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疫霉侵染下辣椒幼苗消减文库的构建与初步分析
  • ISSN号:1673-5102
  • 期刊名称:植物研究
  • 时间:2011
  • 页码:95-99
  • 分类:Q3-3[生物学—遗传学]
  • 作者机构:[1]井冈山大学生命科学学院,吉安343009
  • 相关基金:国家自然科学基金(31060263); 江西省自然科学基金资助项目(2009GQN0077); 江西省教育厅科技计划项目(GJJ10202); 井冈山大学校级课题(JZ09016)
  • 相关项目:辣椒应答疫霉侵染的ERF转录因子的分子特性及功能研究
中文摘要:

为了分离和鉴定辣椒中疫霉诱导基因,以高抗疫霉病辣椒品种L11为材料,以接种辣椒疫霉菌的幼嫩叶片为处理(tester),以未接种自然生长的幼嫩叶片为对照(driver),利用抑制性消减杂交技术(suppression subtractive hybridization,SSH)构建了疫霉侵染下辣椒幼苗的消减文库。从消减文库中随机挑取30个阳性克隆,提取质粒进行PCR鉴定,显示插入片段大小大部分集中在200~1 000 bp之间,文库质量良好。随机挑取40个克隆进行测序,共获得35个有效EST序列。经Blastx分析表明:有30个EST与GenBank中其他序列有同源性,5个EST为未知功能序列。已知功能的EST序列分别编码NAC转录因子、丝氨酸/苏氨酸蛋白激酶、P450单加氧酶、叶绿素a/b结合蛋白、谷胱甘肽转移酶、几丁质酶等,这些蛋白涉及抗病信号传递、抗氧化作用、转录调控及光合作用等多种生理过程。本研究为抗病基因克隆和系统研究疫霉侵染下辣椒基因的表达奠定了重要的理论基础。

英文摘要:

To identify the Phytophthora capsici stress induced genes of pepper,a high P.capsici resistant pepper cultivar was used as main material in this study.With cDNA from pepper seedlings treated with P.capsici as the tester and cDNA from this plant in normal growth as the driver,we constructed subtracted library using suppression subtractive hybridization(SSH).Thirty positive clones were picked out randomly from SSH library,plasmid PCR suggested that the inserts were between 200-1 000 bp and library was suitable for the following work.Forty clones were randomly selected and sequenced,35 differential gene fragments were obtained.Blastx analysis showed that 30 ESTs have high homology with known genes in GenBank,5 ESTs was unknown function genes.The obtained known function ESTs code NAC transcription factor protein,Serine/threonine protein kinase,cytochrome P450 monooxygenase,chlorophyll a b-binding protein,glutathione s-transferase,chitinase,respectively.These expressed genes are involved in disease-resistant signal transduction,antioxidative stress,transcriptional regulation,photosynthesis,physiological processes.The research established a basis for cloning stress resistance genes and further studying genes expression in Capsicum annuum L.seedlings under P.capsici stress.

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期刊信息
  • 《植物研究》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国教育部
  • 主办单位:东北林业大学
  • 主编:付玉洁
  • 地址:哈尔滨市和兴路26号东北林业大学
  • 邮编:150040
  • 邮箱:duanxihua828@163.com
  • 电话:0451-82190611
  • 国际标准刊号:ISSN:1673-5102
  • 国内统一刊号:ISSN:23-1480/S
  • 邮发代号:14-77
  • 获奖情况:
  • 国内外数据库收录:
  • 被引量:13014