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鉴定与从卡氏花生渗入到栽培花生中的线虫抗性基因紧密连锁的随机扩增多态性DNA(RAPD)、序列特征化扩增区域(SCAR)和限制性片段长度多态性(RFLP)标记。

Identification of RAPD, SCAR, and RFLP markers tightly linked to nematode resistance genes introgressed from Arachis cardenasii into Arachis hypogaea.

作者信息

Garcia G M, Stalker H T, Shroeder E, Kochert G

机构信息

Department of Crop Science, North Carolina State University, Raleigh, 27695-7629, USA.

出版信息

Genome. 1996 Oct;39(5):836-45. doi: 10.1139/g96-106.

DOI:10.1139/g96-106
PMID:8890516
Abstract

Two dominant genes conditioning resistance to the root-knot nematode Meloidogyne arenaria were identified in a segregating F2 population derived from the cross of 4x (Arachis hypogaea x Arachis cardenasii)-GA 6 and PI 261942. Mae is proposed as the designation for the dominant gene restricting egg number and Mag is proposed as the designation for the dominant gene restricting galling. The high levels of resistance in GA 6 were introgressed from A. cardenasii and, therefore, a search to identify A. cardenasii specific RAPD markers that are tightly linked to these resistance genes was conducted utilizing bulked segregant analysis. One RAPD marker (Z3/265) was linked at 10 +/- 2.5 (SE) and 14 +/- 2.9 cM from Mag and Mae, respectively. The marker was mapped to linkage group 1 at 5 cM from Xuga.cr239 in the backcross map in an area where introgression from A. cardenasii had previously been reported. This fragment was cloned and used to generate a pair of primers that specifically amplified this locus (sequence characterized amplified region, SCAR) and as a RFLP probe. Their close linkage with the resistance genes will be useful in marker-based selection while transferring nematode resistance from introgression lines into elite breeding lines and cultivars. The Z3/265 marker associated with the genes Mae or Mag was not found in other highly resistant Arachis species (Arachis batizocoi or Arachis stenosperma), in progenies of interspecific crosses with A. cardenasii that were moderately resistant, or in the resistant A. hypogaea lines PI 259634 and PI 259572. These represent the first molecular markers linked with a resistant gene in peanut and the first report of two physiological responses to nematode attack associated with two genetic factors.

摘要

在由四倍体(花生×卡氏花生)-GA 6与PI 261942杂交产生的分离F2群体中,鉴定出两个决定对根结线虫南方根结线虫抗性的显性基因。建议将Mae作为限制卵数的显性基因的命名,将Mag作为限制虫瘿形成的显性基因的命名。GA 6中的高水平抗性是从卡氏花生导入的,因此,利用混合分组分析法,开展了一项寻找与这些抗性基因紧密连锁的卡氏花生特异性RAPD标记的研究。一个RAPD标记(Z3/265)分别与Mag和Mae连锁,距离分别为10±2.5(标准误)和14±2.9厘摩。在回交图谱中,该标记被定位到第1连锁群,距离Xuga.cr239 5厘摩,该区域此前曾报道有卡氏花生的导入。该片段被克隆,并用于生成一对特异性扩增该位点的引物(序列特征性扩增区域,SCAR),并用作RFLP探针。它们与抗性基因的紧密连锁在将线虫抗性从导入系转移到优良育种系和品种时,对基于标记的选择将是有用的。与Mae或Mag基因相关的Z3/265标记在其他高抗花生品种(巴氏花生或狭叶花生)、与卡氏花生种间杂交产生的中度抗性后代或抗性花生品系PI 259634和PI 259572中均未发现。这些代表了花生中与抗性基因连锁的首批分子标记,以及与两个遗传因素相关的对线虫攻击的两种生理反应的首次报道。

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