山羊草属杀配子染色体2C特异SCAR标记的建立

草业学报 ›› 2011, Vol. 20 ›› Issue (4): 305-310.

山羊草属杀配子染色体2C特异SCAR标记的建立

徐国辉¹, 郭长虹^1*, 于洪涛¹, 陈静¹, 丛雯雯¹, 远藤隆²

1.哈尔滨师范大学生命科学与技术学院黑龙江省分子细胞遗传与遗传育种重点实验室, 黑龙江哈尔滨 150025;
2.日本京都大学农学部植物遗传学实验室,日本京都 606-8502

收稿日期:2010-05-10 出版日期:2011-04-25 发布日期:2011-08-20
通讯作者: E-mail:kaku2008@hotmail.com
作者简介:徐国辉(1980-),女,黑龙江哈尔滨人,在读博士。E-mail:xugh520@163.com
基金资助:
科技部国际科技合作项目(2009DFA32470),黑龙江省科技攻关项目(GA08B104),973前期研究专项(2011CB111500),黑龙江省高校科技创新团队研究计划(2010TD10)和哈尔滨师范大学科技创新团队研究计划(KJTD-2011-2)资助。

Establishment of a gametocidal chromosome 2C specific SCAR marker

XU Guo-hui¹, GUO Chang-hong¹, YU Hong-tao¹, CHEN Jing¹, CONG Wen-wen¹, ENDO T R²

1.Key Laboratory of Molecular and Cytogenetics, Heilongjiang Province, Department of Biology, Harbin Normal University, Harbin 150025, China;
2.Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan

Received:2010-05-10 Online:2011-04-25 Published:2011-08-20

摘要/Abstract

摘要： 用40条10碱基随机引物,对普通小麦“中国春”、“中国春”-杀配子染色体2C二体附加系、柱穗山羊草进行RAPD分析,筛选到1个杀配子染色体2C特异引物OPF03。从“中国春”-杀配子染色体2C二体附加系中克隆了该DNA片段。测序结果表明,该片段长1 496 bp,与大麦1个cDNA的同源性为92%。根据OPF03₁₄₉₆的序列设计了1对SCAR引物2C-F₅₈₆、2C-R₅₈₆,对普通小麦“中国春”、“中国春”-杀配子染色体2C二体附加系、柱穗山羊草、二倍体长穗偃麦草、“中国春”-长穗偃麦草7E二体附加系等材料进行了SCAR分析,在“中国春”-杀配子染色体2C二体附加系、柱穗山羊草中扩增出了586 bp的片段,而在普通小麦“中国春”、二倍体长穗偃麦草、“中国春”-长穗偃麦草7E二体附加系中没有该产物,初步证明此标记可以用于杀配子染色体2C的检测。进一步用该对SCAR引物对“中国春”-杀配子染色体2C二体附加系与“中国春”-长穗偃麦草7E二体附加系的杂种F₁代、F₂代进行了分析,结果在全部F₁代以及部分F₂代材料扩增出了目的片段,进一步证明了此SCAR标记可以用来检测小麦背景下的杀配子染色体2C,为小麦背景中杀配子染色体2C的快速跟踪检测提供了新途径。

Abstract: A total of 40 decamer oligonucleotides were used as primers to perform random amplified polymorphic DNA (RAPD) analysis on common wheat CS (Chinese spring), CS-2C disomic addition line, Aegilops cylindrica. Of these primers, one could amplify a specific DNA fragment in CS-2C disomic addition line and A. cylindrica. The fragment OPF03₁₄₉₆ was cloned and sequenced; Sequence data searching in GenBank showed that this fragment was 92% homologous to a cloned cDNA sequence from Barley. Based on the sequence of OPF03₁₄₉₆, a pair of SCAR (sequence characterized amplified region) primer 2C-F₅₈₆, 2C-R₅₈₆ was designed. SCAR analyses were performed on CS, CS-2C disomic addition line, A. cylindrica, and Thinopyrum elongatum (2X) CS-7E disomic addition line. The amplification results of SCAR primer showed that the SCAR₅₈₆ marker appeared in the CS-2C disomic addition line and A. cylindrica but not in CS, T. elongatum (2X) CS-7E disomic addition line. Furthermore, SCAR analyses were also performed on F₁ and F₂ progenies between CS-2C and CS-7E disomic addition lines. PCR amplification results showed that the SCAR₅₈₆ marker was in all of the F₁ progenies between CS-2C and CS-7E and in some of the F₂ progenies. The SCAR₅₈₆ marker can be used for detecting the gametocidal chromosome 2C in common wheat backgrounds.

中图分类号:

徐国辉, 郭长虹, 于洪涛, 陈静, 丛雯雯, 远藤隆. 山羊草属杀配子染色体2C特异SCAR标记的建立[J]. 草业学报, 2011, 20(4): 305-310.

XU Guo-hui, GUO Chang-hong, YU Hong-tao, CHEN Jing, CONG Wen-wen, ENDO T R. Establishment of a gametocidal chromosome 2C specific SCAR marker[J]. Acta Prataculturae Sinica, 2011, 20(4): 305-310.

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