| Verticillium wilt and Fusrium oxsporum f. sp. vansinf ectum of cotton are two kinds of fungi diseases which endanger the vascular tissue,distributed in the whole country of planted cotton areas. They endanger the cotton plant by blocking plant conduit and producing phytotoxin or other ways. In the cotton seedling stage they can cause the dead seedling,and in the cotton growth of middle and later stage they can cause cotton buds and bolls to come off, and leading to production and quality seriously declined. To prevent and cure these diseases, cultivating and spreading the transgenic and disease-resistant varieties of cotton is a simple and effective way.Harpins are heat-stable,Gly-rich,protease-K-sensitive proteins and produced by the plant pathogenic bacteria. It can induce HR in several non-host plant species and it is possibile to be an immportant pathopoiesis agent for the plant pathogenic bacteria. In addition, Harpins can induce plant to produce disease-resistant and insect-resistant and increasing production,improving the quality. The establishment of the cotton transfered-hpa1xoo gene regeneration using Agrobacterium -mediated method.In the experiment, using cotton Zhong35 as receptor for the transformation of species,and using Agrobacterium-mediated method to transform the entire hpalxoo gene and the deletion CD fragment of hpalxoo gene into the cotton explants of stem tip and hypocotyl explant. The hpalxoo gene comes from rice bacterial blight bacteria(Xanthomonas. oryzae pv. oryzae). To study the impact of tissue culture and transformation of the relevant factors in order to establishe a suitable Agrobacterium-mediated stem tip and hypocotyl genetic transformation system. The main findings are as follows:Established the cotton transforming system with the stem tips for the receptor. Through the study of impact of transformation frequency related factors, and the cephalosporins Adriamycin (Cef) inhibitory concentration, Kanamycin (Km) screening concentration, Agrobacterium strains compared screening, and geined the hpa1xoo transformation strains, and PCR amplificated the needed gene. The result shows that the cotton stem tip can be used the good materials as receptor and geined the Agrobacterium tumefaciens mediate of the stem tips transforming system: stem tips with hypocotyl without pre-training 0.5 cm dip-dyed under OD600 = 0.3-0.35 liquid of Agrobacterium bacilli disseminated about 15-20 min, Co-culture 48-56 h; choosing the training to 500 mg / L Cef for inhibitory concentration, 150mg/LKm for the highest screening concentration; The results of this comparison of two strains of the infection hpa1xoo conversion rate higher than the CD.Agrobacterium transformation using embryonic callus, just to select the appropriate state of cell division. We can get a large number of independent transformant, and it can be used the differentiation cultivation after two to four times screening, and we can gein the regenerated plants after 5 to 6 months, the cycle of transgenic cotton are greatly shortened. The experimen is based on this method, but we did not get the regeneration of transgenic plants. There are so many affected factors, such as: Genotype, Plant hormone ratio, The choice of medium, Km selection pressure, Agrobacterium concentration.The identification of the disease-resistant cotton phenotype in the cells and tissues of trans-hpa1xoo gene cotton .The experiment was also conducted to identifiy the disease-resistance of the transed hpa1xoo gene cotton's cell suspension and was tested the Verticillium bacteria-induced trans-hpa1xoo cotton to cause the Micro defense-allergic reaction. The main results are as follows:Culturing cotton cell suspension can produce the relative same of genotype and the quickly splited cell groups, and can be easily added the inducing-substance, while can be simplify the allergic reactions of pathogen-host interaction. Through the cell suspension of cotton and Verticillium wilt and Fusrium oxsporum f. sp. vansinf ectum strains were co-cultured, using the microscope verify the survival of the suspension of cotton after 3h, 6h, 9h, 12h, 24h culturing, and the disease resistance of T-33,T-34 transgenic cotton which transformed Harpin protein are better and under the same conditions the receptor 35 (Z35) the mortality rate is low nearly 10﹪.To use the acupuncture inoculation to leaves, adopted the transfering encode of the HarpinXoo hpa1xoo gene cotton as the material, the interaction of female parent species and the cotton varieties of Verticillium, and adopted the cytology staining methods to microscopic observation of the organization of cotton leaves will produce the microHR or other cytology disease-resistant phenotypes to Verticillium wilt. The experiment results prove that,the transgenic cotton transformed Harpin protein, there is an obvious MicroHR between the oxidation zone and the cells of wound, the non-transgenic cotton without this phenomenon, it show that the Verticillium wilt of cotton inspired the basal defense responses of the transgenic cotton.
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