不同水稻<i>tms5</i>突变位点对雄性不育起点温度的影响

吴明基; 刘华清; 陈建民; 付艳萍; 杨绍华; 宋亚娜; 林艳; 王锋

doi:10.19303/j.issn.1008-0384.2023.07.001

不同水稻tms5突变位点对雄性不育起点温度的影响

Critical Sterility-inducing Temperature of Rice Affected by tms5 Mutation Site

摘要

摘要:
目的研究不同水稻tms5突变位点对雄性不育起点温度的影响，探讨不育起点温度遗传调控途径。
方法在水稻TMS5的6个外显子上设计11个CRISPR/Cas9基因编辑靶点，依次命名为T501 — T511，构建相应载体转化粳稻品种日本晴和籼稻品种明恢86，获得各靶点的tms5移码突变体。田间自然高温及人工气候箱（设日平均22 、24和28 ℃ 3种温度）条件下分析tms5突变体的花粉碘染及自交结实率，鉴定不育起点温度。
结果粳稻日本晴tms5突变体的不育起点温度高于28 ℃，籼稻明恢86的不同tms5突变体不育起点温度为22～28 ℃。此外，同一遗传背景下，通过T501靶点编辑产生的tms5-1移码突变体不育起点温度均显著高于T502靶点的tm5-2突变体，其他位点上的tms5突变体育性特征与tm5-2突变体并无差异。基因表达量分析表明，tms5-1突变体幼穗Ub_L40基因表达量显著低于tm5-2突变体的表达量。
结论水稻tms5突变体不育起点温度不仅受遗传背景的影响，tms5基因突变位点不同也会影响不育起点温度，特别是T501位点与其余位点突变体间不育起点温度差异显著，为研究水稻tms5两系不育起点温度的分子机理及遗传调控网络提供了新思路。

Abstract:
Objective Effects of tms5 mutation site and related genetic regulatory factors in rice on the critical sterility-inducing temperature (CSIT) of the plant were investigated.
Method Eleven CRISPR/Cas9 sequentially numbered T501 through T511 were designed to target 6 exons of tms5 to create mutants of Nipponbare (NIP, Oryza sativa ssp. Geng) and Minghui 86 (MH86, O. sativa ssp. Xian). Pollen male fertility and seed setting rate of the mutants were monitored under long day and high temperature field conditions in early August in Fuzhou (NHT) or at 22, 24, and 28 ℃ in phytotrons to identify the CSIT.
Result The CSIT of the NIP mutants were higher than 28 ℃, while that of the MH86 mutants between 22 ℃ and 28°C. The tms5-1 T501 mutant had a significantly higher CSIT than the genetically identical tms5-2 counterparts T502, but the other 9 tm5 and the tm5-2 mutants did not differ on it. Expressions of the 3 Ub_L40 in young panicles were lower in the tms5-1 than the tm5-2 mutants of either rice varieties.
Conclusion It appeared that the CSIT of tms5 mutants was affected by the genetic factors as well as the mutation site as the tms5-1 mutant T501 was shown significantly differed from the others.

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