甘肃省小麦条锈病主要流行小种的RAPD分析及Su11-4小种SCAR标记建立

草业学报 ›› 2010, Vol. 19 ›› Issue (6): 180-186.

甘肃省小麦条锈病主要流行小种的RAPD分析及Su11-4小种SCAR标记建立

孟亚雄,仲军,马小乐,李葆春,赖勇,司二静,尚勋武,王化俊^*

甘肃省干旱生境作物学重点实验室甘肃省作物遗传改良与种质创新重点实验室甘肃农业大学农学院,甘肃兰州 730070

收稿日期:2010-07-12 出版日期:2010-06-25 发布日期:2010-12-20
作者简介:孟亚雄(1977-),男,甘肃会宁人,助理研究员,在读博士。E-mail:yxmeng1@163.com
基金资助:
科技部支撑计划子项目(2GS064-A41-001-06),甘肃省农业生物技术研究与应用开发项目(GNSW-2007-01)和甘肃省重大专项(0801NKDA013)资助。

RAPD analysis of Puccinia striiformisf. sp tritici in Gansu and SCAR marker establishment for Su11-14

MENG Ya-xiong, ZHONG Jun, MA Xiao-le, LI Bao-chun, LAI Yong, SI Er-jing,
SHANG Xun-wu, WANG Hua-jun

Gansu Provincial Key Laboratory of Aridland Crop Science;
Gansu Key Lab of Crop Improvement
and Germplasm Enhancement;
College of Agronomy, Gansu Agricultural
University, Lanzhou 730070, China

Received:2010-07-12 Online:2010-06-25 Published:2010-12-20

摘要/Abstract

摘要： 本研究应用RAPD标记技术对甘肃省小麦条锈菌主要流行的8个生理小种进行多态性标记分析,旨在寻找小麦条锈菌不同生理小种的特异性标记,共选用220条10碱基随机引物进行筛选,其中有147条可得到稳定清晰的扩增条带。研究结果显示,通过使用147条引物对甘肃省流行的8个条锈菌的生理小种进行RAPD分析,发现各致病小种间遗传变异丰富,其中引物S301在条中33号中扩增得到约507bp的特异条带;引物S39在条中32扩增得到长度约183bp的特异条带;引物S36在Hybrid46-8扩增得到约510bp的特异条带;引物S2140在Su11-4扩增得到约317bp的特异条带。另外,本研究还对扩增得到的特异片段进行回收并进行测序分析,其中依据小麦条锈菌生理小种Su11-4特异条带的测序结果设计特异引物,成功将其转化为对Su11-4小种特异的SCAR标记,这对不同条锈菌生理小种的快速准确检测具有重要意义。

Abstract: Present in this study, random amplified polymorphic DNA(RAPD) was used to analyze polymorphisms of eight wheat yellow rust pathotypes from Gansu, and the specific RAPD markers among pathotypes were characterized. Out of 220 random 10 bp length primers were analyzed, 147 primers were giving distinct amplification bands. The genetic variations among pathotypes of wheat yellow rust were abundance. A specific 507 bp length DNA fragment was obtained from CY33 via PCR amplification with primer S301; a specific 183 bp length DNA fragment was obtained from CY32-1 with primer S39; a specific 510 bp length DNA fragment was obtained from Hybrid46-8 with primer S36; a specific 317 bp length DNA fragment was obtained from Su11-4 with primer S2140, respectively. The DNA fragment obtained from Su11-4 with primer S2140 was cloned and sequenced, specific forward and revise primers were designed for Su11-4 and the sequence-characterized amplified region SCAR marker was developed as well. The results indicate that a molecular detection system for Puccinia striiformisf.sp tritici could be established by mean of searching for specific RAPD fragments among pathotypes. The results also suggest that development of SCAR marker would be a important approach to accurately detect pathotype Su11-4 of Puccinia striiformisf. sp tritici.

中图分类号:

孟亚雄,仲军,马小乐,李葆春,赖勇,司二静,尚勋武,王化俊. 甘肃省小麦条锈病主要流行小种的RAPD分析及Su11-4小种SCAR标记建立[J]. 草业学报, 2010, 19(6): 180-186.

MENG Ya-xiong, ZHONG Jun, MA Xiao-le, LI Bao-chun, LAI Yong, SI Er-jing,
SHANG Xun-wu, WANG Hua-jun. RAPD analysis of Puccinia striiformisf. sp tritici in Gansu and SCAR marker establishment for Su11-14[J]. Acta Prataculturae Sinica, 2010, 19(6): 180-186.

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