Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (16): 3344-3353.doi: 10.3864/j.issn.0578-1752.2013.16.005

• PLANT PROTECTION • Previous Articles     Next Articles

Analysis of Biological Phenotypes and Molecular Cloning of T-DNA Integration Flanking Sequences of Ustilaginoidea virens Mutant Strain B-726

 HUANG Lei1,2, YU Mi-na1, HU Jian-kun1,2, YU Jun-jie1, YIN Xiao-le1, NIE Ya-feng1, CHEN Zhi-yi1, LIU Yong-feng1   

  1. 1.Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014
    2.College of Life Science, Nanjing Agricultural University, Nanjing 210095
  • Received:2013-02-04 Online:2013-08-15 Published:2013-04-26

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Abstract: 【Objective】 The objective of this study is to analyze the phenotypes and T-DNA integration flanking sequence of a mutant strain B-726 and to clone a novel gene which normal expression is disrupted by T-DNA insertional event, understand the role of the gene in U. virens molecular pathogenic process, and to develop a new strategy for controlling rice false smut. 【Method】 Biological phenotypes of B-726 were analyzed by testing growth rate, sporulation ability and pathogenicity. The copy number of T-DNA inserted in B-726 was identified by Southern blot. The flanking sequence of T-DNA was cloned using TAIL-PCR and the completed gene sequence in the flanking was cloned by RACE-PCR. The gene expression was detected by RT-PCR. 【Result】 Phenotypic analysis of B-726 showed that the pathogenicity of B-726 was significant reduced, the sporulation ability and growth rate were declined. Genomic Southern bolt analysis confirmed that B-726 was a single T-DNA insertional event. The T-DNA insertion site was in the 344 bp upstream of a gene named Uvt-726. RT-PCR analysis confirmed that Uvt-726 transcriptions were expressed in B-726 significantly decreased compared to P1. 【Conclusion】 A novel gene maybe associated with pathogenicity of U. virens was cloned, and these results imply that the Uvt-726 might play an important role during the pathogenic process of U. virens.

Key words: Ustilaginoidea virens , T-DNA insertion mutant , pathogenicity , gene clone

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