Genetic Transformation of Cotton with the Harpin-Encoding Gene hpaXoo of Xanthomonas oryzae pv. oryzae and Evaluation of Resistance Against Verticillium Wilt

  • Weiguo MiaoEmail author
  • Jingsheng Wang
Part of the Methods in Molecular Biology book series (MIMB, volume 1902)


The soilborne fungal pathogen Verticillium dahliae Kleb causes Verticillium wilt in a wide range of crops including cotton (Gossypium hirsutum). To date, most upland cotton varieties are susceptible to V. dahliae, and the breeding for cotton varieties with the resistance to Verticillium wilt has not been successful. Hpa1Xoo is a harpin protein from Xanthomonas oryzae pv. oryzae which induces the hypersensitive cell death in plants. When hpa1Xoo was transformed into the susceptible cotton line Z35 through Agrobacterium-mediated transformation, the transgenic cotton line (T-34) with an improved resistance to Verticillium dahliae was obtained. Here, we describe the related research approach, such as Western blot, Southern blot, immuno-gold labeling, evaluation of resistance to Verticillium dahliae, and how to detect the micro-hypersensitive response and oxidative burst elicited by harpinXoo in plant tissue.

Key words

Transgenic cotton hpa1Xoo Verticillium dahliae Western blot Southern blot Immunocytological localization 


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Authors and Affiliations

  1. 1.College of Plant ProtectionHainan UniversityHaikouPeople’s Republic of China
  2. 2.College of Plant ProtectionNanjing Agricultural UniversityNanjingPeople’s Republic of China

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