| 摘要: |
| 以感病×抗病的葡萄种间杂交组合白玉霓×塘尾的F1代及塘尾自交一代33个单株为试验材料,采用BSA法和RAPD技术,对155个随机引物进行筛选,从中获得了1个与中国野生葡萄抗白腐病基因连锁的RAPD标记OPP09-760.用Wizard DNA Clean-up System试剂盒回收纯化RAPD遗传标记OPP09-760,连接于pGME-T-Easy Wector上并克隆测序,得到OPP09-760的全序列,其长度为766 bp.根据两端序列设计特异PCR扩增引物作为合成中国野生葡萄抗白腐病检测探针的基础,用于检测葡萄抗白腐病种质种和分子标记辅助育种. |
| 关键词: 中国野生葡萄 白腐病 RAPD 克隆 测序 |
| DOI： |
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| 基金项目:国家自然科学基金资助项目(39770525,39970524)；高等学校博士点基金资助项目(980701) |
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| Cloning and sequencing of RAPD maker linked to white rot disease resistant gene in wild grapes native to China |
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| Abstract: |
| Bulk segregant analysis (BSA),randomly amplified polymorphic DNA (RAPD) methords were used to tag the white rot resistant genes of grape molecular markers,parents and their 33 individuals of F1 and F1 progeny resulting from a cross 91-5 between Tangwei (vitis davidii),white rot-resistant and Ugni Blanc (V.vinifera),white rot-susceptible.Among 155 Operon primers giving distinct band patterns,one RAPD marker OPP09-760 was tightly linked to a major gene resistant to Coniothyrium diplodiella.OPP09-760 fragment was reclaimed by Wizard DNA Clean-up system from electrophoresis gel,and cloned in T-easy Vector,then sequenced from two sides.The DNA fragment OPP09-760 was actually 766 bp.It is suggested that the sequence of RAPD marker OPP09-760 might be used as a basis for synthesizing the specific PCR primers and the probe for detecting grape white rot-resistant in disease resistant breeding and molecular marker assisted selection (MAS). |
| Key words: Chinese wild Vitis white-rot disease RAPD cloning sequencing |
