Functional characterization of two type-1 diacylglycerol acyltransferase (DGAT1) genes from rice ( Oryza sativa ) embryo
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Functional characterization of two type‑1 diacylglycerol acyltransferase (DGAT1) genes from rice (Oryza sativa) embryo restoring the triacylglycerol accumulation in yeast Rupam Kumar Bhunia1 · Kshitija Sinha1 · Kirti Chawla1 · Vinay Randhawa2 · Tilak Raj Sharma1 Received: 28 January 2020 / Accepted: 13 October 2020 © Springer Nature B.V. 2020
Abstract Key message Two OsDGAT1 genes showed the ability to restore TAG and LB synthesis in yeast H1246. Alterations in the N-terminal region of OsDGAT1-1 gene revealed its regulatory role in gene function. Abstract Accumulation of triacylglycerol (TAG) or oil in vegetative tissues has emerged as a promising approach to meet the global needs of food, feed, and fuel. Rice (Oryza sativa) has been recognized as an important cereal crop containing nutritional rice bran oil with high economic value for renewable energy production. To identify the key component involved in storage lipid biosynthesis, two type-1 diacylglycerol acyltransferases (DGAT1) from rice were characterized for its in vivo function in the H1246 (dga1, lro1, are1 and are2) yeast quadruple mutant. The ectopic expression of rice DGAT1 (designated as OsDGAT1-1 and OsDGAT1-2) genes restored the capability of TAG synthesis and lipid body (LB) formation in H1246. OsDGAT1-1 showed nearly equal substrate preferences to C16:0-CoA and 18:1-CoA whereas OsDGAT1-2 displayed substrate selectivity for C16:0-CoA over 18:1-CoA, indicating that these enzymes have contrasting substrate specificities. In parallel, we have identified the intrinsically disordered region (IDR) at the N-terminal domains of OsDGAT1 proteins. The regulatory role of the N-terminal domain was dissected. Single point mutations at the phosphorylation sites and truncations of the N-terminal region highlighted reduced lipid accumulation capabilities among different OsDGAT1-1 variants. Keywords Rice (Oryza sativa) · Type-1 diacylglycerol acyltransferases (DGAT1) · Triacylglycerol (TAG) · Lipid bodies · Phosphorylation
Introduction Vegetable oil-producing crops have been one of the most dynamic parts of agriculture. Worldwide, over the next thirty years, demand for vegetable oil is expected to be doubled Kirti Chawla and Vinay Randhawa have contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11103-020-01085-w) contains supplementary material, which is available to authorized users. * Rupam Kumar Bhunia [email protected]; [email protected] 1
Plant Tissue Culture and Genetic Engineering, National Agri-Food Biotechnology Institute (NABI), Mohali, Punjab 140306, India
Department of Biochemistry, Panjab University, Chandigarh 160014, India
2
with increasing requirements for food, feed and fuel. Rice is one of the most important crops for mankind as it is a source of dietary energy for nearly half of the world’s population. Apart from starch, outer brown layer of rice grains contain lipids (3–4%), which are used to produce rice bran oil (Liu 2011). In India, harvesti
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