| 紫花苜蓿铝胁迫抑制消减文库的构建和初步分析 |
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| 引用本文： | 夏卓盛,吴跃明,李新伟,李进军,李海庆.紫花苜蓿铝胁迫抑制消减文库的构建和初步分析[J].农业生物技术学报,2007,15(5):805-809. |
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| 作者姓名： | 夏卓盛 吴跃明 李新伟 李进军 李海庆 |
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| 作者单位： | 1. 浙江大学动物科学学院,动物分子营养学教育部重点实验室,杭州,310029;浙江省舟山市畜牧兽医局,舟山,316000
2. 浙江大学动物科学学院,动物分子营养学教育部重点实验室,杭州,310029 |
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| 摘 要： | 为了分离和鉴定铝诱导基因，以紫花苜蓿小冠品种为对象，以经过40μM铝离子胁迫的紫花苜蓿幼根为实验方，未胁迫的为驱赶方，用抑制消减杂交法（SSH）构建了一个紫花苜蓿铝胁迫消减文库。菌落PCR显示插入片段在250bp到1200bp之间，文库质量良好。通过对文库阳性克隆随机测序和BLASTX分析表明,文库中包含了许多在铝胁迫中发挥作用的基因，涉及到抗氧化作用、信号的传导、发育和能量代谢等多种生理过程，比如过氧化物酶、钙依赖蛋白激酶、蔗糖转化酶、蛋白磷酸脂酶调节因子等。而且许多EST序列与其他生物性和非生物性胁迫诱导相关。为了解紫花苜蓿的耐铝机制和抗性相关基因的克隆奠定了基础。
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| 关 键 词： | 抑制消减杂交 紫花苜蓿 铝胁迫 表达序列标签 |
| 文章编号： | 1006-1304-（2007）05-0805-05 |
| 收稿时间： | 2007-01-30 |
| 修稿时间： | 2007-03-16 |
Construction and Primary Analysis of Alfalfa Suppression Subtractive Library Influenced by Aluminum Stress |
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| XIA Zhuo-sheng,WU Yue-ming,LI Xin-wei,LI Jin-jun,LI Hai-qing.Construction and Primary Analysis of Alfalfa Suppression Subtractive Library Influenced by Aluminum Stress[J].Journal of Agricultural Biotechnology,2007,15(5):805-809. |
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| Authors: | XIA Zhuo-sheng WU Yue-ming LI Xin-wei LI Jin-jun LI Hai-qing |
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| Institution: | 1.College of Animal Sciences, Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Zhejiang University, Hangzhou 310029, China; 2.Zhoushan Animal Husbandry and Veterinary Bureau of Zhejiang, Zhoushan 316000, China |
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| Abstract: | To identify the aluminum stress induced genes of alfalfa, a forward subtractive cDNA library was constructed from AL-treated and non-treated seedling roots by suppression subtractive hybridization. Colony PCR suggested that the iniserts were between 250bp~1200bp and the library was suitable for the following work. 40 clones were randomly selecteed, secquenced and obtained 36 unique ESTs. BLASTX show that these ESTs including antioxidative stress, signal transduction, development and energy metaboly, liake peroxidase, calcium-dependent protein kinase, neutral/alkaline invertas, protein phosphatase regulator ect. Many ESTs related to other stresses were also found in our study and nine ESTs had no homology from NCBI. |
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| Keywords: | suppression subtractive hybridization alfalfa aluminum stress expressed secquence tags |
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