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ACTA HORTICULTURAE SINICA ›› 2012, Vol. 39 ›› Issue (5): 923-930.

• Vegetables • Previous Articles     Next Articles

Cloning,Characterization and Real-time RT-PCR Analysis of a Key Gene beta-Amyrin synthase for Saponin Biosynthesis in Barbarea vulgaris

WEI  Xiao-Chun, ZHANG  Xiao-Hui, WU  Qing-Jun, WANG  Hai-Ping, SHEN  Di, QIU  Yang, SONG  Jiang-Ping, LI  Xi-Xiang   

  1. (Institute of Vegetables and Flowers,China Academy of Agricultural Sciences,China Academy of Agricultural Sciences,Beijing Research Station of Vegetable Crop Gene Resource and Germplasm Enhancement Ministry of Agriculture,Beijing 100081,China)
  • Online:2012-05-25 Published:2012-05-25

Abstract: G-type Barbarea vulgaris R.Br. which producesaponins is a rare insect resistant vegetable in Cruciferae. A beta-amyrin synthase gene which codes a key enzyme of saponin biosynthesis pathway was cloned from Barbarea vulgaris R.Br. using RT-PCR and RACE(Rapid Amplification of cDNA Ends)method,and named as Bv-beta-AS. The Bv-beta-AS contained a coding sequence of 2 289 bp(GenBank accession number:JQ172795),which encoded 762 amino acids. The gene of 4 107 bp from genomic DNA was further cloned,which contained 15 introns embedded in its coding sequences. The deduced protein of this gene contained the conserved characteristic motif of beta-amyrin synthase -DCTAE and QW. And the  high similarity of this gene with the beta-amyrin synthase from other plants indicated it was a homologue of beta-amyrin synthase. Multiple sequence alignments and phylogenetic tree analyses showed that Bv-beta-AS was clustered with Arabidopsis beta-amyrin synthase and showed the highly similarity(74%) in amino acid sequences. The expression of Bv-beta-AS under the diamondback moth(Plutella xylostella) attack was determined by real-time RT-PCR. The results showed that Bv-beta-AS gene expression was up-regulate within 12 h of diamondback moth attack,but decline tendency appeared after 12 hours. Both the sequences analysis results and the real-time RT-PCR data indicated that Bv-beta-AS may be an important candidate gene of saponin biosynthesis pathway. d

Key words: Barbarea vulgaris, Plutella xylostella, saponin, beta-amyrin synthase, cloning, real-time RT-PCR, insect resistant