| 利用差异显示技术克隆小麦抗白粉病相关基因的研究 |
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| 引用本文： | 孔凡晶,陈孝,马有志,辛志勇.利用差异显示技术克隆小麦抗白粉病相关基因的研究[J].麦类作物学报,2004,24(1):11-14. |
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| 作者姓名： | 孔凡晶 陈孝 马有志 辛志勇 |
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| 作者单位： | 1. 中国农业科学院作物育种栽培研究所,北京,100081;国土资源部盐湖资源与环境开放实验室,北京,100037
2. 中国农业科学院作物育种栽培研究所,北京,100081 |
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| 基金项目： | 国家863计划项目(Z-17-04-01)。 |
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| 摘 要： | 对小麦—簇毛麦抗病易位系进行差异显示分析,以期获得小麦抗白粉病基因的分子克隆。根据已克隆植物抗病基因的保守域,设计了简并引物,与锚定引物组合,对抗病诱导的小麦抗白粉病易位系进行了差异显示分析,共获得10个差异片段,其中两个片段R3-1和8C1.3-6Northern杂交为阳性。将这两个片段克隆后进行了测序,序列分析结果为两个新序列,GenBank登录号分别为AF498271和AF498272。R3-1没有发现同源性较高的植物基因序列,发现8C1.3-6与Sphenostylisstenocarpa 类几丁质酶基因有同源性。
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| 关 键 词： | 小麦白粉病抗性基因 简并引物 差异显示技术 Northern分析 |
| 文章编号： | 1009-1041(2004)01-0011-04 |
| 修稿时间： | 2003年1月13日 |
Cloning of Genes Related to Wheat Powdery Mildew Resistance by Differential Display |
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| KONG Fan-jing.Cloning of Genes Related to Wheat Powdery Mildew Resistance by Differential Display[J].Journal of Triticeae Crops,2004,24(1):11-14. |
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| Authors: | KONG Fan-jing |
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| Institution: | KONG Fan-jing~ |
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| Abstract: | In order to clone the genes relatd to wheat powdery mildew resistance, different display analysis of H. Villosa-T. aestivum traslocation line were reported in this paper. The (degenerate) oligonucleotide primers were designed according to cloned plant disease resistance genes. The differential display were carried out by (using) anchor primer and (degenerate) oligonucleotide primers. 10 differential displayed cDNA fragments were (collected) and cloned among leaves induced with Erysiphe. graminis f. sp. tritici of 0 h, 24 h and 48 h later. (Among) them, two clones R3-1 and 8C1.3-6 showed positive signals in Northern blot. R3-1 is 840 bp and 8C1.3-6 is 587 bp. BLAST analysis showed R3-1 had no higher homology sequences in GenBank. 8C1.3-6 had (identity) of 100% with Sphenostylis stenocarpa class III chitinase between 15-56bp. The GenBank accession (numbers) of R3-1 and 8C1.3-6 were AF498271 and AF498272 respectively. |
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| Keywords: | Wheat powdery mildew resistance related genes Degenerate oligonucleotide primers Differential display Northern blot analysis |
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