 Sisymbrium irio L. Brasicaceae Khubkalan, khakasi Herb (London Rocket) 169 Sitaria pumila Poaceae Band Kangni (Bottle Herb annual

Serial Species Botanical Name Family Name local Name (English Growth habit No name) Grass)

170 Skimmia laureola Rutaceae Nazar panra Shrub

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171 Solanum nigram L. Solanaceae Kachmach, Maku Herb 172 Solanum pseudo-capsicum Solanaceae Jerusalem Cherry Shrub 173 Solanum surrattense Solanaceae Kindiari, Mohkri Herb 174 Solanum xanthocarpum Solanaceae Kamtakari Herb 175 sonchus asper L. Asteraceae Sontati (Spiny Herb sowthistle) 176 Sorghum halepense L. Poaceae lawar, baru (Cuba grass) Herb perennial 177 Stellaria media Caryophyllaceae Olalai (Winter weed, Herb chick weed) 178 Tagetes minuta Asteraceae Cone marigold, black Herb mint 179 Taraxicum officinale Asteraceae Dandelion, hand dubal Herb 180 Tecomella undulate Bibnoniaceae Rohida, darkht anare Tree shaitan (desert teak) 181 Themeda triandra Poaceae Red grass, red oat grass Herb 182 Thlaspi arvense L. Brasicaceae Penny cress Herb 183 Tinospora cordifolia Minispremaceae Gilo Vine 184 Trianthema Aizoaceae Horse purslane Herb portulacastrum L. 185 Tribulus terrestris L. Zygophyllaceae Tirkundi, bhakhra Herb prostrate

186 Trifolium repens L. Papilionaceae Shaftal Herb 187 Urtica dioica Urticaceae Jalbang, bichu buti pahari Herb (stinging plant)

188 Urtica dubia Urticaceae Bichu buti dashti, Herb jalbang (Roman nettle) 189 Urtica pilulifera Urticaceae Bechu buti, jalbang Herb (Roman nettle) 190 Vaccaria hispanica Caryophyllaceae China rockle, cow herb Herb 191 Verbascum thapsus Scrophulariaceae Gidar tambako Herb (Common mullein) 192 Verbena officinalis L. Verbenaceae Karenta, pamukh Herb 193 Veronica anagallisaquatica Scruphulariaceae Blue water speed well Herb

194 Veronica persica Scruphulariaceae Persian soeed well Herb

195 Vicia sativa L. Papilionaceae Bakla Herb 196 Vitex negundo L. Verbenaceae Sanbhaalo, nirgunda Shrub 197 Withania somnifera Solanaceae Asgand Shrub

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198 Xanthium strumarium Asteraceae Katula (Cocklebur) Herb 199 Zanthoxylum armatum Rutaceae Dambara, thimer Shrub Serial Species Botanical Name Family Name local Name (English Growth habit No name) 200 Zizyphus mauritiana Rhamnaceae Indian jujube, ber, Shrub 201 Zizyphus nummularia Rhamnaceae Wild ber (Indian jujube) Shrub 202 Zizyphus vulgaris Rhamnaceae Makhranay, wild ber Tree (hilly jujube) Appendix 1.2 List of Medicinally Important Plant Species Identified at Khanpur Valley. Scientific Family local Name Gro Part Therapeutic value/Medicinal importance S. name (English wth used N name) habit

1 Acacia Memosacea Phulai, palosa Tree Pods, Its gum is restorative. Leaves are used in backache of modesta e (Acacia) leaves women after delivery and chronic stomach problems.

2 Adhatoda Acanthacea Baikar, Shru Root, The plant is used in bronchitis, heart troubles, asthma, vasika e waikar b leavesfl fever, tumours, diseases of the mouth, useful in straguary (Malaber nut owers and leucorrhoea. tree) 3 Ajuga Labiatae Karvi buti / Herb whole A decoction of the leaves is useful in diabetes, bractiosa Ratti Buti plant hypertension, fever, blood purification, malaria and (Ajuga) stomach pain. The juice of the root is used in the treatment of diarrhoea and dysentery. 4 Amaranthus Amarantha Chulai, Herb whole The plant is cooling, laxative, diuretic, stomachic, viridis L. ceae putkanda, plant antipyretic, blood diseases, leprosy, bronchitis, rat-bite shalkhay and piles. The root lessens the menstrual flow. 5 Artimisia Asteraceae Tharkha, Herb whole Infusion of the plant is used as purgative and as cure for bivifolia (composita Choree saroch plant earache. Smoke is known to be good for burns. e) 6 Asparagus Liliaceae Shahghandal Herb whole The tuberous roots are used as demulcent, toxic and in gracilis (Halyun) plant diarrhoea, dysentery and general debility. 7 Avena sativa Poaceae Jangali jao, Herb whole The seeds are nerve tonic, stimulant, antispasmodic and L. Jai (Wild oat) plant as tonic in spermatorrhoea and insomnia. It useful for heart muscles and for bladder and urethras. 8 Bauhinia Fabaceae Kachnar Tree Flowers The bark of plant is sweet, appetizing, cooling, astringent variegata and to the bowels, used in piles, cures biliousness, cough, leaves asthma, diseases of the blood, ulcers, vaginal discharges, cough, eye diseases and liver complaints. 9 Berberis Berberidac Kwaray, Shru Root The plant is used in cure of liver disorders, abdominal lycium eae simlo, b bark, disorders, skin diseases, cough and ophthalmic. Plant kashmal, leavesfl bears hypoglycaemic, ant carcinogenic and antipyretic ziar largay ower&f properties. The fruits are rich source of vitamins, ruit minerals, antioxidants, anthocyanin etc and used in juices and jams. 10 Boerhavia Nyctagina Chalaray Herb whole The leaves have a sharp taste and are appetizer and used diffusa ceae runner type plant in eye wounds and pain of the joints. The seeds are toxic, (itset) and root expectorant, carminative and useful in muscular pain, lumbago scabies, hasten delivery. The root is well known for its diuretic properties. It is also very good expectorant.

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11 Calatropis Asclepidac Spalwaka, Shru Leaves The hot leaves are applied to abdomen to cure the pain procera eae Mundar b inside. The plant is very bitter, heating, laxative, (Akk) anthelmintic, relieves stranguary, cures ulcers, the ash act as an expectorant. The plant cures leprosy, ulcers, tumours, piles, disease of the spleen, the liver and the abdomen.

Scientific Family local Name Gro Part Therapeutic value/Medicinal importance S. name (English wth used N name) habit

12 Cannabis Cannabace Bhang Herb Young The plant is narcotic drug and is used for malaria, black sativa L. ae Leaves water fever, blood poisoning, anthrax and dysentery. The flowers juice of the leaves is applied on the hand to remove dandruff and vermin. 13 Carthamus Asteraceae Jewled Herb Seed Oil from its seed is used for dressing of ulcers. oxycantha distaff thistle spiny leave d 14 Cassia absus Caesalpinia Chaksu, Herb Seed Its seed is bitter, blood-purifier, astringent, stimulant, L. ceae jasmeejaz shrub diuretic and is used for leucoderma, ringworm, venereal type ulcers and other skin diseases. 15 Cassia fistula Fabaceae Amaltas Tree Leaves, The root is useful in skin diseases, leprosy, tuberculosis, flowers and glands. The leaves are laxative, antipyretic, anti root ulcers and cure ringworms. The flowers are cooling, astringent, cure biliousness. 16 Celtis Canabbace Wild Falsa Tree Leaves, Its leaves and fruit are used in the treatment of australis ae (Hack berry) fruits amenorrhoea, diarrhoea, dysentery and peptic ulcers. 17 Chenopodiu Chenopodi Bathu (Wild Herb whole The plant improves the appetite, oleaginous, m album L. aceae spinach, plant anthelmintic, diuretic, aphrodisiac, tonic, eye diseases, lambs quarter) throat troubles, useful in biliousness, abdominal pain, diseases of the blood, heart and the spleen. 18 Conyza Asteraceae Paleet Herb whole The herb is used in dysentery, diarrhoea and bonariensis (Asthma plant haemorrhage. weed) 19 Datura alba Solanaceae Mangaz Herb whole Its flowers and seed are used useful in treatment of skin butay, (devils plant eruptions, colds, and nervous disorders. trumpit and metel)

20 Datura Solanaceae Kala datura Herb whole The juice of the leaves is a good substitute for the stramonium (jimson plant BellaDona. An extract made from the seeds is a good L. weed) mydriatic, and the leaves are used as emollient and supportive. 21 Euphorbia Euphorbiac chatri dodak, Herb whole The leaves and stems are febrifuge and vermifuge. The helioscopia L. eae gandi buti, plant root is anthelmintic. The plant has anticancer properties. zahar buti The milky sap is applied externally to skin eruptions. (Sun spurge,) The oil from the seeds has purgative properties.

22 Euphorbia Euphorbiac lechosa (Wild Herb whole leaves are taken as a purgative and laxative to treat heterophylla eae poinsettia, plant stomach-ache & constipation, and to expel intestinal L. cruel plant) worms. A leaf decoction is used as a wash to treat skin problems like fungal diseases and abscesses.

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23 Euphorbia Euphorbiac Dhudi, Herb whole The juice of the plant is given in dysentery, fever, colic, hirta L. eae doddak plant and the milk is applied to dysentery warts. A decoction (Asthma is used in asthma and chronic bronchial infection. In weed, spurge) Indonesia it is used in dengue fever The plant is broadly used in anti worms, bowel complaints and cough. 24 Fagonia Zygophylla Dhamasa, Herb whole The plant cures fever, dysentery, urinary discharges, indica ceae Dhaman, (woo plant erysipelas, typhoid, alexipharmic, reduces tumours, and Thand(Fago dy purifies the blood. The plant is acrid and bitter, cooling, nia) stem) emmenagogue, good for liver troubles, in chronic bronchitis, asthma, spitting of blood, ophthalmic and toothache. The bark is used in scabies. 25 Ferula Apiaceae Landes, Tree Latex Pulp of pod is used in childcare. Used in condiments; its assafoetida Hing, (Devils (Pulp of poultice for wound cure; carminative in hysteria and dung) Pods) epilepsy.

Scientific Family local Name Gro Part Therapeutic value/Medicinal importance S. name (English wth used N name) habit

26 Ficus hispida Moraceae Injire dashti Tree Fruit and Expectorant; used to remove Kidney stone; to remove (wild fig) leaves, obstructions of the liver and Spleen latex

27 Ficus Moraceae Wild fig, Tree Fruit and Latex is traditionally used in ulcers and gout. Leaves are variegata green fruited leaves, utilized in cancer, tumours and dermatitis. fig latex

28 Foeniculum Apiaceae Kaga Herb whole Carminative; aromatic, stomach diseases, its decoction is vulgare plant good for eyesight. 29 Fragaria Rosaceae Zmakay Herb whole Fruits are consumed fresh or used in the preparation of orientalis totan (wild plant jams and confectionary products. It is used as digestive strawberry) tonic and bones strengthening 30 Fumaria Fumariacea Papra, shahtra Herb whole It is considered as laxative, diuretic, alterative, toxic, officinalis e plant diaphoretic and febrifuge. Used in Jaundice, as coolant and blood purifier 31 Grewia Malvaceae Bihul, Tree whole The ripe fruit is useful for cooling, digestible, toxic; optivia bhimal, plant aphrodisiac allays thirst and burning sensation, cures (wild falsa) inflammation, heart and blood disorders, fever and consumption. It is also good for troubles of the throat, helps removal of dead foetus. The bark cures biliousness, removes troubles and burning in vagina. 32 Lycium Solanaceae Thornless Shru Red It is used to nourish kidneys and cure other deficiency barbarum berberes b fruits syndromes. As a traditional use, lycium fruit is best known as an aid to vision, anti aging agent and a remedy for diabetes. 33 Mallotus Euphorbiac Kambela Tree Leaves Leaves are bitter, cooling and appetizer. Fruit is heating, philippensis eae (red kamala) flowers Purgative, anthelmintic, detergent, maturant, and carminative, alexiteric and useful in cure of bronchitis, fruits abdominal diseases and against spleen enlargement. 34 Malvastrum Malvaceae Sticky Herb whole It is used in traditional medicine as an antiinflammatory, coromendeli mallow plant analgesic, antidysenteric. It is useful in diabetes. anum 35 Matricaria Asteraceae Babona Herb whole Chamomile plant is locally used as analgesic, chamomilla plant antiinflammatory, antispasmodic, anodyne, carminative, diaphoretic, laxative, stomachic, sedative and as tonic. It bears calming and soothing characteristics.

312

36 Medicago Fabaceae Shpeshtaray Herb whole It is considered as vitamin D supplement and used in the denticulata (Wild clover) plant preparation of drugs for the treatment of immunological disorders, microbial infection and cancer. 37 Melia Meliaceae bakayen, Tree Root The root is useful in vomiting, blood impurities, heart azaderachta bikana, and seed diseases, ulcers, headache, fever and lung complaints. L. darech The oil from the seeds is a brain tonic, good for earache (Persian lilac) and liver disorders.

38 Melilotus Papilionace Ran methi, Herb whole Plant is considered astringent and emollient. It is used as indica ae sinjee plant poultice or plaster for swellings.

39 Mentha Lamiaceae chitta podina Herb whole It is carminative, a cooling medicine. It is used for longifolia (Lamb mint) plant flavouring dishes. It is also used for preparation of local chatni.

40 Mentha Lamiaceae Velanay Herb whole It is famous for ant diarrhoeal and spasmolytic effects. royleana (wild mint) plant Highly useful in stomach problems. 41 Morus nigra Moraceae shahtoot, kala Tree Leaves The fruit is cooling and laxative. The bark is supposed to L. toot (Black and be vermifuge and purgative. The root is considered as mulberry) fruits anthelmintic and astringent.

42 Narcissus Amaryrillid Gul e nargas Herb whole Its oil is used in healing treatments especially in cancer. tazetta L. aceae plant Consumption in larger amount is poisonous.

Scientific Family local Name Gro Part Therapeutic value/Medicinal importance S. name (English wth used N name) habit

43 Nasturtium Brasicaceae Tarmira Herb whole Watercress is locally used as an anti scorbutic, officinale (Water aquat plant depurative, diuretic, expectorant, purgative, cress) ic hypoglycaemic, stimulant, tonic and stomachic. Its leaves juice can be used to cleanse the face from blotches, spots and blemishes. 44 Nirium Apocynace Kaner, Shru Leaves, Its leaves are used in piles diseases. The root is bitter, oleander ae b root good tonic for chronic pain in the abdomen and pain in flowers the joints. A decoction of leaves is recommended to and reduce swellings and oil prepared from the root bark is fruits used for skin diseases and in leprosy. 45 Olea Oleaceae Khona, kao, Tree Leaves The root is a remedy against scorpion sting. Its ash is ferrugineae jangali and useful in rheumatism and diseases of the brain. The zaitoon fruits fruit is appetizer, useful in biliousness, liver (wild olive) complaints, scabies and toothache. Its oil is useful in liver troubles and pain in joints. 46 Oxalis Oxilidaceae Khatti biti, Herb whole The herb is a good appetizer, useful in dysentery and corniculata L. tharoka plant diarrhoea. It also cures skin diseases and fevers. (yellow oxalis) 47 Papaver Papaverace Prickly Herb whole The whole plant is nutritional as well as medicinal. The hybridum L. ae headea plant leaves are rich in vitamins and minerals and also used poppy in digestion. It is a stimulant and expectorant in the treatment of bronchitis. 48 Plantago major Plantaginac Greater Herb whole Leaves are used in chronic dysentery; diarrhoea and eae plantain, plant constipation; grinded seed in vinegar with castor oil is Gule ispagol useful for headache.

313

49 Portulaca Portulacac Warkharay, Herb whole The leaves are sour, bitter, Salish, recommended in oleraceae eae kulfa plant diseases of kidney and the spleen, in stomatitis of (Purslane) children, piles, scabies and burns. The herb is chiefly valued as a refrigerant and alterative pot herb. Leaves are also applied to swellings. 50 Prunus serotina Solanaceae Wild cherry Tree Leaves It is useful for irregular menstruation and debility and following miscarriage. fruits 51 Punica Lythraceae Ananguray, Shru Fruit and Juice is used as tonic in fever. Dried seed is used for granatum anar (Wild b Exocar adding taste. Bark of the root and wood is used for Pomegranate p of killing tapeworms. Also used in diarrhoea and ) fruit dysentery and dyes preparation.

52 Randia Rubiaceae Blackberry Shru Fruit The fruit is taken against dysentery. Leaves are used in Formosa jam fruit, b baths to cure infected sores. wild pomegranate 53 Recinus Euphorbiac Arand (Wild Shru Leaves The decoctions of the leaves are used for stomachache communis L. eae Jatropa, b and and for burns. The seeds oil is useful in leprosy. The Castor bean) fruits root bark is purgative, good in skin diseases. Seed is used in lumbago, boils, piles, ringworm, paralysis, inflammations, asthma, rheumatism, and amenorrhea. 54 Rubus Rosaceae Gurguray Shru Leaves The plant is effective against dysentery, diarrhoea, fructicosus b and haemorrhoids and cystitis. Its leaves are helpful in fruits treatments of wounds, sores, scratches, gum inflammations, ulcers and sore throat. 55 Rumax Polygonace Salumi, Gato Shru Leaves The juice of the plant is considered cooling, aperients histatus ae Tharokay, b and root and to a certain extent diuretic.

56 Rumax Polygonace Chora chitra, Herb whole The leaves and root are used as an astringent longifolius ae Shalkhay plant application in coetaneous disorders. Scientific Family local Name Gro Part Therapeutic value/Medicinal importance S. name (English wth used N name) habit

57 Silybum Asteraceae kandiari, Herb whole It is normally used to stimulate lactation in nursing marianum Aunt katara spiny plant mothers and in the treatments of liver and gallbladder (Milk problems. Thistle) 58 Sinapis arvensis Brasicaceae Jangali Herb whole The plant is used in Bach flower remedies (black sarson, plant depression). Its oil is used against headache and in charlock making soap. (Wild mustard) 59 Skimmia Rutaceae Nazar panra Shru Leaves An essential oil in the leaves is utilized in scenting laureola b and soap. The dried leaves are used as an incense. The fresh shoots leaves are used to prepare garlands for weddings. 60 Solanum Solanaceae Kachmach, Herb whole The berries improve appetite, taste and useful in nigram L. Maku plant diseases of heart. The root bark is laxative, used in (Nightshade) especial diseases of ears, eyes and nose. ly

the berries.

314

61 Solanum Solanaceae Jangli Shru root The root is said to be used as a medicine for horses. It pseudocapsicum bangan b and is used as a remedy for toothache and sore throat. It is (Jerusalem fruits also used for pleurisy and pneumonia. cherry) 62 Solanum Solanaceae Kindiari, Herb whole Root is used in cough, asthma, and catarrh. Fruit and root surrattense mohkri plant are both demulent and expectorant. 63 Stellaria media Caryophyll Olalai Herb whole Used as apitizer and as vegetable aceae (Chick plant weed) 64 Tagetes minuta Asteraceae Wild Herb whole It is used as a remedy for the common cold and stomach marigold plant upset, diarrhoea, and liver diseases. Medicinal tea is prepared from this plant. 65 Urtica pilulifera Urticaceae Bechu buti, Herb whole Used as anti-diabetic, anti-oxidant Jalbang plant and antiinflammatory agent. (Roman nettle) 66 Verbascum Scrophulari Gidar Herb whole Both leaves and flowers are useful in case of pulmonary thapsus aceae tambako plant diseases, bleeding of the lungs, cough and bowels. (Common mullein) 67 Vitex negundo Verbenacea Sanbhaalo, Shru Leaves Its leaf extract possess antioxidant andanti indlamatory L. e Nirgunda b and properties. It is also used in herbal bath soap flowers preparation. 68 Withania Solanaceae Asgand Shru Fruits The leaves are applied to tumors and to tuberculosis somnifera (winter b glands. The tuber is useful in inflammations, cherry) bronchitis, asthma and ulcers. Xanthium Asteraceae Katula Herb Fruit The fruit is considered cooling and effective in the small 69 strumarium (Cocklebur) and seed pox. It is also useful in urinary diseases.

70 Zanthoxylum Rutaceae Dambara, Shru Leaves, It is used in the treatment of toothache, common cold, armatum Thimer b fruit and cough, and fever. Young shoots of thimer are used as seed toothbrushes. Local Chatni and soup are prepared from its fruits. 71 Zizyphus Rhamnacea Makhranay Tree Leaves The bark cures boils, dysentery and diarrhoea. The vulgaris e (hilly jujube) and leaves are bitter, cooling, laxative, diarrhoea, fruits antipyretic, reduce obesity. The ripe fruit is tonic, removes biliousness, burning sensation, thirst, vomiting and blood diseases.

Appendix 1.3

QUESTIONNAIRE:

To determine community preferences of the people of Khanpur Valley.

Site:

315

Village

Respo ndent's

Name:

Age:

Qualification:

Profession:

Major Themes:

1. Most used /preferred local medicinal plant species. • Local name of the species:

• Uses in the Area:

• Which part is collected:

2. Most preferred health problem treated with local plants. Health problems related to:

Reproduction, skin, digestion, nutrition and tonic, respiration, blood purification, bones and joints, brain and nerves, urinary system.

3. Most preferred traditional use / recipe commonly used Tea, paste, fresh, cream, tincture, powder, decoction, juice, cooked.

4. Most preferred plant part used in traditional healthcare.

Flower, shoot, root, rhizome, leaves, fruit, whole plant, bark, seed

316

Appendix 3.1 ANOVA for Proximate Analysis of Adhatoda vasica.

Source DF Moisture DryMatter CrudeProteins CrudeFats Ash CrudeFibers E. Oil NFES NFEE Sites 3 19.36 19.36* 5.52* 1.13* 11.39* 9.04* 1.85* 46.18* 138.67* Reps w/n Sites 8 3.43 3.43 0.69 0.18 0.11 0.10 0.02 1.74 19.33

Seasons 1 127.19* 127.19 * 10.71 * 13.71* 44.34* 10.41* 0.30ns 108.29* 1217.78*

Sites*Seasons 3 3.52ns 3.52ns 2.63 * 1.26 ns 2.13* 0.90* 0.29* 15.33* 142.15* Error 8 0.87 0.87 0.38 0.32 0.28 0.06 0.06 2.13 29.86 *: significant and ns: non-significant, at 5% level of probability.

Appendix 3.2 ANOVA for Elemental Analysis of Adhatoda vasica. Source DF Sodium Potassium Calcium Phosphorus Magnesium Copper Iron Manganese Zinc

Sites 3 0.08ns 1387.22ns 4443.9* 2091.06* 340.9ns 0.02* 1553.3* 23.20* 7.45* Reps w/n Sites 8 0.06 1524.69 280.6 16.81 571.2 0.01 323.0 0.94 0.37

Seasons 1 0.78* 115.28 ns 11.68.2 * 1350.00* 36902.9* 0.61* 30673.5* 356.51* 21.28*

Sites*Seasons 3 0.04ns 4885.89* 25.6 ns 2481.73 * 103.4ns 0.02* 380.3ns 2.23ns 0.99* Error 8 0.02 336.02 179.6 64.59 166.9 0.001 308.2 0.67 0.18 *: significant and ns: non-significant, at 5% level of probability.

Appendix 3.3 ANOVA for Proximate Analysis of Calatropis procera.

Source DF Moisture DryMatter CrudeProteins CrudeFats Ash CrudeFibers E. Oil NFES NFEE Sites 3 10.94ns 10.94ns 1.39* 2.83ns 12.48* 1.78ns 0.61* 15.33* 201.94ns Reps w/n Sites 8 4.61 4.61 0.27 1.13 0.94 0.66 0.07 2.80 76.96

Seasons 1 64.48* 64.48* 20.57* 12.24* 73.95* 42.21* 0.05ns 102.755* 1477.97*

Sites*Seasons 3 1.03ns 1.03ns 1.89* 7.92 * 2.23ns 2.96* 0.14ns 25.48* 377.43* Error 8 0.77 0.77 0.10 0.31 0.81 0.71 0.04 2.62 20.86 *: significant and ns: non-significant, at 5% level of probability. Appendix 3.4 ANOVA for Elemental Analysis of Calatropis procera. Source DF Sodium Potassium Calcium Phosphorus Magnesium Copper Iron Manganese Zinc

Sites 3 12.11* 42485.2* 1784.3 * 358.31* 2378.77* 0.01* 1557.1* 3.62ns 1.97* Reps w/n Sites 8 0.54 578.3 61.7 27.91 509.70 0.001 219.4 2.00 0.33

Seasons 1 18.32* 11550.1* 2240.7 * 1303.90* 2014.83ns 0.02* 14885.2* 100.45* 1.93ns

Sites*Seasons 3 0.21ns 57376.4* 14024.4* 60.83ns* 4127.60* 0.02* 2689.3* 41.93* 6.22* Error 8 0.18 375.3 190.4 19.19 451.97 0.002 405.5 1.07 1.28 *: significant and ns: non-significant, at 5% level of probability. Appendix 3.5 ANOVA for Proximate Analysis of Recinus communis.

317

Source DF Moisture DryMatter CrudeProteins CrudeFats Ash CrudeFibers E. Oil NFES NFEE Sites 3 12.53ns 11.78 ns 1.73ns 1.58ns 9.77* 10.44* 3.92* 19.52* 44.39ns Reps w/n Sites 8 12.27 12.41 0.51 0.88 0.32 0.50 0.14 1.10 34.38

Seasons 1 167.80* 171.47* 22.35* 19.66* 37.42* 35.79* 2.12* 86.34* 2318.9*

Sites*Seasons 3 1.50ns 1.79 ns 2.66* 0.48 * 1.65ns 5.21* 0.56* 0.69ns 21.16* Error 8 4.64 4.33 0.17 0.08 1.41 0.34 0.05 1.52 4.04 *: significant and ns: non-significant, at 5% level of probability. Appendix 3.6 ANOVA for Elemental Analysis of Recinus communis. Source DF Sodium Potassium Calcium Phosphorus Magnesium Copper Iron Manganese Zinc

Sites 3 3.41* 2653.22 * 3057.30* 1218.77ns 3923.86* 0.04* 7204.4* 8.90* 12.20* Reps w/n Sites 8 0.34 162.33 517.33 493.99 45.81 0.002 532.9 0.72 0.35

Seasons 1 8.33* 1093.50* 2803.68* 7898.88* 2412.02ns 0.39* 16140.9* 82.51* 47.88*

Sites*Seasons 3 4.62* 353.44ns 476.00ns 265.47ns 2520.17ns 0.003ns 2360.5* 7.08* 5.34* Error 8 0.05 204.33 478.52 287.30 1134.94 0.004 258.3 0.74 0.35 *: significant and ns: non-significant, at 5% level of probability.

Appendix 3.7 ANOVA for Proximate Analysis of Ajuga bracteosa.

Source DF Moisture DryMatter CrudeProteins CrudeFats Ash CrudeFibers E. Oil NFES NFEE Sites 3 3.80ns 3.80ns 0.59ns 0.46ns 40.10* 1.75* 2.30* 52.39* 115.62* Reps w/n Sites 8 2.01 2.01 0.20 0.38 1.65 0.42 0.16 3.12 23.00

Seasons 1 36.85* 36.85 * 33.91* 19.95* 308.02* 61.28* 0.16* 400.50* 1775.66*

Sites*Seasons 3 3.53ns 0.53ns 3.72* 1.97 * 19.22* 0.47ns 0.52ns 43.10* 149.61* Error 8 0.77 0.77 0.13 0.31 0.39 0.17 0.08 0.71 18.10 *: significant and ns: non-significant, at 5% level of probability.

Appendix 3.8 ANOVA for Elemental Analysis of Ajuga bracteosa. Source DF Sodium Potassium Calcium Phosphorus Magnesium Copper Iron Manganese Zinc

Sites 3 7.80* 25495* 5983.3* 14273.3* 1197.0* 0.04* 582.9ns 37.29* 1.82* Reps w/n Sites 8 0.19 3546 915.7 83.5 124.3 0.01 353.6 0.10 0.27

Seasons 1 15.60* 585781* 26920.6* 20609.6* 47918.4* 0.78* 22149.5* 36.01* 29.04*

Sites*Seasons 3 3.96* 12730* 1589.5* 154.6ns* 1729.7* 0.01* 678.8ns 0.35ns 2.50* Error 8 0.11 1819 248.7 60.3 125.2 0.003 178.9 0.31 0.15 *: significant and ns: non-significant, at 5% level of probability. Appendix 3.9 ANOVA for Proximate Analysis of Euphorbia hirta.

Source DF Moisture DryMatter CrudeProteins CrudeFats Ash CrudeFibers E. Oil NFES NFEE Sites 3 20.55* 20.55* 21.64 * 7.85* 13.82* 8.63* 0.02ns 89.73* 1346.53*

318

Reps w/n Sites 8 1.75 1.75 0.35 0.25 0.81 0.98 0.02 4.14 15.85

Seasons 1 21.93* 21.93* 22.37* 8.19* 12.43* 31.37* 0.37* 279.48* 734.32*

Sites*Seasons 3 1.05ns 1.05ns 2.21* 0.55 * 1.78* 5.74* 0.05* 21.49* 88.21* Error 8 1.11 1.11 0.23 0.12 0.24 0.37 0.01 1.17 12.80 *: significant and ns: non-significant, at 5% level of probability. Appendix 3.10 ANOVA for Elemental Analysis of Euphorbia hirta. Source DF Sodium Potassium Calcium Phosphorus Magnesium Copper Iron Manganese Zinc

Sites 3 1.75* 200.52 * 7552.83* 1197.3* 3401.2* 0.73* 9749.8* 68.72* 1.40ns Reps w/n Sites 8 0.02 36.78 52.79 46.3 702.6 0.09 433.2 3.58 0.80

Seasons 1 0.41* 93.77 ns 3967.82* 17800.8* 23702.0* 5.06* 22625.9* 489.52* 56.83*

Sites*Seasons 3 0.03ns 43.66ns 6065.23* 356.6* 1222.6ns 0.08ns 551.6ns 9.22ns 0.46ns Error 8 0.07 35.84 28.42 75.6 550.7 0.05 327.8 3.43 0.36 *: significant and ns: non-significant at 5% level of probability. Appendix 3.11 ANOVA for Proximate Analysis of Fumaria officinalis.

Source DF Moisture DryMatter CrudeProteins CrudeFats Ash CrudeFibers E. Oil NFES NFEE Sites 3 19.38* 19.38* 0.91ns 3.54* 16.41* 4.03* 0.40* 13.68* 317.89* Reps w/n Sites 8 0.99 0.99 0.52 0.27 0.57 0.16 0.02 0.55 15.87

Seasons 1 47.38* 47.38* 15.58 * 10.01* 62.73* 24.14* 1.63* 102.38* 1102.16*

Sites*Seasons 3 1.64ns 1.64ns 1.10* 0.76 * 3.61* 2.63* 0.16* 4.43* 98.41* Error 8 0.94 0.94 0.12 0.18 0.52 0.06 0.04 0.55 15.81 *: significant and ns: non-significant, at 5% level of probability.

Appendix 3.12 ANOVA for Elemental Analysis of Fumaria officinalis. Source DF Sodium Potassium Calcium Phosphorus Magnesium Copper Iron Manganese Zinc

Sites 3 1.52* 28.6 ns 15988.0* 15288.0* 8877.71* 0.01* 7210.0* 15.17* 1.01* Reps w/n Sites 8 0.21 1356 146.5 100.1 144.24 0.002 256.0 0.33 0.13

Seasons 1 3.89* 697754 * 32465.0* 26162.4* 9452.57* 1.15* 57379.3* 127.42* 14.73*

Sites*Seasons 3 0.67ns 9250* 525.4ns 3244.6* 369.85ns 0.001ns 1700.1ns 4.69* 0.33ns Error 8 0.36 1679 239.8 123.9 76.03 0.008 731.4 0.52 0.22 *: significant and ns: non-significant, at 5% level of probability.

Appendix 4.1 ANOVA for Horticultural Attributes of Adhatoda vasica. Source DF P Height Leaves P-1 Stem Dia Leaf Wt P.Meter-2 Yield P-1 Yield Meter-2 Yield Hac-1

Sites 3 436.17ns 1699.6ns 0.02ns 0.0004ns 0.01* 55.33ns 31.20* 3120.75* Reps w/n Sites 8 151.77 633.0 0.07 0.0006 0.0002 23.05 1.08 107.77

Seasons 1 8599.52* 28126.1* 0.007 ns 0.0009ns 0.0007ns 1756.17* 42.72* 4266.13*

Sites*Seasons 3 47.44ns 190.5ns 0.008ns 0.0003ns 0.001* 4.90ns 4.88* 488.85* Error 8 253.96 353.3 0.14 0.0003 0.0002 16.44 0.64 63.72

319

*: significant and ns: non-significan,t at 5% level of probability.

Appendix 4.2 ANOVA for Horticultural Attributes of Ajuga bracteosa. Source DF P Height Leaves P-1 Stem Dia Leaf Wt P.Meter-2 Yield P-1 Yield Meter-2 Yield Hac-1

Sites 3 4.36ns 3.96ns 0.05ns 0.0004ns 0.34* 35.90* 3592.02* 5.17* Reps w/n Sites 8 3.33 3.33 0.02 0.001 0.01 1.18 117.66 0.94

Seasons 1 522.67* 146.03* 1.17* 0.005* 0.20* 41.00* 4101.37* 10.67*

Sites*Seasons 3 1.92ns 13.20ns 0.008ns 0.002ns 0.001ns 0.35ns 34.75ns 0.12ns Error 8 2.92 2.52 0.17 0.0005 0.01 1.07 107.92 0.13 *: significant and ns: non-significant, at 5% level of probability.

Appendix 4.3 ANOVA for Horticultural Attributes of Calatropis procera. Source DF P Height Leaves P-1 Stem Dia Leaf Wt P.Meter-2 Yield P-1 Yield Meter-2 Yield Hac-1

Sites 3 801.03* 1222.93 * 0.08ns 0.002* 0.07* 303.03* 30300* 99.38* Reps w/n Sites 8 146.26 80.08 0.02 0.0005 0.008 10.39 1039 8.98

Seasons 1 9859.73* 6240.38* 1.26* 0.007* 0.46* 1732.98* 173290* 414.17*

Sites*Seasons 3 392.10ns 70.82ns 0.03ns 0.0005ns 0.02ns 55.61ns 5565ns 6.70ns Error 8 152.88 36.83 0.07 0.0002 0.02 47.23 4724 6.20 *: significant and ns: non-significant, at 5% level of probability.

Appendix 4.4 ANOVA for Horticultural Attributes of Fumaria officinalis. Source DF P Height Leaves P-1 Stem Dia Leaf Wt P.Meter-2 Yield P-1 Yield Meter-2 Yield Hac-1

Sites 3 139.79* 5060* 0.006ns 9.4 E-07ns 0.31* 28.07* 2806.9* 0.92ns Reps w/n Sites 8 23.34 358 0.015 9.5 E-07* 0.03 1.46 146.2 0.38

Seasons 1 4347.04* 372006* 1.31* 5.2 E-04* 0.08ns 984.01* 98402.1* 768.29*

Sites*Seasons 3 12.19ns 3285* 0.01ns 3.9 E-06ns 0.04ns 19.57* 1957.3* 0.47ns Error 8 6.75 390 0.015 1.2 E-06 0.03 2.15 215.1 0.35 *: significant and ns: non-significant, at 5% level of probability.

Appendix 4.5 ANOVA for Horticultural Attributes of Euphorbia hirta. Source DF P Height Leaves P-1 Stem Dia Leaf Wt P.Meter-2 Yield P-1 Yield Meter-2 Yield Hac-1

320

Sites 3 100.15* 413.90 * 0.01ns 2.4E-06ns 0.13* 0.13* 13.25* 0.02ns Reps w/n Sites 8 5.02 42.40 0.007 2.7E -06 0,007 0.009 0.89 0.04

Seasons 1 3874.26* 1109.49* 0.08* 2.4E-05* 0.62 * 0.06* 6.38* 3.99*

Sites*Seasons 3 3.00ns 32.14ns 0.001ns 2.4E-06* 0.04 * 0.008ns 0.79ns 0.006ns Error 8 10.34 16.02 0.002 2.4E-07 0.001 0.005 0.46 0.01 *: significant and ns: non-significant, at 5% level of probability.

Appendix 4.6 ANOVA for Horticultural Attributes of Recinus communis. Source DF P Height Leaves P-1 Stem Dia Leaf Wt P.Meter-2 Yield P-1 Yield Meter-2 Yield Hac-1

Sites 3 10632.6* 1192.6ns 0.26ns 0.09ns 4.3E-04* 171.30* 17126.1* 5071* Reps w/n Sites 8 713.4 609.5 0.21 0.06 2.5E-05ns 5.63 562.9 657

Seasons 1 5667.2* 67204.2* 0.0004ns 0.83* 6.67E-05ns 159.19* 15917.6* 117992*

Sites*Seasons 3 280.9ns 428.2ns 0.20ns 0.05ns 3.33E-05ns 10.81ns 1082.5ns 420ns Error 8 512.9 108.9 0.20 0.03 1.67E-05ns 2.68 267.7 770 *: significant and ns: non-significant, at 5% level of probability.

•

321