Cloning and Functional Verification of Acetyltransferase Gene in Brassica napus

doi:10.7668/hbnxb.20191383

Abstract

Abstract: To explore the effect of acetyltransferase gene on the fatty acid composition of Brassica napus. By using the cDNA of 20-35 d inbred of rapeseed with the lower linolenic acid content, 2 copies(114 and 148) of acetyltransferase gene(AT) were cloned and conducted a bioinformatics analysis.The overexpression vector and interference vector were constructed and transformed into Arabidopsis thaliana. The T₀ seeds were screened by hydomycin and the resistant seedlings were identified by PCR. The T₁ seeds were obtained and the fatty acid composition of T₁ seeds were determined by gas chromatography. The results showed that the protein encoded by 114 was neutral while the protein encoded by 148 was alkaline. The proteins encoded by 114 and 148 were both stable proteins with poor hydrophilicity. They were lipid soluble proteins and located in the cytoplasm without transmembrane structure. The two copies of AT gene, 114 and 148, had an effect on the transformation of saturated fatty acids. In the metabolic pathway of fatty acids, 114 and 148 genes inhibit the transformation from palmitic acid to stearic acid. Overexpression of both 114 and 148 genes leads to an increase in palmitic acid and linoleic acid content, and a decrease in stearic acid content. And it was found that overexpressed 114 genes decreased arachidonic acid content, interfering with the expression of the 148 gene increased in erucic acid content. The results of this study showed that these two copies had a significant effect on improving the fatty acid composition of Brassica napus.

Key words: Brassica napus, Acetyltransferase gene, Gene cloning, Overexpression, RNAi

CLC Number:

S635.03
Q78

CHENG Qian, GUAN Mei, ZHANG Zhenqian, CHANG Tao, TAN Min, GUAN Chunyun. Cloning and Functional Verification of Acetyltransferase Gene in Brassica napus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 36-43. doi: 10.7668/hbnxb.20191383.

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