Map-based cloning of a spotted-leaf mutant gene OsSL5 in Japonica rice
Map-based cloning of a spotted-leaf mutant gene OsSL5 in Japonica rice
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粳稻斑叶突变基因 OsSL5 的图位克隆
DOI：
10.1007/s10725-014-9962-4
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发表时间：
2015-04-01
影响因子：
4.2
通讯作者：
Xue, Da-wei
中科院分区：
文献类型：
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作者：
Ge, Chang-wei;E, Zhi-guo;Xue, Da-wei
利用EMS诱变技术从粳稻云32中筛选到一个突变体，即斑叶5号（sl 5）。从成熟期到晚熟期，s15叶片斑点数逐渐增多，但叶片不干枯、枯萎。sl 5突变体的株高、穗长、一次分支数、二次分支数和千粒重均显著低于云32。遗传分析表明，sl 5为单隐性基因控制。通过图位克隆将OsSL 5定位在第7染色体上的一个40 kb的区间内，两侧为标记MX4和MX5。在该区域共鉴定出4个开放阅读框，其中包括1个SPL 5基因。测序结果表明，OsSL 5基因编码区3,647位G碱基发生突变，变为A。OsSL 5突变体的突变位点与SPL 5突变体的突变位点不同，具有籼稻背景。OsSL 5是一个新的SPL 5等位基因，编码一个剪接因子3b亚基3。QPCR显示，在sl 5突变体中OsSL 5的表达显著低于YUN 32。斑点叶相关基因RLIN 1、SPL 28和SPL 18的表达显著降低，而SPL 7和SL基因的表达显著增加。OsSL 5基因可能在水稻细胞凋亡调控中起重要作用。
A japonica rice mutant, spotted-leaf 5 (sl5), was identified from YUN32 by EMS mutagenesis. The number of spots in leaves increased from maturity to late maturity in sl5, however, the leaves did not dry and withered. The sl5 mutant exhibited significantly lower height, spike length, primary branch number, second branch number, and 1,000-grain weight than YUN32. Genetic analysis shows that sl5 is controlled by a single recessive gene. OsSL5 was mapped into a 40-kb interval flanked by markers MX4 and MX5 on chromosome 7 by map-based cloning. Four ORFs, including one SPL5 gene, were identified in this region. Sequencing reveals that the G base at site 3,647 of the OsSL5 coding region was changed to A. The mutant OsSL5 site was different from that of the SPL5 mutant, with the background of indica rice. OsSL5 is thus a new SPL5 allele which encodes a putative splicing factor 3b subunit 3. QPCR shows that OsSL5 expression in the sl5 mutant is significantly lower than that in YUN32. The spotted leaf-related genes RLIN1, SPL28, and SPL18 expressions were significantly decreased, whereas the SPL7 and SL gene expressions significantly increased. The OsSL5 gene may be important for rice cell apoptosis regulation.