Effect of Chicken Manure and Ammonium Sulphate Fertilizers on Growth and Oil Content of Lemongrass

EFFECT OF CHICKEN MANURE AND AMMONIUM SULPHATE FERTILIZERS ON GROWTH AND OIL CONTENT OF LEMONGRASS

By Amira Yagoub Elmahdi

B.Sc. (Agric.) Sudan University for Science & Technology 1998

A Dissertation submitted to the University of Khartoum in partial fulfillment of the requirements of the Degree of Master of Science in Agriculture (Horticulure)

Department of Horticulture Faculty of Agriculture University of Khartoum

March 2004

1 a

DEDICATION

I dedicate this work To my deer mother, the soul of my father, brothers & sisters And special dedication to my deer husband With love and respect…

ACKNOWLEGDMENT

I would like to express my sincere thanks and deepest appreciation to my supervisor, Prof. Gaafar M. El Hassan, Faculty of Agriculture, University of Khartoum, for his supervision, helpful guidance throughout my study. My deepest thanks are also due to Dr. Awatif Ahmed from the Medicinal and Aromatic Plants Institute; and the members of Chemistry Laboratory for their help during the oil extraction. I'm also indebted to Mr. Elrasheed Ahmed Salim for helpful and printing of the thesis. Thanks are also to Dr. Tag El Sir Ebraheem, the head of Department of Horticulture – College of Agricultural Studies –Sudan University for Science and Technology. Thanks also indebted to those who cooperated with me but not mentioned.

ABSTRACT

This study was carried out to investigate the effect of Ammonium Sulphate and organic fertilizer on herbage yield and oil content of lemon grass (Cymbopogon citratus). The experiment was conducted at the demonstration farm at Khartoum University – Faculty of Agriculture at Shambat during summer season, sowing date of he experiment was 30/3/2003. The treatments of this study were fertilization using pure 5Ton/Fed. And combined fertilizer; Chicken manure, 2.5 Ton/fed with 2 N Ammonium sulphate and control. Fertilizer treatments gave the highest growth and herbage yield but in the second harvest fresh weight in combined fertilizer treatment was better than organic fertilizer. Oil content was the same in the three treatments in the two harvests. But the oil content was decreased in the second harvest compared to the first harvest.

ﻤﻠﺨﺹ ﺍﻷﻁﺭﻭﺤﺔ

ﺃﺠﺭﻴﺕ ﻫﺫﻩ ﺍﻟﺘﺠﺭﺒﺔ ﺒﻬﺩﻑ ﺩﺭﺍﺴﺔ ﺘﺄﺜﻴﺭ ﺍﻟﺴﻤﺎﺩ ﺍﻟﻌﻀﻭﻱ ( ﺍﻟﺩﻭﺍﺠﻥ ) ﻭﻜﺒﺭﻴﺘـﺎﺕ

ﺍﻻﻤﻭﻨﻴﻭﻡ ﻋﻠﻰ ﺇﻨﺘﺎﺝ ﺍﻟﻌﺸﺏ ﻭﻨﺴﺒﺔ ﺍﻟﺯﻴﺕ ﻓﻲ ﻨﺒـﺎﺕ ﺤﺸﻴـﺸﺔ ﺍﻟﻠﻴﻤـﻭﻥ ﺒﺎﻟﺤﻘـل

ﺍﻹﻴﻀﺎﺤﻲ ﺒﻜﻠﻴﺔ ﺍﻟﺯﺭﺍﻋﺔ – ﺠﺎﻤﻌﺔ ﺍﻟﺨﺭﻁﻭ ﻡ ﺒﺸﻤﺒﺎﺕ ﻓﻲ ﻤﻭﺴﻡ ﺍﻟﺼﻴﻑ ﺒﺘـﺎﺭﻴﺦ

30/3/2003ﻡ. ﺍﺸﺘﻤﻠﺕ ﻤﻌﺎﻤﻼﺕ ﺍﻟﺘﺴﻤﻴﺩ ﺒﺈ ﻀﺎﻓﺔ 5 ﻁﻥ ﺴﻤﺎﺩ ﻋﻀﻭﻱ (ﺩﻭﺍﺠـﻥ )

ﻭﺍﻟﻤﻌﺎﻤﻠﺔ ﺍﻟﺜﺎﻨﻴﺔ 2,5 ﻁﻥ ﻟﻠﻔﺩﺍﻥ ﺴﻤﺎﺩ ﻋﻀﻭﻱ ﻤﻊ 2N ﺴﻤﺎﺩ ﻜﻴﻤﻴﺎﺌﻲ (ﻜﺒﺭﻴﺘـﺎﺕ

ﺍﻻﻤﻭﻨﻴﻭﻡ).

ﻤﻌﺎﻤﻼﺕ ﺍﻟﺴﻤﺎﺩ ﺃﻋﻁﺕ ﺃﻓﻀل ﺍﻟﻨﺘﺎﺌﺞ ﻟﻨﻤﺅ ﺍﻟﻨﺒﺎﺕ ﻭﺒﺎﻟﺘﺎﻟﻲ ﺍﻨﺘﺎﺝ ﺍﻟﻌـﺸﺏ،

ﻭﻟﻜﻥ ﻓﻲ ﺍﻟﺤﺼﺩﺓ ﺍﻟﺜﺎﻨﻴﺔ ﺍﻟﻭﺯﻥ ﺍﻟﺭﻁﺏ ﺃﻋﻁﻰ ﺃﻓﻀل ﻨﺘﻴﺠﺔ ﻓﻲ ﻤﻌﺎﻤﻠـﺔ ﺍﻟـﺴﻤﺎﺩ

ﺍﻟﻌﻀﻭﻱ ﻤﻊ ﺍﻟﻜﻴﻤﺎﺌﻲ ﻤﻘﺎﺭﻨﺔ ﺒﺎﻟﻌﻀﻭﻱ.

ﻨﺴﺒﺔ ﺍﻟﺯﻴﺕ ﻓﻲ ﺍﻟﺜﻼﺙ ﻤﻌﺎﻤﻼﺕ ﻤﺘﺴﺎﻭﻴﺔ ﻓﻲ ﻜل ﻤﻥ ﺍﻟﺤـﺼﺎﺩﻴﻥ، ﻭﻟﻜـﻥ

ﻫﻨﺎﻟﻙ ﺍﻨﺨﻔﺎﺽ ﻓﻲ ﻜﻤﻴﺔ ﺍﻟﺯﻴﺕ ﻓﻲ ﺍﻟﺤﺼﺎﺩ ﺍﻟﺜﺎﻨﻲ ﻤﻘﺎﺭﻨﺔ ﺒﺎﻟﺤﺼﺎﺩ ﺍﻻﻭل.

5 Contents Dedication………………………………………………….…………….. i Acknowledgements……………………………………………………… ii English Abstract…………………………………………………….…… iii Acknowledgements……………………………………………………… ii English Abstract………………………………………………………… iii Arabic Abstract………………………………………………………….. iv Contents..………………………………………………………...……… V List of Tables………………………………..…………...... ………...... Viii List of Figures………………………………………………………… viii List of Plates……………………………………………..…………....… ix List of Plates……………………………………………..…………....… ix List of Appendices……………………………………….……….…..…. ix 1. CHAPTER ONE (INTRODUCTION)………………………..………. 1 2. CHAPTER TWO (LITERATURE REVIEW)………………...……… 3 2.1. Plant Botany……………………………………………………...… 3 2.1.1. Botany…………………………………………………………...… 3 2.1.2. Cymbopogon spp………………………………………………….. 4 2.2. Crop establishment………………………………………………….. 6

2.3. Factors effecting oil yield and herbage production in Cymbopogon species……………………………………………………………………. 6 2.3.1. Effect of seasonal variation……………………………………….. 6 2.3. Factors affecting oil yield and herbage production in Cymbopogon species……………………………………………………………………. 6 2.3.1. Effect of seasonal variation……………………………………….. 7 2.3.1. Effect of leaf age…………………………………………………. 7 2.3.3. Effect of Nitrogen fertilizer on growth and oil content…………… 8 2. 4. Effect of Sulphur on growth and development……………………... 9

6 2.5. Effect of organic fertilizer on plant Growth………………………… 10 2.5.2. Chicken manure Fertilizer……………………………………….. 10 2.6. Volatile oil…………………………………………………………. 12 2.6.1. Extraction of volatile oils………………………………………... 14 2.6.2. Chemistry of Cymbopogon citratus oil………………...... 16 2.6.3. The physiochemical properties of lemon grass oil………………. 16 2.7. Uses of Cymbopogon citrates……………………………………… 17 3. CHAPTER THREE (MATERIALS AND METHODS)………….… 19 3.1. Location………………………………………………………….… 19 3.2. Climate……………………………………………..……………… 19 2.3. Soil……………………………………………………………...….. 19 3.4. Source of the plant……………………………………………....…. 19 3.5. Plant management……………………………………………….… 19 3.6. Experimental design……………………………………………….. 20 3.7. Data collection……….…………………………………………….... 24 3.7.1. Growth parameters…………………………………….………… 24 3.7.1.1. Plant height…………………………………………………….. 24 3.7.1.2. Root length…………………………………………………….. 24 2.7.1.3. Number of stools………………………………………………. 24 2.7.1.4. Leaf length……………………………………………………... 24 2.7.1.5. Fresh weight………………………………………………….... 24 2.7.1.6. Dry weight…………………………………………………...... 24 2.8. Determination of oil content (Oil extraction)…………………….... 28 4. CHAPTER FOUR (RESULTS)…………… ……………………..… 30 4.1. Effect of Chicken Manure and Ammonium Sulphate on lemon grass growth parameter……………………………………………………….. 30 4.1.1. Plant height……………………………………………………... 30 4.1.2. Root length………………………………………………………. 30

7 4.1.3. Fresh weight……………………………………………………... 30 4.1.4. Dry weight……………………………………………………….. 35 4.2. Effect of chicken manure and ammonium sulphate on number of stools…………………………………………………………………..... 35 4.3. Effect of chicken manure and ammonium sulphate on leaf length... 39 4.4. Effect of chicken manure and ammonium sulphate on the oil content of lemon grass………………………………………………… 40 5. CHAPER FIVE (DISCUSSION)...... 45 5.1. Effect of chicken manure and ammonium sulphate on growth……………………………………………………………...…… 45 5.1. Effect of chicken manure and ammonium sulphate on oil conten…………………………………………………………..………. 47 6. Suggestions……………………………………………….….. 50 7. References……………………………………………………..…..… 51 8. Appendices………………………………………………………….. 60

LIST OF TABLES

Table (1): Effect of chicken manure and ammonium sulphate on growth parameters……...... ……… 31 Table (2): Effect of chicken manure and ammonium sulphate on number of stools per plant…...... ………. 37 Table (3): Effect of chicken manure and ammonium sulphate on leaf length……………...... ………………… 39

LIST OF FIGURES

Figure (1): Effect of combined and organic fertilizer on height (cm)……...... …………………………plant 32 Figure (2): Effect of combined and organic fertilizer on root length…………………………………...... … 33 Figure (3): Effect of combined and chicken manure on weight (g)……… ………………………..…..…...... fresh 34 Figure (4): Effect of combined and organic fertilizer on dry weight (g)…………...... ………………….. 36

LIST OF PLATES

Plate (1): Control treatment of Lemon grass…...... 21 Plate (2): Combine Fertilizer treatment of Lemon grass plants …. 22 Plate (3): Organic fertilizer treatment of lemon grass plants...... 23 Plate (4): Lemon grass root and shoot (Control)……………...... 25 Plate (5): Lemon grass root and shoot (Combine)……...... 26 Plate (6): Lemon grass root and shoot (Organic)……….……… 27 Plate (7):Lemon grass oil from three fertilizer levels ………...... 29

LIST OF APPENDICES

Appendix (1): Soil and Chicken manure analysis……. 59 Appendix (2): Physio-chemical properties of lemon grass oil...... 60 Appendix (3): Properties of East Indian lemon grass oil……...... 61 Appendix (4):Clevenger apparatus for volatile oil distillation...... 62

12 Chapter One

Introduction

Medicinal and aromatic plants well known since ancient civilizations: Egyptian (1992 B.C.), Indian, Chinese and Japanese; and still many people continue to handle those using primitive methods (Abuzid, 1992). Local herbal medicine around the world is partly attributable to the successes obtained in the People's Republic of China; by using traditional medicine side-by-side with modern medicine, as part of comprehensive health-care program in cities and communes (Ayensu, 1978). Sudan which characterized by vast arable land, available resources and diversification of climate is regarded as one of the richest country. Sudan flora provide different source for variable natural products. Active ingredients obtained from medicinal and aromatic plants either wild or cultivated. Can be used such potential exploited to meet the demand for those natural products either locally and internationally. Medicinal and aromatic plants can be considered as one of the most important plants; especially because their export value (20 % of the total export during 1987). In addition to foreign currency, the Sudan can become self dependent in the pharmaceutical, perfumeries, confectioneries

13 and allied industries as well as being able to supply the foreign markets with finished or semi-finished products. This study was carried out on one of the medicinally important Graminae (Poaceae) plants namely Cymbopogon citratus. It is one of the most important essential oil producing grasses, grown in tropical and subtropical regions. Because of its high citral content, the oil possesses strong lemon odour hence the name lemon grass. The oil of lemon grass is widely used for scenting of soaps, as a detergent and in many kinds of technical products. Larger amount of citral, converted into ionone (a group of very important synthetic aromatics possessing a strong odour) (Guenther, 1950). In Sudan, the grass grows in the southern region, but it is used on a very limited scale for decoration purposes in front of houses and for scenting tea. The objective of this study to explore the effect of ammonium sulphate and chicken manure fertilizer on growth (parameters such as; plant height, root length, number of stools, leaf length, fresh weight, dry weight) and oil content of lemon grass under Shambat conditions.

14 Chapter Two Literature Review 2.1. Plant Botany: 2.1.1. Botany: 1-Common name: Lemon grass 2-Scientific name: Cymbopogon citratus 3-Family: Gramineae (Poaceae) 4-Plant type: Monocot; Herbaceous; Ornamental Grass 5-Origin: Indian 6-Plant height: 4 to 6 feet 7-Plant density: dense 8-Growth rate: fast 9-Foliage: 9-1-Leaf type: Simple 9-2-Leaf margin: entire 9-3-Leaf Shape: linear 9-4-Leaf venation: parallel 9-5-Leaf type and persistence: Fragrant 9-6-Leaf blade length: 18 to 36 inches 9-7-Leaf color: green 9-8-Fall color: red 10-Trunk and Branches: 10-1-Trunk \ bark

15 10-2-branches: typically multi – trunked or clumping stems. 11-Flowers: no flowers (Rarely flower). 12-Fruit: no fruit. 13-Propagation: by means of division of root stock (stools). 14-Mature plant: gave about 50 -70 stools. (Edward F. Gilman, 1999) 2.1.2. Cymbopogon spp. The genus Cymbopogon includes a number of species which produce essential oils of some commercial importance. The genus is a member of the sub-family Panicoideae of the Gramineae , and belongs to the tribe Andropogoneae; It includes between fourty and eighty species of perennial coarse-growing tufted grasses which are widely distributed throughout the tropics, occurring especially as components of the great areas of mesophytic grass land savannah of the old world tropics. Classification of the genus is far from complete since species inter cross readily and may also exist in a variety of forms; in addition some of the cultivated types do not flower making determination of their species difficult. Many species of the genus are highly aromatic due to the presence of essential oils in their stems and leaves: some of these are cultivated in various parts of the tropics for oil extraction, and some commercial oils are obtained from purely wild species.

16 Species in Sudan: 1. Cymbopogon proximus (mahareb) 2. C. nervatus (Naalgrass) 3. C. caesius (Nees) 4.Cymbopogon citrates (Lemon grass ) ((which is categorized into; East India type C. Flexuous and West India type C. citrates)). 5 .C. giganteus 6. C. excavatus 7. C. commutatus 2.1.2.1. Cymbopogon citratus (WEST INDIAN LEMON GRASS): West Indian lemon grass a tufted perennial grass with numerous stiff stems arising from the short rhizome-like root stock, though not so tall or as strongly growing as some of the other cultivated species. It is not found as wild plant, but is found in many parts of the tropics and is cultivated in Jave, Malaya, India, the Belgian Congo, Ceylon, Burma, the West Indies and parts of Central America. Most of the oil entering world trade derives from Indonesia and Guatemala, but the plant grows well in most tropical environments and the oil is distilled on a small scale in several tropical countries. The plant rarely flowers and its propagation is by means of division of the rhizome-like root stook. Harvesting of the leaves takes place several times per year and can be continued for three or more

17 years .Oil content of leaves decrease with age, although the citral content of the oil increase with age citral is the most important constituent of the lemon grass oil and may be present to the extent of 70 percent or more .Citral is aldehyde derivative of terpene, and is extracted in the form of a thin yellow oil (Leslie S. Cobley, 1963). 2.2. Crop establishment: 2.2.1. Soil: Lemon grass can be grown on a wide range of soils but a well drained and fertile soil is more favorable. 2.2.2. Land preparation and planting: The plant rarely flowers and propagation is by means of division of the root stock (stools). The land well prepared, divided into plots. The plant were planted at a rate of three stools per hole at a distance of 60 cm between the plant and 75 cm with in the row (Geunther, 1950). 2.2.3. Harvesting: The plant harvested after 4-5 months from planting, when the plant height 60 - 70 cm above 20 cm from the soil, thus the oil yield depend on plant height. (Singh, 1970). 2.3. Factors effecting oil yield and herbage production in Cymbopogon species: 2.3.1. Effect of seasonal variation: Lemon grass is a perennial crop; grow all round the year in tropical countries.

18 Oil of lemon grass depend on growth and temperature thus the amount of oil increase in worm summer than cold winter (Miyazaki 1969 ) in Sudan Mohammed (1982 ) studied the effect of seasonal variation on the growth and oil yield of Cymbopogon citratus and found that autumn season gave the highest plant growth rate and oil content. in the Philippians, Oliveros and Aureus (1977) found that leaves of wild lemon grass (Cymbopogon citratus ) that was collected during hot dry season rendered the highest amount of oil ranging from 0.31 % to 0.48 % which contain 31.05 % to 45.2 % citral a decline was noticed in the rainy season with yield raging from 0.10 % to 0.36 % . The lowest oil yield was during the cool month ranging between 0.1% and 0.2 with citral content of 25.60 % to 39.14 % most of these studies based on studies done by Miyazaki (1969) were carried to investigate the impact of temperature as indicated by seasonal variation, on plant, oil, content and citral content in open field cultivation. It has been found that the plant is vigorous under high temperature, and the both the oil content, and citral content of oil are high. On the other hand, the fall in temperature invites the lowering in plant growth, oil and citral content. 2.3.2 Effect of leaf age on the oil content. Under high temperature Miyazaki (1959) Miyazaki and Kiyoshi (1992) they found that the oil was higher in young leaves and declines along with advances in age. How-ever, it has

19 been found that the difference in citral content with leaf age, under high temperature was comparatively small. Singh (1970) concluded that the optimum time of harvest for Cymbopogon winterianus was 72 days after growth, although the total weight of leaves was rather small. Harvesting the plant after 4 or 5 month from planting when the plant height 60 - 70 cm thus the oil yield depend on plant height, Rofael and Arnoud (1949) said oil yield of west India lemon grass higher when harvested short ,annual oil yield resulting from harvesting 10, 17 and 19 time during three year of vegetative growth gave 131, 132.3 and 110.5 Ib \ Feddan of oil respectively mount of oil and percentage depend on many factors; plant age, plant species, climate factor, Soil, cultural practice and harvesting time, Although the physical properties and a mount of citral in oil do not effect by the factors. Herat et. al. (1979) found that both the Photosynthetic rate, and oil content, in lemon grass (Cymbopogon citratus) and citronella grass (Cymbopogon nardus) were decreased with the increase in leaf age. Singh et al (1979) studied the effect of age on citronella grass (Cymbopogon winterianus) oil content of leaves decreased in maturity. 2.3.3. Effect of Nitrogen fertilizer on growth and oil content: Lemon grass can grown on a wide range of soils, but a well drained and fertile soil is more favorable

20 Miyazaki (1962) showed that in lemon grass, both plant growth and oil content declined in case of nitrogen deficiency. In Sarawak done by Ng (1972) on the effect of fertilizer on the grass yield, revealed that lemon grass and citronella grass, grown on red yellow podsolic soil increased content, by application of 80Ib.Nitrogen and 40 – 80 Ib K2O. Varying doses of N: P: K had no effect on oil content. Total oil yield increased due to increase in herb yield under different treatments Gubta, et al (1978) in a work done on the effect of fertilizer on the yield of Cymbopogon martini, they found that, the plant height was increased due to balanced application of 80 Kg / hectare each of N: P: K Although it was not reflected on the final yield of oil content. 2. 4. Effect of Sulphur on growth and development: Sulphur application was found to affect the availability of other nutrients to plants. Omer (1970) working on Peanut, reported that application of sulphur resulted in an increase in the up take of phosphorus iron and potassium. Singh (1966) and Tripath (1985) working with wheat reported that sulphur application increased the up take of nitrogen. In addition sulphur reduces soil pH and there by improve the production of excessively alkaline soils (Aulakh and Der, 1976) and (Abdalraaoof, 1996).

21 2.5. Effect of organic fertilizer on plant Growth: Previous study used organic and chemical fertilizer in Cymbopogon proximus (Mahareb) used chicken manure 2.5 Ton / Fed gave the highest rate of growth and herbage production than urea (Abed alla, 2000). Also Ahmed (2000) studied the effect of organic fertilizer and chemical (Urea) in per winkle under Shambat condition used three levels of chicken manure 2.5, 5 and 7.5 Ton / Fed., resulted in significant increase in herbage yield increasing with increase in fertilizer level. Organic fertilizer increase organic matter content of soil (Eltibib et al., 1994) the particular significance of soil organic matter for soil fertility is that it influences so many different soil properties which facilitate aggregation with mineral particles, particularly clays, and in turn modify soil physical structure and influence soil water regime, and it is a source of energy for the soil biota and thus influence many of the biological processes of soil. Thus, soil organic matter itself represents a setoff attributes rather than an entity (Woomer and Swift, 1991). The effectiveness of poultry manure depends on its composition, environmental conditions and the crop characteristics. Generally, chicken manure is considered to have fertilizers and farm Yard manure and it has an appreciable residual effect (Cooke, 1975). Chicken manure contain as much

22 nitrogen as farm yard manure but it richer in phosphorus and potassium (Cooke, 1982) Appendix (1). 2.5.2. Chicken manure Fertilizer: Large amount of manure are produced each year from the growing poultry industry around Khartoum state. The value of this by product in the soil improvement and in crop production is well documented (Mohammed, 1988) and (Gabir, 1994). Chicken manure production and decomposition are influenced by factor viz: chicken type, age, sort and amount of feed and even the climatic conditions during collection, accumulation and storage management. These practices vary widely between farms (Elagib, 1997). The influence of chicken manure as a nitrogen fertilizer, it is value as a source of plant nutrient and its chemical composition were determined by many investigators like Perkin and Barker (1971); Forawi and Elsheikh (1995) and Abdelgani (1997). Fresh poultry dropping contain twice as much nitrogen as FYM, they are richer in phosphorus and contain about as much potassium as FYM. In most soils, about half of the total nitrogen in droppings and deep litter is equivalent to organic nitrogen fertilizer, but the proportion that is immediately useful in composts varies with their composition and maturity. The uric acid in fresh dropping is decomposed by micro organisms to give ammonia which is easily lost if the manure is exposed. Drying fresh droppings quickly by heat stops these

23 losses. Nitrogen is also easily lost from poultry manure taken from houses where cereal straw has been used in deep litter system. Much loss in the house is inevitable, but when deep litter is stored after clearling it should be moistured to prevent ammonia escaping and be protected from leaching by rain (Cooke, 1980). Chicken manure is valuable and known to improve the physical properties such as porosity, pore shape and pore size distribution, nitrate – nitrogen and organic matter of soils (Eltilib et al, 1994). Hence better growth of plants and more production will be obtained. Chicken manure is more or less concentrated organic fertilizer. It is varies according to the way hens are kept and their fed. It contains all the basic nutrient necessary for crops in the form of available compounds (Yagodin, 1984). Chicken manure is comparatively easily mineralizable (Abdelgani, 1997) and is rich in nitrogen and phosphorus and increases soil an available phosphorus, exchangeable potassium, nitrate – nitrogen and organic matter content of the soil (Etilib et al., 1994). It was reported by (Gupta et al. 1988). 2.6. Volatile oil: Volatile oils may occur in different plant organs, for example, Flowers as in rose, barks as in cinnamon, leaves as in lemon grass and peppermint, wood as in Sandal, roots as in

24 Angelic, rhizomes as in ginger, fruits as in bergamot and seeds as in anise. Clause and Tyler ( 1965) stated that monoterpenes and sesquiterpenes are known as volatile oils that may occur in specialized secretary structures, such as glandular hairs as in the family Labiatae or in modified parenchyma cell as in the family Piperaceae. Trease and Evans (1980) reported that volatile oils might exist in free state such as in rose and lavender. However, they occasionally occur in the form of glycosides which under favorable conditions are hydrolyzed in the presence of enzymes as in Jasmine and tuberose. Oil content is a vital item in aromatic plants, and it is focal point to be considered in Cymbopogon spp. Hence it determines the economical value of the plant. Oil content changed according to cultural practices and environmental factors. In Zambia Chisowa, et al. (1998) obtained in 3.0% yield; in Egypt oil contains 0.3- 0.6%, in Brazil Ming, et.al. (1996) they found oil contents 0.42 and 0.13 %. De Silva (1959) found that lemongrass oil yielded about 0.3 - 0.4 %, oil obtained in china Talalag (1964) he found that dried grass yielded 2.1 - 2.5 % and fresh grass yield 0.38 - 0.5, Gonzolo, et al. (1973) showed that essential oil contains 0.35 – 0.40 %, Mouldgill, and Iver (1922) obtained oil 0.8% yield. Rao and Sudborough (1925) obtained oil from the whole plant (0.22- 0.35%), Stalk

25 and leaves (0.16 –0.2%) and flower heads (1- 1.6 %), Chiovend (1906). Oswald (1915) obtained a pale yellow oil in 1- 0 - 1.2 % yield and Ahmed (1982) obtained 0.49 % yield from fresh grass in summer season. 2.6.1. Extraction of volatile oils: There are three methods of distillation; water distillation, water - steam distillation and Direct steam distillation. 2.6.1.1. Water distillation: When this method is employed, the material to be distilled comes in direct contact with boiling water. It may float on the water or be completely immersed, depending upon its specific gravity and the quantity of material handled per charge. The water is boiled by application of heat by any of the usual methods – i.e., direct fire, steam jacket, closed steam coil, or, in a few cases, open or perforated steam coil. The characteristic feature of this method lies in the direct contact it affords between boiling water and plant materials (e.g., powdered almonds, rose and orange blossoms) must be distilled while fully immersed and moving freely in boiling water, because on distillation with injected live steam (direct steam distillation) these materials agglutinate and form large compact lumps, through which the steam can not penetrate (Quenther, 1948). 2.6.1.2. Water and steam distillation: When this second common method of distillation is used, the plant material is supported on a perforated grinder screen

26 inserted some distance above the bottom of the still. The lower part of the still is filled with water, to a level some what below this grid. The water may be heated by any of the methods previously mentioned. Saturated in this case, wet, steam of low pressure rises through the plant material. The typical features of this method are: first that the steam is always fully saturated wet and never super heated; second, that the steam is material is in contact with steam only, and not with boiling water (Geunther, 1948). 2.6.1.3. Steam Distillation: The third method, known as steam distillation or direct steam distillation, resembles the preceding one except that no water is kept in the bottom of the still. Live steam, saturated or superheated, and frequently at pressures higher than atmosphere, is introduced through open or perforated steam coils below the charge, and proceeds upward through the charge above the supporting grid. In so far as the distillation process it self is concerned, and form the purely theoretical point of view, there should be no fundamental difference between these three methods. There exist, how ever, certain variation in practice, and in the practical results obtained, which in some cases are considerable; they depend on the method employed, because of certain reactions which occur during distillation ( Geunther, 1948).

27 2.6.2. Chemistry of Cymbopogon citratus oil: The chemical constituent of Cymbopogon differs from area to another, climate, cultural practices etc. In Zambia chemical composition analyzed by GC and GC -MS which contain sixteen compounds 93.4 % was identified as geraniol 39.0 %, neral 29.4 % and myrecene 18.0 % which represent the major components. Beside minor component such as geranial 1.7% and linalool. (Chesowa et.al, 1996). Torres and Ragadio (1996) they found that Cymbopogon citratus obtained from Philippine analyzed by GC which the main essential oil constituent was citral (69 – 39 %), Geranial, myrecene alpha – and beta. Pinene, ethyl laurate, 1, 8 – cineole [eucalyptol], limonene, citronella, linalool, Caryophyllene, menthol and terpineol. Other of oil of Cymbopogon citratus from Somalia land Rovesri and Variati (1960). They found that it contain 80 % citral, 12 % myrecene 3-4 % dipentene and traces of methylheptanone. 2.6.3. The physiochemical properties of lemon grass oil: The physiochemical properties determine the economical value of essential oil, since there were many specifications was done through out the world, to determine the quality control of the oil. The oil obtained from lemon grass grown in the Sudan (Ahmed 1982) was subjected to physiochemical testing, and compared with standard specification. It was found that it

28 conforms to British standards (B. S 1963) physio – chemical data are given in appendix (2). Gildemeister and Hoffmann reported the following properties of East Indian lemon grass oil Appendix (3). 2.7. Uses of Cymbopogon citratus: Oil of lemon grass is used widely for scenting of soaps, detergents and all kind of technical products. The bulk of the oil, however, is employed for the isolation of citral, which, as such, is used in flavor, Cosmotics and perfumes, or is converted into ionone, a group of very important synthetic aromatics possessing a strong and lasting violet odor. Ahmed et al. (1998) They found that essential oil of Cymbopogon citratus and two plants were antibacterial against Staphococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa, and also essential oil of Cymbopogon citratus and five plants were tested against 20 strains of bacteria and they found that were active on most bacterial tested as 9 strains of Escherichia coli, 8 strains of salmonella, and one strain each of Proteus morganii, Staphylococcus aurens and Shigella sonnei ( Mar - Mar Nyein et al. (1996). Ismail (1994) was found that essential oil of Cymbopogon citratus and three plants antivirus against potato virus Y (PYV) in vitro. Essential oil of Cymbopogon sp. revealed that oils are as effective as chemical oils such as Mylol, which were gave a

29 complete protection against mosquitoes Anopheles Culciferacies, species about 95 – 96 % (Ansari and Razadan, 1995). Wannissorn et al. 1996 they found that essential oil of Cymbopogon citratus obtained from Thailand as antifungal against 35 clinical isolates of 4 dermatophytes (Trichophyton mentagrophytes, T. rubrum. Epidermophyton floccosum and Microsporum gypseum). In Philippines cheroot is used a hot decoction for tooth and internally as diuretic. The leaf is there also applied to the fore head and face to relieve headache. In Indian the oil aids digestion as well as relieves spasms, muscle cramps, sore throats rheumatism, headaches and antiseptic (John Mitchell watt, et al., 1992) and (MARIA Gerdina Breyer – Brand wijk). Citral is main use is in the production of ionone, a synthetic perfume with odour of violets, but more recently it has became increasing by important as a starting point for the synthesis of vitamin A. Lemon grass oil is thus important in medical and pharmaceutical chemistry, apart from its use in the perfume and soup industries (Leslie S. Cobley, 1963).

30 Chapter Three Materials and Methods

3.1. Location: The experiment was carried out in demonstration and research field – Department of Horticulture- Faculty of Agriculture – University of Khartoum. The area lies in latitude 150 40' and longitude 320 32'. 3.2. Climate: The climate can be considered a semi – arid of rainy season from July to September ( 90% of the rain fall concentrated during this period, high temperature 48 0C during summer and as low as 50 C during winter (Oliver, 1965). 2.3. Soil: Shambat soil is heavy clay with pH 7.6 Appendix (1). 3.4. Source of the plant: Cymbopogon citratus "Lemon grass"; Family Graminae (poaceae) plants were raised from stools obtained from fully mature plants grown in the Nursery of the Horticulture Administration of the Federal Ministry of Agriculture – Elmogran Area ( Khartoum – Sudan ). 3.5. Plant management: The land was ploughed, disc – harrowed, leveled and divided into four replicates. Each replicate included three treatments; and the plot area was 3 ×3 m.

31 The plants were planted at the rate of two stools per hole at a distance of 50 cm. within rows and 75 cm apart between rows. Each plot contains 15 plants. Fertilization was conducted by adding chicken manure before planting, while ammonium sulphate was added in three doses; the first one after 45 days from planting and then every 45 days. The plants were watered twice a week during the first two months, after that irrigated once weekly; till harvesting at the six month. Weeding was conducted through out the experiment when necessary. Harvesting was done twice; the first one after the fourth month from planting, the second one was after six month from planting; plate (1), (2) and (3). 3.6. Experimental design: The experimental treatments were laid out in randomized complete block design with four replicates and three treatments each. The three treatments were; Control "no fertilizer", Combine fertilizer "2.5 chicken manure with 2N ammonium sulphate (80 lb) per feddan" and Organic fertilizer "5 tons / feddan chicken manure".

Plate( 1 ) : Control treatment of Lemon grass:

Plate ( 2 ) : Combine Fertilizer treatments of Lemon grass plants:

Plate ( 3 ) : Organic fertilizer of lemon grass plants:

35 3.7. Data collected: 3.7.1. Growth parameters: 3.7.1.1. Plant height: From each plot three plants were selected randomly and tagged. Then plant height was measured from the soil surface to the highest tip of the longest leaf (cm) after each 4 to 6 months from planting. 3.7.1.2. Root length: From each plot three plants were selected randomly and tagged. Then the root length was measured from just below the soil surface to the tip of the roots (in cm) after plant removal. 3.7.1.3. Number of stools: From each plot 9 plants were selected, the number of stools was counted per each plant; every two weeks. 3.7.1.4. Leaf length: From each plot 9 plants were selected, the length of the longest leaf from each plant was measured (cm); every two weeks. 3.7.1.5. Fresh weight: Three plants were selected from each plot; the whole plants (shoot and roots) were weighed; using balance after four month and six month from planting; plates (4), (5) and (6). 3.7.1.6. Dry weight: Three plants were selected from each plot and freshly weighed, put in an electric air – force oven for 65 – 70 0C for 48

Plate (4): Lemon grass root and shoot (Control):

Plate (5): Lemon grass root and shoot (Combine)

Plate (6): Lemon grass root and shoot (Organic):

39 hours. After drying each plant was weighed using balance; after the fourth and six months from planting. 3.7.2. Determination of oil content (Oil extraction): Oil was extracted by water distillation method. 100g of shade dried samples of lemon grass leaves were weighed using sensitive balance. The sample was placed in a flask and the flask was adjusted to glass apparatus for distillation volatile oils higher than water equipped with a condenser Appendix (4). The water in the flask was brought to boil at 70 0C for two hours using electric heating mantle. And the mixture of water vapour and volatile oil were affected to condense and collected as two layers; the volatile oil formed the top layer. The whole system was left for some time (around half an hour) until there was no significant increase in the volume of the oil and it was allowed to cool so that the turbid oil layer become very clear. After distillation was measured and the percentage of oil was determined expressed as volume / weight percentage plate (7).

B A C

Plate(7): Lemon grass oil from three fertilizer levels:

A: Oil from control treatment. B: Oil from combine fertilizer. C: Oil From organic fertilizer.

41 CHAPTER FOUR RESULTs

4.1. Effect of Chicken Manure and Ammonium Sulphate on lemon grass: Effects of chicken manure and ammonium sulphate on growth parameters ;such as plant height ,root length , fresh weight and dry weight during two successive harvest are shown in figures (1),(2),(3) and (4). 4.1.1. Plant height : Addition of Combine fertilizer "2.5 tons of chicken manure / feddan with 2N ammonium" and Organic fertilizer "5 tons of chicken manure / feddan"; to lemon grass showed no significant differences between the fertilizers treatments; during the first harvest and second harvest for plant height (fig. 1) . 4.1.2. Root length: Addition of 2.5 ton / feddan with 2N ammonium sulphate and 5Ton / fed of chicken manure showed no significant differences between treatments on root length during the first and second harvest (figure 2). 4.1.3. Fresh weight: The effect of chicken manure and ammonium sulphate fertilizer on fresh weight shown in figure (3). Addition of both chicken manure 2.5 ton / fed with 2N ammonium sulphate and 5 ton / fed. to lemon grass plants

42 Table (1) : Effect of chicken manure and ammonium sulphate on growth rate and yield:

Harvesting First harvest (4 monthes) Second harvest (6 monthes) Treatments Plant Root Fresh Dry Plant Root Fresh Dry height length weight weight height length weight weight

Control 96.2 17.5 737. 5 205. 5 108.4 18.14 3028.8 736.8

2.5Ton/Fed.Chk.Manr. 93.4 + 18.5 883.3 227.5 11 3.8 19.25 3362.5 967.5 2N (NH4)2 SO4

5 ton / Fed. Chk.Manr. 95.0 18.4 1016.7 308.31 112.9 19.17 3062.3 949.5

L.S.D 4.97 NS 4.65NS 182.8* 39.15** 4.38 ** 2.92 NS 21.61 * 136.1**

• =significant difference (5%) ** =highly significant difference (1%)

43 Figure(1):Effect of combined and organic fertilizer on plant height (cm):

120

100

80 Plant control height 60 combine (Cm) organic 40

0 first harvest second harvest

44 Fig.(2):Effect of combined and organic fertilizer on root length

19.5

18.5 control Root combine Length 18 (Cm) organic

17.5

16.5 first harvest second harvest

45 Fig.(3): Effect of combine and chicken manure on fresh weight (g)

3500

3000

2500 control Fresh 2000 weight combine (g) 1500 organic

1000

500

0 first harvest second harvest

46 increase fresh weight significantly; on the other way there was no significant increase in fresh weight of lemon grass between the two fertilizers during the first harvest; While during the second harvest both fertilizers showed a significant difference . Moreover, combination of fertilizer showed a significant increase over 5 ton chicken treatment. 4.1.4. Dry weight: The effect of chicken manure and ammonium sulphate on dry weight was shown in table (1) and figure (4). Organic fertilizer 5 Ton / Fed clearly enhanced the dry weight than the combination treatment. On the other hand, organic fertilizer was the better treatment as reflected in higher dry weight during the first harvest; while during the second harvest there was highly increment in dry weight resulted from addition of fertilizer's treatments, but there was no significant differences between the two fertilizer treatments. 4.2. Effect of chicken manure and ammonium sulphate on number of stools : Effect of chicken manure and ammonium sulphate shown in Table (2) and figure (5). In the second month (May) addition of fertilizer showed that no significant differences between the two fertilizer's treatments in the second and fourth week; but there was a trend of an increase in fourth week on the number of stools between the two fertilizers treatments.

47 Fig.(4):Effect of combined and organic fertilizer on dry weight(g):

1000

900

800

700 control dry 600 combine weight 500 organic (g) 400

300

200

100

0 first harvest second harvest

Table (2) : Effect of chicken manure and ammonium sulphate on number of stools:

Month May June July August September second Fourth second Fourth second Fourth second Fourth second Fourth Treatments week week week week week week week week week week

Control 1.49 5.97 12.75 23.75 31.92 33.61 39.61 42.47 46.47 51.21

2.5Ton/Fed.Chk.Manr. + 1.00 7.17 14.06 26.39 34.56 7.00 46.96 51.11 59.68 62.86

2N (NH4)2 SO4

5 ton / Fed. Chk.Manr. 1.35 8.91 18.47 31.14 38.2 39.50 46.57 51.32 53.18 57.21

L.S.D 0.55 NS 1.89** 3.66** 1.27** 3.95 * 4.45 * 4.91 * 4.74** 8.82 ** 6.45**

• =significant difference (5%) ** =highly significant difference (1%)

49 Fig.(5):Effect of chicken manure and ammonium sulphate on number of stools:

40 Control Combine 30 Organic

Number of stools of Number 20

0 2nd 4th 6th 8th 10th 12th 14th 16th 18th 20th Weeks

50 In the third month (June) organic fertilizer 5 ton / fed clearly enhanced the number of stools than combination of fertilizer; (2.5) ton / fed with 2N (NH4)2 SO4, during the second and fourth week , with highly significant difference between fertilizer treatments. while during the fourth month (July) in the second week organic fertilizer increased the number of stools than combination fertilizer; showed significant differences between the two fertilizer treatment. But in the fourth week addition of fertilizer showed no significant differences between the two fertilizer treatments but using organic fertilizer there yielded a trend of increase . In the fifth month (August) addition of fertilizer increase the number of stools by both fertilizer treatments; but with no significant difference between the two fertilizers treatments, in the second and fourth week . During the sixth month (September); the last month; during the second and fourth week addition of fertilizer enhanced the number of stools between the two fertilizer treatments; with no significant difference between them. But combination fertilizer showed a trend of an increase on the number of stools reached 62 stools in the sixth month . 4.3. Effect of chicken manure and ammonium sulphate on leaf length: The effect of combine fertilizer and organic fetilizer on leaf length were shown in table (3) figure (6).

51 In the second week of the second month combination fertilizer showed increased significantly on leaf length compared to organic fertilizer, but in the fourth week there was no significant differences between the two fertilizer treatments . During the third month (June) addition of fertilizer showed that no significant difference between both fertilizer treatments, but there was a trend of increased in leaf length of organic fertilizer in the two second and fourth weeks . The fourth week (July) fertilizer's treatments enhanced leaf length, with no significant difference; but there was a trend of increase in organic fertilizer in both weeks . While during the fifth month (August) in the two weeks; the second and fourth; addition of fertilizer showed no significant difference on leaf length between the fertilizer treatments . The sixth month (September); the last month; fertilizer treatments increased the leaf length in both the second and fourth week. A trend of increase in organic fertilizer compared to combine fertilizer in the last week was noticed. 4.4. Effect of chicken manure and ammonium sulphate on the oil content of lemon grass: Effect of fertilizer treatments shown in Table (4). Application of fertilizer showed no significant increase in oil content during both the first and second harvest. The oil yield

52 Table (3) : Effect of chicken manure and ammonium sulphate on number of leaf length:

Month May June July August September second Fourth second Fourth second Fourth second Fourth second Fourth Treatments week week week week week week week week week week

Control 42.3 b 48.2 a 50.6 a 55.6 a 59.8 a 64.8 a 71.9 a 77.6 b 78.9 b 80.2 a

2.5Ton/Fed.Chk.Manr. 48.6 a a a a a a a a a a + 50.2 50.2 55.6 61.5 66.8 75.1 82.8 84.2 85.0 2N (NH4)2 SO4

5 ton / Fed. Chk.Manr. 42.9 b 49.8 a 52.1 a 57.0 a 63.1 a 67.7 a 75.7 a 83. 2a 84. 8 a 86.1ab

L.S.D 4.03 * 7.45NS 5.13NS 5.89NS 4.26NS 3.76NS 3.03NS 3.51* 4.57 ** 4.87NS

• =significant difference (5%) ** = highly significant difference (1%)

53 Fig.(6):Effect of combine manure and organic fertilizer on lemongrass leaf length:

100

80 ) 70

60 Control

50 Combine

40 Organic 30 Leaf length (cm length Leaf

0 2nd 4th 6th 8th 10th 12th 14th 16th 18th 20th Weeks

Table (4): Effect of Combine fertilizer and Organic Fertilizer on the oil content(ml):

Month Treatments First harvest Second harvest 30 / 7 / 2003 30 / 9 /2003

Control 3.11 2.55

Combine fertilizer 3.11 2.55

Organic Fertilizer 3.11 2.55

L.S.D N.S N.S

NS = Non significant differences

55 was better during the fist harvest; 3.11 % ; and decreased to 2.55 % during the second harvest .

56 Chapter five Discussion

The results obtained from this experiment which was conducted in the demonstration farm at Shambat – Faculty of Agriculture – University of Khartoum showed a positive effect of chicken manure and ammonium sulphate on growth and yield components according to the following postulations: 5.1. Effect of chicken manure and ammonium sulphate on growth: The results showed no significant difference between the treatments in plant height and this may resulted from the fact that the plant in the first harvest stage of growth is weak after four month from planting. Since the first leaf emergence of the plant after one month (Ahmed, 1982) and fertilizer decomposition was slow. During the second harvest the significant differences due to activation of nutritional element after the decomposition of chicken manure and also to he addition of ammonium sulphate which exerted a sort of increase as resulting of Gubta, et al. (1978) who found that the plant height was increased as a result of the balance application of 80 Kg/hectare for each N:P:K. The result showed no significant difference in root length between treatments during the first and second harvest. This may be due to the fact that, the roots of lemon grass are a

57 surface roots. And don't go deeper in the soil. Previous studies don’t mention the measurement of roots. Edward (1999) said that the root length can n't be measured. In the first harvest, fertilizer treatments; increased the fresh weight. Addition of chicken manure resulted an increase in herbage. The yield increased with the increase in fertilizer level, this result agrees with (Ahmed, 2000) addition of chicken manure 2.5, 5, 7.5 Ton / Feddan) gave the highest rate of growth and herbage production. Using organic fertilizer (5 Ton / Fed), resulted in a trend of increase in fresh weight. This result may be attributed to the beneficial effect of manure in the soil; this result is in line with Cheung and Wong (1983) who reported that manure is known to provide plant nutrients, improve the soil physical properties and increase aggregate stability and thus might have enhanced the uptake of nutrient. In the second harvest, combined fertilizer gave better production of fresh weight than organic fertilizer (5 Ton / Fed) combined fertilizer contain organic fertilizer (2.5 Ton / Fed.

With 2 N chemical (NH4)2 SO4, 2.5Ton / Fed increased the yield, this result agree with the results obtained by (Abdalla 2000) using organic fertilizer ( 2.5 Ton / Fed ) with cymbopogon proximus ( Mahareeb), gave the highest rate of growth and herbage production than using urea. Ammonium sulphate

58 applied in three split doses (each 45 day ) this mean more nutrient added to the soil. Nitrogen increased the growth and oil contents, this result agree with Miyazaki and Sarawk (1972) they revealed that lemon grass grown on red yellow podsolic soil significantly increase both grass yield and oil content by application of 80 Ib

Nitrogen and 40 – 80 Ib K2O ( varying doses of N : P : K had no effect on oil content). Ammonium sulphate also ( source of sulphur ), was found to affect the availability of other nutrients to the plants, this result agree with Omer (1970) who reported that application of sulphur resulted in an increase in an increase in the up take of phosphorus, iron and potassium, also Singh (1966), Tripath (1985) and Tripath (1984) whose studies on wheat revealed that sulphur application increased the up take of nitrogen, also the addition of sulpher reduced soil pH and thus improved the production of excessive alkaline soils, Aulakh and Dev (1976) and Abd - El Raouf, found that snap bean (reduced 0.3). The results showed significant difference with organic fertilizer (5 Ton/Fed), which increased the dry weight; by using combined fertilizer in the first harvest; and this may be due to the availability of more nutritional element obtained from the chemical fertilizer, after the decomposition of chicken manure, in the second harvest, the highly increase in dry weight associated with organic fertilizer treatments may be due to the

59 decomposition of chemical and organic fertilizer in the soil and these reflected in the growth of the plants. In table (2) the results showed no significant difference on number of stools between fertilizer treatments (except in June month and the second month in July) where significant different existed and this may be due to the amount of organic fertilizer (ammonium sulphate, was not enough to affect the number of stools. In combined fertilizer addition of chemical fertilizer in three split doses enhanced the number of stools until the harvest ( September ) reached 62 stools until the highest number of stools, also high organic fertilizer 5 Ton / Fed gave high number of stools finally reached 57 stools, this results indicated that similar to fertilizer treatments, the number of stools reach 50 - 70 stools at maturity) The results showed no significant difference on leaf length between treatments, addition of fertilizer had no effect on leaf length but it affected fresh weight and number of stools. In fertilizer treatments the leaf length reached 85 – 86 cm. 5.1. Effect of chicken manure and ammonium sulphate on oil content: Summer planting coincide with autumn season, gave higher growth and oil content and this agree with the findings of Miyazaki (1957) who found that the plant was vigorous under high temperature, and the oil and citral contents are high, this

60 result, in turn agree with those reported by Oliveros and Aureus (1977). In the first harvest, high amount of oil was obtained ( 3.11 % ), this result agree with Miyazaki (1959) and Miyazaki and Kiyoshi (1992) they found that under high temperature the oil content was higher in young leaves and declines with the advance in age so in the second harvest. The amount of oil decreased ( 2.55 % ) also Herat et al. (1979) found that the photosynthetic rate, and oil content in lemon grass ( Cymbopogon citratus ) and citronella grass were decreased with the increased in leaf age, Singh et al (1979) studied the effect of age on citronella grass oil and found that the oil contents of leaves decreased in maturity and also Sarawake (1972) and Gubta, et al. (1978) found that fertilizer treatment influenced grass yield more than oil content, a fact which agree with the present results. Also dried grass contained high oil than fresh grass and this agree with the findings of Talalag (1964) who found that dried grass yield 2.1 - 2.5 % than fresh grass 0.38 – 0.5 %. Also the amount of oil ( 3.11% ) agreement with oil obtained in Zambia 3.0% ( Chisowa et al. 1998). From the above discussion we can come to conclude that both 5 tons and 2.5 tons with ammonium sulphate show general a positive increase in growth parameters and yield components. Further more, combined fertilizer (2.5 ton of chicken manure and 2 N ammonium sulphate) showed a significant increase in

61 fresh weight during the second harvest. On the other side, there were no changes in oil content recorded due to fertilizer application. Suggestions

The following suggestions can be considered for Cymbapogon spp. Production improvement and development: 1- Further study on lemon grass spacing under Shambat area have needed, since 50 cm in row spacing lead to establishment of dense plants. 2- Further studies on different levels of organic fertilizer on lemon grass must be conducted to minimize the cost of production by using cheap organic fertilizer. 3- Different combinations of chicken manure with granules and foliar fertilizers are recommended. 4- Further investigations of major and minor components of lemon grass essential oil are recommended. 5- Introduction of other Cymbopogon spp. Are recommended for oil quality and quantity increments and developments. 6- Harvest and post harvest studying of lemon grass oil under Sudan conditions are recommended..

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Appendix (1): Soil and Chicken manure analysis: Samples N % P % K meg / L pH

Soil 0.07 0.025 1.025 7.56

Chicken manure 1.9 0.5 10 % 5.75

Analysis of samples: Dept. Of Soil - F. Of Agric. - U. Of K.

Appendix(2): Physio-chemical properties of lemon grass oil : Characteristics Results B.S. 1963

1- specific gravity 0.898 - 0.901 0.870 - 0.895 at 30 / 300C at 20 / 20 0C 2- optical rotation -10 to +50 at 300 C -30 to +10 at 200 C

3- refractive index 1.481 – 1.484 at 300C 1.483 – 1.489 at 200C

4- Citral content in the 74 % - 89 % Not < 75 % oil as % age

74 Appendix (3): Properties of East Indian lemon grass oil Characteristic Result specific gravity at 150 0.899 to 0.905 exceptionally as 0.895 and as light as 0.911 in other wise good oil Optical Rotation + 10 25' to – 50 0'

Refractive index at 200 1.483 to 85% Aldehyd content: Bisulfite method 70 to 85 % Neutral sulfite method y 65 to 85 %

Solubility Soluble un 1.5 to 3 vol. of 70 % alcohol; occasionally with opalescence or turbidity, due to separation of parafins Source: Gildemeister and Hoffman