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水稻无叶枕突变体的叶夹角较小,叶片直立,在培育叶片直立型水稻新品种中存在潜在价值。本研究对一个新的水稻无叶枕突变体oslg-h进行了鉴定、基因定位和候选基因分析。oslg-h表现为叶枕、叶舌、叶耳缺失,株型紧凑和叶片直立。遗传分析表明,oslg-h受一对隐性基因控制。利用oslg-h/dullar F2群体将该基因定位在第4染色体的Indel12和Indel21之间254 kb范围内,其间包含一个已报道的无叶枕基因OsLG1。对野生型和突变体中OsLG1启动子、5’非翻译区(5’UTR)、开放阅读框和3’非翻译区(3’UTR)测序比对发现:在突变体中OsLG1第2个内含子和3’UTR各有1处插入突变。RT-PCR分析和测序表明,内含子区域的碱基插入突变没有影响该基因的表达和mRNA的剪接。根据3’UTR插入位点设计了一对特异检测该突变位点的Indel标记Indel-U,对苏御糯-5-1/oslg-h F2进行分析,112个无叶枕植株扩增条带均是与oslg-h相似的200 bp带型。353个有叶枕植株扩增出两种带型,一种是与苏御糯-5-1相似的192 bp带型;另一种是与苏御糯-5-1/oslg-h F1相似的192 bp和200 bp带型。以上表明oslg-h无叶枕表型可能是由3’UTR插入突变导致的。Indel-U能直接区分OsLG-HOsLG-H、OsLG-Hoslg-h和oslg-hoslg-h三种基因型,在oslg-h基因分子标记辅助选择和基因聚合育种中具有直接应用价值。
There was a potential value of leaf angle between leaves of non-leaf pillow mutant and upright leaf in cultivating leaf-type erect rice. In this study, a new non-leaf pillow mutant oslg-h was identified, gene mapping and candidate gene analysis. oslg-h performance for the leaf pillow, ligule, lack of leaf ears, plant compact and leaf upright. Genetic analysis showed that oslg-h is controlled by a pair of recessive genes. The oslg-h / dullar F2 population was used to map this gene within 254 kb between Indel12 and Indel21 on chromosome 4, containing a reported non-leaf pillow gene, OsLG1, in between. Sequencing of the OsLG1 promoter, 5 ’untranslated region (5’UTR), open reading frame (ORF) and 3’ untranslated region (3’UTR) in wild type and mutant revealed that in the mutant, Mutants were inserted into each of the 3 ’UTR and the 3’ UTR. RT-PCR analysis and sequencing showed that intron insertion mutation did not affect the gene expression and mRNA splicing. According to the 3’UTR insertion site, a pair of Indel-labeled Indel-U that specifically detected this mutation site was designed to analyze the Suyu Nuo -5-1 / oslg-h F2, and 112 non-leaf pillow plant amplified bands All are 200 bp bands similar to oslg-h. 353 plants with leaf pillow were amplified two bands, one is similar to Su Yu Nuo -5-1 192 bp band; the other is Su Yu Nuo -5-1 / oslg-h F1 similar 192 bp and 200 bp bands. The above shows that oslg-h non-leaf pillow phenotype may be caused by the 3’UTR insertion mutation. Indel-U can directly distinguish the three genotypes of OsLG-HOsLG-H, OsLG-Hoslg-h and oslg-hoslg-h, and has direct application value in molecular marker-assisted selection and gene polymerization breeding of oslg-h gene.