水稻“9311”突变体库的氯酸盐毒性筛选与分析
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摘要
【目的】用氯酸盐筛选水稻(Oryza sativa)突变体库,分析氯酸盐对水稻的毒性。【方法】使用化学诱变剂甲基磺酸乙酯(Ethyl methanesulphonate,EMS)处理水稻材料"9311"种子,构建完成了水稻"9311"突变体库。【结果】通过水培法并利用氯酸盐的特性对诱变材料进行筛选,共筛选了2 361份诱变M2代材料,且与野生型相比获得了40份对氯酸盐表现差异敏感的材料。【结论】研究结果为进一步的材料创建与相关基因克隆提供了基础,也为水稻硝酸盐吸收与转化相关分子机理的研究奠定了基础。
[Purposes]To screen the mutant library of rice(Oryza sativa)was screened by chlorate and investigate the toxicity of chlorate to rice.[Methods]The chemical mutagen,ethyl methanesulphonate(EMS)was used to treat the seeds of rice material"9311"and construct the"9311"rice mutant library.[Findings]The rice mutant library was screened by hydroponics and using the characteristics of chlorate.A total of 2361 mutant materials were screened and 40 lines showed a difference in sensitivity to chlorate compared with the wild type.In addition,the phenotypic differences of the 40 materials screened confirmed further candidate materials by further repeated experiments.[Conclusions]The results provided the basis for our further material creation and related gene cloning,and also laid the foundation for the study of molecular mechanisms related to nitrate uptake and transformation in rice.
[Purposes]To screen the mutant library of rice(Oryza sativa)was screened by chlorate and investigate the toxicity of chlorate to rice.[Methods]The chemical mutagen,ethyl methanesulphonate(EMS)was used to treat the seeds of rice material"9311"and construct the"9311"rice mutant library.[Findings]The rice mutant library was screened by hydroponics and using the characteristics of chlorate.A total of 2361 mutant materials were screened and 40 lines showed a difference in sensitivity to chlorate compared with the wild type.In addition,the phenotypic differences of the 40 materials screened confirmed further candidate materials by further repeated experiments.[Conclusions]The results provided the basis for our further material creation and related gene cloning,and also laid the foundation for the study of molecular mechanisms related to nitrate uptake and transformation in rice.
引文
[1]曲志恒.水稻根系相关性状QTL定位[D].广州:华南农业大学,2016.QU Z H.Mapping of QTL for root-related traits in rice[D].Guangzhou:South China Agricultural University,2016.
[2]马冬雪.水稻空育131苗期耐冷候选基因OsrbcS3的克隆和功能分析[D].哈尔滨:黑龙江大学,2016.MA D X.Cloning and functional analysis of a candidate cold-tolerant gene OsrbcS3 in seedling of rice Kongyu 131[D].Harbin:Heilongjiang University,2016.
[3]于仁波.日本晴/9311水稻重组自交系重要农艺性状QTl定位研究[D].长沙:湖南师范大学,2016.YU R B.Study on QTl mapping of important agronomic characters of Nipponbare/9311 rice recombinant inbred lines[D].Changsha:Hunan Normal University,2016.
[4]符德保,李燕,肖景华,等.中国水稻基因组学研究历史及现状[J].生命科学,2016(10):1113-1121.FU D B,LI Y,XIAO J H,et al.The history and current status of rice genomics research in China[J].Chinese Bulletin of Life Science,2016(10):1113-1121.
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[8]罗文泊.粗糙脉孢菌物理-化学诱变及分子验证[D].长春:吉林农业大学,2005.LUO W B.Physical-chemical mutagenesis and molecular validation of rough neurospora[D].Changchun:Jinlin Agricultural University,2005.
[9]张长正,刘进平,时涛,等.木薯地毯草黄单胞菌转化子T-DNA插入模式分析[J].热带生物学报,2016,7(2):246-252.ZHANG C Z,LIU J P,SHI T,et al.T-DNA insertion patterns for the genome of Xanthomonas axonopodis pv.Manihotis transformants[J].Journal of Troplical Biology,2016,7(2):246-252.
[10]叶俊.水稻“9311”突变体的筛选和突变体库的构建[D].杭州:浙江大学,2006.YE J.Screening of rice“9311”mutant and Construction of mutant library[D].Hangzhou:Zhejiang University,2006.
[11]杨媛媛.玉米EMS突变体库创建及opaque和CENH3基因的TILLING分析[D].成都:四川农业大学,2013.YANG Y Y.Construction of EMS mutant library and TILLING analysis of gene opaque and CENH3 in maize[D].Chengdu:Sichuan Agricultural University,2013.
[12]倪婷.拟南芥硝态氮调控突变体的筛选与鉴定[D].泰安:山东农业大学,2016.NI T.Screening and identification of nitrate regulatory mutant in Arabidopsis[D].Taian:Shandong Agricultural University,2016.
[13]MACKOWN C T.小麦硝酸盐吸收同化与氯酸盐的耐性[J].麦类作物,1997(1):28-32.MACKOWN C T.Absorption and assimilation of wheat nitrate and chloride tolerance[J].Tritical Crops,1997(1):28-32.
[14]黄旭明,陆洁梅,王惠聪.氯酸盐在龙眼生产上的运用和研究现状[J].中国南方果树,2004,33(3):29-30.HUANG X M,LU J M,WANG H C.The application and research status of chlorate in longan production[J].South China Fruits,2004,33(3):29-30.
[15]宋景荣,徐文静,赵曦,等.球孢白僵菌硝酸还原酶缺陷型突变体的筛选与鉴定[J].黑龙江农业科学,2010(5):3-6.SONG J R,XU W J,ZHAO X,et al.Effect of different plant growth retardants on conservation in vitro plantlets of potato[J].Heilongjiang Agricultural Sciences,2010(5):3-6.
[16]ODRISCOLL C.IRGSP cracks genome for rice[J].European Chemical News,2005,83(2160):28.
[17]汪仁银.优质中籼稻新品种:扬稻6号[J].农业科技通讯,2001(2):34.WANG R Y.High quality medium japonica rice variety:Yangdao 6[J].Famous and new improved varieties,2001(2):34.
[18]顾佳清,张智奇,周音,等.EMS诱导水稻中花11突变体的筛选和鉴定[J].上海农业学报,2005,21(1):7-11.GU J Q,ZHANG Z Q,ZHOU Y,et al.Screening and identification of mutants induced from rice Zhonghua 11(Oryza sativa L.subsp.Japonica)by EMS[J].Acta Agriculturae Shanghai,2005,21(1):7-11.
[19]肖娟,严欢欢,杨永清,等.不同品种水稻苗期硝态氮吸收与利用效率差异的筛选及研究[J].植物生理学报,2016,52(12):1941-1949.XIAO J,YAN H H,YANG Y Q,et al.Screening and research of different rice(Oryza sativa)varieties based on nitrate absorption and utilization in seedlings[J].Plant Physiology Journal,2016,52(12):1941-1949.
[20]DUAN D D,ZHANG H M.A single SNP in NRT1.1B,has a major impact on nitrogen use efficiency in rice[J].Science China Life Sciences,2015,58(8):827-828.
[21]FAN X,TANG Z,TAN Y,et al.Overexpression of a pH-sensitive nitrate transporter in rice increases crop yields.[J].PNAS,2016,113(26):7118-7123.
[22]郑静.一份水稻白化突变体的遗传分析及其基因定位[D].成都:四川农业大学,2008.ZHENG J.Genetic analysis and gene mapping of a rice albino mutant[D].Chengdu:Sichuan Agricultural University,2008.
[23]魏溪涓.水稻闭花授粉突变体fod的突变基因克隆与功能分析[D].金华:浙江师范大学,2015.WEI X J.Cloning and function analysis of a cleistogamy mutant in rice(Oryza sativa)[D].Jinhua:Zhejiang Normal University,2015.
[24]徐小金.稻瘟病抗性基因资源的挖掘及其育种利用[D].金华:浙江师范大学,2016.XU X J.Exploitation and breeding utilization of blast resistance gene resources in rice[D].Jinhua:Zhejiang Normal University,2016.
[25]王雪颖,米丽娜,杨铮,等.基因功能的研究方法[J].中国疗养医学,2017,26(2):131-133.WANG X Y,MI L N,YANG Z,et al.Gene function research methods[J].Chinese Journal of Convalescent Medicine,2017,26(2):131-133.
[26]刘峰.烟草激活标签突变体库的构建与分析[D].北京:中国农业科学院,2014.LIU F.Construction and analysis of tobacco activation label mutant library[D].Beijing:Chinese Academy of Agricultural Sciences,2014.
[27]彭慧.水稻不育系Y58S辐射诱变典型突变体农艺性状分析[D].南昌:江西农业大学,2014.Typical analysis of agronomic traits mutant rice sterile line Y58Sradiation mutagenesis[D].Nanchang:Jiangxi Agricultural University,2014.
[28]PRIETO R,FERNáNDEZE.Toxicity of and mutagenesis by chlorate are independent of nitrate reductase activity in Chlamydomonas reinhardtii[J].Molecular&General Genetics,1993,237(3):429-438.
[29]林振武,陈敬祥.硝酸还原酶作为作物育种的生化指标的研究[J].河北农业大学学报,1987,10(3):104-110.LIN Z W,CHEN J X.Study on nitrate reductase activity as a biochemical criterion in plant breeding[J].Journal of Hebei Agricultural University,1987,10(3):104-110.
[2]马冬雪.水稻空育131苗期耐冷候选基因OsrbcS3的克隆和功能分析[D].哈尔滨:黑龙江大学,2016.MA D X.Cloning and functional analysis of a candidate cold-tolerant gene OsrbcS3 in seedling of rice Kongyu 131[D].Harbin:Heilongjiang University,2016.
[3]于仁波.日本晴/9311水稻重组自交系重要农艺性状QTl定位研究[D].长沙:湖南师范大学,2016.YU R B.Study on QTl mapping of important agronomic characters of Nipponbare/9311 rice recombinant inbred lines[D].Changsha:Hunan Normal University,2016.
[4]符德保,李燕,肖景华,等.中国水稻基因组学研究历史及现状[J].生命科学,2016(10):1113-1121.FU D B,LI Y,XIAO J H,et al.The history and current status of rice genomics research in China[J].Chinese Bulletin of Life Science,2016(10):1113-1121.
[5]高志勇,谢恒星,王志平,等.植物突变体库的作用及构建研究进展[J].作物杂志,2016(6):16-19.GAO Z Y,XIE H X,WANG Z P,et al.Progress on the function and construction of plant mutant library[J].Crops,2016(6):16-19.
[6]耿金鹏.电离辐射对玉米种质改良的应用研究[D].天津:河北工业大学,2014.GENG J P.Study on application of ionizing radiation to maize germplasm improvement[D].Tianjin:Hebei University of Technology,2014.
[7]杨瑰丽,陈莹,郭涛,等.碳离子束辐照水稻诱变效应及突变体的筛选[J].华南农业大学学报,2018,39(2):29-33.YANG G L,CHEN Y,GUO T,et al.Mutagenic effects of carbon ion beam irradiation on rice and screening for induced mutants[J].Journal of South China Agricultural University,2018,39(2):29-33.
[8]罗文泊.粗糙脉孢菌物理-化学诱变及分子验证[D].长春:吉林农业大学,2005.LUO W B.Physical-chemical mutagenesis and molecular validation of rough neurospora[D].Changchun:Jinlin Agricultural University,2005.
[9]张长正,刘进平,时涛,等.木薯地毯草黄单胞菌转化子T-DNA插入模式分析[J].热带生物学报,2016,7(2):246-252.ZHANG C Z,LIU J P,SHI T,et al.T-DNA insertion patterns for the genome of Xanthomonas axonopodis pv.Manihotis transformants[J].Journal of Troplical Biology,2016,7(2):246-252.
[10]叶俊.水稻“9311”突变体的筛选和突变体库的构建[D].杭州:浙江大学,2006.YE J.Screening of rice“9311”mutant and Construction of mutant library[D].Hangzhou:Zhejiang University,2006.
[11]杨媛媛.玉米EMS突变体库创建及opaque和CENH3基因的TILLING分析[D].成都:四川农业大学,2013.YANG Y Y.Construction of EMS mutant library and TILLING analysis of gene opaque and CENH3 in maize[D].Chengdu:Sichuan Agricultural University,2013.
[12]倪婷.拟南芥硝态氮调控突变体的筛选与鉴定[D].泰安:山东农业大学,2016.NI T.Screening and identification of nitrate regulatory mutant in Arabidopsis[D].Taian:Shandong Agricultural University,2016.
[13]MACKOWN C T.小麦硝酸盐吸收同化与氯酸盐的耐性[J].麦类作物,1997(1):28-32.MACKOWN C T.Absorption and assimilation of wheat nitrate and chloride tolerance[J].Tritical Crops,1997(1):28-32.
[14]黄旭明,陆洁梅,王惠聪.氯酸盐在龙眼生产上的运用和研究现状[J].中国南方果树,2004,33(3):29-30.HUANG X M,LU J M,WANG H C.The application and research status of chlorate in longan production[J].South China Fruits,2004,33(3):29-30.
[15]宋景荣,徐文静,赵曦,等.球孢白僵菌硝酸还原酶缺陷型突变体的筛选与鉴定[J].黑龙江农业科学,2010(5):3-6.SONG J R,XU W J,ZHAO X,et al.Effect of different plant growth retardants on conservation in vitro plantlets of potato[J].Heilongjiang Agricultural Sciences,2010(5):3-6.
[16]ODRISCOLL C.IRGSP cracks genome for rice[J].European Chemical News,2005,83(2160):28.
[17]汪仁银.优质中籼稻新品种:扬稻6号[J].农业科技通讯,2001(2):34.WANG R Y.High quality medium japonica rice variety:Yangdao 6[J].Famous and new improved varieties,2001(2):34.
[18]顾佳清,张智奇,周音,等.EMS诱导水稻中花11突变体的筛选和鉴定[J].上海农业学报,2005,21(1):7-11.GU J Q,ZHANG Z Q,ZHOU Y,et al.Screening and identification of mutants induced from rice Zhonghua 11(Oryza sativa L.subsp.Japonica)by EMS[J].Acta Agriculturae Shanghai,2005,21(1):7-11.
[19]肖娟,严欢欢,杨永清,等.不同品种水稻苗期硝态氮吸收与利用效率差异的筛选及研究[J].植物生理学报,2016,52(12):1941-1949.XIAO J,YAN H H,YANG Y Q,et al.Screening and research of different rice(Oryza sativa)varieties based on nitrate absorption and utilization in seedlings[J].Plant Physiology Journal,2016,52(12):1941-1949.
[20]DUAN D D,ZHANG H M.A single SNP in NRT1.1B,has a major impact on nitrogen use efficiency in rice[J].Science China Life Sciences,2015,58(8):827-828.
[21]FAN X,TANG Z,TAN Y,et al.Overexpression of a pH-sensitive nitrate transporter in rice increases crop yields.[J].PNAS,2016,113(26):7118-7123.
[22]郑静.一份水稻白化突变体的遗传分析及其基因定位[D].成都:四川农业大学,2008.ZHENG J.Genetic analysis and gene mapping of a rice albino mutant[D].Chengdu:Sichuan Agricultural University,2008.
[23]魏溪涓.水稻闭花授粉突变体fod的突变基因克隆与功能分析[D].金华:浙江师范大学,2015.WEI X J.Cloning and function analysis of a cleistogamy mutant in rice(Oryza sativa)[D].Jinhua:Zhejiang Normal University,2015.
[24]徐小金.稻瘟病抗性基因资源的挖掘及其育种利用[D].金华:浙江师范大学,2016.XU X J.Exploitation and breeding utilization of blast resistance gene resources in rice[D].Jinhua:Zhejiang Normal University,2016.
[25]王雪颖,米丽娜,杨铮,等.基因功能的研究方法[J].中国疗养医学,2017,26(2):131-133.WANG X Y,MI L N,YANG Z,et al.Gene function research methods[J].Chinese Journal of Convalescent Medicine,2017,26(2):131-133.
[26]刘峰.烟草激活标签突变体库的构建与分析[D].北京:中国农业科学院,2014.LIU F.Construction and analysis of tobacco activation label mutant library[D].Beijing:Chinese Academy of Agricultural Sciences,2014.
[27]彭慧.水稻不育系Y58S辐射诱变典型突变体农艺性状分析[D].南昌:江西农业大学,2014.Typical analysis of agronomic traits mutant rice sterile line Y58Sradiation mutagenesis[D].Nanchang:Jiangxi Agricultural University,2014.
[28]PRIETO R,FERNáNDEZE.Toxicity of and mutagenesis by chlorate are independent of nitrate reductase activity in Chlamydomonas reinhardtii[J].Molecular&General Genetics,1993,237(3):429-438.
[29]林振武,陈敬祥.硝酸还原酶作为作物育种的生化指标的研究[J].河北农业大学学报,1987,10(3):104-110.LIN Z W,CHEN J X.Study on nitrate reductase activity as a biochemical criterion in plant breeding[J].Journal of Hebei Agricultural University,1987,10(3):104-110.