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利用CRISPR/Cas9创建osarf7突变体及其农艺性状调查 被引量:2
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作者 李兆伟 孙聪颖 +4 位作者 零东兰 曾慧玲 张晓妹 范凯 林文雄 《中国水稻科学》 CAS CSCD 北大核心 2022年第3期237-247,共11页
【目的】探究OsARF7基因在水稻生长发育中的生物学功能及其对水稻农艺性状的影响。【方法】利用CRISPR/Cas9技术,在粳稻中花11背景下对OsARF7进行定向编辑,获得OsARF7基因突变植株,并考查其农艺性状。【结果】获得22株T0代转基因株系,经... 【目的】探究OsARF7基因在水稻生长发育中的生物学功能及其对水稻农艺性状的影响。【方法】利用CRISPR/Cas9技术,在粳稻中花11背景下对OsARF7进行定向编辑,获得OsARF7基因突变植株,并考查其农艺性状。【结果】获得22株T0代转基因株系,经PCR扩增潮霉素基因鉴定出20株阳性植株。提取T2代转基因植株的基因组DNA,通过对OsARF7的2个编辑位点区域DNA片段进行PCR扩增及测序分析,筛选到15种突变类型的纯合基因型植株。基因表达分析显示,osarf7突变体的OsARF7和生长素转运关键基因OsPIN1b、Os PIN2和OsLAX2的表达量均显著低于野生型。田间调查发现,与野生型相比,osarf7突变体的分蘖成穗率显著降低,其中,分蘖数比野生型增多的osarf7突变体,有效穗数基本保持不变,而分蘖数与野生型相近的osarf7突变体,有效穗数少于野生型,最终表现为无效分蘖明显增多,分蘖成穗率降低。【结论】OsARF7基因与生长素调控水稻分蘖发生相关,并影响分蘖芽的生长发育及成穗;OsARF7基因突变会导致迟发分蘖芽不能正常发育成穗;粳稻OsARF7突变体的创建对揭示OsARF7基因生理功能和探索生长素促控分蘖发生与成穗机理等具有积极意义。 展开更多
关键词 水稻 基因编辑 osarf7 CRISPR/Cas9 生长素
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Nuclear translocation of OsMADS25 facilitated by OsNAR2.1 in reponse to nitrate signals promotes rice root growth by targeting OsMADS27 and OsARF7 被引量:1
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作者 Junyu Wu Shuaiqi Yang +8 位作者 Nana Chen Qining Jiang Linli Huang Jiaxuan Qi Guohua Xu Lisha Shen Hao Yu Xiaorong Fan Yinbo Gan 《Plant Communications》 SCIE CSCD 2023年第6期329-341,共13页
Nitrate is an important nitrogen source and signaling molecule that regulates plant growth and development.Although several components of the nitrate signaling pathway have been identified,the detailed mechanisms are ... Nitrate is an important nitrogen source and signaling molecule that regulates plant growth and development.Although several components of the nitrate signaling pathway have been identified,the detailed mechanisms are still unclear.Our previous results showed that OsMADS25 can regulate root development in response to nitrate signals,but the mechanism is still unknown.Here,we try to answer two key questions:how does OsMADS25 move from the cytoplasm to the nucleus,and what are the direct target genes activated by OsMADS25 to regulate root growth after it moves to the nucleus in response to nitrate?Our results demonstrated that OsMADS25 moves from the cytoplasm to the nucleus in the presence of nitrate in an OsNAR2.1-dependentmanner.Chromatin immunoprecipitation sequencing,chromatin immunoprecipitation qPCR,yeast one-hybrid,and luciferase experiments showed that OsMADS25 directly activates the expression of OsMADS27 and OsARF7,which are reported to be associated with root growth.Finally,OsMADS25-RNAi lines,the Osnar2.1 mutant,and OsMADS25-RNAi Osnar2.1 lines exhibited significantly reduced root growth compared with the wild type in response to nitrate supply,and expression of OsMADS27 and OsARF7 was significantly suppressed in these lines.Collectively,these results reveal a new mechanismby which OsMADS25 interacts with OsNAR2.1.This interaction is required for nuclear accumulation of OsMADS25,which promotes OsMADS27 and OsARF7 expression and root growth in a nitratedependent manner. 展开更多
关键词 OsMADS25 OsNAR2.1 OsMADS27 osarf7 nitrate signaling rice root
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