蒙古冰草组织培养再生体系建立及其dreb基因功能分析-establishment of tissue culture regeneration system of agropyron mongolicum and analysis of dreb gene function.docxVIP

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蒙古冰草组织培养再生体系建立及其dreb基因功能分析-establishment of tissue culture regeneration system of agropyron mongolicum and analysis of dreb gene function.docx

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蒙古冰草组织培养再生体系建立及其dreb基因功能分析-establishment of tissue culture regeneration system of agropyron mongolicum and analysis of dreb gene function

The Establishment of Agropyron Mongolicum Keng Regeneration System and Functional Analysis of DREB Gene Abstract Agropyron mongolicum Keng not only has a cold, drought and other outstanding features, and contains excellent resistance gene. In this study,the regeneration system was Establishment of Agropyron mongolicum Keng, the DREB gene was separation and the functional was studied.The results as follows: Establishment of Regeneration System of Agropyron mongolicum Keng Agropyron mongolicum Keng Shoot tip as the most suitable explants for callus induction ,the optimal medium of callus induction was MS+2,4-D 1.5mg/L+6-BA 0.1mg/L ,the induction rate of 84.9%.The optimal medium of differentiation was MS+6-BA 2.0mg/L+NAA 0.5mg/L with the differentiation rate of 53.2%. The medium of rooting was MS+NAA 0.5mg/L with rooting rate of 100%. Genetic transformation of Agropyron mongolicum Keng DREB gene The Agropyron mongolicum Keng DREB gene inserted between the vector pCAMBIA2300-35S-OCS promoter and terminator,The plant Expression Vector was constructed of DREB Gene of PCAMBIA2300-MDREB. The plant expression vector of PCAMBIA2300-MDREB was transformed into Agrobacterium tumefaciens LB4404, infecting the tobacco leaf discs,30 transgenic tobacco plants were selection by resistance regeneration culture.15 positive plants ware determined by PCR,7 plants appeared PCR-Southern hybrid spectrum belt , and six transgenic tobacco by RT-PCR validation results show that the Agropyron mongolicum Keng DREB gene expression at the transcriptional level. The resistant function analysis of transgenic tobacco Transgenic tobacco leaves wilting degree lower than the wild-type tobacco under drought stress and the Turgor-Pressure in Leaves HYPERLINK /dict_result.aspx?searchword=%e6%81%a2%e5%a4%8damp;tjType=sentenceamp;styleamp;t=recovery recovery faster than wild-type tobacco after watering. Transgenic tobacco membrane relative conductivity is lower than wild-type tobacco and the SOD activity ,