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利用分离群体分组分析法对水稻温敏雄性不育基因进行分子作图。

Molecular mapping of a thermosensitive genetic male sterility gene in rice using bulked segregant analysis.

出版信息

Genome. 1997 Apr;40(2):188-94. doi: 10.1139/g97-027.

Abstract

The thermosensitive genetic male sterility (TGMS) system is considered to be a more efficient alternative to the cytoplasmic male sterility (CMS) system for hybrid rice. An F2 population from a cross between a TGMS mutant line (IR32364TGMS) and IR68 was used to map the TGMS gene tms3(t). Fertile and sterile bulks were constructed following the classification of F2 plants into true breeding sterile, fertile, and segregating fertile plants based on F3 family studies. From the survey of 389 arbitrary primers in bulked segregant analysis, four RAPD markers were identified in which three, OPF182600, OPB19750, and OPAA7550, were linked to tms3(t) in repulsion phase and one, OPAC3640, was linked to tms3(t) in coupling phase. The tms3(t) gene was flanked by OPF182600 and OPAC3640 on one side and by OPAA7550 and OPB19750 on the other side. All four markers were low-copy sequences and two of them (OPF182600 and OPAC3640) detected polymorphism when the markers were used to probe the genomic blots. Subsequently, OPAC3640 was mapped to the short arm of chromosome 6 using a mapping population available at IRRI. However, no RFLP markers from this region showed linkage to tms3(t) owing to the lack of polymorphism between the parents. All RAPD fragments were cloned and partially sequenced from both ends. Thus, PCR primers can be designed to develop PCR markers for marker-assisted breeding to facilitate the transfer of tms3(t) from one genetic background to another.

摘要

温敏雄性不育(TGMS)系统被认为是杂种水稻中比细胞质雄性不育(CMS)系统更有效的替代方法。利用 TGMS 突变体系(IR32364TGMS)与 IR68 的杂交 F2 群体,对 TGMS 基因 tms3(t)进行了定位。根据 F3 家系研究,将 F2 植株分为纯合不育、可育和分离可育植株,构建了可育和不育群体。在对 389 个随机引物进行的混合分离分析中,鉴定出 4 个 RAPD 标记,其中 3 个标记 OPF182600、OPB19750 和 OPAA7550 与 tms3(t)在相斥阶段连锁,1 个标记 OPAC3640 与 tms3(t)在相引阶段连锁。tms3(t)基因一侧被 OPF182600 和 OPAC3640 侧翼,另一侧被 OPAA7550 和 OPB19750 侧翼。所有 4 个标记均为低拷贝序列,其中 2 个标记(OPF182600 和 OPAC3640)在用于探测基因组印迹时检测到多态性。随后,利用 IRRI 提供的作图群体,将 OPAC3640 定位到第 6 号染色体的短臂上。然而,由于亲本之间缺乏多态性,该区域的任何 RFLP 标记都与 tms3(t)没有连锁。从两端克隆并部分测序了所有 RAPD 片段。因此,可以设计 PCR 引物来开发用于标记辅助选择的 PCR 标记,以促进 tms3(t)从一个遗传背景转移到另一个遗传背景。

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