Population Characteristics, Habitat Preference and Bark Harvest Potential of Daphne Bholua in Madane Protected Forest, Gulmi, Nepal

Home , Daphne (plant), Daphne bholua, Daphne papyracea, Lokta paper

Population Characteristics, Habitat Preference and Bark Harvest Potential of Daphne bholua in Madane Protected Forest, Gulmi, Nepal

A Dissertation Submitted for the Partial Fulfillment of the Master Degree in Botany

Submitted by Bikram Jnawali Roll No: Bot 108/071 T.U. Regd. No: 5–249–3–2010 Batch: 2071/73

Plant Systematics and Biodiversity Conservation Unit

Central Department of Botany

Institute of Science and Technology

Tribhuvan University

Kirtipur, Kathmandu, Nepal

April 2019

RECOMMENDATION

It is hereby recommended that Bikram Jnawali, M.Sc. Botany final semester student of ‘Plant Systematics and Biodiversity Conservation Unit’ at Tribhuvan University, Institute of Science and Technology, Kirtipur, Kathmandu has carried out the research work entitled Population Characteristics, Habitat Preference and Bark Harvest Potential of Daphne bholua in Madane Protected Forest, Gulmi, Nepal under my supervision. The entire work is based on the field work performed by him and brings out some useful findings in the field of plant science.

As per my knowledge, this work has not been submitted for any other academic degree. I, therefore recommend this dissertation to be accepted for the partial fulfillment of the requirement of Master’s Degree in Botany at the Insititute of Science and Technology, Tribhuvan University.

………………………………….. (Supervisor) Suresh Kumar Ghimire, Ph.D. Associate Professor

Central Department of Botany

Tribhuvan University, Nepal

Date 25th April, 2019

LETTER OF APPROVAL

The M.Sc. dissertation entitled “Population Characteristics, Habitat Preference and Bark Harvest Potential of Daphne bholua in Madane Protected Forest, Gulmi, Nepal” submitted by Bikram Jnawali has been accepted for the partial fulfillment of Master’s Degree in Botany (Plant Systematics and Biodiversity Conservation Unit)

Expert Committee:

……………………………… …………………………… (Supervisor) (Head of the Department) Suresh Kumar Ghimire, Ph.D. Ram Kailash Prasad Yadav, Ph. D. Associate Professor Professor Central Department of Botany Central Department of Botany Tribhuvan University, Nepal Tribhuvan University, Nepal

…………………………….. ……………………………… (External Examiner) Internal Examiner Uddhab Raj Khadka, Ph. D. Anjana Devkota, Ph.D. Associate Professor Associate Professor Central Department of Environment Science Central Department of Botany Tribhuvan University, Nepal Tribhuvan University, Nepal Date ………………………..

ACKNOWLEDGEMENTS

I would like to express the deepest appreciation to my respected supervisor Dr. Suresh Kumar Ghimire, Associate Professor, Central Department of Botany for providing me with invaluable suggestions, competent guidance, continuous encouragement and valuable advices throughout the preparation of this dissertation. My thanks to him will always be counted less in front of his support and all the motivations.

I am grateful to Prof. Dr. Ram Kailash Prasad Yadav, Head of Central Department of Botany for his administrative support during study work. My warmest thanks are due to Prof. Mohan Siwakoti and Prof. Mohan Panthi, former Head, Central Department of Botany for their valuable suggestions, encouraging words, and necessary help during my study. I am thankful to Prof. Dr. Bijaya Pant, Prof. Dr.Sangeeta Rajbhandary, Dr. Bharat Babu Shrestha, Dr. Chitra Bahadur Baniya and Dr. Chandra Prasad Pokharel for their support and all the motivations during my study. Similarly, I am thankful to all the respected teachers and staff members of Central Department of Botany.

I would like to express sincere thanks to Department of Forest, Government of Nepal, for granting me the permission for this study. I would like to thank the officials of District Forest Office Gulmi, Madane Protrcted Forest Banjhkateri for providing permission to conduct my research work and their valuable help during filed visit. I am also thankful to Mr. Chabilal Sharma, Mr. Dayanidhi Bhusal, Ms. Ramita Bhusal and all the local peoples of Madane Protected Forest for providing necessary help, suggestions and information about the study site.

My special thanks go to Mr. Mukti Ram Poudel for providing me the much–needed knowledge about data collection, data entry and data analysis throughout my study. I would like to thank Dr. Birendra Karna and Dr. Lila Nath Sharma for sharing their knowledge and experience regarding to the research. Acknowledgements also goes to Mr. James Siddartha Lucas, Mr. Yagya Raj Paneru, Ms. Sunita Shrestha, Ms. Pratikshya Chalise, Mr. Suresh Paneru for support and suggestions.

I also extend my special thanks to all the friends and colleagues for their active help and support from beginning to the completion of this research work. In particular, I would like to take this opportunity to admire my friends Mr. Ajay Neupane, Ms.

iii

Sabana Perveen, Mr. Ganesh Joshi, Ms. Shristhi Nepal for their never–ending support, invaluably constructive criticism and friendly advice during the entire work.

My thanks go to Mr. Basudev Poudel who helped me to prepare GIS map. I would also like to thank my friends, Ms. Prativa Paneru, Ms. Ashika Bhusal, Mr. Subash Nyaupane, Mr. Hari G.M., Ms. Illina Bajracharya, Ms. Mamita Shakya, Ms. Basanti Bhatt, Ms. Pooja Joshi, and Ms. Sushila Sharma for their help and support. My words of thank is also due to my senior and junior friends of Central Department of Botany.

Nobody has been more important to me in the pursuit of this project than the members of my family. I would like to thank my parents; whose love and guidance are with me in whatever I pursue. They are the ultimate role models. Most importantly, I wish to thank my loving and supportive parents Mr. Rajendra Gyawali, Mrs. Bhagwati Gyawali, Ms. Bandana Gyawali and Ms. Brasha Gyawali for their unending support and inspiration.

……………………… Bikram Jnawali Date

ACRONYMS AND ABBREVIATIONS

ANOVA Analysis of variance

GoN Government of Nepal

RDA Redundancy Analysis

DCA Detrended Correspondence Analysis

MPF Madane Protected Forest

NR Near Threatened

IUCN International Union for Nature Conservation

HMG His Majesty’s Government

KATH National Herbarium and Plant Laboratories

TUCH Tribhuvan University Central Herbarium, Kathmandu

SE Standard Error

SOC Soil Organic Carbon

NTFP Non–Timber Forest Product

SPSS Statistical Program for Social Science

US$ United State Dollar

ITC International Trade Center

FSRO Forestry Services

ABSTRACT

Unsustainable harvest of non–timber forest products (NTFPs) for commercial purpose may cause severe impacts to the harvested populations. Therefore, there is a need for studies pertaining to their sustainable management. Sustainability of NTFP harvesting depends upon plant performance and growth strategy. Daphne bholua Buch. – Ham. Ex D. Don is a potential non–timber forest product (NTFP), whose bark is extracted for making Nepali handmade paper. Studies pertaining to its bark potential, habitat characteristics and population status are limited in Nepal. We evaluated its habitat characteristics, bark potential and regeneration potentialities along an elevation gradient in Madane Protected Forest (MPF). Altogether 108 plots (5×5) m were established along three elevation sites each of 200–meter elevation with three canopy types: close, semi–close and open. Total elevation range of the species was 1900– 2500 m. The mean density of D. bholua was maximum in the highest elevation band (2300–2500 m) that received less disturbance. A density was obtained minimum in the lowest site (1900–2100 m) that received highest disturbance. The semi–close canopy condition with slightly acidic soil with high nutrient and moisture supports higher growth and regeneration of Daphne as compared to open and close canopy. which are negative to either side of close and open canopies. High bark mass was found in higher elevation sites with semi–close canopy, which was followed by open and close canopy. Good representation of lower size classes followed by higher size classes indicated the good natural regeneration. In majority of cases, regeneration was limited with root suckers and coppice outgrowth. The flower production and fruit set were very poor at lowest site. In the study area, bark of Daphne bholua is only harvested for local use and yet to be commercialized. However, D. bholua populations at lower elevation band were affected greatly by grazing and trampling effects and also by developmental activities. The present stock of of D. bholua bark in MPF is in good condition with potential for sustainable harvest and management, which could serve as best income generating resource to the local people.

Key words: Madane Protected Forest, Daphne bholua, bark, handmade paper

Contents

RECOMMENDATION ...... i

ACKNOWLEDGEMENTS ...... iii

ACRONYMS AND ABBREVIATIONS ...... v

ABSTRACT ...... vi

CHAPTER ONE: INTRODUCTION ...... 1

1.1 General Background ...... 1

1.2 Daphne bholua ...... 4

1.3 Rationale ...... 6

1.4 Objectives ...... 7

CHAPTER TWO: MATERIALS AND METHODS ...... 8

2.1 Study Area, Madane Protected Forest...... 8

2.1.1 Physiography ...... 8

2.1.2 Climate ...... 9

2.1.3 Flora and fauna ...... 10

2.1.4 Socio-culture and Economy ...... 11

2.2. Study Species ...... 11

2.2.1. Taxonomy and morphology ...... 12

2.2.2. Distribution ...... 12

2.2.3. Habitat ...... 13

2.2.4. Use potential ...... 13

2.3. Methods ...... 14

2.3.1 Reconnaissance ...... 14

2.3.2 Population sampling ...... 15

2.3.3 Data collection ...... 16

2.3.4 Laboratory analysis ...... 18

2.3.5 Data analysis ...... 19

CHAPTER THREE: RESULTS ...... 22

vii

3.1 Habitat Characteristics of Daphne bholua ...... 22

3.1.1 Biophysical (substrate) variables ...... 22

3.1.2 Associated species composition ...... 23

3.2 Variation in Density and Size Class Distribution of Daphne ...... 27

3.3. Population Structure...... 30

3.4 Variation in Growth–Related Traits ...... 32

3.5 Total and Harvestable Dry Bark Mass ...... 34

CHAPTER FOUR: DISCUSSION ...... 37

4.1 Habitat characteristics of D. bholua ...... 37

4.2 Density and size class distribution ...... 40

4.3 Variation in Growth–Related Traits and Regeneration ...... 42

4.4 Total and Harvestable Dry Bark Mass ...... 44

CHAPTER FIVE: CONCLUSION & RECOMMENDATIONS ...... 46

5.1 Conclusion ...... 46

5.2 Recommendations ...... 47

References ...... 48

Appendix I. Correlation among different biophysical variables recorded in three sites...... 63

Appendix II. Percentage plots experiencing certain type of disturbance on three different altitudi–nal bands...... 64

Appendix III. List of all plant species found on the study area with their respective family, fre–quency and abbreviation ...... 65

Appendix IV. Density (number of individuals per 25 m2) of Daphne bholua in three elevation bands in MPF. Data shown are mean ± S.E...... 68

Appendix V. Growth related traits of Daphne in Three sites. Values shown are mean ± SE . 69

Appendix VI. Letter of Permission ...... 70

PHOTO PLATES ...... 71

viii

CHAPTER ONE: INTRODUCTION

1.1 General Background Elevation gradients are complex involving many different co-varying factors like topography, soil, and climate (Austin et al. 1996). Elevation strongly influences the length of the growing season and the availability of soil moisture nutrients (Namgail et al. 2010; Singh et al. 2014). Plant species growing along the elevation gradient may show variations in the structure of their populations and in traits related to their life history (Kim and Donhohue 2011). The elevation gradient is commonly explained by the climatic factors, productivity, and other energy-related factors (Turner et al. 1987; Wright et al. 1993). Components of climate and local environment (e.g., temperature, precipitation, seasonality and disturbance regime) vary along the elevation gradients and ultimately may create the variation in overall plant performance (Lomolino 2001).

Temperature–elevation gradient is probably one of the most continuous ecological gradients, which affects the vegetation composition and plant population performance (Bhattrai and Vetaas 2003). The elevation ranging from 1000 m to 3500 m in central Nepal showed good ecological conditions for subtropical to temperate species (K.C. et al. 2010).

Shrubs are the main components in the majority of the global biomes (Archibold 1995), play an important role to form much of the vegetation in tropical savannas, arid regions, Mediterranean ecosystems, and polar and high mountain tundra’s. They are also frequently found in the understory and canopy gaps (Denslow et al. 1990). Behind the successful and wide distribution of shrubs, Götmark et al. (2016) proposed three successful models. Firstly, larger photosynthetic tissue in bark and stem, the larger epidermis (bark), and a larger area for sprouting, and faster production of twigs and canopy. Secondly, multiple stems in shrubs ensure future survival and growth if one or more stems die. Lastly, structural traits of short shrub stem improve survival compared to tall tree stems. Due to the global distribution of shrubs, they are important for climate control, soil stabilization and production, ecosystem water balance, carbon uptake and storage, and for many associated species such as grazing and browsing mammals (Filazzola and Lortie 2014)..

Climatic and geographic variations in Nepal, ranging from the tropical climate of Tarai to alpine tundra of high Himalaya, have endowed the country with the vast treasure of non–timber forest products (Gautam 2006). The term non–timber forest product (NTFP) was coined by de Beer and McDermott in 1988 and used it as an alternative to the dismissive epithet ‘minor forest products’ to encompasses all biological materials other than timber which are extracted from forests for human use. Ros–Tonen et al (1995) define non–timber forest products (NTFPs) as "all tangible animal and plant products other than industrial timber, which can be collected from the forest for subsistence and for trade". Other terms used for a similar meaning are non–wood forest products (NWFPs), minor forest products (MFPs), forest products (FPs), a by-product of forests (Uprety et al. 2016). Since the last decade, the NTFPs has gained much importance and concern (Ticktin 2004; Gautam 2006). The growing awareness about NTFPs is not only for the role they play in the subsistence economy but also for their potential and a real contribution to the economies of many developing countries (FAO 1998).

Human over centuries has been obtaining a significant portion of their subsistence needs in the form of medicine, food, timber, and other useful products from these globally extracted NTFPs. NTFPs also include non–consumptive services to mankind, such as ecological/environmental, cultural and religious, and tourism and recreation services (Walter 1998). Mostly used NTFPs are fruits, nuts, seeds, spices, medicinal plants, and plant parts such as leaves, barks, resins, essential oils, latex, ornamental plants, rattan and bamboo (Ticktin 2004). Harvesting of NTFPs is considered to have a lesser impact on the forest ecosystem than timber harvesting. NTFPs can provide an array of social and economic benefits particularly to the local community and can, therefore, be an important component of forest ecosystem management (Duong 2008). In some rural hilly areas of Nepal, it contributes up to 50% of total annual family income (Gautam 2006). In the past, NTFPs was given secondary importance than timber. NTFPs were referred to as ‘minor’ because revenue generation from them as compared with timber was relatively low. But

The diversity of NTFPs is very high in Nepal. According to Ghimire et al. (2008b) nearly 2000 species of plants are considered to be potentially useful in Nepal. About 161 plant-based NTFP species are harvested from wild for trade in Nepal (Subedi 2006). In Nepal, NTFPs have great conservation and economic value (Gauli and

Hauser 2009). These resources are a key source of income and livelihood for many poor people in Nepal (Shackleton et al. 2011; Sunderlin et al. 2005).

Among different plant parts extracted as NTFPs, bark plays a crucial role to reduce the poverty of forest-dependent people, contributing to livelihoods and improvements in lifestyle, income, and healthy life (Sharma et al. 2017). Presence of higher concentrations of several elements makes the bark more useful, with higher metabolic activity than wood (Kenney et al. 1990; Adler et al. 2005). This is the reason why bark is widely used for various purposes. For example, bark fiber from coppiced branches of Tilia cordata is used in Europe to make cordage for fishing and trap construction (Myking et al. 2005). A bark of Betula species can be used as a covering in the making of canoes, as a drainage layer in roofs, and in the manufacture of shoes and backpacks (Adney and Howard 1964). Bark chips are used as mulch to protect soil moisture and nutrients (Adney and Howard 1964; Aronson et al. 2009). The inner bark of some species is also edible; for example, the Sami people of northern Europe use the inner bark of Scots pine (Pinus sylvestris) as a staple food (Zackrisson et al. 2000). Similarly, a bark of Cinnamomum tamala is used in Nepal as a condiment for flavoring food (Subedi and Sharma 2012). Daphne bark is the major raw material for popular Nepali handmade paper (Jeanraund 1984). According to FSRO (1984), the air–dry weight of Daphne bark mass in some districts like Baglung, Dolakha, Gorkha, Myagdi, Nuwakot, Parwat, and Lamjung were ranging from 6.13 to 25.9 kg/ha. Less number of mature populations might be the reason for lesser bark yield (FSRO 1983).

Nowadays, concerns have been expressed for the erosion and degradation of NTFP resources, unavailability of quality raw materials, high and fluctuating prices, improper marketing, and lack of organized cultivation and secretive nature of trade. But still, the share of NTFPs in the export market is significant (Gautam 2006). Recent years have witnessed various challenges and problems related to the sustainable management of NTFPs. Sustainable harvesting of NTFPs is an issue of concern for farmers, traders, middleman, planners and policymakers (Olsen and Larsen 2003). Over-harvesting due to trade pressure, livestock grazing/trampling, forest fire, and habitat destruction is responsible for the depletion of many plant species. Therefore, NTFP species in the face of these challenges need to be conserved and managed properly for their long-term use.

The policy and regulatory environment play a very significant role in all aspects of the trade of NTFPs. Few NTFPs policy in Nepal emphasized the potential role of NTFPs in alleviating poverty, enhancing economic growth and, improving natural resources management (HMGN 2004). Several legal provisions have been included within the Forest Act (1993) and Forest Regulation (1995) of Government of Nepal NTFP sector on behalf of which their use is regulated (GoN/MFSC 2007).

1.2 Daphne bholua Daphne L., a well-known genus comprising about 100 species distributed across Asia, Europe, and North Africa, is one of the largest genera of Thymelaeaceae (ca. 45 genera, 1,000 species; Rogers 2009). Species of Daphne are deciduous or evergreen subshrubs, shrubs or small trees, and are often cultivated as showy ornamentals (Brickell and Mathew 1976; Halda 2001). Daphne bholua locally known as Lokta, Baruwa in Nepal, an important NTFP that grows naturally between 1600 and 4000 m in Nepal (Ghimire and Nepal 2007; Khadgi et al. 2013; Sharma et al. 2017). It is an erect 1 to 3 m tall shrub. Out of five, two species of Daphne are common in Nepal, D. bholua and D. papyracea (Ghimire and Nepal 2007). Both species produce strong fibers in their bark, used for making handmade Nepali paper. Because of its higher fiber density, D. bholua is mostly preferred than Daphne papyracea (Paudyal 2004; Kharal et al. 2011; Khadgi et al. 2013).

Daphne bholua is commercially threatened species in Nepal. Conservation assessments in central Nepal based on the IUCN criteria for 153 woody plant species, assigned Daphne bholua under near threatened (NT) category (Adhikari et al. 2017). Other studies also revealed that Daphne bholua populations in different parts of Nepal are under threat of depletion (Ghimire et al. 2008; Adhikari et al. 2017). The natural status (national and global) of Daphne bholua is not known. The bark is collected from natural forests for trade with permission of District Forest Offices, paying the government a royalty of Rs.5 per kg (Pyakurel and Baniya 2011). Harvesting of Daphne bholua bark for local paper manufacturing has considerable potential for generating employment and providing income in hilly areas (ANSAB 2009). Along with domestic sales, handmade paper used to prepare various value-added products, which have strong outside Nepal (Banjara 2007; ITC and GoN 2016). The increasing

4 income and trade value of Daphne bholua has pushed this species towards vulnerability (Jeanrenaud 1984; Subedi 2006; Gurung 2007).

Nepali handmade paper is known for its unique quality, strength, durability, and resistance to insects (Jeanrenaud 1984) and different from those produced in China, India, the Philippines, and Thailand (ITC and GoN 2016). Enterprises engaged in this sector produce a diverse range of products and bulk is exported, directly or through visiting tourists. Handmade paper sector scores on several socioeconomic goals. The livelihood of an estimated 55,000 families depends on Daphne collection. About 300 registered small and medium-sized enterprises (SMEs) are producing handmade paper products both in remote mountain and urban areas in Nepal (NESS 2016). Approximately 80% of the entrepreneurs and workforce in this sector are women, and all the value addition are retained in the country (ITC and GoN 2016). Nepal Trade International Strategy (ITC and GoN 2016) identified handmade paper and paper products as priority sectors with high potential for both export development and contributing to inclusive growth. The traditional production technology used for papermaking is itself a unique selling point.

D. bholua is available in 2,910,848 hectares of forest in 55 districts of Nepal (FSRO 1984). Thirty districts are currently producing handmade paper in Nepal (GoN and ITC 2016). The most important handmade paper production districts are, Taplejung, Panchthar, Solukhumbhu, Sankhuwasabha, Terhathum, Okhaldhunga, Bhojpur, Khotang, Jhapa, Dolakha, Ramechhap, Sindhupalchok, Dhading, Gorkha, Rasuwa, Jumla, Kaski, Myagdi, Parbat, Baglung, Bajhang, Bajura, Baitadi, Rukum, Rolpa, Jajarkot, Dailekh and Achham. According to an estimate every year 110,481 ton of Lokta is available in Nepal from 2.91 million hectares of land and about 1,000 ton is collected annually producing 330 ton of paper (FSRO 1984). No similar survey has been conducted since then. The most recent data published by the Ministry of Forest and Soil Conservation (MoFSC) shows that Lokta collection is gradually decreasing in Nepal (MoFSC 2003, 2007, 2009, 2010, 2011). Also, Lokta has started to be collected illegally in national forests and thus does not appear in official statistics. There were a few successful attempts in Terhathum district to grow Lokta in farms. Local community farming of Lokta requires government intervention in terms of land allocation and technical support.

The major export destinations for Nepali paper and paper products are the United States, United Kingdom, France, Germany, Japan, Australia, Singapore, Netherlands, Italy, Denmark, Switzerland, and China. Exports are concentrated in a few destinations, with the United States representing 41.7 % and the first four countries representing 76.8 % of total exports from Nepal in 2015. Germany and Japan are other important markets for paper and paper products from Nepal (ITC GoN and 2016).

Increasing demand and unsustainable use have created pressure on the economically valued NTFPs species such as Daphne bholua. Thus, sustainable harvesting, processing, and value addition mechanisms of such NTFPs species need to be established.

Daphne bholua requires a special type of ecological conditions including specific plant association, which are being depleted by unsustainable bark harvesting. Undersized and immature plants are harvested in haphazard ways which lead to a decrease in quality of products and regeneration of Daphne species for future. This study aims to evaluate the conservation status of Daphne bholua with focus on its habitat characteristics, population performance and bark harvesting potential along an elevation gradient in Madane Protected Forest (MPF) in central Nepal.

1.3 Rationale The relationship between plant population performance and various environmental factors along the Himalayan gradient has been not well studied. During the past two decades, the utility of NTFPs has emerged with particular interest because their exploitation has been considered to be less destructive to the ecosystem than timber harvesting and other forest uses, and the potential income from NTFPs could be considerably higher.

Natural population of Daphne bholua are under threat of depletion due to unsustainable harvesting, trade pressure, livestock grazing/trampling, forest fire, and habitat destruction. Study pertaining to the habitat characteristics and bark potential are limited in Nepal and Madane Protected Forest Gulmi. There is uncertainty about the bark potential of D. bholua due to limited research on this important plant. Therefore, commercially threatened NTFPs facing these challenges need to be

6 conserved and managed properly for their long-term use. This study aims to study population performance and bark harvesting potential of understory shrub Daphne bholua along elevation and microhabitat gradient in Central Nepal.

1.4 Objectives This study is aimed to evaluate the habitat characteristics, population performance, and bark harvesting potential of Daphne bholua along an altitudinal and environmental gradient of in Madane Protected Forest Gulmi, Nepal.

The specific objectives are

 to assess the habitat characteristics, distribution pattern and population density of D. bholua along environmental gradients in MPF.

 to assess the main environment variables that govern the regeneration and population structure of D. bholua and

 to access its growth–related traits and bark harvesting potential along the elevational gradient in MPF.

CHAPTER TWO: MATERIALS AND METHODS

2.1 Study Area, Madane Protected Forest 2.1.1 Physiography Madane Protected Forest (MPF) is located in Gulmi district, federal-state 5, western Nepal. MPF feeds most of the lowermost area of Gulmi, Arghakanchi, Baglung and Pyuthan districts by water. Therefore, it is popularly known as the water tower of Lumbini Zone. MPF lies in a distance of 30 km (west) from the district headquarters, Tamghas. It covers an area of 13,761 ha and ranges in elevation from 975 m (basin of Ghamir) to 2690 m (peak of Hawangdii). MPF has been declared, recently in 2011, as a Protected Forest according to the provision under forest act 1993 of Government of Nepal (DoF 2011). It has been considered as an important habitat with high biodiversity and also holds high socio-cultural importance.

Majority of the users of MPF depend directly or indirectly on this forest for daily life (DoF 2011). A total of 56 community forests and 8 leasehold forests come under this protected forest. MPF has been classified as (i) fringe area (connected to the human settlement) (ii) protected or core zone and (iii) impact zone (nearby area outside the protected forest boundary). Most of the area in MPF is covered by forest (Table 1).

Table 1, Land use classification of Madane Protected Forest, in Gulmi district, Nepal (DoF 2011). Land use type Area (hectares) Percentage Dense forest 5315.12 38.62 Agriculture and human settlement area 7238.99 52.61 Grassland 40.59 0.29 Rivers and ponds 99.57 0.72 Open forest area 1066.73 7.75 Total 13,761 100

(i)

(ii) (iii)

Figure 1. Map showing study area inside Madane Protected Forest in Gulmi district, Nepal. (i) Map of Nepal showing Gulmi district, (ii) Map of Gulmi district showing the location of MPF, (iii) Map of Madane protected Forest (MPF) showing the study area.

2.1.2 Climate

The area has a wet summer and dry winter climate. Analysis of climatic data recorded by Department of Hydrology and Meteorology from 2007 to 2016 of the nearest meteorological station in the district headquarters Tamghas, showed mean annual precipitation of 1917.95 mm with the highest precipitation recorded in June to August (Figure 2). An average minimum temperature of 4.5°C was recorded in January and the maximum temperature of 27.87°C in June.

200 600 Av max temp 180 Av min temp 500 160 Av ppt

140 )

c 400 o 120

100 300

80 Temperature( 200 Precipitation(mm) 60

40 100 20

0 0 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Month

Figure 2. Twelve years (2007–2016) average minimum (Av min temp) and maximum temperatures (Av max temp) and average precipitation (Av ppt) recorded at the weather station in Gulmi (1450m asl), Gulmi, Nepal (Source Department of Hydrology and Meteorology, Government of Nepal, Kathmandu).

2.1.3 Flora and fauna

Madane protected forest supports subtropical to temperate vegetation. Major forest types found in MPF are Schima–Castanopsis and lower temperate oak forest (DoF 2011). Schima–Castanopsis forest occurs between 1600 m and 2000 m. At higher elevation, the forest is dominated by Quercus sp and Rhododendron sp. The major flora found in MPF are Castanopsis indica, Schima wallichii, Daphne bholua, Quercus lanata, Rhododendron arboreum, Pinus wallichiana, Prunus cerasoides, Alnus nepalensis, Engelhardtia spicata, Daphniphyllum himalayense, Myrica esculenta, and Lyonia ovalifolia. This forest also harbors important medicinal plants, such as Dactylorhiza hatagirea, Swertia nervosa, Bergenia ciliate and Paris polyphylla. Twenty–four species of wild animals are recorded in MPF (DoF 2011). MPF is regarded as a biological corridor for wild animals, such as Himalayan Thar (Hemitragus jemlahicus), barking deer, (Muntiacus muntajak), and common leopard (Panthera pardus) (DoF 2011).

2.1.4 Socio-culture and Economy

The total human population of MPF is 31,278 with 6,102 households, directly or indirectly depending on this forest (DoF 2011). The population density (people per sq. km) in MPF was reported at 227/sq. km. The Madane peak is religiously significant for Hindus, people from Gulmi and neighbor districts visit Malika Mandir, located at top of the Madane Peak, in every local festival. Major ethnicities are Brahmin, Kshetri, Magar, Thakuri, Sanyashi, and Dalit. Agriculture is the main occupation supplemented by small business and foreign employment. About 84% of people are engaged in agriculture (GoN 2011). The chief agricultural products are maize, rice, millet, and potatoes. Most of the people depend on the forest resources of MPF for livelihood.

2.2. Study Species Daphne bholua Buchanan–Hamilton ex D. Don, is a large shrub. Plants from the genus Daphne are small bushes or short trees with sparse branches (Halda 2003). Taxonomy of the genus Daphne is very complex and complicated because of the existence of a great number of species and subspecies (Halda 2003). About 95 species of Daphne have been recorded all over the world (Yinzheng et al. 2007). Four species and two varieties of Daphne have been reported from Nepal (Press et al. 2000). Species of Daphne are commonly called as Lokta in Nepali. It is also known by Lokato, Seto Lokta, Baruwa, Seto Baruwa, and Kagaje. But only two species are more common in Nepal, which are Daphne bholua Buch–Ham. ex D. Don, and Daphne papyracea Wall. ex G.Don (Ghimire et al. 2008). Both the species are major income generating non–timber forest products of Nepal (GoN 2009). Daphne bholua is differentiated into two varieties Daphne bholua Var bholua and Daphne bholua Var glacialis (Press et al. 2000).

The life span of both the species of Daphne bholua has been reported to be more than 60 years (NSCFP 2001). Daphne propagates both sexually and asexually. Seed viability is very short and often limited to less than a week; therefore, the plant is difficult to propagate by seeds (Ghimire and Nepal 2007). Wind and insects (bees, flies, and lepidoptera) are major means of pollination. The plant shows extensive vegetative propagation by suckers. These suckers can arise within the periphery of 5m from the taproot and more than eight suckers can arise from one adventitious root.

Regeneration occurs in the proportion of 25 percent by seeds and 75 percent by root suckers (Jeanrenaud 1984). Coppices arise from the cut stem and are less vigorous than plants that have originated from seeds or suckers (Jeanrenaud and Thompson 1986).

2.2.1. Taxonomy and morphology

The species found in the study area has been identified as Daphne bholua var. bholua Buch. –Ham.ex D. Don. Daphne bholua var bholua is an evergreen shrub, stem creeping or erect, 1–4 m tall sometimes up to 6 m. Branches are brownish, becoming dark brown suffused with purple, sparsely hirsute, soon glabrescent. Leaves are alternate, clustered at stem apex; leaf blade dull dark green, narrowly elliptic or oblong–lanceolate, thinly leathery, both surfaces glabrous, base broadly cuneate, margin sometimes slightly revolute and/or undulate, apex acute, rarely acuminate or obtuse. Inflorescences are terminal or axillary, 5–20 flowered; peduncle almost absent; bracts paired, caducous, broadly lanceolate or oblong-ovate, apex caudate. Flowers are very sweet-scented, borne in terminal rounded stalkless clusters. Calyx purplish red or red at least abaxially; tube cylindric, Stamens 8, filaments ca. 0.5 mm; anthers ca. 2 mm. Ovary cylindric–pyriform, ca. 4 mm, glabrous, shortly stipitate; style ca. 0.5 mm; stigma capitate. Drupe black, ovoid, Flowering January to March, fruiting April to May (Yinzheng et al. 2007).

2.2.2. Distribution In Nepal, Daphne bholua is found in temperate moist conifer and broadleaf forests from east to west between 1800 and 3600 m asl. As compared to western Nepal, the species extends to higher altitudes in eastern Nepal then in the western; where the annual precipitation is high and the timberline is also correspondingly higher (Jeanrenaud 1984). However, an altitudinal range for var. bholua has been reported between 2000 and 2900 m and for var. glacilis distribution range has been reported between 2100 and 3100 m altitude (Press et al 2000). Geographically, Daphne bholua var bholua is distributed in the Himalaya (Nepal to Arunachal Pardesh), North East India and West China; whereas, var. glacilis is endemic to Himalaya (Nepal, Sikkim) (Press et al. 2000). It flowers from February to March and then produces black fruits in April. Due to the masses of intoxicatingly fragrant flowers in winter, it is popular

12 as a garden flower in Europe and America since 1982 with the name of Daphne bholua “Jacqueline Postill” (Buffin 2005).

2.2.3. Habitat

Daphne bholua is the winter flowering shrub which blooms in response to reduced light levels, a period of cold or vernalization and in response to short day length (Buffin 2005). Daphne bholua generally grows as an understory shrub and a shade tolerant species. Being a shade demanding shrub Daphne bholua often grows gregariously in the moist conifer and broadleaf forests of the temperate Himalaya. It generally favors sites with medium to light crown–cover dominated by Oak (Quercus semecarpifolia), Rhododendron arboreum, Tsuga dumosa and Abies spp. (Jeanrenaud, 1984) and are found scattered or absent in open forest, pasture or in forest dominated by Pinus wallichiana, Cedrus deodara and Picea smithiana (Forestry services, 1984). The plant also grows well in quite deep shade (Wilson 1983). Daphne bholua survives a wide range of soil types but is often observed growing on moist acidic sites rich in organic humus layer with well-drained sandy loam soil (Ghimire et al. 2008a). It likes plenty of moisture in the growing season (Phillips 1989). In view of the bitterness and other chemical properties of the plant, it is not useful as cattle fodder nor can it be used as fuel, as burning it produces a bad smell and a lot of smoke (Mark 2007). It regenerates asexually through root suckers and sexually through seeds.

2.2.4. Use potential

Medicinal and Other Uses The fibrous bark is used to make fiber or cordage (Kunwar 2014). The plant is also used medicinally, for example, stem pith of Daphne bholua is used as purgative and febrifuge (CSIR 1986). Locally, bark decoction is also useful in fever. Root and bark juice is considered as anthelmintic and used to treat intestinal worms, to cure intestinal disorder as a result of disturbances due to spiritual power and as an anti- leech agent (Ghimire and Nepal 2007; Kunwar 2014). The extracts of the Daphne species have been shown to possess biologically active secondary metabolites, such as coumarins, biflavanoids and daphnane-type diterpene ester (Mark 1997). Phenolics from the flowers and buds have purgative, diuretic and expectorant effects (Nikaido et al. 1987), while a di–terpene compound isolated from

Daphne spp. is reported to exhibit anti-leukaemic property (Kasai et al. 1981). Daphne species contain toxic resins, which often cause nausea and nasal irritation to people involved in its harvesting (Agnihotri et al. 2010)

Papermaking The invention of paper can be regarded as the beginning of the knowledge era in human history. Even before the paper was invented, people drew objects and characters on cave, walls, rocks and dry leaves. It was believed that paper was first invented in China about 2000 years ago (Banjara 2007). Since the 12th century, traditional handmade Lokta paper has been produced in the hills of rural Nepal (Banjara 2007, Biggs 2009). Lokta paper has historically been used in Nepal for all Government documents and religious texts. Even today, most Nepalese have their birth certificate (Janmadarta) and land ownership papers (Lalpurja) written on handmade paper. The biggest demand for handmade paper previously came from Buddhist monasteries as they printed, wrote and drew teaching of Buddha on Lokta handmade paper. More recently, tourism has created a demand for Lokta paper and paper products, including stationery, lampshades, wallpaper, and wrapping paper. Many urban Nepalese now use greeting cards, visiting cards, and invitation card made from handmade paper. The modern handmade paper industry in Nepal started in the 1980s with the establishment of Bhaktapur craft Printers, a UNICEF supported project (Jeanrenaud, 1984). This company was created to provide employment opportunities for the people in Baglung, Parbat and Gorkha districts. The handmade paper industry in Nepal currently employs about 4000 families in rural areas in papermaking and another 2500 people in Kathmandu in paper product making (Banjara 2007).

2.3. Methods 2.3.1 Reconnaissance

Permissions to carry out this research were obtained from the Ministry of Forest and Soil Conservation, Department of Forest, Government of Nepal and from District Forest Office Gulmi (Appendix VII). The reconnaissance of the site was undertaken in the month of May 2017. During this period, suitable sites were identified through informal group discussions and observation of the area. Information on availability,

14 distribution, importance, harvesting, and use of NTFPs were collected through informal discussion with the officials of Madane Protected Forest, District Forest Office (Tamghas) and the local inhabitants. In MPF D. bholua was not harvested for commercial paper making but is locally used for making cordage. Considering the future contribution of D. bholua to local livelihood and national economy, the species was selected for ecological and biological study and resource assessment. This species will be a new source of income to local people in the study area. Both Daphne bholua and Daphne papyracea were found in the study area. D. papyracea was confined to lower elevation; whereas, D. bholua was found from 1900 m to 2500 m. After proper identification during the flowering period, we selected Daphne bholua var bholua for the study. The southern aspect was chosen for sampling because the northern aspect was inaccessible due to the steepness.

2.3.2 Population sampling

The field sampling was done in the month of January 2018. A systematic random sampling approach was used. The whole distribution range of D. bholua from 1900 to 2500 m was considered. Covering the wider habitat heterogeneity, the study area was stratified into three elevational sites, top (Malika 2300–2500 m), mid (Bhainse Mela 2100–2300 m) and low (Uttise 1900–2100 m). Detail characteristics of three sites are given in Table 2. Malika is located in the core area of forest and is far from the human settlement (4 hr. by walk); hence, characterized by very low anthropogenic disturbance, whereas the site Bhainse Mela lies at mid-elevation of the forest (2 hr. by walk) and receive moderate anthropogenic disturbance due to infrequent and irregular human visit. Uttise falls on the fringe area (Protected Forest near human settlement), which received high disturbance, where local resident visits frequently for collecting firewood, fruits, vegetables and to harvest fodder.

Table 2. Elevation, aspect, and slope of different sites in the study area

Study sites along Elevation m No. of plot Aspect (°) Slope (°) elevation gradient Top [Malika (MLK)] 2300–2500 18 95–155 SW 8–33 Mid [Bhaise mela (BHM)] 2100–2300 18 105–140 SE 11–34 Low [Uttise (UTS)] 1900–2100 18 185–260 SW 21–33

In each elevation band, three canopy categories were used as microsite variability (i) open (having <25% tree canopy coverage; assigned here as ‘open canopy’); (ii) semi– close (having >25% < 75% tree canopy coverage; assigned here as ‘semi-close canopy’); and (iii) close (having >75% tree canopy coverage; assigned here as ‘close canopy’) (Shrestha 2012). In each elevational site, three horizontal transects were made at 50 m elevation intervals. In each site, 18 plots (six in each open, semi–close and close canopy) of size 10 m x 10 m were randomly laid. Randomly two diagonal subplots (5 m x 5 m) were studied for recording plant density, and vegetative and reproductive characters of D. bholua (Figure 3). Similar method was used by Dutta and Devi (2013), to study the shrubs. Associated species of tree and shrub layer were also recorded from those subplots. The plots were at least 30 m distance from forest trails, maintaining inter plot distance of at least 20 m.

Figure 3. Sampling layout of 10 x 10 m plots divided into four subplots where the shaded ones were studied for the account of D. bholua.

2.3.3 Data collection

D. bholua shows extensive clonal growth supported by adventitious buds on the root (Herman and Kester 1976 in Jeanrenaud, 1984). Superficially, large plants appear to be a single individual, but on closer inspection it can be composed of several clones of

16 roots, suggesting clonality (Jeanrenaud, 1984). Due to the extensive clonality distinguishing individuals of D. bholua was almost impossible, thus we focused our study at ramet level.

Ramets of D. bholua were categorized under four size classes based on stem girth measured at 20 cm above ground level (Table 2). We assumed here, seedlings arise from both sexual and vegetative means (through root sucker and root collar) origin (Brune et al, 2003; Shrestha 2012). Adult plants were distinguished in two categories adult 1 (having girth 4 ≤ 8 cm), and adult 2 (having girt ≥ 8cm) (Table 3).

Table 3. Different life stages of D. bholua based on stem girth.

Life Stages Explanation Seedling Plant with <1 cm girth (arising sexually or vegetatively) Juvenile ramet Plant with 1 ≤ 4 cm girth Adult 1 Plant with 4 ≤ 8 cm girth Adult 2 Plant with ≥ 8cm girth Stem girth measured at 20 cm above ground, both sexual and vegetative origin (Ghimire and Nepal 2007)

Juvenile and adult plants occurring at each sampling subplots were measured/counted for its height, stem diameter, crown radius, number of main branches, number of coppice growth and number of floraloutputs (number of buds and flowers). The number of buds and flowers were averaged and calculated as total floral output per individual. Fruits did not observe during the filed work because the sampling was done prior to the fruiting season. For adults, bark thickness was measured to calculate the bark potential for papermaking. Stem diameter and bark thickness were measured with the help of a Vernier caliper by peeling the small section of bark from 20 cm above the ground and the stem height and crown radius with the help of meter tape. Altogether, 54 soil samples were collected from the center point of (10 × 10 m) plot from a depth of 15 cm using a soil digger. The soil samples were air dried in shade and stored in airtight plastic bags until laboratory analysis at Central Department of Botany Kirtipur, Kathmandu, Nepal. Soil samples were analyzed for pH, moisture and organic matter content by using standard protocol (Gupta 2002). Altogether, 108 plots were sampled in three sites. Topographical features, such as latitude, longitude, and altitude of each plot were recorded by Gramin etrex GPS and slope and aspect by

17 clinometer. Biophysical variables were recorded in the subplot which included the surface cover (%) of shrubs, herbs, grasses, lichens, bryophytes, bare ground, and litter. In addition, bryophyte and lichen on a rock (%) and under vascular plants were separately recorded in each subplot studied.

A human disturbance was recorded, in each subplot studied directly by observing animal droppings, grazing, trampling, harvesting, and fire. These were recorded as categorical variables starting from 0 to 4 (0 – no disturbance, and 4 – very high disturbance) (Kala and Dubey 2012).

In the two subplots studied in each plot, presence/absence of associated plant species was recorded. A voucher specimen of each species on the state of either flowering or fruiting or both were collected. Identification of those specimens was done by following standard literature (Press et al. 2000, Polunin and Stainton 1984, Stainton 1988, Shrestha 2018), expert consultation and comparing with specimen housed at TUCH and KATH. Herbarium specimens prepared are deposited at TUCH.

2.3.4 Laboratory analysis Thus, collected soil from the field was brought to the lab and moisture content was measured. The soil was air dried for a week in green house. Soil was grinded and sieved through sieve as per the requirement of the analysis. Soil moisture, soil organic carbon and soil pH were determined. Details of the methods used for the analysis are given below.

2.3.4.1 Soil moisture Ten gm fresh soil sample was taken in pre weighed crucible with lid and oven dried at 105°C for 48 hours. The samples were cooled and then weighed. Soil moisture was calculated as the percentage of fresh mass (Zoebel et al., 1987).

2.3.4.2 Soil pH For soil pH, ten grams of soil was taken into a 100ml beaker and 25 ml distilled water was added to it. Thus, prepared solution was stirred for 5 minutes and kept for half an

18 hour. Finally, pH of the solution was measured using a pocket pH meter with accuracy of ± 1.

2.3.4.3 Soil Organic Carbon (SOC) Soil Organic Carbon was estimated by using standard Walkely and Black method following Gupta (2002). During the procedure in few of the samples since the amount of organic carbon was higher, the amount of soil used for the measurement was reduced to half than the amount stated in the method. For organic carbon, 0.5 g of soil was taken into conical flasks of 250 ml and 5 ml of K2Cr2O7 was added to it with a pipette. Then 10 ml of conc. H2SO4 was added to the mixture, stirred for a while and kept for 30 minutes for digestion. After 30 minutes, 100 ml of distilled water was added to the mixture and 5ml of phosphoric acid was added to it. Then 0.5ml of diphenyl amine indicator was added to the mixture. The color of mixture turns blue violet. Then the mixture was titrated with ferrous ammonium sulphate till the blue violet color changes to green and the amount of ferrous ammonium sulphate consumed by the soil sample was noted. Similar process was repeated for a blank solution without soil as control. Amount of carbon present in soil was calculated by using following formula:

Where,

N= normality of ferrous ammonium sulphate

Finally, Soil organic carbon was calculated by multiplying estimated organic carbon by a factor 1.3 based on assumption that there is 77% recovery of organic matter in this procedure and Organic Carbon measure was obtained as:

Organic carbon = estimated organic carbon % × 1.3

2.3.5 Data analysis

Habitat characteristics of Daphne bholua were evaluated in terms of variation in physical/topographic variables and by analyzing patterns of associated species diversity and composition along the elevation gradient. Both the parametric and non–

19 parametric statistical tests were performed to analyze the data. Non–parametric tests were performed if the data (even after transformation) did not meet normality and homogeneity of variance. One–way analysis of variance (ANOVA) was used when the data met assumptions of the parametric test (i.e., normal distribution and homogeneity of variance) to detect differences in the subset of habitat variables and plant traits among three sites (Underwood, 1997). The effect of elevation and tree canopy cover were tested separately for three sites through nested ANOVA using elevation as the main effect (fixed factor) and canopy nested within elevation as the nested factor (Burne 2003), Tukey’s HSD test at 5% level of significance was used for multiple comparisons.

2.3.5.1 Habitat association In order to understand the relationship between the species and environment, Detrended correspondence analysis (DCA), an indirect gradient analysis, was carried out. It was used to study the distribution of sample units and species in environmental space (Šmilauer 2003). The value of DCA first axis was estimated to be 2.477. Redundancy analysis (RDA) was employed to explain the species–environment relationship. RDA is a direct gradient analysis (ter Brank 1986) used to access the relationship between the species and the environment. In RDA, different environmental parameters such as elevation, slope, disturbance, litter cover, rock and scree cover, and tree canopy cover, was carried out to understand the relationship between the species and environment. Shrub cover, forbs cover, grass cover, bryophyte below vascular plants, lichen below vascular plants were not used because they were confounding variables, highly correlated with tree cover. Further, their effect on species composition was not significant, as tree cover can influence under canopy species. CANOCO version 4.5 (ter. Braak and Smilauer 2002) was used to perform DCA and RDA ordinations.

2.3.4.2 Density and size distribution Density and population structure (the relative proportions of seedling, young sprout, juvenile, and adult to total density) of Daphne bholua were analyzed for each elevation band and three canopy types (open, semi-close close canopy). Variation in population density among elevation bands and canopy types was compared using one–way ANOVA. Multiple comparisons of the proportion of seedling, young

20 sprouts, juvenile and adult life stages in three different elevational sites were done using one–way ANOVA.

2.3.4.3 Vegetative and reproductive trait variation The vegetative (adult plant height, adult stem diameter, adult bark thickness, adult crown radius and number of main branches per plant) and reproductive (number of flowers and buds as total reproductive output per individual) traits were computed among three sites using one–way ANOVA. Similarly, ms excel 2016, IBM SPSS statistics version 20 (Underwood, 1997) and CANOCO for windows 4.5 (ter. Braak and Smilauer 2002) were used throughout the process of data analysis.

2.3.4.4 Dry mass of inner bark The dry mass of inner bark (bast) of Daphne bholua was calculated by using the following regression equation (Ghimire and Nepal 2007).

LnY= 2.165+2.052LnD20 Where Y is the total dry mass of the inner bark of Daphne

D20 is the diameter of Daphne at height 20 cm above the ground Using this formula total dry bark mass was estimated from all adult plants (adult 1 and adult 2) having height ≥ 50 cm, and having girth > 4cm. Whereas, the harvestable dry mass of inner bark was calculated from the adult 2 only having girth more than 8 cm (Ghimire and Nepal 2007; Khadgi et al. 2013).

CHAPTER THREE: RESULTS

3.1 Habitat Characteristics of Daphne bholua 3.1.1 Biophysical (substrate) variables

The environmental variables (mean ± SE) recorded in plots along the elevational sites are given in Table 4. The results showed that plots sampled in Uttise were highly disturbed (having a higher percentage of plots with a higher level of disturbance) and disturbance decreased as the elevation increased from low to top elevational sites (Appendix I). The sampling plots in three sites did not differ in terms of tree canopy cover (Kruskal–Wallis test χ2 = 1.3, p = 0.5) and shrub cover (Kruskal–Wallis test χ2 = 2.09, p = 0.35). The three–sites differed in terms of ground vegetation cover (Table 4). The Bhainse Mela site was found with a significantly high percentage of bryophyte below vascular plant (30.27 ± 1.32), and bryophyte on a rock (18.83 ± 1.11) as compared to the other sites. Similarly, the Malika was found with a significantly high amount of litter cover (46.11 ± 2.88). However, litter depth (5.33 ± 0.16 cm) and litter cover (36.88 ± 2.07) were low in Uttise (Table 4). In contrary, bryophyte on rocks (19.52 ± 1.09) and grass cover (16.55 ± 1.36) were significantly high in Uttise. The total forb cover was found to be almost similar in three sites.

The correlation between different biophysical variables are given in Appendix I. Litter cover, bryophyte on vascular plant and rock cover were positively correlated with elevation; whereas, disturbances, lichen on rock and grass cover were negatively correlated with elevation. Tree cover was positively correlated with litter cover and negatively correlated with grass cover, grazing, and trampling. Forb cover was positively correlated with grass cover and grazing and trampling, and negatively with litter cover and bryophyte on the soil. Litter cover, litter depth and bryophyte on soil were negatively correlated with disturbance component; whereas, positively correlated with forbs cover, bryophyte on a vascular plant, and lichen on the rock (Appendix I).

The substrate of Daphne was slightly acidic and nearly uniform in all three sites (overall pH 5.3 ± 0.3). The elevation–wise analysis of soil organic matter showed higher value in Malika and Bhainse Mela (2.10 ± 0.97, 2.11 ± 0.87) respectively as

22 compared to Utttise (1.40 ± 0.72). Similar, the trend for soil moisture content (Table 4)

Table 4. Biophysical (substrate) variables (mean ± SE) recorded in three different elevation bands in MPF.

Biophysical variables Malika (High) Bhainse Mela (Mid) Uttise (Low) Overall mean χ2

b c Elevation (m) 2485.61 ± 4.71a 2347.61 ± 6.78 2036 ± 9.12 2289.74 ± 18.62 95.16** (2300–2500) m (2100–2300) m (1900–2100) m

Slope (°) 41 ± 2.41a 39.11 ± 1.79a 49.05 ± 1.28b 43.05 ± 1.15 17.62**

(95 – 155) SE (105 – 140) SE (185 – 260) SW

Tree cover (%) 57.94 ± 4.76a 56.94 ± 5.30a 53.88 ± 4.5a 56.25 ± 29.0 1.30#

Litter depth (cm) 9.86 ±0.46a 10.13 ± 0.46a 5.33 ± 0.16b 8.44 ± 0.31 60.16**

Forbs cover (%) 12.00 ± 2.16a 15.66 ± 2.12a 19.30 ± 1.12b 15.65 ± 1.10 18.80**

Grass cover (%) 8.86 ± 0.91a 11.27 ± 1.50a 16.55 ± 1.36 b 12.23 ± 0.79 14.44**

Shrub cover (%) 27.27 ± 2.92a 21.77 ± 1.98a 22.72 ± 1.99a 23.92 ± 14.13 2.09#

Litter cover (%) 46.11 ± 2.88a 42.86 ± 3.32a 36.88 ± 2.07ab 41.95 ± 1.64 7.79*

Lichen on rock (%) 7.30 ± 0.66 a 12.55 ± 1.05b 15.02 ± 1.31b 11.62 ± 0.673 23.94**

Bryophyte on rock (%) 17.25 ± 2.31a 18.83 ± 1.11b 19.52 ± 1.09b 18.53 ± 0.92 6.18*

Bryophyte on VP* (%) 28.63 ± 0.82a 30.27 ± 1.32a 24.25 ± 1.52b 27.72 ± 0.76 12.63*

Soil pH 5.16 ± 0.59a 5.29 ± 0.56b 5.50 ± 0.4c 5.3 ± 0.3 20.44**

Soil organic matter (%) 2.10 ± 0.97a 2.10 ± 0.87b 1.40 ± 0.72c 1.86 ± 0.61 35.91**

Soil moisture (%) 13.26 ± 0.27a 11.07 ± 0.20b 10.18 ± 0.32 c 11.50 ± 2.0 42.85** Values associated with same superscript letter are not statistically significant (multiple comparison based on Mann–Whitney U test). (for overall habitat variables, n = 108, and for soil variables, n = 54) ** –significant at p < 0.001, * – significant at P < 0.05, # – non–significant. VP* – Vascular plant

3.1.2 Associated species composition

Altogether, 107 plant species (belonging to 57 families and 98 genera), associated with Daphne bholua were identified in the study area (Appendix III). Asteraceae was the dominant family which comprised of seven species, followed by Ericaceae (5 species belonging to 3 genera), Lamiaceae (5 species belonging 3 genera), Lauraceae (4 species belonging 2 genera), Rosaceae (4 species belonging to 2 genera) and Fagaceae (3 species belonging to 2 genera). Among the species associated with Daphne bholua, combining all plots, Rhododendron arboreum (85.18%), Quercus semecarpifolia (81.48%), Quercus lanata (55.55%), Symplocos theifolia (55.55%), Lindera pulcherrima (51.85%), Polystichum aculeatum (48.14%) Wikstroemia canescens (42.59%), and Lyonia ovalifolia (40.74%) showed high frequency of occurrence (Appendix III).

The RDA ordination explained the relationship between samples, species and environment variables. Forward selection and Monte Carlo permutation tests revealed that disturbance was the only significant variable (F=2.19, p =0.003) governing the species composition (Table 5). Summary of the Monte Carlo permutations tests showing the relative importance of environmental variables on the species composition is given in Table 5.

Table 5. The relative importance of environmental variables on species composition based on RDA analysis. The statistically significant (P<0.05) variables were obtained using Monte Carlo test with 9999 permutations.

Environmental variables Abbreviation F p Disturbance dis_Tram 2.19 0.0027 Slope SLP 1.516 0.0642 Rock and scree RCK 1.222 0.1883 Litter cover Lt_cov 1.045 0.3738 Tree cover TR 0.971 0.4857 Elevation ELE 0.737 0.6924

Table 6. Summary of RDA ordination (total inertia = 3.894, sum of all canonical eigen values = 0.999).

Axes Parameters 1 2 3 4 Eigenvalues 0.107 0.044 0.021 0.017 Species–environment correlations 0.955 0.627 0.787 0.767 Cumulative percentage variance of species data 10.7 15.1 17.2 18.9 of species–environment relation, 38.6 54.4 61.8 67.9

The DCA ordination calculated a lower gradient length (2.477 SD units) for the DCA first axis, indicating the linear relationship among species along the main gradient. The RDA ordination clearly explained that the axis I was correlated with disturbance (extracted as PCA component) which explains about 38.6 variances in the species– environment data and 10.7 % in the species data.

Axis II Axis

Axis I

Figure 4. RDA biplots for sample plots and environmental variables; numerical values denote the plot number, (Elev = elevation, Tr_can = tree cover, Ltr = litter cover, Slp = slope, Cop = number of coppices, Dist = disturbance, Rck_scr = Rock and scree, Dap_den = Density of Daphne

The RDA second axis represented a gradient related to topographical and habitat variables such as tree canopy, elevation, and rock scree. The second axis explained 15.1 variances in species data. The eigen–value for RDA second axis was comparatively lower (0.044) than the first axis (Table 6).

Axis II

Axis I

Figure 5. RDA biplots for species and biophysical variables (Elev = elevation, Tr_can = tree canopy cover, Ltr = litter cover, Slp = slope, Cop = number of coppice, Dist = disturbance, Rck_scr = Rock and scree, Dap_den = Density of Daphne. abbreviation of species is given in Appendix III).

The sample by environment ordination diagram (Figure 4) showed that the most sampling plots from Uttise fall towards highly disturbed sites (plots 37 to 54) where the coppice growth was also seen along the disturbance gradient which can be seen towards the positive right–hand side of the diagram (RDA first axis), indicating high harvesting and low density of Daphne bholua (Figure 4). The most samples of Malika and Bhanise Mela tended to distribute along the less disturbed area and in higher elevation along with the high litter cover (RDA second axis) (Figure 4).

Species, such as Hedyotis scandens (Hed_sca), Tetrastigma serrulatum (Tet_ser), Polystichum aculeatum (Pol_acu) Thalictrum punduanum (Tha_pun), Garuga pinnata (Gar_pin) Imperata cylindrica (Imp_cyl), Flemingia strobilifera (Fle_str), and Pogostemon benghalensis (Pog_ben) seemed to be evenly distributed along the gradient. (Figure 5). Species, like Dichroa fabrifuga (Dic_fab), Ageratina adenophora (Age_ade), Leucosceptum canum (Leu_can) and Vibernum erubescens (Vib_eru), Viola biflora (Vio_bif), Castanopsis tribuloides (Cas_tri) and Plantago

26 depressa (Pla_dep), preferred plots along the higher disturbance level where the density of Daphne was low (Figure 5).

The density of different life stages of D. bholua was found higher in the plots where the species such as Rhododendron arboreum (Rho_arb), Quercus lanata (Que_lan), Quercus semecarpifolia (Que_sem), Wikstroemia canascens (Wik_can) and Pieris formosa (Pie_for) were found. In contrast, plants such as Anaphilis triplinervis (Ana_tri), Schima wallichii (Sci_wal), Castanopsis indica (Cas_ind), Castanopsis tribuloides (Cas_tri), Osyris wightiana (Osy_wig) and Pteris wallichiana (Pte_wal) showed negative association with the Daphne bholua (Figure 5).

3.2 Variation in Density and Size Class Distribution of Daphne Total plant density of different life stages varied widely among the three different elevational sites. The density of seedlings, juvenile, adult 1 and adult 2 individuals in the entire study area was found to be (mean ± SE) 1.75 ± 0.35, 0.94 ± 0.77, 0.85 ± 1.24 and 0.83 ± 1.22 individuals per 25 m2 plot respectively (Table 7). Total density was highest in Malika and lowest in Uttise. The density of seedling and juvenile was high in Malika and Bhainse Mela than that of Uttise. The density of adult plants both (adult1 and adult 2) were highest in Bhainse Mela (mid-elevation site) and Malika and lowest in Uttise.

Table 7. Density (number of individuals per 25 m2 of different size classes; (seedling, Juvenile, Adult 1 and Adult 2) of D. bholua in three elevation sites in MPF.

Malika Bhainse Mela Uttise Life stages/ Sites F p (high) (mid) (low) Seedling 2.72 ± 0.52a 2.08 ± 0.32a 0.47 ± 1.16b 10.27 <0.001 Juvenile 1.22 ± 0.12a 1.11 ± 0.13a 0.5 ± 0.93b 10.83 <0.001 Adult 1 1.08 ± 0.24a 1.22 ± 0.22a 0.25 ± 0.08b 7.71 0.001 Adult 2 1.11 ± 0.21a 1.19 ± 0.24a 0.19 ± 0.06b 8.38 <0.001

Total 6.13 ± 0.67a 5.61 ± 0.31a 1.33 ± 0.18b 35.34 <0.001 F and p values based on one–way ANOVA, followed by Tukey HSD multiple comparison test. Values associated with same superscript letter were not statistically significant (p>0.05). n = 108 for all life stages.

In Malika and Bhainse Mela, the density of all life stages was highest in close canopy plots, followed by a semi-close and lowest in open canopy plots (Table 8). On the contrary, in Uttise the density of all life stages was highest in semi-close canopy type and lowest in open canopy type. The close canopy contains a higher density of seedlings in Malika and Bhainse Mela; whereas, semi-close favors more seedlings in Uttise (Appendix IV). Adult density was high in semi-close and close canopy in all the plots from three sites (Appendix IV).

Table 8. Total density (number of individuals per 25 m2) of Daphne bholua in plots from three canopy coverage types in MPF.

Malika Bhainse Mela Uttise Canopy cover types Overall (High) (Mid) (Low) Open (<25%) 4.50 ± 0.48a 4.58 ± 0.49a 1.07 ± 0.21b 3.33 ± 0.36

Semi-close (25 <75%) 6.66 ± 0.96a 5.91 ± 0.46a 1.75 ± 0.35a 4.97 ± 0.48

Close (>75%) 7.20 ± 1.68a 6.33 ± 0.55a 1.33 ± 0.37a 4.77 ± 0.75

Overall average 6.13 ± 0.67 5.61 ± 0.31 1.33 ± 0.18 Values associated with same superscript letter were not statistically significant (p>0.05). n = 108.

Total D. bholua density (no. of individual per 25 m2) was highest in the semi–close canopy (4.97 ± 0.48) and lowest in open canopy (3.33 ± 0.30) type (Table 8). The density of seedling and juvenile were high in close and semi–close canopy whereas, adult 1 and adult 2 were high in the semi–close canopy (Table 9, AppendV).

Table 9. Density (number of individuals per 25 m2) of different size classes; (seedlings, juveniles, adult 1 and adult 2) in different canopy coverage type.

Open Semi-close Close Life stages/Canopy F p (<25%) (25<75%) (>75%) Seedling 1.16 ± 0.20a 1.83 ± 0.32a 2.27 ± 0.55a 2.07 0.13 Juvenile 0.80 ± 0.11a 1.13 ± 0.12 a 0.88 ± 1.47a 1.85 0.16

Adult 1 0.83 ± 0.18a 0.88 ± 0.22a 0.83 ± 0.21a 0.02 0.97

Adult 2 0.52 ± 0.61a 1.1 ± 0.23a 0.80 ± 0.21a 2.52 0.08

Total 3.33 ± 0.36a 4.97 ± 0.48a 4.77 ± 0.75a 2.55 0.08 F and p values based on one–way ANOVA, followed by Tukey HSD multiple comparison test. Values associated with same superscript letter were not statistically significant (p>0.05). n = 108.

The result of nested ANOVA revealed significant effects of elevation in D. bholua density (Table 10). Canopy, on the other hand, had a significant effect only on total plant density (F = 4.69, p = 0.011) (Table 10).

Table 10. Results of nested ANOVA showing the effects of elevation (fixed factor) and canopy (nested with elevation) on plant traits of Daphne in three elevational bands.

Variables Factors Df F P Total Daphne density Elevation 2 36.74 <0.001 Canopy 2 4.69 0.011 Elevation (canopy) 4 0.95 0.435 Seedling density Elevation 2 10.57 <0.001 canopy 2 2.44 0.092 Elevation(canopy) 4 1.91 0.113 Juvenile density Elevation 2 9.97 0.001 Canopy 2 1.58 0.209 Elevation (canopy) 4 0.45 0.771 Adult 1 density Elevation 2 6.92 <0.002 canopy 2 0.02 0.975 Elevation(canopy) 4 0.29 0.881 Adult 2 density Elevation 2 7.75 <0.001 canopy 2 2.21 0.115 Elevation(canopy) 4 0.70 0.589 df, F and p value based on nested ANOVA

3.3. Population Structure The overall elevation wise proportion of seedlings, juvenile, adult 1 and adult 2 of Daphne bholua in all plots were 0.389, 0.257, 0.264, and 0.193 respectively (Figure 6). Hence, size class distribution of D. bholua plants showed almost reverse ‘J’ shaped pattern with the proportions of seedling exceeding the proportions of juvenile and adult stages in all the plots from Malika and Bhainse Mela. (Figure 6 d, h). Uttise had nearly bell-shaped pattern characterized by a dominance of intermediate-sized plants, with few juveniles and few adults (Figure 6; i, j, k, l).

The overall population proportion in three different canopy coverage also showed reverse ‘J’ shaped pattern in open and semi–close canopy (Figure 6; m, n). In close canopy type, the proportions of seedlings and adult 1 were higher than juvenile and adult 2 (Figure 6; o).

The proportion of seedlings were very high in compared to other life stages in close canopy except in Uttise (Figure 6; c, g, k, o). The proportion of adult 2 was comparatively lower than adult 1 in open canopy from all sites (Figure 6; a, e, i, m).

Figure 6. Population proportion of D. bholua at three different study sites (i) MLK= Malika, (ii) BHM= Bhainse Mela, (iii) UTS= Uttise with different canopy coverage type (open, semi–close, close) and overall structure combining all canopy types for each site.

3.4 Variation in Growth–Related Traits The three study sites differed in terms of seven out of eight growth–related traits studied. Total floral output (flower + bud) per individual did not vary significantly (F = 2.17, p = 0.120) (Table 11).

Table 11. Growth–related traits of D. bholua in Three sites. Values shown are mean ± SE.

Growth–related traits Malika (High) Bhainse Mela (Mid) Uttise (Low) F P

Plant height (cm) 139.37 ± 10.97a 123.80 ± 5.17a 58.94 ± 3.77b 33.81 <0.001

Crown radius (cm) 51.76 ± 3.97a 45.50 ± 2.71ab 36.26 ± 1.69a 7.02 <0.001 Stem diameter at 20 2.96 ± 0.27a 2.12 ± 0.11b 1.67 ± 0.22b 9.28 <0.001 cm height (mm)

No. of branch per plant 12.08 ± 1.22a 7.61 ± 0.45b 6.55 ± 0.07b 23.65 <0.001 Floral output 110.44 ± 12.76a 99.27 ± 8.79a 82.47 ± 5.85a 2.17 0.120 (no. of flowers + buds) Dry bark mass 3.23 ± 0.19a 2.71 ± 0.08b 1.136 ± 0.16c 50.69 <0.001 gm per plant

Bark thickness (mm) 2.25 ± 0.17a 1.75 ± 0.11a 0.867 ± 0.10b 22.49 <0.001

Coppice growth (per plant) 0.83 ± 0.2a 0.22 ± 0.15a 3.51 ± 0.30b 13.82 <0.001 Value of F and p based on one–way ANOVA. Data were log transformed to meet the normality and homogeneity of variance. Values associated with the same superscript letter are not statistically significant (p>0.05). Multiple comparisons based on Tukey HSD test. n = 108 for all growth-related traits.

However, growth traits, like adult plant height, number of branches per plant, bark thickness, dry bark mass, stem diameter, and crown radius varied significantly among the three sites. Plant height of D. bholua was high in Malika (139.37 ± 10.97 cm), followed by Bahinse Mela (123.80 ± 5.17 cm) and Uttise (58.94 ± 3.77 cm). Stem diameter, bark thickness, and dry bark mass were lower in Uttise than those in Malika and Bhainse Mela. The plants in the harvested site (i.e, Uttise) were observed with profuse coppice growth (1.50 ± 0.30 per plant) as compared to least or no harvested site; i.e, Bhainse Mela (0.22 ± 0.15 per plant) and Malika (0.83 ± 0.20 per plant). Malika (3.23 ± 0.19 gm per plant) contained more than double dry bark mass than that of Uttise (1.136 ± 0.16 gm per plant) (Table 11). Hence, D. bholua showed low growth performance in Uttise in terms of all growth parameters studied; but it showed high growth performance in Malika and Bhainse Mela (Table 11).

Table 12. The result of nested ANOVA showing the effects of elevation (fixed factor) and canopy (nested with elevation) on plant traits of D bholua in three elevational sites.

Variables Factors Df F p Plant Height (cm) Elevation 2 35.27 <0.001 canopy 2 2.94 0.050 Elevation(canopy) 4 1.82 0.110 No. of branches/plant Elevation 2 23.05 <0.001 canopy 2 1.22 0.290 Elevation(canopy) 4 1.94 0.940 Crown Radius (cm) Elevation 2 7.27 0.010 canopy 2 0.12 0.880 Elevation(canopy) 4 2.12 0.080 Stem Diameter (mm) Elevation 2 8.58 <0.001 canopy 2 0.18 0.830 Elevation(canopy) 4 0.58 0.670 Bark–thickness (mm) Elevation 2 22.43 <0.001 Canopy 2 0.7 0.290 Elevation(canopy) 4 1.13 0.940 Dry bark mass gm per plant Elevation 2 43.55 <0.001 canopy 2 0.74 0.420 Elevation(canopy) 4 0.43 0.730 Total floral output per plant Elevation 2 2.12 0.120 (flower + bud) canopy 2 3.59 0.030 Elevation(canopy) 4 1.24 0.290 df, F and p–value based on nested ANOVA

Elevation also proved itself as the most important variable in influencing the growth of D. bholua revealing its significance with most growth-related traits (Table 12). Plant height (F=35.27, p = 0.001), stem diameter (F = 8.58, p = 0.001), and bark thickness (F=22.43, p= 0.001) were greatly varied with elevation (Table 12). Similarly, tree canopy covers significantly influenced plant height (F = 35.27, p = 0.001) the total floral output (flower + bud) of D. bholua in the study area (F = 3.59, p = 0.03) (Table 9). However, stem diameter, bark thickness, and total dry bark mass failed to exhibit any such relationship with tree canopy cover. No significant effect was seen when the canopy was nested with elevation (Table 12).

Table 13, Study of growth–related traits according to different canopy coverage types. Values shown are mean ± SE.

Growth–related traits Open (< 25%) Semi–close (25 <75%) Close (>75%)

Plant height (cm) 102 ± 7.2 124 ± 11.2 95 ± 8.4

Crown radius (cm) 43.7 ± 3.41 44.5 ± 2.7 45.2 ± 3.1

Stem diameter (mm) 2.2 ± 0.2 2.3 ± 0.1 2.2 ± 0.2

No. of branch per plant 9.9 ± 0.9 8.61 ± 0.8 7.7 ± 0.8

Floral output per plant 92.5 ± 11.3 116.9 ± 10.2 82.6 ± 5.8

Dry bark mass (gm per plant) 2.3 ± 0.2 2.5 ± 0.2 2.2 ± 0.1

Bark thickness mm 1.5 ± 0.1 1.9 ± 0.1 1.6 ± 0.2

Coppice growth per plant 1.8 ± 0.5 1.0 ± 0.3 1.8 ± 0.5 n = 108 for all growth-related traits.

Growth–related traits in accordance with three canopy types showed that semi–close canopy preferred most of the growth traits, i.e, plant height, stem diameter, total floral output bark thickness and dry bark mass (Table 13, Appendix V). Crown radius and stem diameter of D. bholua did not vary in different canopy coverage types (Table 13).

3.5 Total and Harvestable Dry Bark Mass The average dry bark mass of total (adult 1 and adult 2); and harvestable (adult 2; having girth >8cm); D. bholua was 71 kg/ha and 56 kg/ha respectively (Figure 7). Total inner bark mass of adult D. bholua was highest in Malika (40 kg/ha) and lowest in Uttise (12 kg/ha). Similarly, harvestable dry bark mass (adult 2) was also high in Malika (36 kg/ha) followed by Bhainse Mela (14 kg/ha) and Uttise (7 kg/ha) (Figure 7).

Figure 7. Total and harvestable dry bark mass of Daphne bholua along three study sites.

Overall inner bark mass of adult Daphne bholua (adult 1 and adult 2) and harvestable (adult 2) was high in semi-close canopy i.e., 28 kg/ha and 23 kg/ha respectively (Figure 8). Open and close canopy contained a similar mass of total dry bark mass (Figure 10). In Bhainse Mela, the total dry mass was similar in all three canopy types. In case of Malika and Uttise, semi–close canopy contained high total and harvestable bark mass. (Figure 8).

Figure 8. Total and harvestable dry bark mass of Daphne bholua along three study sites with different canopy coverage (Open, semi–close, and close)

CHAPTER FOUR: DISCUSSION

4.1 Habitat characteristics of D. bholua Habitat conditions and quality are important regulatory factors and plant populations of understory shrub species (Brosofske et al. 2001; Volis et al. 2005; Schroeer et al. 2008) and variation. Habitat characteristics can influence species composition, interaction, and for long–term persistence (Pavolvic 1994; Murphy and Lovett–Doust 2004; Young et al. 1996). However, the response of plant towards habitat conditions vary depending on the life history strategy, life span, population distribution, and size of the targeted species (Pavlovic 1994; Brys et al. 2005; Yamada et al. 2007; Ghimire et al. 2005).

D. bholua is shade tolerant, understory shrub prefers moist places with low biotic disturbances, the presence of tree falls gap with relatively low litter accumulation (Ghimire and Nepal 2007, Khadgi et al. 2013 Chapagain and Rai 2014). It prefers semi–close and close canopy (Table 7) in the southeast to southwest facing slopes in MPF from 1900–2500 m. The associated habitat species of Malika (2300–2500 m) and Bhainse Mela (2100–2300 m) site were Rhododendron arboreum, Quercus semicarpifolia, Quercus lanata, Pieris formosa, Wikstroemia canascens, and Lyonia ovalifolia. A similar association was found on previous studies (Jeanrenaud 1984, Dutt 1994, Ghimire and Nepal 2007, Pyakurel and Baniya 2011; Khadgi et al 2013). But in Uttise (1900–2100 m) the associated species were different from upper two sites which were Lindera pulchhirema, Symplocos theifolia and Gaultheria fraganentisima (Figures 4,5). Whereas, D. bholua did not prefer to grow with the association of Castanopsis tribuloides, Vibernum erubescens, Rubus elipticus, Dichroa fabrifuga, Budleja asiatica (Figures 4,5).

The density of Daphne stand is dependent on several ecological and biotic factors; highest densities occurring between 2100 and 2800 m on north-facing slopes (FSRO 1984, Amatya Forestry services 1983). On east and west facing slopes, plant exhibits stands of medium density whereas in drier south-facing slopes usually displays a scattered population pattern (Jeanrenaud 1984). The increased plant density of Daphne of all life stages towards higher elevation (Malika and Bhainse Mela) as compared to a lower elevation (Uttise) in MPF might have been influenced by the substrate type (Figure 6, Table 7). Lower elevation in the study area was dominated

37 by rocky and sloppy substrate type. Also, the population of those areas were affected by distance to the settlement walking trails towards Malika Mandir and Ashram. Therefore, high anthropogenic disturbance and rocky and sloppy habitat towards lower elevation support a fewer number of D. bholua plant compared to highly disturbed, open southwest facing slopes of MPF. It is also due to that the lack of good association and vegetation types which favors the proper growth (Figure 6, Table 7).

D.bholua appears to thrive on a wide range of soil types but generally favors moist sites with rich organic humus layer overlying well-drained sandy loams or brown earth (Jeanrenaud 1984; Pyakurel and Baniya 2011; Khadgi et al. 2013). The overall enhanced growth influenced by elevation and can probably be attributed to various parameters, such as disturbance, temperature, and soil nutrient availability which changes along with elevation (Korner 1999; Lomolino 2001; Bhatterai et al. 2004). The value of physiochemical parameters varied along the elevation gradient. Soil organic carbon is the main terrestrial carbon pool (Batjes 1996) which increase the water holding capacity and availability of nutrients (Salter and Williams 1969). The amount of soil organic carbon in present study was high and similar in Malika and Bhainse mela than Uttise (Table 4). Dense forest contributes a lot of organic matter to the soil in the form of leaves, twigs, stems, flowers, and fruits which after decomposition result in the formation of organic carbon and release different nutrients (Sharma and Sharma 2004 as cited by Iqbal et al. (2013). The amount of leaf litterfall was more in Malika and Bhainse mela than in Uttise. The highest value of SOC was found in Malika which showed that elevation had a direct effect on SOC (Table 4). This might be also due to thick litter cover and slow decomposition rate. High soil nutrients not only indicate well–preserved moisture regime but also suggest potential humus provisioning (Dutt 1994), which helps in maintaining the chemical and physical conditions necessary for healthy adjustment in plant growth (Kimmins 1997).

Wide range of winter–flowering shrubs including D. bholua generally require a pH of 5.5 to 7.5 is optimum for proper growth (Buffin 2005). We found a similar range of pH 5.3 in present study area (Table 4). Similarly, previous studies also showed that Daphne prefers acidic soil, reading of pH 5.5 in Quercus stand in Makawanpur, Central Nepal (Jeanrenaud 1984). In the case of a cultivated plant, it does well in both acidic and alkaline conditions even growing on every chalky soil (Brickell and

Mathew 1976 cited by Jeanrenaud 1984). Some studies showed that Himalayan Daphne thrives in both acidic and alkaline conditions as long as they receive sufficient moisture during the growing season (Buffin 2005). Although, the edaphic properties (soil characters) were explaining the altitudinal effect on plant traits. At species, population and community levels, it is difficult to determine which altitudinal properties are most governing. Future work should attempt to define environmental constraints on the species distribution recorded here.

The tree canopy cover is known to be one of the most important factors characterizing the habitat conditions (Jeanrenaud 1984; Dutt 1994; Ghimire et al. 2008b; Khadgi et al. 2013), hence cover can directly affect the population density of D. bholua. It was observed that tree cover was highest in the upper two elevational sites Malika and Bhainse mela which was recorded as core forest of MPF (DoF 2011). These sites were far from the nearest village and had a dense canopy coverage with very old aged trees (Table 4). Numerous studies have indicated that patterns of understory vegetation and physical environment (e.g., temperate, soil nutrients and moisture availability) were directly related to canopy openness (Chavez et al. 2004; Barbier et al. 2008). Forest canopies provide microhabitats for plants, including mosses, herbs, and shrubs (Schroeer et al. 2008; Brosofske et al. 2001), and influence the composition of understory species, seedling regeneration, and microclimate conditions (Chavez et al. 2012; Barbier et al. 2008). A high degree of diversity of the canopy supports high microsite variability, and consequently, diverse understory vegetation (Song et al.2014; Mestre et al. 2017).

Shrub cover was found lower in Bhainse Mela and Uttise which indicated that local people cut more shrub for firewood and fodder. The forbs and grass cover tend to increase with the decrease in elevation such that the highest value was noted in Uttise and lowest in Malika (Table 4). This might be due to lower isolation on the forest floor (Rahman et al. 2009), more rocky area, decrease in available moisture and nutrients, presence of barren land and high anthropogenic disturbances (Bhattarai et al. 2014). Elevation and canopy are the influential site characteristics affecting the growth and other traits of several plant species with clonal growth strategy (Dutt 1994; Gracia and Retana 2004).

4.2 Density and size class distribution The key environmental factors for the growth of D. bholua are shade, humus, and moisture. Despite its shade tolerance, studies showed that light can have a remarkable influence on the growth and regeneration of Daphne bholua (Dutt 1994). The present study demonstrated that a high density of D. bholua at Malika and Bhainse mela (higher elevation) rather than in Uttise (lower elevation) indicated that D. bholua prefers higher elevational habitat (Figure 6).

The overall density of D. bholua in the MPF increased from the lowest elevation to the highest elevation (Figure 6). This increase in density along with elevation might be due to lower disturbance and more favorable conditions, that prevailed at a higher elevation (Table 10) (also discussed in 4.1). There was a gradual decrease in density of D. bholua with an increase in disturbance toward lower elevation; Uttise (as discussed in Khadgi et al. 2013). This may be due to two factors, harvest, and habitat. First, since Uttise is the site that is most accessible to villagers, it experiences the greatest amount of harvest for medicinal purposes (Ghimire et al. 2005; Gubhaju and Ghimire 2009). Consequently, reproductive individuals are more likely to be extracted, thereby contributing to their lower abundances. Secondly, since Uttise is at a lower elevation, which further limits the survivorship of D. bholua plants to reproductive age. Seed germination rate is found to be very low in natural habitat. At the meantime, reduced number of adults in Uttise exhibiting lower vegetative propagation by root sucker (Campbell 1980, Ghimire and Nepal 2007). Not only that they are generally sparse in more open forest and in pasture land (Jeanrenaud 1984). Further, the plant population at Uttise was affected by some large landslides and development activities such as road construction for the Butwal – Saljhandi – Dhorpatn corridor.

A higher environmental disturbance was also correlated with a higher number of coppice growth at Uttise (Table 9,10). Adult plants were harvested from the Uttise for local use, harvested adults’ plants were showing coppice growth, indicates the positive influence of harvesting on the sprouting/coppicing ability of the species. The level of disturbance and harvesting might promote coppice growth in case of harvested and damaged plant silviculture (Sawadogo et al. 2002). The rate of coppice growth is very low in Bhainse Mela and Malika due to low harvesting of the adult

40 plant. Although some older D. bholua plants sampled, showed coppice growth which was started to defoliated from the top.

The positive correlation of D. bholua density with soil nutrient concentrations in present study sites suggested that nutrient availability was an important factor affecting plant density, thereby supporting results from earlier studies (Gentry 1988; Ashton 1989; Aiba & Kitayama 1999; Givnish 1999). The observed pattern of high density at a higher and mid-elevation site was might be the reason for resource heterogeneity at heterogenous habitat (Homeier et al. 2010).

Present study area showed the highest contribution of seedlings followed by juvenile and adults to the population may be the result of maximum adults were reproducing well by both sexual and vegetative means of reproduction and the maximum seedlings grown to juveniles. In present study, the number of seedlings was relatively higher than the number of juveniles and adults. It can be concluded that vegetative reproduction (sprouting) plays a crucial role in the population of D. bholua. The seed viability of D. bholua was reported to be very short. Further, heavy weevil affecting the seed maturation and limiting the viable seed set (Campbell 1980, Ghimire and Nepal 2007). Similarly, Jeanraund (1984) also mentioned that the fruit of D. bholua is very attractive to frugivore birds which also causing the reduction on mature fruits per plant.

The combining effect of elevation and canopy was less noticeable. However, individually elevation and canopy had an effect on the sprouting ability of D. bholua (Table 10). The density of D. bholua varied along the elevation as well as between canopy types. This is possibly due to microenvironmental heterogeneity created by the tree and shrub canopy affect species composition, frequency and cover percentage of the individual species (Chavez et al. 2010; Barbier et al. 2008). In MPF the higher plant density was found at semi–close and close canopy coverage whereas lower in the open canopy (Table 8). Previous studies showed that D. bholua prefer medium forest crown–cover and usually avoiding sites with dense crown cover and large open areas (Jeanrenaud 1984 Khadgi et al. 2013). In the case of D. bholua, a minimum of 30% canopy shade profile is essential for the wellbeing of the species (Branney 1994; Jeanrenaud 1998; Ghimire and Nepal 2007).

Certain plants require a period of cold (vernalization) to initiate flowering. Many winter–flowering plants, such as D. bholua, flower both in response to vernalization and short–day length (short–day) and they will not flower unless they have been exposed to cold temperatures, usually recorded between 0 to 5°C (Buffin 2005; Mestre et al. 2017). Previous literature supported that the semi-close canopy is the good habitat for the plant. In present study total, Daphne population was higher on semi–close followed by close and least in open canopy type (Table 8). The reason behind those habitats might be related to the high plant performance at lower temperature and high moisture. The seedlings and juvenile stages were high in close canopy, that might be the reason for less disturbance in close canopy and high amount of moisture supports for seed germination and seedling recruitment (Appendix V).

4.3 Variation in Growth–Related Traits and Regeneration Despite the short elevation gradient (1700–2300 m) covered in this study, elevation showed a differential effect on growth-related traits in three sites (Table 11). Values of almost all the growth-related traits (plant height, crown radius, stem diameter, number of branches per plant, bark thickness, dry bark mass) were lower at Uttise (lower elevation). This is due to the more unfavorable condition such as, nutrient limiting, high anthropogenic disturbances and more competition which reduces the plant vigor.

Coppice branch growth was high in Uttise (Table 11) due to the higher rate of harvesting for local uses by nearby villagers. Moderate livestock grazing does not have a negative effect on stool sprouting after plant harvesting (Sawadogo et al. 2002). This finding indicated that the sustainability of such populations could be maintained by regulated silviculture. Livestock grazing and fire regimes are used to promote coppice growth in Sudanian Savannah silviculture (Sawadogo et al. 2002). During the harvesting period, several studies strongly recommend that the plant individuals should be cut down with >2.5cm diameter, above 20cm from the ground level. This supports healthy regeneration from the cut stem within six years (Jeanrenaud and Thomson 1986); however, plants grown from seeds require almost eight years to attain maturity (FSRO 1984, Ghimire and Nepal 2007). In contrast, bark peeling from the standing stem and whole–plant harvest is unsustainable, as these processes affect the meristematic tissue so that growth of coppice branch is

42 greatly checked and the whole plant may die (FSRO 1984; Jeanrenaud 1984; Ghimire and Nepal 2007).

Bark thickness and dry bark mass increased with the elevation as the more adult plant were sampled from the Bhainse mela and Malika with minimum coppice outgrowth. Vigorous coppice growth might directly cause a decrease in fitness of the individuals, which could detrimentally affect the quality and yield of bark harvested for papermaking (Iwasa and Kubo 1997; Bond and Midgley 2001).

The floral output (Flowers and buds) were higher in Malika and Bhainse mela than in Uttise. The increased elevation is known to favor flowering phenology, which may explain the lack of flowers at Uttise at the time of survey (January). Moreover, several large sized adult individual plants were observed as flowering contained more branches and more number of inflorescence producing on the individual plant (as discussed in Given 1975). D. bholua is moderately shade tolerating plant but too much-limiting light penetration is also not favorable as it reduces reproductive output (FSRO 1984).

Most of the growth-related traits prefer semi–close canopy including plant height, crown radius, floral output, and bark thickness (Table 13). The present study supports that certain level of canopy gap is needed for the better performance of the plant, as also reported in previous studies (Jeanrenaud 1984; Dutt 1994; Peterson et al. 2007) (Appendix VI).

The success of regeneration can be predicted on the basis of current population structure, growth performance, and reproductive output potential (Guedje et al. 2003). Presence of sufficient proportion of seedlings, juveniles, and reproductive adults in a given population of a species indicates successful regeneration and long–term persistence (Saxena and Singh 1985; Saikai and Khan 2013).

Overall, age structure combining all elevational site was reverse J–shaped, with the number of seedlings exceeding the number of juveniles, and adults indicating a stable age structure. This pattern in all three elevational sites suggested good natural regeneration potential with high levels of recent recruitment (Cousins et al. 2014). A similar demographic pattern was also revealed in the previous study of Daphne spp. carried out in the eastern part of Nepal (Ghimire and Nepal 2007).

Ecologists often use size distribution to indicate the health of the population (Zhang et al. 2015). Two factors likely influence this demographic pattern, low harvest, and high recruitment. First, since the sites are located in a government–protected forest, mature D. bholua individuals are unlikely to be harvested for medicine or paper. Secondly, D. bholua reproduces through root suckers, and from seed and the study area holds suitable ecological conditions for the growth and development with mild disturbance.

Elevation more strongly influences age structure than canopy cover (Table 10). Malika and Uttise had almost reverse ‘J’ shaped pattern in all canopy types while in Uttise, the nearly bell-shaped pattern indicates, a low germination rate and recent past adult harvest. Overall, semi-close canopy had almost reversed ‘J' shaped pattern with good regeneration, while in the close canopy the number of juveniles was less, it may be related to the increasing effect of limiting light. Variations in population size and density within a species distribution may result from numerous factors, such as disturbance gradient between sites (Helm and Witkowski 2012) and habitat degradation and fragmentation (Witkowski and Lamont 1997).

Shade–tolerant species are relatively abundant as seedlings and juvenile because seedlings and juveniles are persistent, with most surviving and growing slowly in deep shade (Wright et al. 2003). In MPF the lower size classes of D. bholua were dominant than the adults. Since the scope of this study was limited to one–year, further research that incorporating demographic modeling over several years is essential (Singh et al. 1986; Cousin et al. 2014; Garcia 2003) to assess whether the populations of D. bholua at MPF, Gulmi are increasing or decreasing.

4.4 Total and Harvestable Dry Bark Mass D. bholua is slow growing and long-lived plants (total life span is more than 60 years, (Jeanraund 1984, NSCFP 2001) and matures only after 8 years of age (Ghimire and Nepal 2007). Thus, harvesting of a plant before maturity stage (8 years) has been considered to be unsustainable. In the case of D. bholua the best harvesting season is June to October (Ghimire and Nepal 2008b). The mean total, as well as harvestable bark mass, were; 6850 and 3340 kg/ha, in Lamjung district (Khadgi et al. 2013). The dry harvestable bark of Daphne spp. in various community managed forests of

Kanchenjunga Conservation Area (KCA) ranged from 3.06 to 56.7 kg/ha with the mean of 33.74 kg/ha19 (Ghimire and Nepal 2007).

The harvestable dry mass of the bark of D. bholua in the present study site is varied from the value reported by Ghimire and Nepal (2007) and Khadgi et al. (2013). In MPF total dry bark mass and harvestable bark mass were 71 kg/ha and 50 kg/ha respectively. This disparity is because of the difference in the harvesting intensity. While in KCA, the D. bholua has been harvested periodically, there was no commercial harvest in the present study area. Therefore, Malika and Bhainse Mela have a better stock of the bark than the Uttisse (also discussed in 4.2 and 4.3 section).

Similarly, semi-close canopy supports the more D. bholua bark and dry bark mass as well 28kg/ha and 23kg/ha respectively, where open and close canopy holds the similar dry bark mass (also discussed in 4.2, 4.3 and 4.4).

Calculation based on the International Trade Centre and Government of Nepal 2016; the study area has the potential to produce a total of 16.73kg paper/ha from a harvestable mass (only adult 2) of D. bholua. In monetary terms study area supported 1,414 US$/ha (Exchange rate of April 2019) from the harvestable mass of D. bholua (based on natural 20–gram paper). D. bholua of MPF is awaiting sustainable harvesting. If financial and technical assistance is provided to the local community on harvesting and entrepreneur support to papermaking then this could be an attractive source of income to the local livelihood.

CHAPTER FIVE: CONCLUSION & RECOMMENDATIONS

5.1 Conclusion D. bholua an understory shrub, generally occurs in a semi–close and close canopy having tree canopy in association with Rhododendron arboreum, Quercus semecarpifolia, Lyonia ovalifolia, Pieris formosa, Polystichum aculeatum, and Wikstroemia canescens in MPF. High density of D. bholua of all life stages at Malika and Bhainse mela (high elevation) rather than in Uttise (lower elevation) indicates that the plant prefers higher elevation habitat generally in (2100–2500 m). Comparatively, the disturbance was slightly higher in Uttise than the other two sites. D. bholua prefers slightly acidic soil of pH 5.3, with the soil organic matter content of 1.86 % and soil moisture of 11.50 % in MPF.

Both elevation and canopy cover are strong indicators of microclimate in sites occupied by D. bholua populations. Overall, the stage structure was reverse ‘J’– shaped, with high number of seedlings followed by juvenile and adults indicating a good natural regeneration. Two factors that likely influence this demographic pattern are lower harvest level and moderate canopy cover. Elevation gradient influences the plant performance of D. bholua. The semi-close forest canopy positively supports growth-related traits of D. bholua and its regeneration, the values which were negative in plots with close and open canopies. Plant height, number of branches, crown radius, stem diameter, bark thickness were higher in upper and mid elevational sites, except for coppice growth; that is pronounced in lower site. The stock of D. bholua bark is well preserved at MPF, Gulmi specially in higher elevation (2100- 2500m). The harvestable stock of Lokta bark in MPF is 56 kg/ha and it can produce 16.73 kg paper/ha, which supports 1414 US$/ha.

5.2 Recommendations

Present study indicated a good stock of Daphne bark harvest for future. However, a rigorous investigation on multiple year–wise comparison of plant population along the environmental gradient is suggested for harvest sustainability and long–term management.

There is uncertainty about the volume of Daphne (Lokta) collection nationwide and this is aggravated by concern about overharvesting in some regions. There is a need to prepare a long-term national plan for the sustainable management of Lokta resources in forests. Likewise, harvesting techniques must improve while providing processors with upgrade quality. Similarly, research on improved cultivation techniques for the regeneration of Daphne should be discovered and shared with producers. A proper survey covering entire MPF should be carried out in order to verify the actual bark potential of Lokta, current production, and harvesting practices.

Present study showed that MPF holds very good stock of D. bholua bark. Training about sustainable harvesting and paper making should be provided to the local people for the production of unique, popular, durable, and versatile handmade paper and paper products, which will contribute to economies of local people and whole nation.

References

Adler, A., Verwijst, T. and Aronsson, P. 2005. Estimation and relevance of bark proportion in a willow stand. Biomass and Bioenergy, 29: 102–113.

Adney, T. and Howard, I.C. 1964. The Bark Canoes and Skin Boats of North America., Smithsonian Institution. Washington DC, USA.

Agnihorti, S., Wakode, S. and Agnihorti, S. 2010. An overview on anti–inflammatory properties and chemo–profiles of plants used in traditional medicine. Indian Journal of Natural Products and Resources, 1: 150–167.

Aiba, S. and Kitayama, K. 1999. Structure, composition and species diversity in an altitude–substrate matrix of rain forest tree communities on Mount Kinabalu, Borneo. Plant Ecology, 140: 139–157.

ANSAB (Asia Network for Sustainable agriculture and Bioresources) 2009. Challenges and Opportunities for Nepal’s small and Medium Forest Enterprises (SMFEs). Food and Agriculture Organization of the United Nations (FAO), Rome, Italy.

Archibold, O.W. 1995. Ecology of World Vegetation. London, Chapman and Hall.

Aronson, J., Pereira, J.S. and Pausas, J.G. 2009. Cork Oak Woodlands on the Edge, Conservation, Adaptive Management, and Restoration. Washington, DC, Island Press.

Ashton, P.S. 1989. Species richness in tropical forests. In L.B. Holm–Nielsen, I.C. Nielsen and H. Balslev (eds.), Tropical Forests, Botanical Dynamics, Speciation and Diversity. Academic Press INC, London, UK. pp. 241–251.

Austin, M.P., Pauses, J.G and Nicholls, A.O. 1996. Pattern of tree species richness in relation to environment in south–western New South Wales Australia. Australian Journal of Ecology, 21: 154–164.

Banjara, G.B. 2007. Handmade Paper in Nepal, Upgrading with Value Chain Network. German Technical Cooperation/Private Sector Promotion. Kathmandu, Nepal.

Barbier, S., Gosselin, F. and Balandier, P. 2008. Influence of tree species on understory vegetation diversity and mechanisms involved: A critical review for temperate and boreal forests. Forest Ecology Management, 254: 1–15.

Batjes, N.H. 1996. Total carbon and nitrogen in the soils of the world. European Journal of Soil Science. 47: 151–163.

Beals, E.W. 1969. Vegetational change along altitudinal gradients. Science, 165, 981– 985.

Bhattarai, P., Bhatta, K.P., Chettri R. and Chaudhary, R.P. 2014. Vascular plant species richness along elevation gradient of the Karnali river Valley, Nepal Himalaya. International Journal of Plant and Environmental Sciences, 4: 114–126.

Bhattarai, K.R. and Vetaas, O.R. 2003. Variation in plant species richness of different life forms along a subtropical elevation gradient in the Himalayas, east Nepal. Global Ecology & Biogeography, 12, 327–340.

Bhattarai, K.R., Vetaas, O.R. and Grytnes, J.A. 2004. Fern species richness along a central Himalayan elevational gradient, Nepal. Journal of Biogeography, 31: 389–400.

Biggs, S. and Messerchmiat, D. 2005. Social responsibility in the growing handmade paper industry of Nepal. World Development, 33: 1821–1843.

Brickell, C.D. and Mathew, B. 1976. Daphne, The genus in the wild and in cultivation. Biology, 10: 1– 67.

Brosofske, K.D., Chen, J., and Crow, T.R., 2001. Understory vegetation and site factors, implications for a managed Wisconsin landscape. Forest Ecology and Management, 146: 75–87.

Bond, W.J. and Midgley, J.J. 2001. Ecology of sprouting in woody Plants, The persistence niche. Trends in Ecology and Evolution, 16: 45–51.

Branney P. 1994. Community Forestry Guidelines Series. Guidelines for Managing Community Forests in the Koshi Hills (2nd Edition). Nepal–UK Community Forestry Project.

Brown, S., Iverson, L. R., Prasad, A. and Liu, D. 1993. Geographical distributions of carbon in biomass and soils of tropical Asian forests. Geocarto International, 8: 45–59.

Brown, J.H. 1984. On the relationship between abundance and distribution of species. American Naturalist, 124: 255–279.

Brys, R.H., Jacquemyn. P., Endels, G.D. Blust and M. Hermy 2005. Effects of habitat deterioration on population dynamics and extinction risks in previously common perennial herb, Primula veris. Journal of Applied Ecology, 41: 1080–1091.

Buffin, M.W. 2005. Winter Flowering Shrubs. Timber Press, Inc. Oregon, U.S.A.

Burlakoti, C. and Kunwar, R.M. 2008. Medicinal Plants in Nepal, An Anthology of Contemporary Research. Ecological Society (ECOS). Kathmandu, Nepal.

Burne, H.M., Yatesb, C.J. and Ladd, P.G. 2003. Comparative population structure and reproductive biology of the critically endangered shrub Grevillea althoferorum and two closely related more common congeners. Biological Conservation, 114: 53–65.

Campbell, M.W. 1980. Plant propagation for reforestation in Nepal. Nepal–Australia Forestry Project, Department of Forestry, A.N. U.

Chapagain, S.P., Rai, J.K., and Pathak, A., 2014. Value Chain Analysis of Forest Products in Koshi Hill Districts of Nepal, Challenges and Opportunities for Economic Growth. In, Rai and Chapagain (eds.), Lokta (Daphne Bholua), Widely Commercialized but Limited Investments for Sustainable

Conservation and Management. ForestAction Nepal and RRN, Kathmandu, Nepal. pp. 173–195.

Chavez, V. and Macdonald, S.E. 2012. Partitioning vascular understory diversity in mixed wood boreal forests, the importance of mixed canopies for diversity conservation. Forest Ecology Management, 271: 19–26.

Chávez, V and Macdonald, S.E. 2010. The influence of canopy patch mosaics on understory plant community composition in boreal mixed wood forest. Forest Ecology and Management, 259: 1067–1075.

Condit, R., Sukumar, R., Hubbell, S.P. and Foster, R.B. 1998. Predicting population trends from size distributions, a direct test in a tropical tree community. The American Naturalist, 152: 496–509.

Condit, R., Sukumar, R., Hubbell, S.P., and Foster, R.B. 1998. Predicting population trends from size distributions, A direct test in a tropical tree community. The american naturalist, 152.

Cousins, S.R., Witkowski, E.T.F. and Pfab, M.F. 2014. Elucidating patterns in the population size structure and density of Aloe plicatilis, a tree aloe endemic to the Cape fynbos. South Africa South African Journal of Botany, 90: 20–36.

Department of Forest 2011. Madane Protected Forest Management Work Plan. Government of Nepal Ministry of Forest and Soil conservation (Nepali).

Department of Forest 2016. Madane Protected Forest an Introduction. Government of Nepal Ministry of Forest and Soil conservation (Nepali).

Denslow, J.S., Schultz, J.C., Vitousek, P.M.,and Strain, B.R. 1990. Growth responses of tropical shrubs to tree fall gap environments. Ecology, 71: 165–179.

Duong, N.H. 2008. The role of Non–Timber Forest Products in livelihood strategies and household economics in a remote upland village in the upper river basin, the Phuong. Journal of Science and development, 88–89.

Dutta, G. and Devi, A. 2013. Plant diversity, population structure, and regeneration Status in disturbed trobical forests in Assam, Northen India. Journal of Forestry Research, 24: 715–720.

Dutt, P. 1994. Lokta Daphne species. The Supply situation in the Basantapur Area. Project Report, Nepal– UK Community Forestry Project, Coordinators Office, Kathmandu, Nepal.

Filazzola, A., and Lortie, C.J. 2014.A systematic review and conceptual Frame work for the mechanistic pathways of nurse plants. Global Ecol. Biogeography, 23: 1335–1345.

Forest Act. 1993. Ministry of Forest and Soil Conservation. Development Project, HMG/USAID, Kathmandu.

Forestry Services (FSRO). 1983. Daphne Forest Survey of Hatiya Forest Range, Dhaulagiri Forest range, Division, Baglung, Forestry Services. Submitted to UNICEF/Nepal. Kathmandu, Nepal.

Forestry Services (FSRO)1984. Management plan for sustained harvest and utilization of Daphne (Lokta) from the Upper Rahughat river forest and vicinity in Myagdi distruct, Nepal. (Report to SECID/RCUP) Forestry Services Kathmandu, Nepal.

Forestry Services (FSRO). 1984. Management plan for sustained harvest and utilization of Daphne (Lokta) from the Kalinchok and Sailung forest areas, conservation and development objectives? Ecological Economics, 39: 437– 447.

Garcia, M.B. 2003. Demographic viability of a relict population of the critically endangered plant Borderea chouardii. Conservation Biology, 17: 1672–1680.

Garg, A. and Rogers, S.Z. 2011. A palynological investigation of Daphne papyracea and Daphne bholua (Thymelaeaceae) In India. Journal of the Botanical Research Institute of Texas, 5.

Gauli, K. and Hauser, M. 2009. Commercial Management of Non–timber Forest Products in Nepal's Community Forest User Groups. International Mountain Society, 29: 298–307.

Gautam, J.C. 2006. A Report on Compilation and prioritization of Ten Important NTFPs of Nepal for Commercial Promotion through private Sector Investment. Federation of Nepalese Chambers of Commerce and Industry Agro Enterprise Center (AEC/FNCCI).

Gentry, A.H. 1988. Changes in plant community diversity and floristic composition on environmental and geographical gradients. Annals of Missouri Botanical. Garden, 75: 1–34.

Ghimire, S.K., Pyakurel, D., Nepal, B.K., Sapkota, I.B., Parajuli, R. R. and Oli, B. R. 2008a. A Manual of NTFPs of Nepal Himalaya (Gair Kastha Ban Paidawar Digdarshan). WWF Nepal, Kathmandu, Nepal. (In Nepali)

Ghimire, S.K., Sapkota, I.B., Oli, B.R. and Parajuli–Rai, R. 2008.b Non–Timber Forest Products of Nepal Himalaya, Database of Some Important Species Found in the Mountain Protected Areas and Surrounding Regions. WWF Nepal, Kathmandu, Nepal.

Ghimire, S.K., McKey, D. and Aumeeruddy–Thomas, Y. 2005. Conservation of Himalayan medicinal plants, harvesting patterns and ecology of two threatened species, Nardostachys grandiflora DC. and Neopicrorhiza scrophulariiflora (Pennell) Hong. Biological Conservation, 124: 463–475.

Ghimire, S.K. and Nepal, B.K. 2007. Developing a Community–based Monitoring System and Sustainable Harvesting Guidelines for Non–Timber Forest Products (NTFP) in Kanchanjunga Conservation Area (KCA), East Nepal. WWF Nepal Program, Kathmandu, Nepal.

Given, D.R. 1975. Conservation of rare and threatened plant taxa in Newzealand– some principles. Proceeding of the Newzealand Ecological Society, 22: 1–6.

Givnish, T.J. 1999. On the causes of gradients in tropical tree diversity. Journal of Ecology, 87: 193–210.

GoN/MoFSC. 2007. Hamro Kalpabrikshya. Ministry of Frest and Soil Conservation, Department of Forest, Babarmahal, Kathmandu.

GoN/MoFSC/DoF. 2010. Country Report– Nepal, State of Forestry in Nepal. A Synopsis Report Department of Forests. Kthmandu, Nepal.

Government of Nepal. 2010. Nepal Trade Integration Strategy Kathmandu. Ministry of Commerce and Supplies. Kathmandu, Nepal.

Götmark, F. Götmark, E. and Jensen, A.M. 2016. Why be a Shrub? A basic model and hypotheses for the adaptive values of a common growth form. Frontiers in Plant Science, 7: Article 1095.

Gracia, M. and Retana, J. 2004. Effect of site quality and shading on sprouting patterns of holm oak coppices. Forest Ecology and Management, 188: 39–49.

Grey–Wilson, C. and Matthews, V. 1983. Gardening on walls. London, Collins. pp 1– 7.

Gubhaju, M.R. and Ghimire, S.K. 2009. Diversity and population Status of Non– Timber forest Products (NTFPs) in community forest of Dovan, Palpa. Journal of Natural History, 24, 22–47.

Guedje, N.M., Lejoly, B.A., Nkongmeneck, B.A. and Jonkers, W.B.J. 2003. Population Dynamics of Gracinia lucida (Clusiaceae) in a Cameroonian Atlantic Forests. Forest Ecology and Management., 117: 231–241.

Gupta, P. K. 2002. Methods in Environmental Analysis of Water, Soil and Air. Second edition. Agrobios India.

Halda, J. J. 2003. Some taxonomic problems in the genus Daphne L. Acta Musei Richnoviensis Sect. Nat. 6: 195–233.

Halda, J. J. 2001. The genus Daphne. SEN, Dobré (Czech Republic).

Hamilton, A.C. and Perrott, R.A. 1981. A study of altitudinal zonation in the montane forest belt of Mt. Elgon, Kenya/Uganda. Vegetation, 54: 107–125.

Hammet, A.L. 1993. Non–timber Forest Products, Profits and Panacea? Timber Forest Products, Kathmandu, Nepal.

Helm, C.V and. Witkowski, E.T.F. Forest. 2012. Characterising wide spatial variation in population size structure of a keystone African savanna tree. Ecology and Management, 263: 175–188.

Homeier, J., Siegmar, W., Breckle, S. G., Rollenbeck, R. T. and Christoph, L. 2010. Diversity, forest structure and productivity along altitudinal and topographical gradients in a species–rich Ecuadorian Montane Rain Forest. Biotropica, 42: 140–148.

Iqbal, K., Bhat, J.A., Pala, N.A. and Negi, A.K. 2013. Physio–chemical, properties of soils under the vegetation along and away of the Khoh river in Garhwal, Himalaya. Indian Journal of Forestry, 36: 205–212.

ITC and GoN. 2017. Nepal national Sector Export Strategy, Handmade Paper and Paper Products (2017–2021). International Trade Center and Government of Nepal.

Iwasa, Y. and Kubo, T. 1997. Optimal Size of Storage for Recovery after Unpredictable Disturbances. Evolutionary Ecology, 11: 41–65.

Jeanrenaud J.P. 1984. Daphne (Lokta), Some Silvicultural Considerations. A paper presented at the Department of Forest, Government of Nepal and UNICEF Lokta (Daphne) Forestry Policy and Planning Workshop–Seminar. 1–19.

Jeanrenaud, J.P. 1984. Lokta (Daphne spp.) and craft paper–making Nepal. A report on the current status based on a literature view and preliminary Field observations, FSRO Kathmandu Nepal.

Jeanrenaud, J.P. and Thompson, I. 1986. Daphne (lokta), bark biomass production management implications for paper making in Nepal. Commonwealth Forestry Review, 65: 117–130.

Kala, C.P. and Dubey, Y. 2012. AnthropogenicDisturbances and Status of Forest and Wildlife in the Dry deciduous Forest of Chattishgarh state in India. Journal of Forestry Research, 23: 45–52.

Kasai, R., Lee, K.H. and Huang, H.C. 1981. Genkwadaphnin, a potent anti–leukaemic diterpene from Daphne genkwa. Phytochem, 20: 2592–2594.

KC, M., Phoboo, S. and Jha P.K. 2010. Ecological Study of Paris polyphylla SM. Ecoprint, 17: 87–93.

Kenney, W.A., Sennerby–Forsse, L. and Layton, P. 1990. A review of biomass quality research relevant to the use of poplar and willow for energy conversion. Biomass, 21: 163–188.

Khadgi, N., Shrestha, B.B., and Siwakoti, M. 2013. Resource assessment and habitat analysis of Daphne bholua in Bhujung of Annapurna Conservation Area, Central Nepal. Research Journal of Agriculture and Forestry Sciences, 1: 1– 9.

Kharal, D.K., Oli, B.N. and Poudel, I. 2011. Distribution and availability of raw materials for production of Nepali handmade paper from Daphne species in Darchula district, Nepal. Banko Jankari, 21: 24–30.

Kimmins, J. P. 1997. A Foundation for Sustainable Management. Forest Ecology Second Edition, Prentice Hall, New Jersy, USA.

Kim, E. and Donohue, K. 2011. Demographic, developmental and life–history variation across altitude in Erysimum capitatum. Journal of Ecology, 99: 1237–1249.

Ko¨rner, C. 1999. Alpine plant life. Springer. Verlag, Berlin.

Leps, J. and Smilauer, P. 2003. Multivariate Analaysis of Ecological Data using CANACO. Cambridge University Press. United Kingdom.

Lomolino, M.V. 2001. Elevation gradients of species–richness, historical and prospective views. Global Ecology and Biogeography, 10: 3–13.

Marks, T.R. 1997. Micropropagation of Daphne L. In, Bajaj Y.P.S. (eds) High–Tech and Micropropagation VI. Biotechnology in Agriculture and Forestry., Berlin, Heidelberg. Springer, 40: 113-130.

Mestre, L., Manríquez, M.T., Soler, R., Herrera, A. H., Pastur, G. M. and Lencinas, M. V. 2017. The influence of canopy–layer composition on understory plant diversity in southern temperate forests. Forest Ecosystems, 4: 6.

Moral, R.del 1972. Diversity patterns in forest vegetation of the Wenatchee mountains, mountain in Oman. Journal of Biogeography, 28: 299–309.

Murphy, H.T. and Lovett–Doust, J. 2004. Context and connectivity in plant metapopulation and landscapes mosaics, does the matrix matter? Oikos, 105: 3–14.

Myking, T., Hertzberg, A. and Skrøppa, T. 2005. History, manufacture and properties of lime bast cordage in northern Europe. Forestry, 78: 65–71.

Namgail, T., Rawat, G.S., Mishra, C., and Herbert, H.T.P. 2010. Biomass and diversity of dry alpine plant communities along altitudinal gradients in the Himalayas. Journal of Plant Resources, 125: 93–101.

Nikaido, T., Ohmoto, T. and Sankawa, U. 1987. Inhibitors of adenosine 32032,52032–cyclic monophosphate phosphodiesterase in Daphne genkwa Sieb. et Zucc. Chemistry Pharmaceutical Bulletin, 35: 675–681.

Olsen, C.S. and Larsen, H.O. 2003. Alpine medicinal plant trade and Himalayan mountain livelihood strategies. The Geographical Journal, 169: 243–254.

Oosttermeijer, J.G.B., Luitjen, S.H., Klenova, Z.V. and Den, Jis, J.C.M. 1998. Relationships between population and habitat characteristics and reproduction of the rare Gentiana pneumonathe L. Conservation Biology, 12: 1042–1056.

Paudel, A., Subedi B.P., Gyawali, S., Thapa G.K. and Sharma, M.B. 2011. Value Chain Analysis of Non–Timber Forest Products in Baglung district, Banko Jankari, 19: 33–41.

Paudyal, A.S. 2004. Sustainability of local handmade paper enterprises. Journal of Forest and Livelihood, 4: 64–69.

Pavlovic, N.B. 1994. Disturbance–dependent persistence of rare plants, anthropogenic impacts and restoration implications. In, M.L. Bowles, C.J. Whelan (eds), Restoration of Endangered Species Conceptual Issues, Planning and Implementation. Cambridge. University Press. New York, U.S.A.

Peterson, D.W., Reich, P.B. and Wrage, K.J. 2007. Plant functional group responses to fire frequency and tree canopy cover gradients in Oak Savanas and woodlands. Journal of Vegetation Science, 18: 3–12.

Polunin, O. and Stainton, A. 1984. Flowers of the Himalaya. Oxford University Press.

Press, J.R., Shrestha, K.K. and Sutton, D.A. 2000. Annotated Checklist of the Flowering Plants of Nepal. The Natural History Museum, London, UK and Central Department of Botany, Tribhuvan University, Kathmandu, Nepal.

Purohit, A., Maikhuri, R.K., Rao, S.K. and Nautiyal, S. 2001. impact of bark removal on survival of Taxux baccata L. (Himalayan Yew) in Nanda Devi biosphere reserve, Garhwal Himalaya, India. Current Science, 81: 586–590.

Pyke, A., David., and Zamora B.A. 1982. Relationships between Overstory Structure and Understory Production in the Grand Fir /Myrtle boxwdod habitat type of Northcentrai Idaho. Journal of Range Management, 35: 769–773.

Pyakurel, D. and Baniya, A. 2011. NTFPs, Impetus for Conservation and Livelihood support in Nepal. A Reference Book on Ecology, Conservation, Product Development and Economic Analysis of Selected NTFPs of Langtang Area in the Sacred Himalayan Landscape. WWF Nepal.

Rahman, M.M., Nishant, A. and Vacik, H. 2009. Anthropogenic disturbances and plant diversity of the madhupur sal forest (Shorea robusta C.F. Garetn.) of Bangladesh. International Journal of Biodiversity Science and Management, 5: 162–173.

Rogers, S.Z., Garg, A. 2011. A Palynological Investigation of Daphne Papyracea and Daphne Bholua (Thymelaeaceae) In India. Journal of the Botanical Research Institute of Texas 5(2).

Ros–Tonen, M., Dijkman, W. and Lammerts van Bueren, E. 1995. Commercial and sustainable extraction of Non–timber Forest Products towards a policy and management orient research strategy. The Tropenbos Foundation, Wageningen, the Netherland.

Saikia, P. and Khan, M.L. 2013. Populaation structure and regeneration status of Aquilaria malaccenis Lam. in homegardens of Upper Assam, Northeast, India. Tropical Ecology, 54: 1–13.

Salter, P.J. and Williams, J.B. 1969. The moisture characteristics of some Rothamsted, Woburn and Saxmundham soils. The Journal of Agricultural Science, 73: 155–158.

Sawadogoa, L., Nygård, R. and Pallo, F. 2002. Effects of livestock and prescribed fire on coppice growth after selective cutting of Sudanian savannah in Sudanian Faso. Annals of Forest Science, 59: 185–195.

Saxena, A.K. and Singh, J.S. 1985. Tree population structure of certain Himalayan forest association and implications concerning their future. composition.Vegetation, 58: 61–69.

Scholes, M.A. 1988. A population study of Aloe peglerae in habitat. South African Journal of Botany, 54: 137–139.

Schroeer, A.E., Hendrick, R,L., and Harrington, T.B. 1999. Root, ground cover, and litterfall dynamics within canopy gaps in a slash pine (Pinus elliottii Engelm.) dominated forest. Ecoscience, 6: 548–555.

Shackleton S., Delang C.O. and Angelsen A. 2011. From subsistence to safety nets and cash income, exploring the diverse values of non–timber forest products for livelihoods and poverty alleviation. In, Non–Timber Forest Products in the global context (S. Shackleton, C. Shackleton and P. Shanley, eds.), Springer–Verlag, Amsterdam, Netherlands. pp 55–82.

Sharma, R.P., Bhandari, S.K. and BK, R.B. 2017. Allometric bark biomass model for Daphne bholua in the Mid-Hills of Nepal. International Mountain Society, 37: 206–215.

Sharma, U.R. and Das, P.K. 2004. Reviewing current issues and prospects of Non– Timber Forest Products (NTFPs) sub–sector development in Nepal. In Bhattarai and Karki (editors), Proceeding of the National Workshop on "Local Experience–Based National Strategy for Organic Production and Management of MAPs/NTFPs in Nepal" held at Kathmandu, Nepal 27–28 Feb 2004. Published by IDRC/ MAPPA. pp. 87–96.

Shrestha, K.K., Bhattarai, S. and Bhandari, P. 2018. Handbook of Flowering Plants of Nepal (Volume 1). Gymnosperm and Angiosperms, Cycadaceae– Betulaceae). Scientific Publishers, Jodhpur India.

Shrestha, S. 2012. Growth Performance of Daphne bholua in Relation to Harvesting and Ecological Conditions in Langtang National Park, North–Central Nepal. [M.Sc thesis]. Kathmandu: Central Department of Botany, Tribhuvan University.

Singh, J.S., Singh, S.P. and Gupta, S.R. 2014. Ecology Environmental Science and Conservation. S. Chand and Company Pvt. Ltd, Ramnagar, New Delhi.

Singh, S.P., Tewari, J.C., Yadav, S. and Ralhan, P.K. 1986. Population structure of tree species in forests as an indicator of regeneration and future stability. Proc. Plant Science, 96: 443–455.

Smith E. 2011. Ecological relationships between overstory and understory vegetation in Ponderosa Pine Forests of the Southwest. Report prepared for the Kaibab National Forest by the Nature Conservancy. pp 1–16.

Song, B., Chen, J. and Williams, T.M. 2014. Spatial Relationships between Canopy Structure and Understory Vegetation of an Old–Growth Douglas–Fir Forest. Forest Research, 3: 1–12.

Stainton, A. 1998. Flowers of Himalaya, A supplement. Oxford University Press, New Delhi.

Subedi, B. 2006. Linking Plant–Based Enterprises and Local Communities to Biodiversity Conservation in Nepal Himalaya. Adroit Publishers, New Delhi, India.

Subedi, M. and Sharma, R.P. 2012. Allometric biomass models for bark of Cinnamomum tamala in mid–hill of Nepal. Biomass and Bioenergy, 47: 44– 49.

Sunderlin, W.D., Angelsen, A., Belcher, B., Burgers, P., Nasi, R., Santoso, L. and Wunder, S. 2005. Livelihoods, forests, and conservation in developing countries, an overview. World Development, 33: 1383–1402.

Ticktin, T. 2004. The ecological implication of harvesting non–timber forest products. Journal of Applied Ecology, 41: 11–21.

Turner, J.R., Gatehouse, C.M. & Corey, C.A. 1987. Does solar energy control organic diversity? Butterflies, moths and British climate. Oikos, 48: 195–205.

Underwood, A.J. 1997. Experiments in Ecology, Their Logical Design and Interpretation Using Analysis of Variance. Cambridge University Press, Cambridge.

Uprety, Y., Poudel, R.C., Chaudhary, R.P., Oli, B.N., Bhatta, L.D. and Baral, S.P. 2016. Sustainable Utilization and Conservation of Non–timber Forest Products, Major Species of Kailash Sacred Landscape Nepal. Ministry of Forests and Soil Conservation (MoFSC), Government of Nepal; Research Centre for Applied Science and Technology (RECAST), Tribhuvan University; and International Centre for Integrated Mountain Development (ICIMOD). Kathmandu, Nepal.

Volis, S., Bohrer, G., Oosttermeijer, J.G.B. and Tienderen, P.V. 2005. Regional consquences of local Population demography and genetics in relation to habitat management in Gentiana pneumonathe L. Conservation Biology, 19: 357–367.

Walter, S. 1998. The Utilization of Non–Timber Forest Products in the Rainforests of Madagaskar, A Case Study. Plant Research and Development, 47/48: 121– 144.

Whittaker, R.H. 1977. Evolution of species diversity in land communisties. Evolutionary Biology, 10: 1-67.

Witkowski, E.T.F., Hamont, B.B., 1997. Does the rare Banksia goodii have inferior vegetative, reproductive or ecological attributes compared with its widespread co–occurring relative B. gardneri? Journal of Biogeography, 24: 469–482.

Wright, S.J., Muller-Landau, H.C., Condit, R. and Hubbell, S.P. 2003. Gap– dependent recruitment, realized vital rates, and size distributions of Tropical Trees. Ecology, 84: 3174–3185.

Yamda, T.P., Zuidema, P.A., Itoh, A., Yamakura, T., Ohkubo T., Kanzankis, M. and Asthon P.S. 2007. Strong habitat preference of tropical rain forest tree does not imply large diffrences in population dynamics across habitat. Journal of Ecology, 95: 332–342.

Yinzheng, W., Gilbert, M.G., Mathew, B. and Brickell C.D. 2007. DAPHNE Linnaeus, Sp. Pl. 1: 356. 1753. Flora of China, 13: 230–245.

Young, A., Boyle, T. and Brown, T. 1996. The population genetic consequences of habitat fragmentation for plants. Trends in Ecology and Evaluation, 11: 413– 418.

Zackrisson, O., €Ostlund, L., Korhonen, O., Bergman, I. 2000. The ancient use of Pinus sylvestris L. (Scots pine) inner bark by Sami people in northern Sweden, related to cultural and ecological factors. Vegetation History and Archaeobotany, 9: 99–109.

Zhang, Jianwei., Young, D.H., Oliver W.W and Fiddler G.O. 2015. Effect of overstorey trees on understorey vegetation in California (USA) Ponderosa pine plantations. Forestry, 89: 91–99.

Appendix I. Correlation among different biophysical variables recorded in three sites.

can Elv slp trcan canht ltdp trcov shcov fbcov grcov brs lhsl ltcov Rkcov brg lhrk lhvs brrk brvs dist Canopy (can) 1

Elevation (Elv) –0.01 1 – Slope (slp) 0.1 .289** 1 Tree canopy (trcan) .933** 0.083 0.069 1 canopy height – (canht) –0.135 .374** .263** –0.119 1 litter depth (Ltdp) .378** .645** –.198* .480** –.228* 1 Tree cover (trcov) .862** 0.1 0.108 .928** –0.077 .495** 1 Shurb cover – – – (shcov) .341** 0.088 .242* .318** 0.173 –0.121 .283** 1 Forbs cover – – – – – (fbcov) .389** .264** –0.121 .485** 0.034 .394** .504** 0.056 1 Grass cover – – – – – (grcov) .253** .452** 0.042 .389** .195* .561** .410** .190* .413** 1 Bryophyte on – soil (brs) 0.099 .269** –0.006 0.101 –0.131 .319** 0.098 0.03 .280** –.224* 1 Lichen on soil (lhsl) –0.077 –0.134 –0.011 –0.049 .371** –0.169 –0.051 0.162 –0.148 .325** –0.002 1 Litter cover – – (Ltcov) .434** .307** 0.091 .558** –0.108 .460** .582** –.194* .715** .718** –0.095 –.216* 1 Rock cover – (rkcov) –0.109 .215* –0.176 –0.067 –0.156 0.119 –0.104 –.217* 0.099 –0.169 –0.188 .295** –0.047 1 Bare ground – – (Brg) –.196* –0.188 –0.02 –.232* 0.156 .251** –.228* .376** 0.004 .371** –0.063 .355** .396** –0.188 1 Lichen on rock – – – – – – (lhrk) .388** .444** 0.089 .402** .349** .519** .373** .196* .209* .501** –0.166 .365** .393** –0.093 .203* 1 Lichen on vascular plant – – – – – (lhvs) .318** –0.057 .201* .359** 0.031 .250** .322** .208* 0.185 .315** 0.163 0.062 .330** –0.147 0.103 .498** 1 Bryophyte on – – – rock (brrk) –.210* –0.077 .202* .269** 0.117 .340** .254** 0.185 0.002 .290** –0.098 0.117 –.192* .240* 0.089 .291** .259** 1 bryophyte on vascular plant (brvs) –0.043 .344** 0.09 –0.001 –.234* .246* 0.069 .200* –0.158 –.207* 0.062 –0.101 0.168 0.129 –0.047 –0.03 .309** 0.06 1 Disturbance – – – – – – – – factor(dist) .519** .532** –0.016 .598** .433** .642** .606** 0.12 .597** .555** .266** .218* .660** 0.009 .332** .445** .193* 0.164 .344** 1 ** Correlation is significant at the 0.01 level (2tailed). * Correlation is significant at the 0.05 level (2– tailed).

Appendix II. Percentage plots experiencing certain type of disturbance on three different altitudi–nal bands. % plot experiencing certain type of disturbance Band Disturbance type 0 1 2 3 4 Top Band Animal Droppings 13.9 52.8 27.8 5.6 0.0 Grazing 41.7 50.0 8.3 0.0 0.0 Trampling 16.7 52.8 22.2 8.3 0.0 Harvesting 55.6 27.8 11.1 5.6 0.0 Fire 77.8 22.2 0.0 0.0 0.0

Mid Band Animal Droppings 27.5 38.9 33.3 0.0 0.0 Grazing 65.0 16.0 8.9 5.0 5.1 trampling 30.6 22.2 2.2 2.8 0.0 Harvesting 0.0 0.0 5.6 0.0 0.0 Fire 0.0 0.0 0.0 0.0 0.0

Low Band Animal Droppings 8.3 40.0 27.8 20.0 4.0 Grazing 0.0 44.4 30.6 13.9 11.1 trampling 2.8 11.1 52.8 33.3 0.0 Harvesting 2.0 50.0 33.3 5.6 9.1 Fire 77.8 22.2 0.0 0.0 0.0

Appendix III. List of all plant species found on the study area with their respective family, fre–quency and abbreviation Frequency S.No. Plant Species Abbrevation Family (%) 1 Aconogonum molle (D. Don) H. Hara Aco_mol Polygonaceae 16.667 Ageratina adenophora (Spreng.) King & H. 2 Age_ade Asteraceae 31.481 Rob. 3 Alnus nepalensis D. Don Aln_nep Betulaceae 7.215 4 Anaphilis controta (D. Don) Hook. F. Ana_con Asteraceae 14.815 5 Anaphilis triplinervis (Sims) C.B. Clarke Ana_tri Asteraceae 14.815 6 Androsace primuloides D. Don And_pri primulaceae 7.407 7 Arundinaria nepalensis Trin., Gram. Pan Aru_nep Poaceae 25.926 8 Berberis asiatica Roxb.ex. DC. Ber_asi Berberidaceae 37.037 9 Berberis glaucocarpa Stapf Ber_gla Berberidaceae 11.111 10 Brassiopsis polycantha (wall.) Banerjee Bra_pol Araliaceae 11.111 11 Buddleja asiatica Lour. Bud_asi Scrophulariaceae 9.259 12 Calanthe sylvatica ( Thouras ) Lindl. Cal_syl Orchidaceae 14.815 13 Carex condensata Nees Car_con Cyperaceae 20.37 14 Castanopsis indica (Roxb.) Miq. Cas_ind Fagaceae 7.407 15 Castanopsis tribuloides (Sm.) A. DC. Cas_tri Fagaceae 5.556 Cinnamomum tamala (Buch.–Ham.) Ness & 16 Cin_tam Lauraceae 9.259 Eberm 17 Clematis acuminata DC. Cle_acu Ranunculaceae 24.074 18 Clematis buchananiana DC Cle_buc Ranunculaceae 12.963 19 Coelogyne cristata Lindl. Coe_cri Orchidaceae 22.222 20 Colebrookea oppositifolia Sm. Col_opp Lamiacea 5.556 21 Daphne papyreceae Wall. Ex Steud. Dap_pap Thymalaceae 12.963 Daphniphyllacea 22 Daphniphyllum himalayense (K. Rosenthal) Dap_him 5.556 e 23 Desmodium microphyllum(Thunb.) DC. Des_mic Fabaceae 14.815 24 Dichroa fabrifuga Lour. Dic_fab Hydragenaceae 9.259 25 Digitaria cilliaris (Retz.) Koeler Dig_cil Poaceae 12.963 26 Digitaria longiflora (Retz.) Pers. Dig_lon Poaceae 14.815 27 Dipsacus inermis Wall. Dip_ine Dipsacaceae 16.667 28 Drepanostachym flacatum (Ness) Keng F. Dre_fla Poaceae 11.111 29 Drymaria cordata (L.) Willd.ex Roem & Scolt Dry_cor Caryophyllaceae 1.852 30 Elaeagnus parvifolia Wall. Ex Royale Ela_par Elagnaceae 31.481 Eregeron multiradiatus (Lindl. Ex DC.) Benth 31 Ere_mul Asteraceae 11.111 .ex CB Clarke 32 Eurya acuminata DC. Eur_acu Theaceae 24.074 33 Evernia prunastri (L.)Ach. Eve_pru Parmeliaceae 7.407 34 Flemingia strobilifera (L.) W. T. Aiton Fle_str Fabaceae 11.111 35 Fragaria nubicola Lindl. Ex Lacaita Fra_nub Rosaceae 14.815 36 Fraxinus Floribunda Wall. Fra_Flo Oleaceae 12.963 37 Galium aparine L. Gal_apa Rubiaceae 22.222 38 Gaultheria fragrantissima Wall. Gau_fra Ericaceae 20.37

Frequency S.No. Plant Species Abbrevation Family (%) 39 Gentiana depressa L. Gen_dep Gentianaceae 4.238 40 Geranium wallichianum (D.Don) ex Sweet Ger_wal Geraniaceae 20.37 41 Girardiana diversifolia (Link) Friss Gir_div Urticaceae 12.963 42 Gnaphalium affine D. Don Gna_aff Asteraceae 24.074 43 Hedyotis scandens Roxb. Hed_sca Rubiaceae 27.778 44 Hemiphragma heterophyllum Wall. Hem_het Scrophulariaceae 33.333 45 Hypericum cordifolium DC. Hyp_cor Clusiaceae 25.926 46 Ilex dipreyana Wall. Ile_dip Aquifoliaceae 31.481 47 Imperata cylindrica (L.) P. Beauv Imp_cyl Poaceae 31.481 48 Juncus articulatus L. Jun_art Poaceae 1.852 49 Lepisorous bicolor (Takeda) Ching, Bull Lep_bic Polypodiaceae 16.667 50 Leucosceptum canum Sm. Leu_can Lamiacea 7.407 51 Lindera pulcherrima (Ness) Hook.f. Lin_pul Lauraceae 51.852 52 Litsea cubeba (Lour.) Press. Lit_cub Lauraceae 14.815 53 Lyonia ovalifolia (Wall.) Drude Lyo_ova Ericaceae 40.741 54 Maesa chisia Buch. –Ham ex D. Don Mae_chi Primulaceae 24.074 55 Mahonia napaulensis DC. Mah_nap berberidaceae 16.667 Nephrolepidacea 56 Nephrolepis cordifolia (L.) Press. Nep_cor 14.815 e 57 Onychium japonicum (Thunberg) Kunze Ony_jap Pteridaceae 11.263 58 Oplismenus compositus (L.) P. Beauv. Opl_com poaceae 12.963 59 Osbekia stellata Buch.–Ham.ex D. Don Osb_ste Melastomataceae 14.815 60 Osyris wightiana Wall/ ex Wight Osy_wig Santalaceae 12.963 61 Persicaria nepalensis (Meisn.) Miyabe Per_nep Polygonaceae 18.519 62 Phyllanthus niruri L. Phy_nir Phyllanthaceae 20.37 63 Pieris formosa (Wall.) D. Don Pie_for Ericaceae 18.519 64 Plantago depressa Willd. Pla_dep Plantaginaceae 11.111 65 Pogostemon benghalensis (Brum.f. Kuntze) Pog_ben Lamiacea 11.111 66 Polygonatum griffithii Baker Pol_gri Asparagaceae 7.407 67 Polystichum aculeatum (L) Schott. Pol_acu Dryopteridaceae 48.148 68 Potentilla lineata Trevir. Pot_lin Rosaceae 29.63 69 Prunus cerasoides D.Don Pru_cer Rosaceae 18.519 70 Pteris wallichiana J. Agardh Pte_wal Pteridaceae 38.889 71 Pyracantha crenulata (D. Don) M. Roem. Pyr_cre Rosaceae 12.963 72 Quercus lanata Sm. Que_lan Fagacea 55.556 73 Quercus semecarpifolia Sm. Que_sem Fagacea 81.481 Randia tetrasperma (Roxb.) Benth. & Hook.f.ex 74 Ran_tet Rubiaceae 12.963 Brandis 75 Reinwardtia indica Dumort Rei_ind Linaceae 22.222 76 Rhododendron arboreum Sm. Rho_arb Ericaceae 85.185 77 Rosa laevigata Michx. Ros_lae Rosaceae 12.963 78 Roscoea alpina Royle Ros_alp Araceae 14.815 79 Rubia manjith roxb. Ex Flming Rub_man Rubiaceae 25.926

Frequency S.No. Plant Species Abbrevation Family (%) 80 Rubus ellipticus Sm. Rub_ell Rosaceae 16.667 81 Rubus rosifolius Smith Rub_ros Rosaceae 27.778 82 Rumex nepalensis Spreng. Rum_nep Polygonaceae 11.111 83 Sarcococca coriaceae (Hook.) Sweet Sar_cor Buxaceae 33.333 84 Saurauia napaulensis DC. Sau_nap Saurauiaceae 14.815 85 Schima wallichii (DC.) Korth Sch_wal Theaceae 7.407 86 Scurulla parasitica L. Scu_par Loranthaceae 22.222 87 Semecarpus anacardium L.f. Sem_ana Anacardiaceae 7.407 88 Senecio cappa Buch–Ham.ex D.Don Sen_cap Asteraceae 16.667 89 Senecio cappa Buch–Ham.ex D.Don Sen_cap Asteraceae 6.549 90 Smilax ovalifolia Roxb. Smi_ova smilaceae 38.889 91 Swertia nervosa (G. DON) Griseb Swe_ner Gentianaceae 9.259 92 Symplocos theifolia D. Don Sym_the Symplocaceae 55.556 93 Taraxacum officinale F. H. Camus Tar_off Asteraceae 11.111 94 Tetrastigma serrulatum (Roxb.) planch. Tet_ser Vitaceae 33.333 95 Thalictrum punduanum wall Tha_pun Ranunculaceae 35.185 96 Thunbergia coccinea Wall. Ex D. Don Thu_coc Acanthaceae 7.407 97 Thymas linearis Benth Thy_lin Labiateae 9.259 98 Unknown 1 Unk_1 7.407 Vaccinium retusum (Griff.) Hook. F. ex C. B. 99 Vac_ret Ericaceae 35.185 Clarke 100 Vibernum cylindricum Buch– Ham.ex. D. Don Vib_cyl Sambucaceae 14.815 101 Vibernum erubescens Wall. Ex Dc. Vib_eru Sambucaceae 33.333 102 viola biflora L. vio_bif Violaceae 24.074 103 Wikstroemia canescens Wall.ex Mesin Wik_can Thymalaceae 42.593 104 Zanthoxyllum oxyphyllum Edgew. Zan_oxy Rutaceae 9.259

Appendix IV. Density (number of individuals per 25 m2) of Daphne bholua in three elevation bands in MPF. Data shown are mean ± S.E.

Malika (2300–2500) m Bhainse Mela (2100–2300) m Uttise (1900–2100) m

open semi close Average open semi close Average Open semi close Average

Seedling 1.66 ± 0.41 3.00 ± 0.62 3.50 ± 1.37 2.72 ± 0.52 1.50 ± 0.28 1.75 ± 0.59 3.00 ± 0.67 2.08 ± 0.32 0.33 ± 0.01 0.66 ± 0.25 0.041 ± 0.14 0.47 ± 1.16

Juvenile 1.08 ± 0.14 1.25 ± 1.79 1.33 ± 0.30 1.22 ± 0.12 1.00 ± 0.21 1.33 ± 0.33 1.00 ± 0.21 1.11 ± 0.13 0.33 ± 0.14 0.58 ± 0.14 0.58 ± 0.19 0.50 ± 0.93

Adult 1 1.00 ± 0.36 1.25 ± 0.49 1.00 ± 0.40 1.08 ± 0.24 1.33± 0.33 1.08 ± 0.41 1.25 ± 0.44 1.22 ± 0.22 0.16 ± 0.11 0.33 ± 0.14 0.25 ± 0.17 0.25 ± 0.08

Adult 2 0.75 ± 0.25 1.16 ± 0.40 1.41 ± 0.45 1.11 ± 0.21 0.75 ± 0.32 1.75 ± 0.49 1.08 ± 0.39 1.19 ± 0.24 0.08 ± 0.01 0.33 ± 0.14 0.16 ± 0.11 0.19 ± 0.06

Total 4.50 ± 0.48 6.66 ± 0.96 7.20 ± 1.68 6.12 ± 0.91 4.58 ± 0.49 5.91 ± 0.46 6.33 ± 0.55 5.61 ± 1.17 0.91 ± 0.14 1.75 ± 0.35 1.33 ± 0.37 0.99 ± 0.32

Appendix V. Growth related traits of Daphne in Three sites. Values shown are mean ± SE Malika (2300–250) m Bhainse Mela (2100– 2300) m Uttise (1900–2100) m Growth related traits open semi close Average open semi close Average open semi close Average

Plant height (cm) 123 ± 8 178 ± 25 118 ± 18 139 ± 10 126 ± 9 129 ± 10 115 ± 7 123 ± 5 67 ± 8 64 ± 6 59 ± 6 58 ± 3b

Crown radius (cm) 54 ± 7 45 ± 6 57 ± 6 51 ± 3 40 ± 4 53 ± 4 42 ± 5 45 ± 2 30 ± 2 35 ± 2 36 ± 2 36 ± 1

Stem diameter (mm) 2.6 ± 0.4 2.8 ± 0.3 3.0 ± 0.6 2.9 ± 0.2 2.1 ± 0.2 2.0 ± 0.1 2.1 ± 0.2 2.1 ± 0.1 1.7 ± 0.2 1.5 ± 0.4 1.5 ± 0.3 1.6 ± 0.2

No. of branch per plant 13.2 ± 2.1 12.2 ± 2.0 11 ± 2.1 12.0 ± 1.2 8.1 ± 0.6 8.2 ± 0.9 6.5 ± 0.6 7.61 ± 0.4 6.2 ± 2.0 2.9 ± 0.6 3.6 ± 0.5 6.5 ± 0.7

Floral output 86.± 19.5 140 ± 14.7 75.8 ± 14.0 110 ± 12.7 88 ± 4.5 109 ± 8.5 97 ± 7.12 99.2 ±8.79 55.7 ± 4.5 87.24± 7.65 107± 9.5 82.4± 5.5

Dry bark mass per plot (gm) 3.5 ± 0.3 3.1 ± 0.4 3.1 ± 0.3 3.2 ± 0.1 8.0 ± 0.6 8.2 ± 0.9 6.5 ± 0.6 2.7 ± 0 1.5 ± 0.3 1.0 ± 0.2 1.1 ± 0.2 1.1 ± 0.1

Bark thickness (mm) 2.4 ± 0.4 1.4 ± 0.3 2.4 ± 0.3 2.2 ± 0.1 1.7 ± 0.2 1.6 ± 0.2 2.0 ± 0.2 1.7 ± 0.1 1.0 ± 0.3 0.7 ± 0.1 0.4 ± 0.1 0.8 ± 0.1

Coppice growth 1.1 ± 0.6 0.2 ± 0.1 1 ± 0.6 0.8 ± 0.2 0.4 ± 0.4 0.2 ± 0.2 0.3 ± 0.3 0.2 ± 0.1 4 ± 1.9 3.8 ± 1.4 2.3 ± 0.9 1.5 ± 0.3

Appendix VI. Letter of Permission

PHOTO PLATES

Documenting information about of D. bholua at study area.

Documenting the status of D. bholua in the study plots

Visiting and documenting the handmade paper making traditions in Ilam, Jhapa, Panchthar and Sindhupalchoke Nepal.