Molecular genetical analysis of the two locus self incompatibility in the grasses (Poaceae)

禾本科植物两个位点自交不亲和性的分子遗传学分析

• 批准号: 12660005
• 负责人: KAKEDA Katsuyuki
• 金额: $ 2.24万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660005/
• 关键词: Self-incompatibility; S gene; AFLP; mRNA fingerprinting; Barley; Hordeum bulbosum; Poaceae; Linkage map; 形質転換; Z遺伝子; RFLP; 細胞間認識; Hordeum; 花粉

The self-incompatibility (SI) in the grasses (Poaceae) is gametophytically controlled by two unlinked loci, Sand Z, The aim of this project is to establish the genetic basis of the SI in wild barley, Hordeum bulbosum (2x) and to clone the SI gene(s). The basic genetical analysis involves crossing experiments of plant lines, determination of SI genotypes, production of large mapping populations, identification of DNA markers closely linked to the SI loci, etc. Toward the,, cloning of the S gene, the AFLP-based mRNA fingerprinting (AMF) method was employed to search S genotype-specific transcripts from pistils and anthers at anthesis. By the AMF analysis, dozens of S specific fragments were detected from both the pistil and anther transcripts. The fragments were cloned and subjected to the RFLP analysis to examine linkage to the S locus. Several clones have so far been confirmed for the close linkage to the S locus. The S-linked clones were used to construct a detailed genetic map around the S locus. No recombination was found between the S locus and some of the clones, indicating that those clones are candidates of the S gene. The expression pattern and sequence polymorphism of the candidate clones should be analyzed in detail to examine whether they are the S gene or not. It is also necessary to develop the transformation system for H bulbosum to demonstrate that the candidate gene is the determinant of S specificity

禾本科植物的自交不亲和性（self-incompatibility，SI）受两个非连锁位点Sand Z的配子体控制。本研究旨在建立野生大麦（Hordeum bulbosum，2x）自交不亲和性的遗传基础，并克隆SI基因。基本遗传分析包括植物系的杂交试验、SI基因型的确定、大作图群体的产生、与SI基因座紧密连锁的DNA标记的鉴定等。在S基因的克隆方面，采用基于AFLP的mRNA指纹（AMF）方法从雌蕊和花药中寻找S基因型特异转录本。通过AMF分析，从雌蕊和花药转录本中检测到几十个S特异片段。克隆片段并进行RFLP分析以检查与S位点的连锁。到目前为止，已经证实了几个克隆与S位点紧密连锁。利用S连锁克隆构建了S位点附近的详细遗传图谱。在S基因座和一些克隆之间没有发现重组，表明这些克隆是S基因的候选者。候选克隆的表达模式和序列多态性应进行详细分析，以检查它们是否是S基因。还需要进一步建立球孢菌的转化体系，以证明候选基因是S特异性的决定基因

项目成果

K.Kakeda: "A self-compatible mutant S allele conferring a dominant negative effect on the functional S alelle in Ipomoea trifida."Sexual Plant Reproduction. 13. 119-125 (2000)

K.Kakeda：“一种自交亲和突变体 S 等位基因，对三裂叶薯的功能性 S 等位基因产生显性负面影响。”有性植物繁殖。

伊吹敏郎: "Hordeum bulbosumにおけるAMF法を用いた自家不和合性遺伝子座転写産物の解析"育種学研究. 3(別冊2). 197 (2001)

Toshiro Ibuki：“使用 AMF 方法分析大麦的自交不亲和性基因座转录本”育种研究 3（特刊 197（2001））。

掛田克行: "チオレドキシン様遺伝子の塩基配列を用いたHordeum属の系統分化の解析"育種学研究. 3(別冊2). 289 (2001)

Katsuyuki Kakeda：“使用硫氧还蛋白样基因的核苷酸序列分析大麦属的谱系分化”育种研究3（特刊289）。

T.Ibuki: "Analysis of transcripts from the self-incompatibility loci of Hordeum bulbosum by using the AMF method"Breeding Science. Vol.3 (Suppl.2). 197 (2001)

T.Ibuki：“使用 AMF 方法分析球根大麦自交不亲和基因座的转录本”《育种科学》。

K.Kakeda: "A self-compatible mutant S allele conferring a dominant negative effect on the functional S allele in Ipomoea trifida"Sexual Plant Reproduction. 13. 191-196 (2000)

K.Kakeda：“自交亲和突变体 S 等位基因对三裂叶薯中的功能性 S 等位基因产生显性负面影响”有性植物繁殖。