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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (05): 725-732.doi: 10.3724/SP.J.1006.2015.00725

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

Cloning and Expression Analysis of Defensin Genes from Brassica napus

ZHENG Xiao-Min,GUO Nan,GAO Tian-Shu,GONG Hui-Ming,ZHANG Tao   

  1. College of Life Sciences / Chongqing Key Laboratory of Plant Molecular Biology Adaptation to the Environment, Chongqing Normal University, Chongqing 401331, China
  • Received:2014-09-22 Revised:2015-02-06 Online:2015-05-12 Published:2015-03-13
  • Contact: 张涛, E-mail: zht2188@126.com

Abstract:

Plant defensins show a broad spectrum of antimicrobial activity, which not merely have antifungal activity, antibacterial activity, proteinase inhibitory activity and insect amylase inhibitory activity, but play roles in regulating plant growth and development as well. According to the sequences of B. rapa defensin genes, specific primers were designed to clone five B. napus defensin genes. The lengths of cDNA sequence were between 325 and 461 bp, with 177 to 243 bp of open reading frames (ORFs), encoding polypeptides of 58 to 80 amino acid residues. The amino acid sequences of plant defensins showed a big difference, but the six to eight conservative cysteine residues contained were stable. All the five cloned B. napus defensin genes contained a conservative Knot1 function domain. Phylogenetic analysis showed that BnPDF2.1, BnPDF2.3, BnPDF2.5 and Arabidopsis thaliana PDF2 were clustered into a group, which indicates they may have a protease inhibitory activity. RTFQ PCR analysis indicated that B. napus defensin genes were expressed in various organs, but the levels of expression were obviously different. The higher expression appeared in the bud and leaf, following silique. During the flowering stage, 1 mmol L–1 SA was used to treat B. napus for 2 hours, which caused the expression levels of defensin genes to be increased in varying degrees in the stem, bud, and silique, but decreased in the leaf, and even no remarked change in the root.

Key words: Brassica napus, Plant defensin, Gene cloning, Expression analysis, Salicylic acid

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