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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (12): 1779-1786.doi: 10.3724/SP.J.1006.2016.01779


Cloning and Expression Analysis of a Functional Gene GbVWR Induced by Verticillium dahliae in Gossypium barbadense

ZHANG Li-Jia**,ZHANG Yan**,RONG Wei,YANG Jun,ZHANG Gui-Yin,WU Li-Qiang,LI Zhi-Kun,WU Jin-Hua,MA Zhi-Ying,WANG Xing-Fen*   

  1. North China Key Laboratory for Crop Germplasm Resources of Education Ministry / Key Laboratory for Crop Germplasm Resources of Hebei / Agricultural University of Hebei, Baoding 071001, China
  • Received:2016-03-14 Revised:2016-06-20 Online:2016-12-12 Published:2016-06-27
  • Contact: Wang Shengfen, E-mail: cotton@hebau.edu.cn E-mail:646560324@qq.com
  • Supported by:

    This study was supported by the Natural Science Foundation of Hebei Province (C2013204141), the Special Research Found for the Doctoral Program of Higher Education (20131302120002), and the Science and Technology Support Project of Hebei Province (14226308D).


Verticillium dahliae is a destructive, soil-borne fungal pathogen that causes severe losses in cotton yield and fiber quality. Mining functional genes related to resistance against V. dahliae will benefit efforts to genetically improve crop plants. In this study, we identified a gene that involved in cotton defense against V. dahliae based on screening the full-length cDNA library and suppression subtractive hybridization library (SSH) induced by V. dahliae in Gossypium barbadense and Gossypium. hirsutum, respectively. Sequence analysis indicated that there was no any annotation in NCBI database, and we named the sequence from G. barbadense as GbVWR. We characterized GbVWR gene and analyzed its expression. The full length cDNA of GbVWR was 520 bp including a 198 bp open reading frame (ORF), encoding 65 amino acid residues. Bioinformatic analyses suggested that GbVWR belonged to secretory protein and tis theoretical isoelectric point was 5.32. Using pET-32a(+) as a fused expression vector, a recombinant plasmid pET32a-GbVWR was constructed. The recombinant protein was induced in Escherichia coli BL21 (DE3) with 1.0 mmol L–1 IPTG then GbVWR could express about 7 kD protein in E. coli BL21 (DE3). In addition, diverse cis-acting promoter elements involved in fungal elicitor response, hormone response, wound-response, and flavonoid biosynthetic gene regulation were discovered in the promoter region of GbVWR. qPCR analysis showed that expression level of GbVWR was the highest in roots, and was significantly induced by V. dahliae. Besides, GbVWR could also be induced by SA, ET and GA treatments, respectively. In conclusion, GbVWR is a new functional gene, which involved in multiple signal pathways in cotton defense response to Verticillium wilt.

Key words: Gossypium barbadense, Verticillium wilt resistance, GbVWR, Clone, Gene expression

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