Transgenic Cotton pp 223-246

Part of the Methods in Molecular Biology book series (MIMB, volume 958)

Genetic Transformation of Cotton with a Harpin-Encoding Gene hpaXoo Confers an Enhanced Defense Response Against Verticillium dahliae Kleb

Protocol

Abstract

The soil-borne 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.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

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

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