Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (17): 3506-3514.doi: 10.3864/j.issn.0578-1752.2012.17.006

• PLANT PROTECTION • Previous Articles     Next Articles

Root and Leaf Infection as Revealed by Autofluorescent Reporter Protein GFP Labeled Bipolaris sorokiniana in Wheat

 YAN  Li-Min, WANG  Xiao-Ming, XU  Rong-Qi, DONG  Fang-Yang, LI  Hong-Jie   

  1. 1.河北科技师范学院生命科技学院，河北秦皇岛066004
    2.中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程，北京100081
    3.中国农业科学院生物技术研究所，北京 100081
  • Received:2011-12-29 Online:2012-09-01 Published:2012-03-05

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Abstract: 【Objective】The objective of this study is to label Bipolaris sorokiniana, which is the causal agent of common root rot and leaf spot blotch of wheat (Triticum aestivum L.), with green fluorescent protein (GFP) for studying infection of B. sorokiniana on wheat roots and leaves, and to establish a direct and non-destructive method for in vivo observing interaction between pathogen and host plant. 【Method】 Gene gfp was transferred into B. sorokiniana strain Bs-1 via Agrobacterium tumefaciens-mediated transformation (ATMT). Based on analysis of fluorescence expression, PCR verification, genetic stability, growth parameters, and metabolism of extra cellular enzymes, a transformed strain Bs-GFP that resembled its wild-type strain, was chosen to investigate infection of B. sorokiniana on roots and leaves of wheat cultivar Aikang 58. 【Result】Bright green fluorescence was observed in hyphae and conidia of the transformant Bs-GFP. The amplification of the diagnostic bands with the gene-specific markers indicated that gfp was integrated into the genome of B. sorokiniana. Analysis of genetic stability and growth parameters demonstrated that gfp was inherited normally, and the growth rate and metabolism of extracellular enzymes were comparable to the wild type strain. The GFP-labeled strain Bs-GFP was able to incite symptoms on both underground and upperground parts of wheat plants, and the quantities of colonization (in terms of colony forming unit) in roots and basal stem tissues of wheat plants by Bs-GFP was similar as those by Bs-1. 【Conclusion】The B. sorokiniana strain expressing GFP obtained via ATMT can be used to directly monitor infection and colonization of the fungus on wheat. The results from this study will facilitate better understanding of the interactions between B. sorokiniana and wheat, as well other cereal crops.

Key words: Agrobacterium tumefaciens, Bipolaris sorokiniana (teleomorph Cochliobolus sativus), GFP (green fluorescence protein), genetic transformation

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