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利用NGS辅助的BSA策略在水稻中鉴定耐碱候选基因

Identification of alkali-tolerant candidate genes using the NGS-assisted BSA strategy in rice.

作者信息

Sun Jian, Wang Jingguo, Guo Wei, Yin Tianjiao, Zhang Shuli, Wang Liang, Xie Dongwei, Zou Detang

机构信息

Key Laboratory of Germplasm Enhancement, Physiology and Ecology of Food Crops in Cold Region, Chinese Ministry of Education (Northeast Agricultural University), Harbin, 150030 China.

Biotechnology Institute of Heilongjiang Academy of Agricultural Sciences, Harbin, 150086 China.

出版信息

Mol Breed. 2021 Jul 16;41(7):44. doi: 10.1007/s11032-021-01228-x. eCollection 2021 Jul.

Abstract

UNLABELLED

Rice () is a saline-alkali-sensitive crop. Saline-alkali environments can seriously affect the growth, development, and yield of rice. The mechanisms of salt tolerance and alkali tolerance in rice are different; thus, it is very important to study and explore the alkali-tolerant gene loci to improve the saline-alkali tolerance of rice varieties. In this study, the rice varieties Dongnong 425 (DN425) and Changbai 10 (CB10) and a hybridized recombinant inbred line (RIL) population were used as materials to be irrigated with NaCO solution under field test conditions. A resistant pool (R-pool) and a sensitive pool (S-pool) were constructed by selecting the lines with extremely high and extremely low 1000-grain weight (TGW), respectively, from the RIL population under alkali treatment. Four candidate TGW regions on chromosomes (Chr.) 2 and 3 were associated using the bulked segregant analysis (BSA) strategy assisted by next-generation sequencing (NGS) technology (NGS-assisted BSA). Using the linkage analysis, QTL- in the candidate region was mapped within a range of 116 Kb between the SSR marker RM13592 and the Indel marker Indel3 of Chr. 2, which contained 18 predictive genes. The BSA sequencing results showed that contained a nonsynonymous substitution mutation SNP (nsSNP), leading to the transformation of a residue from arginine (cGg) to glutamine (cAg); thus, was inferred to be the candidate gene of . The results of the qRT-PCR analysis also confirmed this. This paper provides important information for the rapid and accurate identification of the alkali-tolerant gene loci in rice.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s11032-021-01228-x.

摘要

未标注

水稻()是一种对盐碱敏感的作物。盐碱环境会严重影响水稻的生长、发育和产量。水稻的耐盐和耐碱机制不同;因此,研究和探索耐碱基因位点对于提高水稻品种的盐碱耐受性非常重要。在本研究中,以水稻品种东农425(DN425)和长白10(CB10)以及一个杂交重组自交系(RIL)群体为材料,在田间试验条件下用NaCO溶液灌溉。通过在碱处理下从RIL群体中分别选择千粒重(TGW)极高和极低的株系构建抗性池(R池)和敏感池(S池)。利用下一代测序(NGS)技术辅助的混合分离分析(BSA)策略(NGS辅助BSA)关联了第2和第3染色体上的四个候选TGW区域。通过连锁分析,候选区域内的QTL - 被定位在第2染色体的SSR标记RM13592和Indel标记Indel3之间116 Kb的范围内,该区域包含18个预测基因。BSA测序结果表明,包含一个非同义替换突变SNP(nsSNP),导致一个残基从精氨酸(cGg)转变为谷氨酰胺(cAg);因此,推断为的候选基因。qRT - PCR分析结果也证实了这一点。本文为快速准确鉴定水稻耐碱基因位点提供了重要信息。

补充信息

在线版本包含可在10.1007/s11032 - 021 - 01228 - x获取的补充材料。

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