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Agrobacterium tumefaciens-Mediated Transformation of the Causative Agent of Valsa canker of Apple Tree Valsa mali var. mali

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Abstract

Valsa mali var. mali (Vmm), which is the causative agent of Valsa canker of apple tree, causes heavy damage to apple production in eastern Asia. In this article, we report Agrobacterium tumefaciens-mediated transformation (ATMT) of Vmm and expression of gfp (green fluorescent protein) in this fungus. The transformation system was optimized to a transformation efficiency of approximately 150 transformants/106 conidia, and a library containing over 4,000 transformants was generated. The tested transformants were mitotically stable. One hundred percent hph (hygromycin B phosphotransferase) integration into Vmm was identified by PCR and five single-copy integration of T-DNA was detected in the eighteen transformants by Southern blot. To our knowledge, this is the first report of ATMT of Vmm. Furthermore, this library has been used to identify genes involved in the virulence of the pathogen, and the transformation system may also be useful to the transformation of other species of the genus Valsa.

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Acknowledgments

The authors thank Fengming Song (Zhejiang University, China) for providing the pBIG2RHPH2-GFP-GUS plasmid; Jinrong Xu (Purdue-NWAFU Joint Research Center, Northwest A&F University, China) for providing A. tumefaciens SK1044 harboring the binary vector pSK1044; undergranduates Ms. Dianru Wang, Mr. Jie Zhou, and Mr. Decheng Ren for their help in the collection of transformants; Ms. Na Song, a doctoral student, for microscopic observation; and Ms. Jing Gao for testing the hygromycin B sensitivity of Vmm. The authors also thank Dr. Jinrong Xu and Dr. Xiaoyu Qiang for correcting the grammatical and spelling errors as well as critical reading of the manuscript. This study was funded by the National Natural Science Foundation of China (No. 31171796), Specialized Research Fund for the Doctoral Program of Higher Education (20120204110002), and the Special Fund for Agro-scientific Research in the Public Interest of China (nyhyzx201203034-03).

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Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, 712100, People’s Republic of China

    Yang Hu, Qingqing Dai, Yangyang Liu, Na Song, Xiaoning Gao, Zhensheng Kang & Lili Huang

  2. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Life Science, Northwest A&F University, Yangling, 712100, People’s Republic of China

    Zhe Yang

  3. Fachgebiet Phytopathologie, Institut für Phytomedizin, Universität Hohenheim, 70599, Stuttgart, Germany

    Ralf Thomas Voegele

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  1. Yang Hu
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Correspondence to Lili Huang.

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Hu, Y., Dai, Q., Liu, Y. et al. Agrobacterium tumefaciens-Mediated Transformation of the Causative Agent of Valsa canker of Apple Tree Valsa mali var. mali . Curr Microbiol 68, 769–776 (2014). https://doi.org/10.1007/s00284-014-0541-8

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