Cloning and Functional Analysis of Promoter of Diacylglycerol Acyltransferase Gene GmDGAT1A in Soybean

doi:10.7668/hbnxb.20190821

Abstract

Abstract: Diacylglycerol acyltransferase (DGAT) is a key enzyme in the biosynthesis of triacylglycerol,which plays an important role in the synthesis and accumulation of triacylglycerol. In order to study the molecular mechanism of DGAT gene expression regulation in soybean,the promoter of GmDGAT1A (promoter-GmDGAT1A,pGmDGATIA) was cloned from the variety Kefeng 1 by using PCR amplification,and its function was analyzed by transforming Arabidopsis thaliana and GUS tissue localization. The results showed that:With leaf DNA as template,the 2 192 bp promoter fragment of GmDGAT1A was successfully cloned from the variety Kefeng 1. Sequence analysis showed that p GmDGAT1A not only had the basic cis-acting elements such as TATA-box and CAAT-box,but also contained a number of cis-acting elements responsive to light,gibberellin and abscisic acid. The plant expression vector pCAMBIA1381Z-p GmDGAT1A was successfully constructed using GUS as a reporter gene and transformed into wild type Arabidopsis thaliana to obtain transgenic plants. The 2 192 bp band could be amplified from the transgenic Arabidopsis thaliana plants by PCR,indicating that a transgenic line containing p GmDGAT1A was obtained. GUS histochemical staining of transgenic Arabidopsis thaliana seedlings showed that the veins and roots were deeply stained,but the root tips of main roots and lateral roots were not stained. For mature Arabidopsis thaliana plants,enhanced GUS staining was observed in roots,veins and diaphragms and funiculi of siliques,while the stems and seeds were not stained. These suggested that p GmDGAT1A -driven GUS was mainly expressed in roots,veins and diaphragms and funiculi of siliques. In conclusion,the cloned promoter of GmDGAT1A has the activity,can drive the expression of downstream target gene,and may be used in transgenic breeding.

Key words: Soybean, Diacylglycerol acyltransferase (DGAT), Promoter, Oil, GUS

CLC Number:

S565.03
Q78

CHAO Maoni, HU Xigui, ZHANG Jinyu, WANG Runhao, WEN Qingyu, SUN Xinkai, HUANG Zhongwen. Cloning and Functional Analysis of Promoter of Diacylglycerol Acyltransferase Gene GmDGAT1A in Soybean[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 27-34. doi: 10.7668/hbnxb.20190821.

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