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当前位置: 首页 > 文章 > 山葡萄‘通化-3’抗寒相关基因SSH文库的构建及分析 果树学报 2015 (4) 546-554
Position: Home > Articles > Construction and analyses of SSH libraries of cold resistance related genes in Vitis amrensis accession‘Tonghua-3' Journal of Fruit Science 2015 (4) 546-554

山葡萄‘通化-3’抗寒相关基因SSH文库的构建及分析

作  者：
米宝琴;毛娟;申鹏;陈佰鸿;郝燕
单  位：
甘肃农业大学农学院;甘肃省农科院果树研究所;甘肃农业大学园艺学院武威市林业综合服务中心甘肃省农业科学院林果花卉研究所
关键词：
山葡萄;‘红地球’;SSH;抗寒蛋白;差异表达
摘  要：
【目的】克隆山葡萄抗寒相关基因和分析葡萄的抗寒机理。【方法】利用抑制性消减杂交技术(SSH),选用抗寒性强的山葡萄株系‘通化-3’和抗寒性弱的欧洲葡萄品种‘红地球’分别作Driver和Tester,建立了ASSH(山葡萄)与SSSH(‘红地球’)2个c DNA表达文库。【结果】差异表达克隆测序后经DNAMAN比对,从657个克隆中筛选出了70个Unigenes;通过BLAST比对分析,发现在ASSH文库中含有与山葡萄相关的r RNA基因内含子编码的归巢核酸内切酶、衰老相关蛋白质、细胞色素P450、抗冻蛋白、防御素样蛋白、热休克蛋白等相关的EST序列;SSSH文库中含有与‘红地球’相关的衰老蛋白。此外还发现了14个未知蛋白的ESTs序列。【结论】筛选出了与山葡萄抗寒性相关的AFPs、Haps、防御素样蛋白、细胞色素P450加单氧酶等多个相关的ESTs,以及14个未知蛋白序列,也可能参与山葡萄的抗寒代谢。
译  名：
Construction and analyses of SSH libraries of cold resistance related genes in Vitis amrensis accession‘Tonghua-3'
作  者：
MI Baoqin;MAO Juan;SHEN Peng;CHEN Baihong;HAO Yan;YANG Rui;College of Horticulture,Gansu Agricultural University;Wuwei Forestry Comprehensive Service Center;Institute of Fruit and Floriculture Research,Gansu Academy of Agriculture Science;
关键词：
Vitis amurensis;;Red Globe;;SSH;;Cold resistance protein;;Differential expression
摘  要：
【Objective】To clone the related- genes involved in response to cold stress of V. amrensis andanalyse cold stress mechanism of grape.【Methods】With the suppressive subtraction hybridization(SSH)technique,the pros and cons of two differentially expressed c DNA libraries ASSH(V. amurensis‘Tonghua-3') and SSSH(V. vinifera L.‘Red globe') were constructed using V. amurensis accession‘Tonghua-3'with strong cold resistance and V. vinifera L.‘Red Globe'with weak cold resistance as Driver and Tester.【Results】After sequencing the clones expressing differentially and comparing DNAMAN,we selected 70 unigenes from the 657 clones;After blasting their sequences,found some ESTs regulating r RNA gene in-tron-encoded homing endonuclease,aging-related proteins,cytochrome P450,antifreeze proteins,de-fensin- like protein,heat shock proteins and others involved in ASSH library of V. amurensis;In theSSSH c DNA library,we found aging-related proteins in‘Red Globe'cultivar. In addition,we also foundthe fourteen unknown protein of ESTs.【Conclusion】Screened AFPs,HSPs,defensin-like protein,Cyto-chrome P450 monooxygenase of ESTs related to the cold resistance in V. amurensis and the fourteen un-known protein sequences may also be involved in cold metabolism of V. amurensis.

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