烟草耐低温胁迫早花突变体材料的筛选
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摘要
烟草(Nicotiana tabacum L.)是重要的经济作物,能够极大地增加偏远山区农民的收入,同时为国家创造大量的税收。但是烟叶种植生产过程中,容易受到低温胁迫的影响,从而会使烟草发生提早开花现象,而烟草提早开花会极大地影响烟草的产量与质量,给烟农造成重大的经济损失。目前主要通过农艺措施来减少烟草提早开花造成的损失,但这需要耗费大量的人力、物力、财力。因此培育耐低温胁迫早花的烟草品种是解决该问题的最佳方案。与以往在分子生物学层面解析基因功能不同,本研究通过EMS突变体筛选获得9份长势较好且对低温胁迫不敏感的烟草材料,该方法获得的烟草材料不含有转基因元件,因此可以直接作为育种材料应用与烟草抗早花品种选育。
Tobacco(Nicotiana tabacum L.) is an economic crop of importance, which increases the incomes of farmers in remote mountainous areas and also creates a lot of tax revenues for the country. However, tobacco is susceptible to low temperature stress during the process of planting, which results in early flowering. The early flowering has a considerable effect on the tobacco yield and quality and brings heavy losses of economy to farmers. At present, agronomic measures are mainly used to reduce the losses from tobacco early flowering, but it takes a lot of manpower, material and financial resources. Therefore, the best method to resolve this problem is to cultivate new tobacco cultivars that can be resistant to low temperature stress and early flowering. Different from the previous studies on gene function analysis at molecular biology level, this study screened the EMS mutants and obtained nine tobacco materials that grew well and were insensitive to low temperature stress. The tobacco materials obtained by this method did not contain any transgenic elements, which therefore could directly be used as materials for tobacco breeding of cultivars that are resistant to early flowering.
Tobacco(Nicotiana tabacum L.) is an economic crop of importance, which increases the incomes of farmers in remote mountainous areas and also creates a lot of tax revenues for the country. However, tobacco is susceptible to low temperature stress during the process of planting, which results in early flowering. The early flowering has a considerable effect on the tobacco yield and quality and brings heavy losses of economy to farmers. At present, agronomic measures are mainly used to reduce the losses from tobacco early flowering, but it takes a lot of manpower, material and financial resources. Therefore, the best method to resolve this problem is to cultivate new tobacco cultivars that can be resistant to low temperature stress and early flowering. Different from the previous studies on gene function analysis at molecular biology level, this study screened the EMS mutants and obtained nine tobacco materials that grew well and were insensitive to low temperature stress. The tobacco materials obtained by this method did not contain any transgenic elements, which therefore could directly be used as materials for tobacco breeding of cultivars that are resistant to early flowering.
引文
An X.L.,Cai Y.L.,Wang J.G.,Wang G.Q.,and Sun H.Y.,2003,Mutagenic effects of EMS on maize inbred line,Yumi Kexue(Journal of Maize Sciences),11(3):74-75(安学丽,蔡一林,王久光,王国强,孙海燕,2003,甲基磺酸乙酯(EMS)对玉米自交系诱变效应的研究,玉米科学,11(3):74-75)
Blümel M.,Dally N.,and Jung C.,2015,Flowering time regulation in crops-what did we learn from Arabidopsis,Curr.Opin.Biotechnol.,32:121-129
Cui X.,Liang Y.,and Wang L.H.,2013,Effect of different concentrations of chemical mutagen EMS on tomato seed germination,Beifang Yuanyi(Northern Horticulture),(14):28-30(崔霞,梁燕,王玲慧,2013,不同浓度化学诱变剂甲基磺酸乙酯对番茄种子发芽的影响,北方园艺,(14):28-30)
Chang A.X.,Guo L.J.Liu D.,Luo C.G.,Wang L.S.,Feng Q.F.,and Wang L.,2015,Tobacco rapid backcross improvement mediated by Arabidopsis flowering gene FT,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(13):2508-2517(常爱霞,郭利杰,刘旦,罗成刚,王林嵩,冯全福,王兰,2015,早花基因(FT)介导的烟草快速回交改良研究,中国农业科学,48(13):2508-2517)
Guo L.J.,2015,A method for improving tobacco rapid backcross mediated by Arabidopsis flowering gene FT,Thesis for M.S.,Henan Normal University,Supervisor:Wang L.,pp.1-65(郭利杰,2015,早花基因(FT)介导的烟草快速回交育种技术研究,硕士学位论文,河南师范大学,导师:王兰,pp.1-65)
Han Y.Y.,Ma Y.Q.,Li D.Z.,and Xu Z.Q.,2015,The influences of Arabidopsis Pri-miR156a on flowering time of Nicotiana tabacum,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(2):350-356(韩瑶瑶,马燕勤,李典珍,徐子勤,2015,拟南芥Pri-miR156a基因对烟草开花时间的影响,基因组学与应用生物学,34(2):350-356)
Jin L.,Jin Y.,Zhou J.H.,Yang Y.H.,Yang H.Q.,Huang C.J.,Duan Y.Q.,and Chen H.W.,2007,Effects of low temperature in seeding stage on floral differentiation and development of flue-cured tobacco,Zhongguo Yancao Kexue(Chi nese Tobacco Science),28(6):1-5(金磊,晋艳,周冀衡,杨宇虹,杨虹琦,黄成江,段玉琪,陈恒旺,2007,苗期低温对烤烟花芽分化及发育进程的影响,中国烟草科学,28(6):1-5)
Li D.,Lewis R.S.,Jack A.M.,Dewey R.E.,Bowen S.W.,and Miller R.D.,2012,Development of CAPS and dCAPSmarkers for CYP82E4,CYP82E5v2 and CYP82E10 gene mutants reducing nicotine to nornicotine conversion in tobacco,Molecular Breeding,29(3):589-599
Li H.J.,Pu X.B.,Zhang J.F.,Huang C.,Song K.X.,Jiang J.,He P.,Li P.,and Jiang L.C.,2012,Agronomic traits and molecular detection of Brassica napus L.generation induced by EMS,Henongxue Bao(Journal of Nuclear Agricultural Sciences),26(2):245-249(李浩杰,蒲晓斌,张锦芳,黄驰,宋克先,蒋俊,何平,李平,蒋梁材,2012,甘蓝型油菜EMS诱变后代农艺性状观察及分子检测,核农学报,26(2):245-249)
Li Y.Y.,2011,Research on cold induced early flowering and homologous cloning of MADS-box genes in tobacco,Thesis for M.S.,Chinese Academy of Agricultural Sciences,Supervisor:Wang Y.Y.,pp.1-59(李元元,2011,低温诱导烟草早花研究与烟草MADS-box基因的同源克隆,硕士学位论文,中国农业科学院,导师:王元英,pp.1-59)
Liu C.W.,Guo S.P.,and He Y.Y.,2012,Effect of low temperature stress on early flowering of different flue-cured tobacco varieties with different seedling-age,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),24(3):132-134(刘成伟,郭仕平,何余勇,2012,低温胁迫对不同品种和苗龄烤烟早花发生的影响,江西农业学报,24(3):132-134)
Liu M.T.,2014,Construction and characterization of EMS mutant library of Jingu 21,Thesis for M.S.,Shanxi Agricultural University,Supervisor:Li H.Y.,pp.1-42(刘美桃,2014,晋谷21 EMS突变体库的构建与特征分析,硕士学位论文,山西农业大学,导师:李红英,pp.1-42)
Pakdeechanuan P.,Teoh S.,Shoji T.,and Hashimoto T.,2012,Non-functionalization of two CYP82E nicotine N-demethylase genes abolishes nornicotine formation in Nicotiana langsdorffii,Plant Cell Physiol.,53(12):2038-2046
Wan X.Q.,Yan P.Q.,Li L.J.,Kang H.,and Guo Z.K.,2005,Study on the qualitative determination of genetically modified tobacco,Zhongguo Yancao Xuebao(Acta Tabacaria Sinica),11(2):24-27(万秀清,颜培强,李丽杰,康宏,郭兆奎,2005,转基因烟草定性检测方法的研究,中国烟草学报,11(2):24-27)
Wang F.,Xu B.,Yang Z.L.,Lin Y.H.,He G.H.,Chen S.,Qing M.J.,and Sang X.C.,2011,Identification and analysis of EMS-induced dwarf mutants in rice,Henongxue Bao(Journal of Nuclear Agricultural Sciences),25(2):197-201(王峰,徐飚,杨正林,凌英华,何光华,陈胜,卿明敬,桑贤春,2011,EMS诱变水稻矮生资源的鉴定评价,核农学报,25(2):197-201)
Wang X.Y.,2009,Analysis of factors correlated with earlyflowering induced by low temperature in tobacco,Thesis for M.S.,Henan Agricultural University,Supervisor:Liu W.Q.,pp.1-33(王小彦,2009,低温诱导烟草早花相关因子的分析,硕士学位论文,河南农业大学,导师:刘卫群,pp.1-33)
Yang Q.,2011,Study on potato salt-tolerant variants of EMS inducing and selection,Thesis for M.S.,Gansu Agricultural University,Supervisor:Wang D.,and Zhang F.,pp.1-43(杨乾,2011,马铃薯耐盐突变体的EMS诱变和筛选,硕士学位论文,甘肃农业大学,导师:王蒂,张峰,pp.1-43)
Yu H.Q.,Yin J.,Huang C.J.,Li Y.P.,Shu Q.Y.,and Liu Y.,2016,Study on the suitable mutation dose ofγrays for tobacco seeds and pollen,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(10):2749-2755(于海芹,印洁,黄昌军,李永平,舒庆尧,刘勇,2016,烟草种子和花粉的γ射线适宜诱变剂量探究,分子植物育种,14(10):2749-2755)
Blümel M.,Dally N.,and Jung C.,2015,Flowering time regulation in crops-what did we learn from Arabidopsis,Curr.Opin.Biotechnol.,32:121-129
Cui X.,Liang Y.,and Wang L.H.,2013,Effect of different concentrations of chemical mutagen EMS on tomato seed germination,Beifang Yuanyi(Northern Horticulture),(14):28-30(崔霞,梁燕,王玲慧,2013,不同浓度化学诱变剂甲基磺酸乙酯对番茄种子发芽的影响,北方园艺,(14):28-30)
Chang A.X.,Guo L.J.Liu D.,Luo C.G.,Wang L.S.,Feng Q.F.,and Wang L.,2015,Tobacco rapid backcross improvement mediated by Arabidopsis flowering gene FT,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(13):2508-2517(常爱霞,郭利杰,刘旦,罗成刚,王林嵩,冯全福,王兰,2015,早花基因(FT)介导的烟草快速回交改良研究,中国农业科学,48(13):2508-2517)
Guo L.J.,2015,A method for improving tobacco rapid backcross mediated by Arabidopsis flowering gene FT,Thesis for M.S.,Henan Normal University,Supervisor:Wang L.,pp.1-65(郭利杰,2015,早花基因(FT)介导的烟草快速回交育种技术研究,硕士学位论文,河南师范大学,导师:王兰,pp.1-65)
Han Y.Y.,Ma Y.Q.,Li D.Z.,and Xu Z.Q.,2015,The influences of Arabidopsis Pri-miR156a on flowering time of Nicotiana tabacum,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(2):350-356(韩瑶瑶,马燕勤,李典珍,徐子勤,2015,拟南芥Pri-miR156a基因对烟草开花时间的影响,基因组学与应用生物学,34(2):350-356)
Jin L.,Jin Y.,Zhou J.H.,Yang Y.H.,Yang H.Q.,Huang C.J.,Duan Y.Q.,and Chen H.W.,2007,Effects of low temperature in seeding stage on floral differentiation and development of flue-cured tobacco,Zhongguo Yancao Kexue(Chi nese Tobacco Science),28(6):1-5(金磊,晋艳,周冀衡,杨宇虹,杨虹琦,黄成江,段玉琪,陈恒旺,2007,苗期低温对烤烟花芽分化及发育进程的影响,中国烟草科学,28(6):1-5)
Li D.,Lewis R.S.,Jack A.M.,Dewey R.E.,Bowen S.W.,and Miller R.D.,2012,Development of CAPS and dCAPSmarkers for CYP82E4,CYP82E5v2 and CYP82E10 gene mutants reducing nicotine to nornicotine conversion in tobacco,Molecular Breeding,29(3):589-599
Li H.J.,Pu X.B.,Zhang J.F.,Huang C.,Song K.X.,Jiang J.,He P.,Li P.,and Jiang L.C.,2012,Agronomic traits and molecular detection of Brassica napus L.generation induced by EMS,Henongxue Bao(Journal of Nuclear Agricultural Sciences),26(2):245-249(李浩杰,蒲晓斌,张锦芳,黄驰,宋克先,蒋俊,何平,李平,蒋梁材,2012,甘蓝型油菜EMS诱变后代农艺性状观察及分子检测,核农学报,26(2):245-249)
Li Y.Y.,2011,Research on cold induced early flowering and homologous cloning of MADS-box genes in tobacco,Thesis for M.S.,Chinese Academy of Agricultural Sciences,Supervisor:Wang Y.Y.,pp.1-59(李元元,2011,低温诱导烟草早花研究与烟草MADS-box基因的同源克隆,硕士学位论文,中国农业科学院,导师:王元英,pp.1-59)
Liu C.W.,Guo S.P.,and He Y.Y.,2012,Effect of low temperature stress on early flowering of different flue-cured tobacco varieties with different seedling-age,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),24(3):132-134(刘成伟,郭仕平,何余勇,2012,低温胁迫对不同品种和苗龄烤烟早花发生的影响,江西农业学报,24(3):132-134)
Liu M.T.,2014,Construction and characterization of EMS mutant library of Jingu 21,Thesis for M.S.,Shanxi Agricultural University,Supervisor:Li H.Y.,pp.1-42(刘美桃,2014,晋谷21 EMS突变体库的构建与特征分析,硕士学位论文,山西农业大学,导师:李红英,pp.1-42)
Pakdeechanuan P.,Teoh S.,Shoji T.,and Hashimoto T.,2012,Non-functionalization of two CYP82E nicotine N-demethylase genes abolishes nornicotine formation in Nicotiana langsdorffii,Plant Cell Physiol.,53(12):2038-2046
Wan X.Q.,Yan P.Q.,Li L.J.,Kang H.,and Guo Z.K.,2005,Study on the qualitative determination of genetically modified tobacco,Zhongguo Yancao Xuebao(Acta Tabacaria Sinica),11(2):24-27(万秀清,颜培强,李丽杰,康宏,郭兆奎,2005,转基因烟草定性检测方法的研究,中国烟草学报,11(2):24-27)
Wang F.,Xu B.,Yang Z.L.,Lin Y.H.,He G.H.,Chen S.,Qing M.J.,and Sang X.C.,2011,Identification and analysis of EMS-induced dwarf mutants in rice,Henongxue Bao(Journal of Nuclear Agricultural Sciences),25(2):197-201(王峰,徐飚,杨正林,凌英华,何光华,陈胜,卿明敬,桑贤春,2011,EMS诱变水稻矮生资源的鉴定评价,核农学报,25(2):197-201)
Wang X.Y.,2009,Analysis of factors correlated with earlyflowering induced by low temperature in tobacco,Thesis for M.S.,Henan Agricultural University,Supervisor:Liu W.Q.,pp.1-33(王小彦,2009,低温诱导烟草早花相关因子的分析,硕士学位论文,河南农业大学,导师:刘卫群,pp.1-33)
Yang Q.,2011,Study on potato salt-tolerant variants of EMS inducing and selection,Thesis for M.S.,Gansu Agricultural University,Supervisor:Wang D.,and Zhang F.,pp.1-43(杨乾,2011,马铃薯耐盐突变体的EMS诱变和筛选,硕士学位论文,甘肃农业大学,导师:王蒂,张峰,pp.1-43)
Yu H.Q.,Yin J.,Huang C.J.,Li Y.P.,Shu Q.Y.,and Liu Y.,2016,Study on the suitable mutation dose ofγrays for tobacco seeds and pollen,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(10):2749-2755(于海芹,印洁,黄昌军,李永平,舒庆尧,刘勇,2016,烟草种子和花粉的γ射线适宜诱变剂量探究,分子植物育种,14(10):2749-2755)