水稻温敏型叶色突变体tsa58的变异检测及转录组分析

陈能刚; 鄢小青; 李欢; 杨占烈; 吴荣菊; 陈锋

doi:10.16590/j.cnki.1001-4705.2023.05.001

水稻温敏型叶色突变体tsa58的变异检测及转录组分析

贵州省农业科学院农作物品种资源研究所

基金项目:

贵州省自然科学基金项目“水稻粒宽新基因GS 114的基因克隆与功能研究”(黔科合基础-ZK[2022]一般243)

贵州省科技支撑计划项目“农作物和食用菌种质资源精准鉴定评价(表型和基因型精准鉴定)”(黔科合支撑[2022]重点025号)

贵州省科技支撑计划项目“农作物种质资源精准鉴定评价(优异基因挖掘与利用)”(黔科合支撑[2022]重点026号)

贵州省科技计划项目“贵州省育种科研基础平台创新能力建设”(黔科合服企[2022]014)

详细信息

作者简介:
陈能刚(1980—),男(汉族),贵州遵义人,博士,副研究员,主要从事种质资源保存利用研究(E-mail:364968121@qq.com)

中图分类号: S511
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出版历程
- 收稿日期: 2022-12-20

Variation Detection and Transcriptome Analysis of a Thermo-sensitive Leaf Color Mutant tsa58 in Rice

Institute of Crop Germplasm Resources, Guizhou Academy of Agricultural Sciences

摘要

摘要: 利用⁶⁰Co辐射诱变贵州地方粳稻种质资源“桥港珍珠米”,筛选获得一个温敏型叶色突变体tsa58（temperature-sensitive albino 58）,该突变体在20℃和30℃条件下培养，幼苗存在显著的叶色差异。在20℃条件下，tsa58叶片呈现完全白化；在30℃条件下，tsa58叶色与野生型无明显差异。在20℃条件下，与野生型相比，tsa58叶片的总叶绿素、叶绿素a、叶绿素b和类胡萝卜素含量降低幅度最大，均达到极显著差异水平；在30℃条件下，与野生型相比，tsa58叶片的总叶绿素、叶绿素a、叶绿素b和类胡萝卜素含量均与野生型无显著差异。为了明确该突变表型的候选基因，采用二代高通量测序技术对突变体tsa58和野生型进行全基因组变异分析。研究发现，在定位区间内的20878044～21473236间发生了INV（倒置）结构变异，其位点21473236位于Os02g0565400的外显子上。转录组分析结果表明，该基因在野生型中存在TSS、TTS、AE等3种剪接类型，并存在5个不同的剪接位点，而突变体tsa58未检测到可变剪接。由于该INV变异导致该基因的可变剪接类型发生变化，因此推测tsa58为Os02g0565400（WSL4）的新等位突变体。转录组差异基因分析表明，突变体tsa58与野生型的差异基因在核糖体最多，为27个，其中由叶绿体基因组编码的基因rps7、rpl32、rpl20和rps16均显著下调，而由细胞核基因组编码的基因中除OsRPL44显著下调外，其余基因均显著上调。
- 水稻 /
- 温敏型叶色突变体 /
- 变异检测 /
- 转录组分析 /
- 辐射诱变
Abstract: The thermo-sensitive mutant tsa58（temperature-sensitive albino 58） was obtained from “Qiao-gang Pearl Rice”, a germplasm resource of japonica rice in Guizhou by ⁶⁰Co radiation. The mutant seedlings of tsa58 showed significant leaf color difference at 20 ℃ and 30 ℃. The leaves of tsa58 showed albino at 20 ℃. However, there was no significant difference in leaf color between tsa58 and wild type at 30 ℃. The contents of total chlorophyll, chlorophyll a, chlorophyll b and carotenoid in the leaves of tsa58 decreased the most at 20 ℃, and all reached the significant difference level compared with the wild type. The contents of total chlorophyll, chlorophyll a, chlorophyll b and carotenoid in leaves of tsa58 were not significantly different from those of the wild type at 30 ℃. In order to identify candidate genes for the mutant phenotype, tsa58 and the wild type were analyzed with whole-genome variation using second-generation high-throughput sequencing technology. It was found that INV（inverted） occurred between 20878044 and 21473236, and the mutation site（21473236） was located in the exon of Os02g0565400. The transcriptome analysis showed that there were three splicing types（TSS, TTS, AE） and five different splicing sites in the wild type, but no anyone type of variable splicing was detected in the mutant tsa58. Due to the INV mutation caused a change in the variable splicing type of this gene, and it was speculated that the tsa58 was a new allele mutants of Os02g0565400（WSL4）. The study of the transcriptome differential gene analysis showed that the tsa58 had a maximum of 27 differential genes in the ribosome rps7, rpl32, rpl20 and rps16 encoded by chloroplast genome were significantly down-regulated, while all genes encoded by nuclear genome were significantly up-regulated except OsRPL44.
- rice /
- thermo-sensitive leaf color mutant /
- variation detection /
- transcriptome analysis /
- radiation

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参考文献(9)

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