NUTRIENT REQUIRMENTS FOR IN VITRO PROPAGATION OF JATROPHA CURCAS (LINN.) ZYGOTIC EMBRYO ON THE BASAL MEDIA OF LINSMAIER AND SKOOG (1965) AND SCHENK AND HILDERBRANDT (1972)

TABLE OF CONTENTS
Title page
Certification Page
Dedication
Acknowledgement
List of Tables
List of Plates
Abstract
Table of contents

CHAPTER ONE: INTRODUCTION

CHAPTER TWO: LITERATURE REVIEW
2.1 Morphology of Jatropha curcas L.
2.2 In vitro Clonal Propagation
2.3 A simple regeneration protocol from stem explants of Jatropha curcas
2.4 Benefits of Jatropha curcas

CHAPTER THREE: MATERIALS AND METHODS
3.1 Site of the Study
3.2 Source of explants
3.3 Preparation of stock solutions for media
3.4 Procedure for Schenk and Hilderbrandt (1972) and Linsmaier and Skoog (1965)
3.5 Media Composition
3.6 Preparation of Stock Solutions of growth regulators
3.7 Culture Media
3.8 Culture media procedures and preparations
3.9 Sterilization techniques and embryo transfer
3.10 Regeneration studies
3.10.1 Per cent sprouting
3.10.2 Sprout rate
3.10.3 Length of shoot produced
3.10.4  Length of root produced
3.10.5  Leaf area of plantlet produced
3.10.6  Number of leaves produced
3.10.7  Fresh weight of plantlets produced
3.11 Experimental Design/Statistical Analysis

CHAPTER FOUR: RESULTS
4.1 Effects of KIN and NAA on per cent sprouting and sprout rate on LS Medium
4.2Effects  of  KIN  and  NAA  on  shoot  length  (cm),  and  root  length  (cm) of plantlets produced on LS Medium after two weeks of Growth
4.3 Effects of KIN and NAA on leaf area (cm2 ) and number of leaves produced on LS Medium after two weeks of Growth
4.4 Effects of KIN and NAA on Fresh weight (g) of plantlets produced on LS Medium after two weeks of Growth
4.5 Effects of KIN and NAA on per cent sprouting and sprout rate on SH Medium after two weeks of Growth
4.6 Effects of KIN and NAA on shoot length (cm) and root length (cm) of plantlets produced on SH Medium after two weeks of Growth
4.7 Effects of KIN and NAA on leaf area (cm2 ) and number of leaves produced on LS Medium after two weeks of Growth
4.8 Effects of KIN and NAA on Fresh weight (g) of plantlets produced on SH Medium after two weeks of Growth

CHAPTER FIVE: DISCUSSION
REFERENCES
APPENDICES


ABSTRACT
An investigation was carried out on the nutrient requirements for the in vitro propagation of Jatropha curcas Linn. employing the basal media of Linsmaier and Skoog (1965) and Schenk and Hilderbrandt (1972) using zygotic embryos as explants. Zygotic embryos were excised from mature seeds and cultured on the two basal

media  in  which  each  was
supplemented  with  an
auxin,  α-naphthaleneacetic  acid
(NAA)   and   a   cytokinin,
6-furfurylaminopurine
(kinetin)   used   singly   and   in

combination at 0 to 3.0 mgl-1 with 3 per cent sucrose as carbon source. The results obtained showed that growth regulators (auxin and cytokinin) did not necessarily enhance plant regeneration (P = 0.05) from the Jatropha curcas embryo explants at the concentrations applied irrespective of the medium employed. LS basal medium was significantly (P < 0.05) superior to SH basal medium in the enhancement of such growth parameters as per cent sprouting, shoot length, root length, number of leaves and plantlet fresh weight while SH basal medium was significantly (P < 0.05) superior to LS medium in the support of sprout rate and leaf area, all in the absence of externally applied growth regulators. It is apparent that at the time of seed maturity, the embryos had acquired the optimal level of endogenous growth regulators necessary of sprouting. The results are discussed in the light of the potential for mass producing J. curcas as a viable alternative to fossil fuels and for other economic purposes.


CHAPTER ONE
INTRODUCTION
The overdependence on fossil fuels in many countries could be a great problem in the years to come as it has been estimated that in less than fifty years, fossil fuels will either be in serious state of depletion or exhaustion (Heller, 1996). In addition, fossil fuel causes heavy pollution, climate change, energy insecurity, rural poverty in developing countries and depletion of ozone layer. Consequently, research efforts have been geared towards finding alternative and cleaner sources of energy. Devanesan et al. (2007) and Ahmed et al. (2012) reported that Jatropha curcas (a bio-diesel plant) could serve as a viable alternative to fossil fuels as its oil is clean (i.e. its oil burns without emitting smoke) and therefore more environment-friendly. Shrivastava and Banerjee (2008) reported that the improvement of in vitro propagation efficiency of the species is very important for its biodiesel production. In addition, it can ensure a steady supply of disease - free plant and large quantity of planting material in and out of season.

Achten et al. (2008) reported that the origin of J. curcas is still speculative but is believed to have originated from Mexico and Central America from where it spread to Bolivia, Argentina, Brazil, Paraguay and Peru. It was introduced to Africa and India by the Portuguese explorers. Sujatha et al. (2005) reported that the species is abundant in the tropics and sub-tropics, not just for its medicinal value but more as a source of biodiesel. Currently, it is cultivated globally (Hanna-Jones and Csurshes, 2008). Openshaw (2000) reported that J. curcas belongs to the family Euphorbiaceaee. It is a perennial shrub that grows up to 6 m high but can attain a height of 8 - 10 m under favourable conditions (Ravi et al., 2004; Makkar and Becker, 2009). It has smooth gray bark, which exudes whitish colored, latex when cut but turns brown when dry. Jatropha curcas has life span of 30 - 50 years (Joker and Jepsen, 2003). The root grows deep into the soil with a taproot that helps it to withstand wind action and water erosion (Khemeladngoen et al., 2011). Senthikumar et al. (2003) reported that the....

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