Study On Plant Regeneration System Of Cotton And Genetic Transformation Of CarNAC5Gene

Cotton is as an important economical crop worldwide. Genetic improvement has been affected by theworld’s attention. In the past century, significant improvments in yield and quality of cotton have beenmade through traditional breeding methods. Due to the lack of efficient cotton somatic embryogenesis andplant regeneration system,transgenic cotton was to rely on the conversion efficiency and genetic stabilityare not ideal pollen tube pathway, seriously restricted the development of the cotton transgenic breedingresearch, therefore the establishment of an efficient, excellent cotton renewable system﹑promote aprerequisite for transgenic technology is widely used in cotton breeding, compared to other crops, cottonlag of the tissue and cell culture of other crops，mainly because of the in the embryogenic callus from callusefficiently converted into﹑embryoid germination rate is low﹑renewable less in variety﹑a higher rate ofdeformity seedlings﹑long regeneration cycle﹑the complexity of gene conversion technologies(conversionrate is low,difficult to repeat)and transferred character variation of genetically modified cotton.Address the above problems, In this study, four cotton cultivars in Xinjiang (xinhai25﹑xinhai16﹑xinluzao39﹑xinluzao42) as experimental materials, somatic embryogenesis and plant regenerationsystemwere studied, establish a stable and efficient system, embryogenic callus of Gossypium hirsutum L.zhongmiansuo44as receptor, agrobacterium-mediated genetic transformation of the NAC transcripts familygeneand obtained regenerated the shoots, expanding renewable cotton cultured in Xinjiang, the main resultsare as follows:1. By using the hypocotyls of Gossypium barbadense L.and Gossypium hirsutum L.as a explant,callus induction period the ratio of hormones, screened the most suitable medium of the callus induction,the result indicates that, callus were effectively produced on the medium with four different hormoneregime, the optimal hormone combination for Gossypium hirsutum L. callus induction was0.1mg/LKT+0.05mg/L2,4-D, callus induction rate of above90%, the best culture media for Gossypiumbarbadense L.callus0.1mg/L KT+0.1mg/L2,4-D，callus induction rate of100%.2. The best embryo callus multiplication medium is MSB adding1.9g/L KNO3,1.8g/L gelrite and0.1g/L Asn﹑0.1g/l Gln; The optimum medium of somatic embryogenesis is MSB medium with1.9g/LKNO3, without NH4NO3,0.1g/L Asn,0.1g/L Gln, and2.0g/L gelrite. Filter paper in the medium padcould promote somatic embryos grow and root system regeneration. The adult seedling medium thecoagulating agent for gelrite better than agar.3. Transgenie plants of NAC transeripts family were obtained by embryogenic callus of Gossypiumhirsutum L. cotton zhongmiansuo44using the agrobaeterium-mediated method. By kanamycinscreened has obtained antibiotic plants，PCR tests showed that CarNAC5gene had been transferred intoregenerated cotton plants; After PEG stress for GUS histochemical staining confirmed that under thecondition of rd29A promoter can induce the expression of GUS gene in transgenic cotton.