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A 9.5-kb deletion in the 1st intron of OsMADS51 enhances temperature sensitivity in rice 被引量:1
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作者 Lei Zhao Haifei Hu +11 位作者 Junyu Chen Chongrong Wang Yibo Chen Hong Li Daoqiang Huang Zhidong Wang Degui Zhou Rong Gong Yangyang Pan Junliang Zhao Liangyong Ma Shaochuan Zhou 《The Crop Journal》 SCIE CSCD 2024年第4期1031-1040,共10页
Temperature is an important environmental factor affecting heading date of rice.Despite its importance,genes responsible for temperature-sensitive heading in rice have remained elusive.Our previous study identified a ... Temperature is an important environmental factor affecting heading date of rice.Despite its importance,genes responsible for temperature-sensitive heading in rice have remained elusive.Our previous study identified a quantitative trait locus qHd1 which advances heading date under high temperatures.A 9.5-kb insertion was found in the first intron of OsMADS51 in indica variety Zhenshan 97(ZS97).However,the function of this natural variant in controlling temperature sensitivity has not been verified.In this study,we used CRISPR/Cas9 to knock out the 9.5-kb insertion in ZS97.Experiments conducted under cotrolled conditions in phytotrons confirmed that deletion increased temperature sensitivity and advanced heading by downregulating the expression level of OsMADS51.One-hybrid assays in yeast,ChIP-quantitative polymerase chain reaction,electrophoretic mobility shift,and luciferase-based transient transactivation assays collectively confirmed that OsMADS51 affects heading date by regulation of heading date gene Ehd1.We further determined that the long non-coding RNA HEATINR is generated from the first intron of OsMADS51,offering an explanation for how the 9.5-kb insertion affects temperature sensitivity.We also found that OsMADS51 was strongly selected in early/late-season rice varieties in South China,possibly accounting for their strong temperature sensitivity.These insights not only advance our understanding of the molecular mechanisms underlying the temperature-responsive regulation of heading date in rice but also provide a valuable genetic target for molecular breeding. 展开更多
关键词 osmads51 INSERTION Long non-coding RNA Oryza sativa
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FCS-Like锌指蛋白OsFLZ18在调控水稻抽穗期中的作用 被引量:3
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作者 马雅美 张少红 +1 位作者 赵均良 刘斌 《中国农业科学》 CAS CSCD 北大核心 2022年第20期3875-3884,共10页
【目的】抽穗期是水稻重要的农艺性状,适时抽穗是确保水稻产量和区域适应性的关键因素。然而,水稻抽穗期调控的基因及其调控网络仍需进一步研究。已知FCS-like锌指蛋白是植物特有且在植物生长发育和逆境响应过程中发挥重要作用的蛋白家... 【目的】抽穗期是水稻重要的农艺性状,适时抽穗是确保水稻产量和区域适应性的关键因素。然而,水稻抽穗期调控的基因及其调控网络仍需进一步研究。已知FCS-like锌指蛋白是植物特有且在植物生长发育和逆境响应过程中发挥重要作用的蛋白家族,但其调控植物开花的功能未知。研究OsFLZs在调控水稻抽穗期中的功能,有助于完善水稻抽穗期调控网络,同时为抽穗期遗传育种提供新的理论基础和基因资源。【方法】根据RGAP数据库公布的基因序列,构建OsFLZ18的过量表达载体和CRISPR-Cas9敲除载体,通过农杆菌介导的遗传转化方法转化日本晴愈伤组织,从而获得相应的水稻转基因植株;利用PCR技术筛选并鉴定OsFLZ18敲除纯合株系;在水稻生长过程中,调查过量表达、敲除植株和日本晴植株的抽穗时间;采用qRT-PCR方法分析OsFLZ18的时空表达模式,昼夜节律性以及OsFLZ18对抽穗期调控基因表达的影响;通过酵母双杂交试验验证OsFLZ18与调控水稻抽穗期关键因子的相互作用。【结果】OsFLZ18的表达无明显组织特异性,在14 d的幼苗中表达量最高,其次是分蘖期的叶鞘和叶片,以及生殖生长阶段的茎和幼穗;成功构建了OsFLZ18-CRISPR载体,并转入粳稻日本晴中,筛选并鉴定获得2个CRISPR敲除纯合突变体。采用表达量较高的2个OE植株(OE-2和OE-3)以及纯合的敲除植株(CRISPR-21和CRISPR-25)进行抽穗期表型观察。结果发现,在广州自然短日照和自然长日照条件下,OE植株均出现明显的晚花表型,但是CRISPR敲除植株的抽穗时间较野生型无明显差异。qRT-PCR结果表明,在人工短日照条件下,与野生型相比,OE-2植株中Ehd1、Hd3a、RFT1的表达量明显被抑制,但Hd1的表达量没有明显的变化。酵母双杂交试验表明OsFLZ18和正向调控水稻抽穗期的转录因子OsMADS51存在互作。同时,OsFLZ18的表达量存在昼夜节律性,在白天表达量低,夜晚表达量高,并在半夜达到最高值。【结论】过量表达OsFLZ18可以导致水稻延迟抽穗。 展开更多
关键词 水稻 抽穗期 FLZ osmads51
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