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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (05): 658-666.doi: 10.3724/SP.J.1006.2016.00658


Cloning and Characterization of Phospholipids:Diacylglycerol Acyltransferase (BnPDAT1) cDNA from Brassica napus L.

TAN Tai-Long,FENG Tao,LUO Hai-Yan,PENG Ye,LIU Rui-Yang,GUAN Chun-Yun   

  1. College of Agronomy, Hunan Agricultural University / National Oilseed Crops Improvement Center in Hunan, Changsha 410128, China
  • Received:2015-11-13 Revised:2016-03-02 Online:2016-05-12 Published:2016-03-11
  • Contact: Guan Chunyun, E-mail: guancy2011@aliyun.com E-mail:ttl205@aliyun.com
  • Supported by:

    This work was supported by the National Nature Science Foundation of China (31401419), Outstanding youth project of Hunan Provincial Education Department (13B051), the National High Technology Research and Development Program of China (863 Program) (2011AA10A104, 2012AA101107), Science Foundation of Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization in Crop (11KFXM08)


Phospholipids:diacylglycerol acyltransferase (PDAT1) is a key enzyme in triacylglycerol (TAG) biosynthesis of plants. In this study, three novel PDAT1 coding sequences (CDSs) were isolated from cDNA of Brassica napus L. cv. Xiangyou 15 seeds, which were mapped to the chromosomes A02, A10, and C09, and designated as BnPDAT1-A02, BnPDAT1-A10, and BnPDAT1-C09, respectively. Three BnPDAT1 CDSs were 1998, 2002, and 2005 bp in length and encoded predicted proteins with 665, 666, and 667 amino acid residues, respectively. BnPDAT1 proteins were predicted to be located on the cell membrane and have a typical PDAT1 conserved domain. Multiple sequence alignments and phylogenetic analysis showed that the deduced amino acid sequences of BnPDAT1 were highly homologous to previously reported PDAT1 in Brassica oleracea, Arabidopsis thalian, and Eruca sativa. Furthermore, the catalytic enzyme activity of the cloned BnPDAT1 genes was confirmed by the yeast complementary experiment. The expression level of BnPDAT1s increased gradually in seed development and reached the maximum from 25 to 30 days after flowering. However, three BnPDAT1 copies were also found to be different in expression pattern.

Key words: Brassica napus L, PDAT1, Gene clone, Yeast complementary experiment

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