苦瓜种子EMS诱变条件分析
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摘要
以江门大顶苦瓜自交系干种子为材料,设置EMS溶液不同浓度(0、0.3%、0.6%、0.9%、1.2%、1.5%)和不同诱变时间(2、4、6 h)处理组合,通过统计苦瓜种子发芽势和发芽率指标以及M0植株表型观察来确定苦瓜种子EMS诱变的适宜条件。结果表明,EMS溶液浓度、诱变时间以及两者的互作均对苦瓜种子发芽势和发芽率具有显著的影响;根据半致死条件原则,初步认为清水浸种8 h,然后利用1.5%或1.2%EMS溶液处理2 h是苦瓜种子EMS诱变的适宜条件;将1.2%或1.5%EMS溶液处理2 h所获得的共129株M0苦瓜幼苗进行田间定植和表型观察,发现苦瓜植株表型突变频率分别为5.12%和7.85%,且表型变异多发生于叶片和果实,表明所筛选的诱变条件可以用于苦瓜种子EMS诱变研究。试验结果为下一步利用EMS诱变技术开展苦瓜突变体库构建以及创制苦瓜新材料奠定了基础。
Taking dry seeds of bitter gourd inbred line ‘Jiangmendading' as materials,setting up combinations of treatments with different EMS concentrations(0,0.3%,0.6%,0.9%,1.2% and 1.5%) and different mutagenesis times(2,4,6 hours),this study determined the suitable conditions for EMS mutagenesis of bitter gourd seed through statistical data of seed germination vigor and rate and observation on M0 plant phenotypes. Results showed that EMS concentration,mutagenesis time and their interaction had significant impact on germination vigor and rate of bitter gourd seed.According to the principles of semi-lethal condition,8 hours of soaking with water followed by 2 hours with 1.5% or 1.2% EMS treatment were the suitable conditions for EMS mutagenesis of bitter gourd seed.Field planting and phenotype observation of 129 M0 plants after 2 hours with 1.2% or 1.5% EMS treatment revealed that the mutation frequency of bitter gourd plant phenotypes were 5.12% and 7.85%,respectively,and the phenotypic variations often occurred in leaf and fruit,indicating that the selected conditions were suitable for studying EMS mutagenesis of bitter gourd seed.These results have laid a foundation for further using EMS mutagenesis technology to construct mutant library and create new bitter gourd materials.
Taking dry seeds of bitter gourd inbred line ‘Jiangmendading' as materials,setting up combinations of treatments with different EMS concentrations(0,0.3%,0.6%,0.9%,1.2% and 1.5%) and different mutagenesis times(2,4,6 hours),this study determined the suitable conditions for EMS mutagenesis of bitter gourd seed through statistical data of seed germination vigor and rate and observation on M0 plant phenotypes. Results showed that EMS concentration,mutagenesis time and their interaction had significant impact on germination vigor and rate of bitter gourd seed.According to the principles of semi-lethal condition,8 hours of soaking with water followed by 2 hours with 1.5% or 1.2% EMS treatment were the suitable conditions for EMS mutagenesis of bitter gourd seed.Field planting and phenotype observation of 129 M0 plants after 2 hours with 1.2% or 1.5% EMS treatment revealed that the mutation frequency of bitter gourd plant phenotypes were 5.12% and 7.85%,respectively,and the phenotypic variations often occurred in leaf and fruit,indicating that the selected conditions were suitable for studying EMS mutagenesis of bitter gourd seed.These results have laid a foundation for further using EMS mutagenesis technology to construct mutant library and create new bitter gourd materials.
引文
陈小凤,黄如葵,黄玉辉,罗海玲,陈振东.2011.苦瓜育种及相关基础研究进展.南方农业学报,42(3):246-249.
程欣,任德勇,马娇,朱晓燕,桑贤春,凌英华,赵芳明,何光华.2013.水稻叶片灰白转黄突变体pyr1的鉴定与基因定位.作物学报,39(6):992-998.
崔竣杰,李波,程蛟文,胡开林.2015.苦瓜苦味物质及其生物合成研究进展.园艺学报,42(9):1707-1718.
崔霞,梁燕,李翠,秦蕾,李云洲.2013.化学诱变及其在蔬菜育种中的应用.西北农林科技大学学报:自然科学版,41(3):205-212.
黄冬福,付文婷,韩世玉,何建文.2015.农作物EMS诱变研究进展.北方园艺,(24):188-194.
黄如葵,黄熊娟,梁家作,黄玉辉,琚茜茜,陈小凤,冯诚诚,陈振东,刘杏连.2015.利用辐射诱变及分子标记辅助筛选进行苦瓜种质创新.南方农业学报,46(3):376-380.
黄亚杰,运广荣,李梅,李长增,肖瑜,张斌.2012.苦瓜遗传育种研究进展.中国蔬菜,(8):11-19.
康保珊,李俊香,刘丽锋,古勤生.2017.甲基磺酸乙酯诱变瓠瓜种子的条件筛选.中国瓜菜,30(2):7-10.
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.
刘忠松,罗赫荣.2010.现代植物育种学.北京:科学出版社.
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.
王晶,娄群峰,魏庆镇,李子昂,付文苑,陈劲枫.2015.长春密刺黄瓜突变体库的构建和部分性状分析.核农学报,29(8):1479-1486.
王平荣,王兵,孙小秋,孙昌辉,万春美,马晓智,邓晓建.2013.水稻白化转绿基因gra75的精细定位和生理特性分析.中国农业科学,46(2):225-232.
王学征,朱娜娜,高清宇,刘璐,张雅楠,栾非时.2015.EMS诱变西瓜种子条件分析.东北农业大学学报,46(7):35-39.
谢圣男,王宏光,杨振,刘春燕,蒋洪蔚,辛大伟,胡国华,陈庆山.2013.大豆绥农14突变体库构建及株高性状分析.核农学报,27(3):307-313.
薛存宝.2016.黄瓜EMS诱变突变体库及TILLING平台的初步构建[硕士论文].哈尔滨:东北农业大学.
张兵.2012.EMS诱导黄瓜突变体的初步研究[硕士论文].泰安:山东农业大学,
赵会芳.2005.辣椒诱变及其诱变机理的研究[硕士论文].杨凌:西北农林科技大学.
周书栋,杨博智,欧立军,刘周斌,吕俊恒,马艳青,邹学校.2017.辣椒突变体库的构建及突变群体表型变异分析.湖南农业大学学报:自然科学版,43(1):31-36.
Abe A,Kosugi S,Yoshida K,Natsume S,Takagi H,Kanzaki H,Matsumura H,Yoshida K,Mitsuoka C,Tamiru M,Innan H,Cano L,Kamoun S,Terauchi R.2012.Genome sequencing reveals agronomically important loci in rice using MutMap.Natutre Biotechnology,30(2):174-178.
Cui J,Cheng J,Nong D,Peng J,Hu Y,He W,Zhou Q,Dhillon N,Hu K.2017.Genome-wide analysis of simple sequence repeats in bitter gourd(Momordica charantia).Frontiers in Plant Science,8:1103.
Cui J,Luo S,Niu Y,Huang R,Wen Q,Su J,Miao N,He W,Dong Z,Cheng J,Hu K.2018.A RAD-based genetic map for anchoring scaffold sequences and identifying QTLs in bitter gourd(Momordica charantia).Frontiers in Plant Science,9:477.
Dhillon N,Sanguansil S,Schafleitner R,Wang Y,Mccreight J.2016.Diversity among a wide Asian collection of bitter gourd landraces and their genetic relationships with commercial hybrid cultivars.Journal of the American Society for Horticultural Science,141(5):475-484.
Fang E F,Ng T B.2013.The bitter fruit with sweet health benefits:a comprehensive synopsis of recent research progress on medicinal properties of Momordica charantia//Fang E F,Ng T B.Antitumor Potential and Other Emerging Medicinal Properties of Natural Compounds.Dordrecht:Springer:315-334.
Kim Y,Schumaker K S,Zhu J K.2006.EMS mutagenesis of Arabidopsis.Methods Molecular Biology,323:101-103.
Lu X,Liu J,Ren W,Yang Q,Chai Z,Chen R,Wang L,Zhao J,Lang Z,Wang H.2018.Gene-indexed mutations in maize(Zea mays).Molecular Plant,11(3):496-504.
Schaefer H,Renner S.2010.A three-genome phylogeny of Momordica(Cucurbitaceae)suggests seven returns from dioecy to monoecy and recent long-distance dispersal to Asia.Molecular Phylogenetics and Evolution,54(2):553-560.
Urasaki N,Takagi H,Natsume S,Uemura A,Taniai N,Miyagi N,Fukushima M,Suzuki S,Tarora K,Tamaki M,Sakamoto M,Terauchi R,Matsumura H.2017.Draft genome sequence of bitter gourd(Momordica charantia),a vegetable and medicinal plant in tropical and subtropical regions.DNA Research,24(1):51-58.
van Wyk B.2015.A review of commercially important African medicinal plants.Journal of Ethnopharmacology,176:118-134.
Yang G,Li S,Wang S.2016.Research progress in enzyme activity and pharmacological effects of ribosome-inactivity protein in bitter melon.Toxin Reviews,35:128-132.
程欣,任德勇,马娇,朱晓燕,桑贤春,凌英华,赵芳明,何光华.2013.水稻叶片灰白转黄突变体pyr1的鉴定与基因定位.作物学报,39(6):992-998.
崔竣杰,李波,程蛟文,胡开林.2015.苦瓜苦味物质及其生物合成研究进展.园艺学报,42(9):1707-1718.
崔霞,梁燕,李翠,秦蕾,李云洲.2013.化学诱变及其在蔬菜育种中的应用.西北农林科技大学学报:自然科学版,41(3):205-212.
黄冬福,付文婷,韩世玉,何建文.2015.农作物EMS诱变研究进展.北方园艺,(24):188-194.
黄如葵,黄熊娟,梁家作,黄玉辉,琚茜茜,陈小凤,冯诚诚,陈振东,刘杏连.2015.利用辐射诱变及分子标记辅助筛选进行苦瓜种质创新.南方农业学报,46(3):376-380.
黄亚杰,运广荣,李梅,李长增,肖瑜,张斌.2012.苦瓜遗传育种研究进展.中国蔬菜,(8):11-19.
康保珊,李俊香,刘丽锋,古勤生.2017.甲基磺酸乙酯诱变瓠瓜种子的条件筛选.中国瓜菜,30(2):7-10.
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.
刘忠松,罗赫荣.2010.现代植物育种学.北京:科学出版社.
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.
王晶,娄群峰,魏庆镇,李子昂,付文苑,陈劲枫.2015.长春密刺黄瓜突变体库的构建和部分性状分析.核农学报,29(8):1479-1486.
王平荣,王兵,孙小秋,孙昌辉,万春美,马晓智,邓晓建.2013.水稻白化转绿基因gra75的精细定位和生理特性分析.中国农业科学,46(2):225-232.
王学征,朱娜娜,高清宇,刘璐,张雅楠,栾非时.2015.EMS诱变西瓜种子条件分析.东北农业大学学报,46(7):35-39.
谢圣男,王宏光,杨振,刘春燕,蒋洪蔚,辛大伟,胡国华,陈庆山.2013.大豆绥农14突变体库构建及株高性状分析.核农学报,27(3):307-313.
薛存宝.2016.黄瓜EMS诱变突变体库及TILLING平台的初步构建[硕士论文].哈尔滨:东北农业大学.
张兵.2012.EMS诱导黄瓜突变体的初步研究[硕士论文].泰安:山东农业大学,
赵会芳.2005.辣椒诱变及其诱变机理的研究[硕士论文].杨凌:西北农林科技大学.
周书栋,杨博智,欧立军,刘周斌,吕俊恒,马艳青,邹学校.2017.辣椒突变体库的构建及突变群体表型变异分析.湖南农业大学学报:自然科学版,43(1):31-36.
Abe A,Kosugi S,Yoshida K,Natsume S,Takagi H,Kanzaki H,Matsumura H,Yoshida K,Mitsuoka C,Tamiru M,Innan H,Cano L,Kamoun S,Terauchi R.2012.Genome sequencing reveals agronomically important loci in rice using MutMap.Natutre Biotechnology,30(2):174-178.
Cui J,Cheng J,Nong D,Peng J,Hu Y,He W,Zhou Q,Dhillon N,Hu K.2017.Genome-wide analysis of simple sequence repeats in bitter gourd(Momordica charantia).Frontiers in Plant Science,8:1103.
Cui J,Luo S,Niu Y,Huang R,Wen Q,Su J,Miao N,He W,Dong Z,Cheng J,Hu K.2018.A RAD-based genetic map for anchoring scaffold sequences and identifying QTLs in bitter gourd(Momordica charantia).Frontiers in Plant Science,9:477.
Dhillon N,Sanguansil S,Schafleitner R,Wang Y,Mccreight J.2016.Diversity among a wide Asian collection of bitter gourd landraces and their genetic relationships with commercial hybrid cultivars.Journal of the American Society for Horticultural Science,141(5):475-484.
Fang E F,Ng T B.2013.The bitter fruit with sweet health benefits:a comprehensive synopsis of recent research progress on medicinal properties of Momordica charantia//Fang E F,Ng T B.Antitumor Potential and Other Emerging Medicinal Properties of Natural Compounds.Dordrecht:Springer:315-334.
Kim Y,Schumaker K S,Zhu J K.2006.EMS mutagenesis of Arabidopsis.Methods Molecular Biology,323:101-103.
Lu X,Liu J,Ren W,Yang Q,Chai Z,Chen R,Wang L,Zhao J,Lang Z,Wang H.2018.Gene-indexed mutations in maize(Zea mays).Molecular Plant,11(3):496-504.
Schaefer H,Renner S.2010.A three-genome phylogeny of Momordica(Cucurbitaceae)suggests seven returns from dioecy to monoecy and recent long-distance dispersal to Asia.Molecular Phylogenetics and Evolution,54(2):553-560.
Urasaki N,Takagi H,Natsume S,Uemura A,Taniai N,Miyagi N,Fukushima M,Suzuki S,Tarora K,Tamaki M,Sakamoto M,Terauchi R,Matsumura H.2017.Draft genome sequence of bitter gourd(Momordica charantia),a vegetable and medicinal plant in tropical and subtropical regions.DNA Research,24(1):51-58.
van Wyk B.2015.A review of commercially important African medicinal plants.Journal of Ethnopharmacology,176:118-134.
Yang G,Li S,Wang S.2016.Research progress in enzyme activity and pharmacological effects of ribosome-inactivity protein in bitter melon.Toxin Reviews,35:128-132.