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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (07): 1000-1008.doi: 10.3724/SP.J.1006.2016.01000

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Analyses of Expression and for BnFAD2 Genes in Brassica napus

LIU Rui-Yang,LIU Fang,GUAN Chun-Yun   

  1. College of Agronomy, Hunan Agricultural University / National Oilseed Crops Improvement Center in Hunan, Changsha 410128, China
  • Received:2016-01-11 Revised:2016-05-09 Online:2016-07-12 Published:2016-05-11
  • Contact: 官春云, E-mail: guancy2011@yahoo.com.cn E-mail:ruiyang_liu2007@126.com
  • Supported by:

    This study was supported by Graduate Innovation Foundation of Hunan (CX2013A012) and the Major State Basic Research Development Program of China (2015CB150200).

Abstract:

The oil containing high oleic acid is high nutritional. In Brassica napus, the fatty acid desaturase gene (FAD2) is the key gene controlling oleic acid content. In this study, the full-length cDNA sequences of three genes located on chromosome A5, C5 and A1 in Brassica napus were cloned and named BnFAD2-A5, BnFAD2-C5 and BnFAD2-A1. The three genes encode proteins with 384, 384 and 136 amino acid residues, respectively. TMHMM was used to predict transmembrane domain and Clust X software was used to analyze the activity center of FAD2 genes. Both of the results showed that BnFAD2-A1 did not have the function of dehydrogenase. The yeast complementary experiment on four genes (including published BnFAD2-C1 gene) showed that the desaturation capability of BnFAD2-A5 gene was next to that of BnFAD2-C5 gene, and both of them were greater than that of BnFAD2-C1 gene. The expression patterns of the four genes were analyzed by using qRT-PCR technique in different tissues and the protein stability of BnFAD2-C1, BnFAD2-A5 and BnFAD2-C5 was analyzed by using Hemagglutinin labeling method. Both of the results revealed that BnFAD2-A5 and BnFAD2-C5 aremajor genes affecting the accumulation of oleic acid in rape seed.

Key words: Brassica napus, BnFAD2, Bioinformation, Protein activity, Protein stability

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