Ectopic expression of AtDGAT1, encoding diacylglycerol O-acyltransferase exclusively committed to TAG biosynthesis, enhances oil accumulation in seeds and leaves of Jatropha.

Devendra Kumar Maravi,Nozomu Sakurai,Hiroyuki Koyama,Sanjeev Kumar,Prabin Kumar Sharma,Yasufumi Kobayashi,Vaibhav V Goud,Lingaraj Sahoo

doi:10.1186/s13068-016-0642-7

Abstract

Background Jatropha curcas is an important biofuel crop due to the presence of high amount of oil in its seeds suitable for biodiesel production. Triacylglycerols (TAGs) are the most abundant form of storage oil in plants. Diacylglycerol O-acyltransferase (DGAT1) enzyme is responsible for the last and only committed step in seed TAG biosynthesis. Direct upregulation of TAG biosynthesis in seeds and vegetative tissues through overexpression of the DGAT1 could enhance the energy density of the biomass, making significant impact on biofuel production.ResultsThe enzyme diacylglycerol O-acyltransferase is the rate-limiting enzyme responsible for the TAG biosynthesis in seeds. We generated transgenic Jatropha ectopically expressing an Arabidopsis DGAT1 gene through Agrobacterium-mediated transformation. The resulting AtDGAT1 transgenic plants showed a dramatic increase in lipid content by 1.5- to 2 fold in leaves and 20–30 % in seeds, and an overall increase in TAG and DAG, and lower free fatty acid (FFA) levels compared to the wild-type plants. The increase in oil content in transgenic plants is accompanied with increase in average plant height, seeds per tree, average 100-seed weight, and seed length and breadth. The enhanced TAG accumulation in transgenic plants had no penalty on the growth rates, growth patterns, leaf number, and leaf size of plants.ConclusionsIn this study, we produced transgenic Jatropha ectopically expressing AtDGAT1. We successfully increased the oil content by 20–30 % in seeds and 1.5- to 2.0-fold in leaves of Jatropha through genetic engineering. Transgenic plants had reduced FFA content compared with control plants. Our strategy of increasing energy density by enhancing oil accumulation in both seeds and leaves in Jatropha would make it economically more sustainable for biofuel production.

Highlights

Jatropha curcas is an important biofuel crop due to the presence of high amount of oil in its seeds suitable for biodiesel production
Generation of AtDGAT1 overexpressing transgenic Jatropha using a constitutive promoter and molecular characterization To investigate the impact of constitutive overexpression of Arabidopsis diacylglycerol acyltransferase 1 (DGAT1) cDNA on TAG accumulation in leaves and seeds of Jatropha, we prepared a 35S::AtDGAT1 construct that consisted of AtDGAT1 fused to gus reporter gene driven by CaMV35S promoter and nptII as plant selection marker (Fig. 1A)
These plants were confirmed by genotyping through genomic DNA Polymerase chain reaction (PCR) using primers specific to AtDGAT1, nptII, and gus

Summary

Introduction

Jatropha curcas is an important biofuel crop due to the presence of high amount of oil in its seeds suitable for biodiesel production. Direct upregulation of TAG biosynthesis in seeds and vegetative tissues through overexpression of the DGAT1 could enhance the energy density of the biomass, making significant impact on biofuel production. Despite the significance of Jatropha seed oil as a potential source for biodiesel, not much research efforts have been made through breeding or transgenic approaches to improve its seed oil content and quality for sustainable biodiesel production. FFAs are involved in sequential acylation of the sn-1, sn-2, and sn-3 positions of glycerol-3-phosphate with acyl-CoA to yield TAG through Kennedy pathway

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Conclusion