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通过辐射诱变籼稻品种明恢86,获得两种短穗突变体,分别命名为rpl1(reduced panicle length 1)和rpl2(reduced panicle length 2)。两突变体都表现为穗变短、枝梗数和穗粒数减少、枝梗缩短,但其再生植株的穗长有所恢复,与野生型的差异明显减少。等位性测验表明,rpl1和rpl2是等位突变。利用M4分离群体和基于高通量测序的分离体混合分析(BSA-seq)方法,发现rpl2中LOC_Os11g12740基因的剪接位点发生了单碱基置换突变。进一步对rpl1测序分析,发现该基因完全缺失。这说明rpl1和rpl2都是LOC_Os11g12740突变引起的,由此证实它就是目标基因。该基因与已报道的SP1是同一个基因,因此rpl1和rpl2是SP1的新等位突变,但二者的表型与已报道的3种短穗突变体并不完全相同,说明不同突变和遗传背景会影响基因功能的表现。本研究进一步验证了SP1的功能,并为深入研究其分子作用机理提供新材料。
Two short-spike mutants were obtained by radiation-induced mutagenesis of indica rice Minghui86, which were named reduced panicle length (rpl1) and reduced panicle length (rpl2), respectively. Both mutants showed shortened spikes, fewer stems and fewer spikelets, and shorter stems. However, the spike length of regenerated plants recovered, with a significant decrease from the wild type. Allelism tests showed that rpl1 and rpl2 are allelic mutations. Based on the BSA-seq method, the splice site of LOC_Os11g12740 in rpl2 was found to be a single-base substitution mutation using M4 segregation population and high-throughput sequencing-based segregation analysis. Further analysis of the rpl1 gene revealed that the gene was completely deleted. This shows that rpl1 and rpl2 are caused by LOC_Os11g12740 mutation, thus confirming that it is the target gene. This gene is identical to the reported SP1 gene. Therefore, rpl1 and rpl2 are new allelic mutations of SP1, but their phenotypes are not exactly the same as the reported three short-spike mutants, indicating that different mutations and genetic Background will affect the performance of gene function. This study further validates the function of SP1 and provides new material for further study of its molecular mechanism of action.