水稻矮化少分蘖基因D26的图位克隆及功能研究
发布时间：2021-02-13 23:19
作为重要的粮食作物,水稻从传统的经验育种向现代精准育种转变的过程中,株型是影响水稻产量最为重要因素之一,它取决于植株高度、分蘖数目、分蘖角度、穗形态等因素。对于水稻株型的机制研究,是短期内难以突破的科学问题和技术难题,其中矮化突变体是非常理想的研究对象,但是调控机制知之甚少。本文在前期研究的工作基础上,对水稻矮化、少分蘖D26突变体进行系统研究,揭示它参与水稻油菜素内酯激素调控的信号通路机制。本文从水稻突变体库中组织培养变异获得了2个农艺性状表现矮化、少分蘖的突变材料D26-1和D26-2,通过图位克隆实现精细定位和进一步的功能分析,揭示了D26突变体矮化表型的遗传机理,并从互作蛋白筛选和转录组测序分析角度,进一步挖掘水稻矮化机制。主要结果如下:D26突变体一个隐性单基因控制表型。利用D26突变体做母本和93-11亲本做父本,构建杂交F2群体,通过425个群体和531个群体将其分别定位在3号染色体短臂上63 kb区间,该区间内包含12个候选基因;分析预测和克隆、测序表明D26基因是一个编码GRAS家族蛋白的转录因子,两个突变体在D26基因540 bp和560 bp...
【文章来源】：华侨大学福建省
【文章页数】：143 页
【学位级别】：博士
【部分图文】：
水稻油菜素内酯相关基因研究网络(Tong,2018)
D26基因分别从水稻籼稻明恢86和D26突变体的c DNA中扩增获得全长,片段回收与pMD18-T连接、测序比对分析。互补实验载体构建:一个含有D26基因启动子、c DNA全长和3"UTR非编码区3563 bp全长片段从中间载体p UPGWPR上由HindIII/Eco RV酶切,与Bam HI酶切p CAMBIA1300载体分别回收,两片段经平端连接,最终构建互补载体p CAMBIAGWP26(见图1)。2.2.8 植物叶片总RNA提取-TRIzol法
转录组测序流程
【参考文献】：
期刊论文
[1]Brassinosteroid-regulated plant growth and development and gene expression in soybean[J]. Wenchao Yin,Nana Dong,Mei Niu,Xiaoxing Zhang,Lulu Li,Jun Liu,Bin Liu,Hongning Tong. The Crop Journal. 2019(03)
[2]OsIDD2, a zinc finger and INDETERMINATE DOMAIN protein, regulates secondary cell wall formation[J]. Peng Huang,Hideki Yoshida,Kenji Yano,Shunsuke Kinoshita,Kyosuke Kawai,Eriko Koketsu,Masako Hattori,Sayaka Takehara,Ji Huang,Ko Hirano,Reynante Lacsamana Ordonio,Makoto Matsuoka,Miyako Ueguchi-Tanaka. Journal of Integrative Plant Biology. 2018(02)
[3]A zinc finger protein, interacted with cyclophilin,affects root development via IAA pathway in rice[J]. Peng Cui,Hongbo Liu,Songlin Ruan,Basharat Ali,Rafaqat Ali Gill,Huasheng Ma,Zhifu Zheng,Weijun Zhou. Journal of Integrative Plant Biology. 2017(07)
[4]OsGRF4 controls grain shape, panicle length and seed shattering in rice[J]. Pingyong Sun,Wuhan Zhang,Yihua Wang,Qiang He,Fu Shu,Hai Liu,Jie Wang,Jianmin Wang,Longping Yuan,Huafeng Deng. Journal of Integrative Plant Biology. 2016(10)
[5]Rapid improvement of grain weight via highly efficient CRISPR/Cas9-mediated multiplex genome editing in rice[J]. Rongfang Xu,Yachun Yang,Ruiying Qin,Hao Li,Chunhong Qiu,Li Li,Pengcheng Wei,Jianbo Yang. Journal of Genetics and Genomics. 2016(08)
[6]TALEN-Mediated Homologous Recombination Produces Site-Directed DNA Base Change and Herbicide-Resistant Rice[J]. Ting Li,Bo Liu,Chih Ying Chen,Bing Yang. Journal of Genetics and Genomics. 2016(05)
[7]水稻小粒基因SG4（D11等位基因）的克隆和功能研究（英文）[J]. 石珍源,饶玉春,徐杰,胡时开,方云霞,余海萍,潘江杰,刘瑞芳,任德勇,王小虎,祝阳舟,朱丽,董国军,张光恒,曾大力,郭龙彪,胡江,钱前. Science Bulletin. 2015(10)
[8]GS6, A Member of the GRAS Gene Family,Negatively Regulates Grain Size in Rice[J]. Lianjun Sun,Xiaojiao Li,Yongcai Fu,Zuofeng Zhu,Lubin Tan,Fengxia Liu,Xianyou Sun,Xuewen Sun,Chuanqing Sun. Journal of Integrative Plant Biology. 2013(10)
[9]Toward understanding genetic mechanisms of complex traits in rice[J]. Wei Hao, Hong-Xuan Lin* National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. 遗传学报. 2010(10)
本文编号：3032729
【文章来源】：华侨大学福建省
【文章页数】：143 页
【学位级别】：博士
【部分图文】：
水稻油菜素内酯相关基因研究网络(Tong,2018)
D26基因分别从水稻籼稻明恢86和D26突变体的c DNA中扩增获得全长,片段回收与pMD18-T连接、测序比对分析。互补实验载体构建:一个含有D26基因启动子、c DNA全长和3"UTR非编码区3563 bp全长片段从中间载体p UPGWPR上由HindIII/Eco RV酶切,与Bam HI酶切p CAMBIA1300载体分别回收,两片段经平端连接,最终构建互补载体p CAMBIAGWP26(见图1)。2.2.8 植物叶片总RNA提取-TRIzol法
转录组测序流程
【参考文献】：
期刊论文
[1]Brassinosteroid-regulated plant growth and development and gene expression in soybean[J]. Wenchao Yin,Nana Dong,Mei Niu,Xiaoxing Zhang,Lulu Li,Jun Liu,Bin Liu,Hongning Tong. The Crop Journal. 2019(03)
[2]OsIDD2, a zinc finger and INDETERMINATE DOMAIN protein, regulates secondary cell wall formation[J]. Peng Huang,Hideki Yoshida,Kenji Yano,Shunsuke Kinoshita,Kyosuke Kawai,Eriko Koketsu,Masako Hattori,Sayaka Takehara,Ji Huang,Ko Hirano,Reynante Lacsamana Ordonio,Makoto Matsuoka,Miyako Ueguchi-Tanaka. Journal of Integrative Plant Biology. 2018(02)
[3]A zinc finger protein, interacted with cyclophilin,affects root development via IAA pathway in rice[J]. Peng Cui,Hongbo Liu,Songlin Ruan,Basharat Ali,Rafaqat Ali Gill,Huasheng Ma,Zhifu Zheng,Weijun Zhou. Journal of Integrative Plant Biology. 2017(07)
[4]OsGRF4 controls grain shape, panicle length and seed shattering in rice[J]. Pingyong Sun,Wuhan Zhang,Yihua Wang,Qiang He,Fu Shu,Hai Liu,Jie Wang,Jianmin Wang,Longping Yuan,Huafeng Deng. Journal of Integrative Plant Biology. 2016(10)
[5]Rapid improvement of grain weight via highly efficient CRISPR/Cas9-mediated multiplex genome editing in rice[J]. Rongfang Xu,Yachun Yang,Ruiying Qin,Hao Li,Chunhong Qiu,Li Li,Pengcheng Wei,Jianbo Yang. Journal of Genetics and Genomics. 2016(08)
[6]TALEN-Mediated Homologous Recombination Produces Site-Directed DNA Base Change and Herbicide-Resistant Rice[J]. Ting Li,Bo Liu,Chih Ying Chen,Bing Yang. Journal of Genetics and Genomics. 2016(05)
[7]水稻小粒基因SG4（D11等位基因）的克隆和功能研究（英文）[J]. 石珍源,饶玉春,徐杰,胡时开,方云霞,余海萍,潘江杰,刘瑞芳,任德勇,王小虎,祝阳舟,朱丽,董国军,张光恒,曾大力,郭龙彪,胡江,钱前. Science Bulletin. 2015(10)
[8]GS6, A Member of the GRAS Gene Family,Negatively Regulates Grain Size in Rice[J]. Lianjun Sun,Xiaojiao Li,Yongcai Fu,Zuofeng Zhu,Lubin Tan,Fengxia Liu,Xianyou Sun,Xuewen Sun,Chuanqing Sun. Journal of Integrative Plant Biology. 2013(10)
[9]Toward understanding genetic mechanisms of complex traits in rice[J]. Wei Hao, Hong-Xuan Lin* National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. 遗传学报. 2010(10)
本文编号：3032729
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