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摘要：水稻是中国第一大粮食作物，其产量占中国粮食产量的40%左右。由稻黄单胞菌稻生致病变种（Xanthomonas oryzae pv. oryzicola，Xoc）引起的水稻细菌性条斑病（bacterial leaf streak，BLS）已发展成为水稻的第四大病害。研究表明Xoc的毒力主要依靠III型分泌系统所分泌的效应物。类转录激活因子（transcription activator-like effectors, Tales）是来自黄单胞菌属的一类III型效应因子。为了解水稻细菌性条斑病菌菌株GD41中含有Tale家族基因的情况，本研究在已构建的GD41 Tale基因组文库的基础上，通过Southern杂交分析，得到111个独立克隆。这111个克隆中大约含有30个不同的Tale家族基因。本研究基本明确了菌株GD41基因组中Tale基因的数量，并为进一步研究菌株GD41中Tale基因的功能奠定基础。28357
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毕业论文关键词：Tale；稻黄单胞菌稻生致病变种；PCR；菌落原位杂交；Southern杂交
The Research to Screen the Tale Genes of Xoc Strain GD41
Abstract：Rice is the largest grain crop which accounts for about 40% of China's grain output. Bacterial leaf streak(BLS) has been increased as the forth disease of rice caused by Xanthomonas oryzae pv. oryzicola (Xoc).It is known that virulence of Xoc depend on the effectors secreted by the type III secretion system(T3SS).And transcription activator-like effectors(Tales), one kind of type III effectors from Xanthomonas, has been proved as a major determinant of virulence in Xoo which is a pathovarity of Xoc . In order to research the role of Tale family genes in bacterial virulence of Xoc strain GD41,this study aims to screen all Tale gene in genomic library of GD41based on the highly conservation between Tale gene sequences. Firstly, we screen the library by polymerase chain reaction (pcr) with the primers which target the conserved 5’region of Xanthomonas oryzaetal. Then we confirm putative clones by Southern blot. Apprpximately we isolate 30 different Tale genes harboring in 111 library clones. This research basically explicit the number of Tale genes in strain GD41,which may provide a solid basis for further studying the function of Tale genes in strain GD41.
Key words:Tale;Xanthomonas oryzae pv. oryzicola;PCR;Colony in situ hybridization;Southern blot
目录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 1
1 材料与方法 3
1.1供试菌株与质粒 3
1.2培养基 3
1.3所用抗生素浓度 3
1.4主要试剂 3
1.5tal基因SphI片段阳性克隆获取 4
1.5.1 热转化 4
1.5.2质粒DNA的提取 4
1.5.3 SphI酶酶切质粒DNA 4
1.5.4 探针的标记及定量标记反应效率 5
1.5.5 菌落原位杂交 6
1.6 阳性克隆的验证 — Southern印迹杂交 8
2 结果与分析 9
2.1探针的获得 9
2.2菌落原位杂交分析菌株GD41文库克隆中的tal家族基因 9
2.3获得含有tal基因片段的阳性克隆 11
2.4酶切Southern杂交分析菌株GD41文库克隆中的tal家族基因 11
3 讨论 12
致谢 15
参考文献： 15
Xoc strain GD41 Tale基因的筛选
引言
水稻是中国的主要粮食作物，其中稻黄单胞菌稻致病变种（Xanthomonas oryzae pv. oryzae，Xoo）水稻黄单胞菌引起的白叶枯病（bacterial leaf blight， BLB）、和稻黄单胞菌稻生致病变种（Xanthomonas oryzae pv. oryzicola，Xoc）引起的细菌性条斑病（bacterial leaf streak，BLS），对稻米的生产安全构成极大危害[1]。