黄瓜突变体库的构建及表型变异的初步研究
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摘要
为构建黄瓜突变体库,利用不同浓度的甲基磺酸乙酯(EMS)溶液在不同处理时间下诱变华南型黄瓜高代自交系6457,并对诱变前后黄瓜表型变异特征进行分析。结果表明,根据诱变后的种子发芽率、成苗率,筛选出1. 5%EMS处理10 h为最佳诱变条件。采用最佳诱变条件处理2 500粒黄瓜种子,构建了包含221个M2株系的黄瓜突变体库。在M1群体中出现叶片皱缩、叶色嵌合、植株多卷须、矮化、黄化、白化、畸形、果实嵌合体等变异性状,表型突变频率达18. 9%;在M2群体中筛选到长瓜、短瓜、心形叶、花萼叶片化、矮化植株等典型的突变体,表型突变频率达11. 7%。本研究结果丰富了黄瓜种质资源,为黄瓜功能基因组学研究和新品种选育提供了一定的理论依据。
In order to produce cucuber mutants. In this study,seeds of the high-generation inbred line 6457 was treated by ethyl methane sulfonate( EMS) solution with different concentrations and times. The phenotypic variation characteristics of cucumber before and after mutagenesis were analyzed. Considering the germination rate and seedling rate,1. 5% EMS for 10 h was selected as the appropriate treatment condition. Then 2 500 seeds of 6457 were treated by1. 5% EMS for 10 h,and a cucumber mutant library contained 221 M2 families was constructed. Variations with abnormal appearance i. e. wrinkled leaf,variegated leaf,plant with multiple tendrils,dwarf,etiolation,albino,deformed and chimera fruit were observed in the M1 population with the mutation frequency of 18. 9%. Numerous valuable mutants were isolated in the M2 population, including long fruit, short fruit, heart-shaped leaves,hermaphrodite flower,dwarfing plant and etc. The mutation frequency in the M2 population was 11. 7%. The mutants obtained in this study enviched in germplasm resources,provide useful mutant materials for cucumber functional genomics research and breeding improvement.
In order to produce cucuber mutants. In this study,seeds of the high-generation inbred line 6457 was treated by ethyl methane sulfonate( EMS) solution with different concentrations and times. The phenotypic variation characteristics of cucumber before and after mutagenesis were analyzed. Considering the germination rate and seedling rate,1. 5% EMS for 10 h was selected as the appropriate treatment condition. Then 2 500 seeds of 6457 were treated by1. 5% EMS for 10 h,and a cucumber mutant library contained 221 M2 families was constructed. Variations with abnormal appearance i. e. wrinkled leaf,variegated leaf,plant with multiple tendrils,dwarf,etiolation,albino,deformed and chimera fruit were observed in the M1 population with the mutation frequency of 18. 9%. Numerous valuable mutants were isolated in the M2 population, including long fruit, short fruit, heart-shaped leaves,hermaphrodite flower,dwarfing plant and etc. The mutation frequency in the M2 population was 11. 7%. The mutants obtained in this study enviched in germplasm resources,provide useful mutant materials for cucumber functional genomics research and breeding improvement.
引文
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[21]张兵.EMS诱导黄瓜突变体的初步研究[D].泰安:山东农业大学,2012
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[24]王春语,朱振兴,李丹,丛玲,张丽霞.高粱EMS诱变及突变体筛选、鉴定[J].生物技术通报,2014(9):78-83
[25]卢银,刘梦洋,赵建军,王彦华,罗双霞,轩淑欣,代双燕,王超硕,申书兴.大白菜突变体库的构建及M2叶片表型变异的研究[J].园艺学报,2014,41(8):1609-1619
[26]骆海波,邹颉,贺从安,李德超,熊秋芳,张小康,张贵生,李世升.甲基磺酸乙酯(EMS)对萝卜早期发育性状的诱变效应[J].基因组学与应用生物学,2013,32(5):608-611
[27]张虹,龙宏周,路国栋,陈任.黑果枸杞多倍体诱导及鉴定[J].核农学报,2017,31(1):59-65
[28]陈宸.EMS诱导黄瓜406突变体的筛选与鉴定[D].长沙:湖南农业大学,2015
[2]Staub J E,Fredric L,Marty T L.Electrophoretic variation in cross compatible wild diploid species of Cucumis[J].Canadian Journal of Botany,1987,65(4):792-798
[3]张国庆,周伟军,姚先伶,张志军.芸薹属植物远缘杂交研究现状[J].山西农业科学,2001(4):25-30
[4]Cuartero J,Bolarin M C,Asins M J.Increasing salt tolerance in the tomato[J].Journal of Experimental Botany,2006,57(5):1045-1058
[5]周立名,王飞,王佳.EMS诱变处理定向筛选猕猴桃耐盐突变体研究[J].西北农业学报,2009,18(5):330-335
[6]胡齐赞,龚亚明,韦顺恋.大白菜花药和游离小孢子培养比较研究[J].浙江农业学报,2007,19(5):352-355
[7]卢银.EMS处理诱发大白菜变异的研究[D].保定:河北农业大学,2011
[8]Shiwa Y,Tanaka S F,Kasahara K,Takayuki H,Yoshikawa H.Whole-genome profiling of a novel mutagenesis technique using proofreading-deficient DNA polymerase[J].International Journal of Evolutionary Biology,2012,860797.doi:10.1155/2012/860797
[9]Chawade A,Sikora P,Brutigam M,Larsson M,VivekanandⅤ,Nakash M A,Chen T,Olsson O.Development and characterization of an oat TILLING-population and identification of mutations in lignin andβ-glucan biosynthesis genes[J].BMC Plant Biology,2010,10(86):86-98
[10]Lochlainn S,Amoah S,Graham N S,Alamer K,Rios J J,Kurup S,Stoute A,Hammond J P,stergaard L,King G J,White P J,Broadley M R.High resolution melt(HRM)analysis is an efficient tool to genotype EMS mutants in complex crop genomes[J].Plant Methods,2011,7(11):43-51
[11]Harloff H J,Lemcke S,Mittasch J,Frolov A,Wu J G,Dreyer F,Leckband G,Jung C.A mutation screening platform for rapeseed(Brassica napus L.)and the detection of sinapine biosynthesis mutants[J].Theoretical and Applied Genetics,2012,124(5):957-969
[12]SiluéS,Diarrassouba N,FofanaⅠJ,Muhovski Y,Toussaint A,Mergeai G,Jacquemin J M,Baudoin J P.Description of Phaseolus vulgaris L.aborting embryos from ethyl methanesulfonate(EMS)mutagenized plants[J].Biotechnologie,Agronomie,SociétéEt Environnement,2013,17(4):563-571
[13]Chen Y L,Liang H L,Ma X L,Lou S L,Xie Y Y,Liu Z L,Chen L T,Liu Y G.An efficient rice mutagenesis system based on suspension cultured cells[J].Journal of Integrative Plant Biology,2013,55(2):122-130
[14]王瑾,刘桂茹,杨学举.EMS诱变小麦愈伤组织选择抗旱突变体的研究[J].中国农学通报,2005,21(12):190-193
[15]卢银.大白菜EMS突变体库的创建与筛选鉴定[D].保定:河北农业大学,2014
[16]Mc Callum C M,Comai L,Greene E A,Henikoff S.Targeted screening for induced mutations[J].Nature Biotechnology,2000,18(4):455-457
[17]Akira A,Shunichi K,Kentaro Y,Satoshi N,Hiroki T,Hiroyuki K,Hideo M,Kakoto Y,Chikako M,Muluneh T,HidekiⅠ,Liliana C,Sophien K,R yohei T.Genome sequencing reveals agronomically important loci in rice using MutMap[J].Nature Biotechnology,2012,30(2):174-179
[18]王峰,徐飚,杨正林,凌英华,何光华,陈胜,卿明敬,桑贤春.EMS诱变水稻矮生资源的鉴定评价[J].核农学报,2011,25(2):197-201
[19]王晶,娄群峰,魏庆镇.长春密刺黄瓜突变体库的构建和部分性状分析[J].核农学报,2015,29(8):1479-1486
[20]薛存宝.黄瓜EMS诱变突变体库及TILLING平台的初步构建[D].哈尔滨:东北农业大学,2016
[21]张兵.EMS诱导黄瓜突变体的初步研究[D].泰安:山东农业大学,2012
[22]李锡香,朱德蔚.黄瓜种质资源描述规范和数据标准[M].北京:中国农业出版社,2005
[23]朱娜娜.EMS诱变西瓜突变体库的构建及表型分析[D].哈尔滨:东北农业大学,2015
[24]王春语,朱振兴,李丹,丛玲,张丽霞.高粱EMS诱变及突变体筛选、鉴定[J].生物技术通报,2014(9):78-83
[25]卢银,刘梦洋,赵建军,王彦华,罗双霞,轩淑欣,代双燕,王超硕,申书兴.大白菜突变体库的构建及M2叶片表型变异的研究[J].园艺学报,2014,41(8):1609-1619
[26]骆海波,邹颉,贺从安,李德超,熊秋芳,张小康,张贵生,李世升.甲基磺酸乙酯(EMS)对萝卜早期发育性状的诱变效应[J].基因组学与应用生物学,2013,32(5):608-611
[27]张虹,龙宏周,路国栋,陈任.黑果枸杞多倍体诱导及鉴定[J].核农学报,2017,31(1):59-65
[28]陈宸.EMS诱导黄瓜406突变体的筛选与鉴定[D].长沙:湖南农业大学,2015