Iqbal Munir, Ijaz Naeem, Timothy P Durrett, Aqib Iqbal and Mian Afaq Ahmad
University of Agriculture Peshawar, Pakistan
Kansas State University, USA
Posters & Accepted Abstracts: J Fundam Renewable Energy Appl
Energy crises and environmental concerns are driving researchers to develop viable alternative fuels from renewable sources. The use of Brassica juncea oil as an alternative fuels suffers from problems such as high viscosity, low volatility and poor cold temperature properties. The seed of Euonymus alatus produces unusual triacylglycerol (TAGs) called acetyl triacylglycerol (acTAGs) where the sn-3 position is esterified with acetate instead of a long chain fatty acid. The enzyme Euonymus alatus diacylglycerol acetyltransferase (EaDacT) present in these plants is an acetyltransferase that catalyzes the transfer of an acetyl group from acetyl-CoA to diacylglycerol (DAG) to produce acetyl TAG (AcTAG). In order to reduce the viscosity of Brassica juncea oil by synthesizing acTAG, we have optimized an efficient and simple agrobacterium mediated floral dip transformation method to generate transgenic Brassica juncea plants with EaDacT gene. A binary vector containing the EaDacT gene under the transcriptional control of a glycinin promoter and with a basta selection marker was transformed into Agrobacterium tumefaciens strain GV-3101 through electroporation and subsequently to B. juncea through floral dip method. The basta resistant putative transgenic plants were further confirmed by PCR. The developed transgenic B. juncea seeds showed altered TAG fatty acid composition with enhanced level of oleic acid (from 41% to 63%) and reduced Erucic acid level (11.18%), which is an ideal composition of fatty acids to be used as biodiesel. The results showed that the Agrobacterium-mediated floral-dip transformation can be a successful strategy to develop transgenic Brassica Juncea having oil with modified fatty acids profile that could directly be used as biodiesel. Further, the developed protocols could be used to accumulate unusual acTAG in B. juncea seed, providing a direct way of biodiesel production from plant oil.
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