NUTRIENT REQUIREMENTS FOR IN VITRO PROPAGATION OF RICINUS COMMUNIS L. ZYGOTIC EMBRYO USING THE BASAL MEDIA OF MURASHIGE AND SKOOG, GAMBORG ET AL. AND SCHENK AND HILDEBRANDT

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ABSTRACT

This study was carried out on the nutrient requirements for the in vitro propagation of Ricinus communis employing the basal media of Murashige and Skoog (1962), Gamborg et al. (1968), and Schenk and Hildebrandt (1972) using zygotic embryos as explants. Zygotic embryos were excised from mature seeds and cultured on the three basal media with 3 per cent sucrose and 8 g/l of agar. Plant growth regulators were not added to the media. This study was done to determine the most suitable basal medium for the growth of R. communis zygotic embryo. The results obtained showed that the three basal media employed supported in vitro regeneration of the embryo explants. The highest mean shoot length (4.450±0.231 cm), the highest mean root length (2.190±0.262 cm), highest mean fresh weight (0.365±0.032 g), highest mean leaf area


(1.999±0.189 cm2), highest mean per cent sprouting (91.660±0.000), and highest mean number of roots (4.600±0.163) were observed on Murashige and Skoog medium whereas the highest mean sprout rate (0.330±0.000) was obtained on Murashige and Skoog and Gamborg et al. media. The embryo explants were able to develop into normal plantlets even in the absence of growth regulators. This may suggest that endogenous hormones in the zygotic embryo were present at an optimal level to support regeneration. Results from this study indicated that Murashige and Skoog basal medium was the best basal medium for the in vitro propagation of Ricinus communis zygotic embryo. The results are discussed in the light of its potential for mass production of Ricinus communis for its economic values.


TABLE OF CONTENT

Title Page
List of plates
List of Tables
Abstract
Table of content

CHAPTER ONE: INTRODUCTION
1.1       Economic importance of R. communis
1.2       Problem Statement

CHAPTER TWO: LITERATURE REVIEW
2.1Botany of R. communis
2.2       Root system
2.3       Foliage system
2.4       Reproductive system
2.5       Pest and Diseases
2.6       In vitro plant propagation
2.7       In vitro propagation of Ricinus communis
2.8       Effects of basal media on in vitro culture of plants
2.9       Tissue culture media
2.10 Explants

CHAPTER THREE: MATERIALS AND METHODS
3.1       Source of explant
3.2       Site of experiment
3.3       Media composition
3.4       Preparation of stock solution of media
3.5       Sterilization techniques and embryo transfer
3.6       Preparation of one liter of basal media (MS, B5 and SH)
3.7       Experimental design
3.8       Statistical analysis
3.9       Regeneration studies
3.9.1    Time course in sprouting
3.9.2    Rate of sprouting of explant
3.9.3    Length of shoot produced
3.9.4    Length of root produced
3.9.5    Leaf area of plantlets
3.9.6    Number of leaves and roots produced
3.9.7    Fresh weight of plantlets

CHAPTER FOUR: RESULTS
4.1 In vitro growth studies of embryo explants of R. communis
4.2 Effects of the three basal media on per cent sprouting and sprout rate of R. communis zygotic embryos after two weeks of growth
4.3 Effects of basal media on shoot and root length of plantlets produced after two weeks of growth
4.4 Effects of basal media on fresh weight and leaf area of plantlets produced after two weeks of growth
4.5 Effects of basal media on number of leaves and roots produced after two weeks of growth

CHAPTER FIVE: DISCUSSION
CONCLUSION
RFERENCES
LIST OF ABBREVIATION
APPENDICES


CHAPTER ONE

INTRODUCTION
Castor oil plant (Ricinus communis L.) is a non food, drought resistant, energy plant gaining attention for producing biofuel as biodiesel in developing countries (Kamrun, 2013). As an oil bearing biomass feedstock, it can ensure an alternative source of energy and reduce the over dependency on fossil fuel (Kamrun, 2013). Ethiopia is considered to be the most probable site of origin of castor oil plant because of the presence of high diversity (Anjani, 2012). However, according to Sujatha et al. (2008) the plant originated from Abyssinia. It is distributed throughout the tropics and the subtropics, and is well adapted to the temperate region (Sujatha et al., 2008).

Oil extracted from castor seeds is of economic importance especially in the chemical industries (Abayeh et al., 1998). The economic importance of this plant has made it necessary that the plant be properly investigated to develop the most reliable method of propagation. Conventional method of cultivation using the seed is limited by problems of seed viability. Propagation through tissue culture techniques employing embryo is necessary to eliminate the limitations of seed germination.

1.1    Economic importance


Ogiri is a food condiment obtained by traditional fermentation of castor seeds in the eastern part of Nigeria. This food condiment provides dietary fiber, energy, mineral and vitamins (Kolapo et al., 2007). At present, the potentials of castor oil have not been fully explored in Nigeria (Oluwole et al., 2012).
Castor seed is an important source of vegetable and medicinal oil and has numerous benefits to humanity. The oil from castor seeds has many industrial applications. Dehydrated oil from castor seeds is used in the paint and varnish industry. The oil is also used in the manufacture of a wide range of sophisticated products like nylon fibers, jet engine lubricants, hydraulic fluids, plastics, artificial leather, and fiber optics (Ogunniyi, 2006).
The seeds contain 40 to 60% oil that is rich in triglycerides mainly ricinolein, a toxic alkaloid ricinine and very toxic albumen called ricin (Zohary, 1987). Oil from castor seed is prescribed for infestation of intestinal worms. In vitro antiviral activity and hypoglycemic activity were also reported from leaf extracts (Khafagi, 2007).

Oil from castor seeds also has great promise in the field of biodiesel production as it is inexpensive and environment friendly (Ogunniyi, 2006). Consequently, there has been a steady increase in the demand of castor oil and its products in the world market due to their renewable nature, biodegradability and eco-friendliness (Ogunniyi, 2006).


Many phytochemicals found in the plant tissue and seeds of castor have potential medicinal uses (Morris, 2004). Castor oil has been used as purgative since ancient times and it is still considered to be a safe and effective laxative (Kalaiselvi et al., 2003). The ricin A-chain has also been linked to antibodies able to target cancer cells while not harming normal cells (Olsnes and Pihl, 1981; Lam et al., 2004). These immunotoxins have been reported to have many potential uses in modern medicine (Scadden et al., 1998; Longo et al., 2000; Sandler et al., 2006). Experimental medicines containing castor oil were also shown to be effective treatments of evaporative dry eye (Khanal et al., 2007)....

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