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PROJECT TOPIC- 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)

PROJECT TOPIC- 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)

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,  -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 subtropics, 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).

PROJECT TOPIC- 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)

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 oil content in the seed ranges from 30 – 50 % while kernel ranges from 45 – 60 %. The leaves are palmate in shape, arranged alternately along the stems and each leaf has 3-5 lobes, 10-15 cm in length and width. J. curcas is recognized by many vernacular names such as, oboboloti (Igbo); lapalapa (Yoruba); butuje (Hausa); jarak budge (Indonesia); kpoti (Togo); physic nut (Australia) and mapuluka (Angola) (Prakash et al., 2007; Wikipedia, 2007). The oil from the seed contains insecticides which protect it from insect attack. Raju and Ezradanam (2002) reported that the species is monoecious and produces male and female flowers in the same inflorescence.

The ratio of male and female flowers ranges from 13:1 to 29:1. Normally, the species flowers only once a year during the rainy season. The male flowers open first and persist until all male buds are exhausted (Achten et al., 2008). The flowers are formed with female flowers usually slightly larger than the male in the hot season and they are unisexual and have sepals up to 18 mm long. Pan and Xu (2010) reported that the female flowers open between the second and sixth day after formation. Each inflorescence is composed of 100 – 300 flowers and after pollination the inflorescence forms a bunch of green ellipsoidal fruits. The fruit is an ellipsoid capsule, 2.5 – 3 cm long and 2.3 cm in diameter and they are produced in winter when the shrub is leafless (Watt and Breyer – Brandwijk, 2001).
Prakash et al. (2007) reported that the ovary is trilocular and hence has three styles and three bifid stigmas which lead to three seeds per capsule. The seed is black and about 18 mm long and 10 mm wide. The weight of seed (per 1000) is about 727 g (Heller, 1996). The seeds (less than 300 mg each) are not all viable, that is the higher the size/weight of the seed, the greater the viability (Hannan –Jones and Csurshes, 2008). The plant begins to produce fruits
4 – 5 months after germination and reaches peck of seed production at about 3 years of age.
The plant grows on well-drained soil with good aeration as well as marginal soil with low nutrient content; the species grows almost in all soils, including sandy, gravelly and saline soil (Pankaji and Divay, 2011; Katwal and Soni, 2003). The leaves are shed during the winter months and it is best adapted to arid and semi-arid conditions. Garg et al. (2011) reported that it has thrived successfully in higher regions of the tropics with an annual rainfall of between
300 mm and 1000 mm. The species tolerates a very light frost (a weather condition in which the temperature falls below freezing point) and is very sensitive to wind (Wiesenhutter, 2003) Heller (1996) reported that the physic nut is planted in the tropics as a hedge (Living fence) around fields, villages and homesteads. It is also an excellent species for agro forestry and also as land boundary mark.

The plant is used to control soil erosion (Warra, 2010).
All parts of the plant; seeds and bark are used in traditional medicine to treat dropsy, paralysis, guinea worm, sores and veterinary purposes (Wiesenhutter, 2003). The leaves of J. curcas contain apigenin, vitexin and isovitexin along with other phytochemicals that enable them to be medicinally used in treating malaria, rheumatic pains, muscular pains and tooth ache (Zewdneh et al., 2011). The latex contains alkaloids known as “Jatrophine” which is these anti-cancerous properties (Kosasi, 1989). Further, the tender twigs of the plant are used for cleaning the teeth while the juice from the leaf is used as an external application for piles (Rajore and Bartra, 2005). In Gabon, the seed powder of J. curcas mixed with palm oil is used as a rodenticide as well as for skin diseases, while dried seeds immersed into palm oil are used as torches (Gubitz, et al., 1999).

The oil from the seed can serve as diesel for vehicle, lamps and generator (Shrivastava and Banerjee, 2008; Purkayastha et al., 2010). In Africa and India, the species is planted for reforestation of eroded areas. Moshood et al. (2011) reported that the species contains the four most important fatty acids which include: palmitic acid (C 16:10), Linoleic acid (C 18:2), Stearic acid (C 18:0) and Oleic acid (C 18:1). Palmitic (C 16:10) and Stearic acid (C 18:0) are the saturated fatty acids with acid content of 15.38% and 6.24% while Oleic acid (C 18:1) and Linoleic acid (C 18:2) are the unsaturated acids of about 40.23% and 36.32% respectively.

Jatropha curcas has a cosmetic potential because the fruit contains viscous oil that can be used for soap making and gives a very good foaming, white soap with positive effects on the skin, partly due to the glycerine content of the soap (Warra, 2012). Some pests and diseases have been reported on Jatropha curcas such as, powdery mildew which damages the leaves and flowers, Alternaria spp that causes premature leaf fall and golden flea beetles, which eat young leaves and shoots (Marieke et al., 2012). 1.5 Due to the multi-potential of the species, Jatropha curcas was chosen for the present study with the following objectives:
 To develop a propagation protocol for J. curcas employing zygotic embryos.

 To investigate the percentage growth rates with respect to different treatment combinations of growth regulators, and two basal media (Linsmaier and Skoog, 1965 and Schenk and Hildebrandt, 1972).
 To determine the best treatment combinations of growth regulators suitable for in vitro multiplication of J. curcas.
The study if successful can provide information for full biotechnological exploitation of the species potential.

PROJECT TOPIC- 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)

 

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