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FOR
IMPROVING
Directorate of Oilseeds Research, Rajendranagar, Hyderabad 500 030, India E-mail: mulpurisujata@yahoo.com
ABSTRACT
This article highlights the role of biotechnological tools in the genetic improvement of Jatropha and castor belonging to the family euphorbiaceae. Ever increasing fuel prices and depletion of fossil reserves demands alternative fuel sources. For a country like India with predominantly rainfed agriculture, biofuels from water demanding crops and food crops is a difficult proposition and hence, tree borne and non-edible oilseed crops are being researched for exploitation as bioenergy crops. Among the potential biofuel crops, Jatropha and Pongamia assumed priority owing to their easy adaptability, growth on harsh environments and use of transesterified oil in diesel engines. Pongamia is a tree species and improvement through conventional breeding and biotechnological tools is a long drawn process. India holds monopoly in castor area and production and can be exploited for production of biodiesel (www.castoroil.in). While castor is valued for the oil and its derivatives in a host of medicinal and industrial products, the utilitarian value of Jatropha oil and its by-products need extensive investigations. The limitations in available germplasm of Jatropha include; lack of knowledge of the genetic base, poor yields on problematic sites, low genetic diversity and vulnerability to wide array of insects and diseases. During the past three years there is an upsurge of activities in jatropha and information on molecular markers, gene sequences, microsatellite database and cloning and characterization of useful genes (curcin, stearoyl-acyl carrier protein desaturase, JcERF for enhanced resistance to salt and frost) is generated. Tissue culture protocols are refined and regeneration through direct adventitious and somatic embryogenesis is reported. Genetic transformation protocols are in place which indicates potential for widening the genetic base through biotechnological tools. Different types of molecular markers are used in assessment of genetic diversity in the germplasm and should pave way for marker assisted breeding. In castor, the major limitations include susceptibility to biotic stresses and presence of the toxic protein ricin in the seed meal which are being addressed through transgenic approach and TILLING, respectively. Castor genome sequencing has been initiated at the Institute for Genomic Research (TIGR) and around 50,000 expressed sequence tags (ESTs) have been produced for gene identification and annotation. Bioengineering of fatty acid metabolism pathway in both these crops has huge scope for downstream processing and value-chain expansion. The current status and future prospects of these technologies for genetic upgradation of Jatropha and castor will be discussed.
Introduction
India imports petroleum crude supplies to meet 70% of the energy requirement and the energy demand in the country is increasing at a rate of 6% annually, which is a major limitation threatening sustainable development. Of the several options available, biofuels from energy crops has attracted attention as they are renewable when compared to the fossil fuels that are finite, combat climate change, and as an alternative for the ever increasing raw petroleum prices. The production of biofuels has to be considered against different aspects such as energy yield per unit area under cultivation, food versus fuel needs, potential for substitution of fossil fuels, cost of production and competitiveness with fossil fuels, overall energy balance and net reduction potential of greenhouse gas emissions, sustainable agricultural practice, competition with food and animal fodder and energy, trade and agricultural policy issues (1). The developed nations have the option for exploitation of food crops for biofuel purpose, but introduction of first-
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Table 1: Fatty acid composition (%) of seed oils of J. curcas and castor
Fatty acid Myristic acid Palmitic acid Stearic acid Arachidic acid Behemic acid Palmitoleic acid Oleic acid Linoleic acid Ricinoleic acid Dihydroxystearic acid Linolenic acid Eicosanoic acid J. curcas oil 0-0.1 9-22 5-8 0-2 0-2 0-1 35-51 27-42 ----Castor oil -1 1 ---3 4.2 89.5 0.7 0.3 0.3
The presence of hydroxyl group and double bonds imparts unique chemical and physical properties that make castor oil a vital industrial raw material and stabilizes the oil against oxidation (9). The fuel specific properties of both the oils are presented in Table 2. Castor oil is the only oil soluble in alcohol and does not require the consequent energy requirement in transesterification as for other vegetable oils. Castor oil has a good shelf life when compared to other vegetable oil and it does not turn rancid when subjected to excessive heat. Castor oil is four times more stable than olive oil. The high viscosity, high water content, higher compressibility than other vegetable oils, reduction of about 10% of hydroxyl and acid values if stored for about 90 days and the premium price of castor oil are the major issues limiting the use of straight castor oil as a fuel for internal combustion engines (10). However, the limit values of viscosity, density and cetane number can be met through transesterification followed by dilution or blending with conventional diesel fuel and other vegetable oils. The key areas of research include development of oleic acid rich castor or production of ricinoleic acid in heterologous systems. It is relatively easy to modify the biosynthetic pathway rather than producing the industrial fatty acids in heterologous systems owing to low level of accumulation of these products in transgenic plants. One such example is the transgenic expression of the fatty acid hydroxylase (Fah12) gene under the control of strong seed specific promoter which resulted in very low levels (<1% in tobacco, 17% in Arabidopsis) of accumulation of ricinoleic acid thus, limiting commercial exploitation (11). Metabolic pathways including those involved in oil accumulation in seeds are conserved across plant species and such information has been utilized to develop designer oil crops. Information on mechanisms in seed-oil accumulation and metabolic engineering of fatty acid profile can be applied for improvement of oil content in J. curcas and oil quality in castor. The expression profiles of genes involved in fatty acid biosynthesis, transport during cellular endosperm development and accumulation of triacylglycerols were investigated in castor (12). Oil content can be increased by introducing/overexpressing the genes. Conversion of ricinoleic rich castor oil to oleic rich castor oil can be accomplished through silencing the fatty acid hydroxylase gene. Glycerine is the coproduct which is in great demand as raw material for cosmetics, medicine and food product industries. Before the economic evaluation of all the various plant parts, commercial development of J. curcas remains a major challenge.
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PETROTECH 2009 11 - 15 JANUARY 2009, NEW DELHI, INDIA Table 2: Fuel specific properties of J. curcas and castor oil
Property Seed oil content (5) Molecular weight Melting point Solidification point Viscosity (dPa.s) Ash content Sulphur Potassium Calorific value (MJ/kg) Iodine value Cetane value (Flammability) Density (g/ml) Flash point Viscosity at 400C (CSt) Acid value Saponification value Kinematic viscosity (400C) mm2s-1 Water content (%) Oxidation stability (lus) J. curcas oil 33-36% ---4.4 <0.01 --38-40 94-102 58.5 0.88-0.92 172-2680C 34.36 1.24-3.8 177-198 --8.39 Castor oil 48-54% 298 50C -120C to -180C 9.5-10.0 <0.02% 0.04% Negligible 39.5 82-90 42 0.956-0.963 2600C ---240-300 0.15-0.30 --
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