Acquirement Of Transgenic Cotton Lines With HhERF2、PeDREB2α And CP4EPSPS Intergrated And Analysis Of Their Resistance To Drought,Salt Stresses And Herbicide

Cotton is a crop plant belongs to the mallow family (Malvaceae) Gossypium of (Gossypium). Cotton is a colossal special commodity related to the agriculture and textile industry, also is original material for defense, medicine, and other aspects of the automotive industry. With the increasingly serious environmental problems in human society, global cotton production is seriously decreased. The main results are listed as follows:(1) The target gene HhERF2and PeDREB2a were inserted into the pCAMBLA2301-muti cloning site, and other elements such as omega、Poly4and NOS were also included in this vector, thus a trivalent vector pCAMBLA2301-Kanamycin-CP4-HhERF2:PeDREB2a was obtained. The Trivalent vector was transformed into Agrobacterium tumefaciens LBA4404using the heat shock method. The trivalent expression vector was transformed into cotton varieties WJ. WSH and WY using the non-tissue culture Agrobacterium-mediated method.(2) In order to effectively transform and screen the transgenic cotton, green house growing system was established, the conditions of the cultural temperature, humidity, light, cotton potted soil ratio were characterized. The herbicide resistance experiment was also performed in green house, by spraying different concentrations of glyphosate to cotton. The wilting index was determined based on the damage symptoms.(3) The TO generation seeds were sown in field. The seeds were harvested from7Tl transgenic plants which confirmed by PCR and glyphosate resistance assay. Tl transgenic plants were self-pollinated strictly, and finally two genetically modified cotton plants were obtained, named WJ17-24and WJ17-25-2.respectively. (4) Observations of the agronomic traits of WJ17-24-2and WJ17-25-2. the results showed that the plant height of transgenic plant was significantly lower than the control plant. And the stem was also shorter as compared to the control stem was also shorter compared the wild type.(5) The transgenic plants performed higher salt-tolerant As compared to the control. The gene expression levels of PeDREB2a and HhERF2were higher under the salt and drought stress condition Comparing to to the control, and the expression levels in flowering and boll stage were higher than in the seedling and boll opening stage.(6) Transgenic and control plants sprayed with different concentrations of herbicides found that the transgenic plants compared with the control has significant herbicide-resistant traits0.4%glyphosate concentration was confirmed to screen the transgenic cotton through statistic analysis by SPSS16.0software one-way ANOVA..Identified by the lethal concentration of1.0%glyphosate concentration of the transgenic plants, glyphosate concentration of0.8%lethal concentration control plants.