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松杨栅锈菌小种特异性DNA片段的SCAR标记转换
  • ISSN号:1002-0470
  • 期刊名称:《高技术通讯》
  • 时间:0
  • 分类:S512.1[农业科学—作物学]
  • 作者机构:[1]西北农林科技大学林学院,杨凌712100
  • 相关基金:国家自然科学基金(30471394)、陕西省自然科学基金(陕自20008C109)、校人才科研启动费(01140503)和长江学者和创新团队发展计划资助项目.
中文摘要:

用SCAR标记技术对松杨栅锈菌生理小种特异性DNA片段进行了转化和检测研究。通过对RAPD随机引物的扩增筛选,发现随机引物BAO118扩增产生的约700bp的DNA片段与松杨栅锈菌小种特异性相关联。将该片段进行回收、TA克隆、双向测序和序列整合后,序列全长685bp,3’端有BAO118随机引物序列。根据整合后序列,用Primerse-lect软件设计了一对包含随机引物序列的简并引物对。用该引物对对供试菌样进行PCR扩增,成功获得了松杨栅锈菌小种685bp的SCAR标记。用该标记进行田间混合孢子菌样、单孢子接种菌样DNA检测,均得到了685bp的DNA片段,不受寄主DNA和孢子菌系类型的影响,具有较好的稳定性和特异性。

英文摘要:

Sequence characterized amplified region (SCAR) markers were used to transfer and verify the DNA fragment related to physiology race of Melampsora larici-populina. A DNA fragment with the length of 700 bp, which was produced by using random amplification polymorphic DNA (RAPD) primer BA0118,was found to be closely related to the race C3 and race C4 in the electrophoresis map. The DNA fragment was recycled, then cloned, sequenced from its two ends, and finally combined. The combined DNA sequence has a length of 685bp. Based on the sequence discovered, one pair of primers with the primer BA0118 sequence at 3' end was designed by using Primerselect software package. The designed primers were used to identify the race-specific SCAR markers successfully from 11 mono-urediniospore isolates by running polymerase chain reaction (PCR). The advanced application with the designed primers showed that the fragment with the length of 685bp can be obtained from the DNA of urediniospores mixture collected from the field and mono-urediniospores cultivated in the laboratory, which indicated the SCAR markers were stable and unique, and were not influenced by the DNA of the host and the types of urediniospore.

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期刊信息
  • 《高技术通讯》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国科学科技部
  • 主办单位:中国科学技术信息研究所
  • 主编:赵志耘
  • 地址:北京市三里河路54号
  • 邮编:100045
  • 邮箱:hitech@istic.ac.cn
  • 电话:010-68514060 68598272
  • 国际标准刊号:ISSN:1002-0470
  • 国内统一刊号:ISSN:11-2770/N
  • 邮发代号:82-516
  • 获奖情况:
  • 《中国科学引文数据》刊源,《中国科技论文统计与分析》刊源
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰文摘与引文数据库,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国英国皇家化学学会文摘
  • 被引量:12178