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从与辣椒(Capsicum annuum L.)抗疫霉病的限制性片段长度多态性(RFLP)连锁的BAC衍生标记。

BAC-derived markers converted from RFLP linked to Phytophthora capsici resistance in pepper (Capsicum annuum L.).

作者信息

Kim Hyoun-Joung, Nahm Seok-Hyeon, Lee Heung-Ryul, Yoon Gi-Bo, Kim Ki-Taek, Kang Byoung-Cheorl, Choi Doil, Kweon Oh Yeol, Cho Myeong-Cheoul, Kwon Jin-Kyung, Han Jung-Heon, Kim Jeong-Ho, Park Minkyu, Ahn Jong Hwa, Choi Soon Ho, Her Nam Han, Sung Joo-Hee, Kim Byung-Dong

机构信息

Department of Plant Science, Seoul National University, Seoul, 151-921, South Korea.

出版信息

Theor Appl Genet. 2008 Dec;118(1):15-27. doi: 10.1007/s00122-008-0873-5. Epub 2008 Sep 16.

Abstract

Phytophthora capsici Leonian, an oomycete pathogen, is a serious problem in pepper worldwide. Its resistance in pepper is controlled by quantitative trait loci (QTL). To detect QTL associated with P. capsici resistance, a molecular linkage map was constructed using 100 F(2) individuals from a cross between Capsicum annuum 'CM334' and C. annuum 'Chilsungcho'. This linkage map consisted of 202 restriction fragment length polymorphisms (RFLPs), 6 WRKYs and 1 simple sequence repeat (SSR) covering 1482.3 cM, with an average interval marker distance of 7.09 cM. QTL mapping of Phytophthora root rot and damping-off resistance was performed in F(2:3) originated from a cross between resistant Mexican landrace C. annuum 'CM334' and susceptible Korean landrace C. annuum 'Chilsungcho' using composite interval mapping (CIM) analysis. Four QTL explained 66.3% of the total phenotypic variations for root rot resistance and three 44.9% for damping-off resistance. Of these QTL loci, two were located close to RFLP markers CDI25 on chromosome 5 (P5) and CT211A on P9. A bacterial artificial chromosome (BAC) library from C. annuum 'CM334' was screened with these two RFLP probes to obtain sequence information around the RFLP marker loci for development of PCR-based markers. CDI25 and CT211 probes identified seven and eight BAC clones, respectively. Nine positive BAC clones containing probe regions were sequenced and used for cytogenetic analysis. One single-nucleotide amplified polymorphism (SNAP) for the CDI25 locus, and two SSRs and cleaved amplified polymorphic sequence (CAPS) for CT211 were developed using sequences of the positive BAC clones. These markers will be valuable for rapid selection of genotypes and map-based cloning for resistance genes against P. capsici.

摘要

辣椒疫霉(Phytophthora capsici Leonian)是一种卵菌病原体,在全球范围内给辣椒种植带来严重问题。辣椒对其抗性由数量性状基因座(QTL)控制。为了检测与辣椒疫霉抗性相关的QTL,利用辣椒(Capsicum annuum)‘CM334’和辣椒‘Chilsungcho’杂交产生的100个F2个体构建了分子连锁图谱。该连锁图谱由202个限制性片段长度多态性(RFLP)、6个WRKY基因和1个简单序列重复(SSR)组成,覆盖1482.3 cM,平均标记间隔距离为7.09 cM。利用复合区间作图(CIM)分析,对来自抗性墨西哥地方品种辣椒‘CM334’和感病韩国地方品种辣椒‘Chilsungcho’杂交产生的F2:3群体进行了辣椒疫霉根腐病和猝倒病抗性的QTL定位。四个QTL解释了根腐病抗性总表型变异的66.3%,三个QTL解释了猝倒病抗性总表型变异的44.9%。在这些QTL位点中,两个位于第5号染色体(P5)上的RFLP标记CDI25和第9号染色体(P9)上的CT211A附近。用这两个RFLP探针筛选辣椒‘CM334’的细菌人工染色体(BAC)文库,以获取RFLP标记位点周围的序列信息,用于开发基于PCR的标记。CDI25和CT211探针分别鉴定出7个和8个BAC克隆。对9个包含探针区域的阳性BAC克隆进行测序,并用于细胞遗传学分析。利用阳性BAC克隆的序列,开发了一个针对CDI25位点的单核苷酸扩增多态性(SNAP)标记,以及两个针对CT211的SSR标记和酶切扩增多态性序列(CAPS)标记。这些标记对于快速选择基因型和基于图谱克隆辣椒疫霉抗性基因具有重要价值。

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