利用高密度遗传图谱挖掘水稻芽期耐盐 QTL

雷 蕾; 孙世臣; 曹良子; 丁国华; 周劲松; 白良明; 洛 育; 夏天舒

文章摘要

雷蕾，孙世臣，曹良子，丁国华，周劲松，白良明，洛育，夏天舒.利用高密度遗传图谱挖掘水稻芽期耐盐 QTL[J].广东农业科学,2024,51(9):18-29

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Excavation of Salt-tolerant QTL in Rice at Bud Stage with High-Density Genetic Map

DOI：10.16768/j.issn.1004-874X.2024.09.002

中文关键词: 高密度 Bin 图谱 QTL 重组自交系耐盐水稻芽期

英文关键词: high-density Bin map QTL recombinant inbred line salt tolerance rice bud stage

基金项目:黑龙江省重点研发计划项目（2022ZX02B04-3）；中国博士后科学基金（2023MD734178）；黑龙江省农业科技创新跨越工程农业科技基础创新优青项目（CX22BS01）

作者	单位
雷蕾，孙世臣，曹良子，丁国华，周劲松，白良明，洛育，夏天舒	黑龙江省农业科学院耕作栽培研究所 / 黑龙江省水稻品质改良与遗传育种工程技术研究中心 / 黑龙江省作物分子设计与种质创新重点实验室

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中文摘要:

摘要：【目的】挖掘水稻芽期耐盐新位点，为进一步开展基因功能分析和盐渍土直播生产提供理论依据。【方法】以耐盐品种‘龙稻 133’和盐敏品种‘彩稻’衍生的 169 个 RIL 群体为试验材料，利用包含 1 107 个高质量多态性 Bin 标记的高密度遗传连锁图谱，对对照和 0.5% NaCl 胁迫下的胚芽鞘长、胚根数、胚根长以及相对胚芽鞘长、相对胚根数和相对胚根长共 9 个性状进行 QTL 定位分析。【结果】高密度遗传连锁图谱共覆盖水稻基因组 2 957.35 cM，标记间的平均距离为 2.67 cM。12 条染色体含有的平均标记数为 92.25 个。描述性统计分析表明，亲本‘龙稻 133’的耐盐性显著高于‘彩稻’，亲本的性状表型值位于 RIL 群体的极值之间，群体表现出超亲分离现象。盐胁迫严重抑制 RIL 群体的胚芽鞘长、胚根数和胚根长，且不同株系受胁迫影响差异较大，各性状基本符合正态分布，具有数量性状的遗传特性。连锁分析共鉴定到 19 个 QTL，贡献率为 2.58%~14.83%，发现 2 个 QTL 区间内存在已克隆的耐盐相关基因，其中 qTCL3 区间内包含 DST，qRRN10 区间内包含 OsMSRA4.1。此外，发现控制盐胁迫下胚芽鞘长的 qTCL10、控制相对胚芽鞘长的 qRCL10 以及控制相对胚根长的 qRRL10 均位于同一 QTL 区间内；控制盐胁迫下胚根长的 qTRL7 和控制相对胚根长的 qRRL7 也位于同一 QTL 区间内。2 个共定位区间内的 25 个候选基因 qRT-PCR 分析结果显示，盐处理后，LOC_Os07g44210、LOC_Os07g44240、LOC_Os07g44250、LOC_ Os07g44350、LOC_Os10g42940 和 LOC_Os10g43060 在‘彩稻’或‘龙稻 133’均显著上调表达。其中，LOC_ Os07g44350 在盐处理后的‘龙稻 133’胚芽鞘和胚根内均极显著上调表达。【结论】发掘 19 个与水稻芽期耐盐相关 QTL，其中包含 2 个共定位位点 qTRL7 和 qTCL10，2 个共定位区间内共有 25 个候选基因。通过 qRT-PCR 分析发现 6 个在盐处理后表达量上调的基因，其中 LOC_Os07g44350 是芽期耐盐的重要候选基因。

英文摘要:

Abstract: 【Objective】Excavation of new salt resistance sites in rice bud stage can provide theoretical basis for further gene function analysis and live production of saline soil.【Method】The QTL localization analysis of 169 RIL populations derived from ‘Longdao133’ and ‘Caidao’ was conducted by using a high-density genetic linkage map containing 1 107 highquality polymorphism Bin markers. Nine traits, including the coleoptile length, radicle number and radicle length under control condition, the coleoptile length, radicle number and radicle length under 0.5% NaCl treatment, and the relative coleoptile length, the relative radicle number and the relative radicle length, were analyzed for QTL localization analysis.【Result】The high-density genetic linkage map covered 2 957.35 cM of the rice genome, and the average distance between markers was 2.67 cM. The twelve chromosomes contained an average number of markers of 92.25. Descriptive statistical analysis showed that the salt tolerance of parental ‘Longdao133’ was significantly higher than that of ‘Caidao’, and the trait phenotypic values of the parents were between the extreme values of the RIL population, and the population showed superaffinity separation. Salt stress severely inhibited the coleoptile length, radicle number and radicle length of the RIL population, and different lines were greatly affected by stress; all traits basically conformed to the normal distribution and had the genetic characteristics of quantitative traits. Linkage analysis identified 19 QTLs that ranged from 2.58% to 14.83%, and identified cloned salt tolerance genes in two QTLs intervals, including DST in the qRTCL3 interval and OsMSRA4.1 in the qRRN10 interval. Moreover, it was found that, the qTCL10 controlling the coleoptile length, qRCL10 controlling the relative radicle length and qRRL10 controlling the relative radicle length under salt stress were located within the same QTL interval. Furthermore, qTRL7, which controlled the coleoptile length, and qRRL7, which controlled relative radicle length under salt stress, were located within the same QTL interval. Analysis results of qRT-PCR of 25 candidate genes within the 2 colocalization intervals showed that LOC_Os07g44210, LOC_ Os07g44240, LOC_Os07g44250, LOC_Os07g44350, LOC_Os10g42940 and LOC_Os10g43060 were significantly upregulated in ‘Caidao’ or ‘Longdao133’ after salt treatment. Among them, LOC_Os07g44350 was significantly up-regulated in both the coleoptile and radicle of ‘Longdao133’ after salt treatment.【Conclusion】A total of 19 QTLs associated with salt tolerance at the bud stage of rice were excavated, including two colocalization sites, qTRL7 and qTCL10, and 25 candidate genes in the two colocalization intervals. Six genes were found to be up-regulated after salt treatment by qRT-PCR analysis, and LOC_Os07g44350 was an important candidate for salt tolerance at the bud stage.

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