摘要
获得转化梨几丁质酶基因Lchi1的烟草,检测组成型表达Lchi1基因烟草是否能提高对梨腐烂病菌(Valsa ceratosperma)的抗性,从而初步确定Lchi1基因的功能,为进一步开展梨抗腐烂病分子育种工作提供参考。构建梨几丁质酶基因Lchi1的植物表达载体,通过农杆菌介导的叶盘法转化烟草,进而对转化的烟草T2代进行PCR及RT-PCR检测。利用离体叶片接种法,更直观地验证转基因烟草对梨腐烂病菌的抗性。Lchi1基因已经整合到烟草基因组中,并在烟草T2代中有效表达,在接种腐烂病菌后转基因烟草较未转基因烟草的抗腐烂病菌能力明显增强。梨几丁质酶基因Lchi1与腐烂病抗性相关联,可作为今后梨的抗腐烂病分子育种研究重要的基因来源。
The study aims to obtain the transgenic tobacco containing Lchi1 which is the chitinase gene, anddetect whether the transgenic tobacco with constitutive Lchi1 gene expression can improve resistance to Valsa ceratosperma, thus to preliminarily determine the function of Lchi1 gene and provide a reference for the furtherresearch on molecular breeding about pear Valsa canker. Plant expression vector with gene Lchi1 wasconstructed and transformed into tobacco by leaf disc mediated with Agrobacterium tumefaciens, andtransgenic tobacco T2 generation was detected by PCR and RT- PCR. The resistance of transgenic tobaccoagainst Valsa ceratosperma was verified more intuitively by isolated leaf inoculation method. Lchi1 wasintegrated into tobacco genome, and it was expressed effectively in the transgenic tobacco T2 generation. Afterinoculating Valsa ceratosperma, the resistance of transformant against Valsa ceratosperma was obviously moreenhanced compared with non transgenic tobacco. In conclusion, Lchi1, which is the pear chitinase gene, isassociated with the resistance of pear Valsa canker, and it will be used as an important genetic source in theresearch of molecular resistance breeding.
出处
《中国农学通报》
2015年第22期184-190,共7页
Chinese Agricultural Science Bulletin
基金
国家高技术研究发展计划(863计划)子课题"梨分子育种与品种创新"(2011AA10020604)
新疆维吾尔自治区果树学重点学科(201007)
关键词
几丁质酶
抗腐烂病
转基因
烟草
chitinase
resistance to pear Valsa canker
transgene
tobacco
