叠氮化钠、甲基磺酸乙酯复合诱变对湖北百合耐盐性的影响
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摘要
以湖北百合鳞片为试材,采用9种不同培养基接种,获得诱导愈伤组织最佳配方为MS+6-BA 1.0mg·L~(-1)+NAA 0.1mg·L~(-1)。以该配方诱导出的愈伤组织为诱变材料,先后分别用NaN_3(1、2、3mmol·L~(-1))处理(3、4、5h)、EMS(0.2%、0.4%、0.6%)处理(1、2、3h)愈伤组织,对诱变处理的愈伤组织分化的不定芽用0.4%NaCl胁迫处理0～12d进行盐胁迫筛选,以期获得复合诱变处理后的最佳耐盐浓度。结果表明:3mmol·L~(-1)的NaN_3处理3h达半致死,诱变率达46.7%,使用0.4%EMS再次诱变3h达半致死,诱变率达53.3%。复合诱变测得的超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、可溶性糖含量以及可溶性蛋白质含量的变化始终高于对照。因此,在盐胁迫复合诱变筛选的湖北百合的新陈代谢水平较高,表现出对盐胁迫的适应性。
Lilium henryi was used as the test material,and 9 different medium were used for inoculation.The optimal formula for inducing callus was MS+6-BA 1.0 mg· L~(-1)+ NAA0.1 mg·L~(-1).According to the formula of the callus induced mutation material,successively with NaN3(1,2,3 mmol·L~(-1))processing(3,4,5 hours),EMS(0.2%,0.4%,0.6%)treatment(1,2,3 hours)callus and differentiation of mutagenic treatment of adventitious buds treated with 0.4%NaCl stress 0-12 days to salt stress screening,in order to obtain the best resistance to salt concentration compound mutation after processing.The results showed that NaN3 treated with3 mmol·L~(-1) was semi-lethal at 3 hours and the mutagenesis rate was 46.7%,and the mutagenesis rate was up to 53.3% with 0.4%EMS.The changes of SOD activity,POD activity,soluble sugar content and soluble protein content in the mutagenesis were higher than those in the control group.Therefore,the metabolism of Lilium henryi was higher,showing its adaptability to salt stress.
Lilium henryi was used as the test material,and 9 different medium were used for inoculation.The optimal formula for inducing callus was MS+6-BA 1.0 mg· L~(-1)+ NAA0.1 mg·L~(-1).According to the formula of the callus induced mutation material,successively with NaN3(1,2,3 mmol·L~(-1))processing(3,4,5 hours),EMS(0.2%,0.4%,0.6%)treatment(1,2,3 hours)callus and differentiation of mutagenic treatment of adventitious buds treated with 0.4%NaCl stress 0-12 days to salt stress screening,in order to obtain the best resistance to salt concentration compound mutation after processing.The results showed that NaN3 treated with3 mmol·L~(-1) was semi-lethal at 3 hours and the mutagenesis rate was 46.7%,and the mutagenesis rate was up to 53.3% with 0.4%EMS.The changes of SOD activity,POD activity,soluble sugar content and soluble protein content in the mutagenesis were higher than those in the control group.Therefore,the metabolism of Lilium henryi was higher,showing its adaptability to salt stress.
引文
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[35]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.
[36]何慧怡,樊丽娜,齐永文,等.EMS诱变甘蔗愈伤组织的初步研究[J].甘蔗糖业,2017(2):1-8.
[2]肖玉菲,陈博雯,覃子海,等.不同因素对大百合鳞茎诱导出芽的影响[J].江西农业学报,2018,30(8):25-28.
[3]丁芳兵,孙伟博,原雅玲,等.百合花色及相关育种研究进展[J].北方园艺,2017(18):170-175.
[4]潘娟.湖北百合组织培养与快繁技术研究[D].重庆:西南大学,2009.
[5]隆世良,蒋宇,郑绍伟,等.岷江上游干旱河谷野生百合资源分布与繁殖力比较[J].四川林业科技,2018,39(2):61-64.
[6]许雯雯,龙松华,邱财生,等.甘蔗诱变育种研究进展[J].热带农业科学,2017,37(8):68-73.
[7]蒋姝,孟金贵,崔光芬,等.植物离体诱变及在百合育种中的应用[J].中国农学通报,2012,28(4):69-73.
[8]冯学金,郭秀娟,杨建春,等.诱变技术在亚麻育种中的应用[J].核农学报,2017,31(7):1310-1316.
[9]颜志勤.三种化学诱变剂在杉木育种中的应用研究[D].福州:福建农林大学,2013.
[10]李波,林浩,邬婷婷,等.NaN3和紫外线复合诱变对苜蓿愈伤组织在干旱和盐胁迫下生长的影响[J].干旱地区农业研究,2018,36(4):187-192.
[11]付凤玲,李晚忱,荣廷昭,等.用γ射线和叠氮化钠诱变的玉米愈伤组织筛选耐旱和雄性不育材料[J].核农学报,2005,19(5):356-359.
[12]刘翔.EMS诱变技术在植物育种中的研究进展[J].激光生物学报,2014,23(3):197-201.
[13]程志锋.小麦近等基因系TcLr10,TcLr21和TcLr44的诱变研究[D].保定:河北农业大学,2008.
[14]SERRAT X,ESTEBAN R,GUIBOURT N,et al.EMSmutagenesis in mature seed-derived rice calli,as a new method for rapidly obtaining TILLING mutant populations[J].Plant Methods,2014,10(1):5.
[15]ROSWITHA B,AXEL P,GüNTER S,et al.Development of a TILLING resource in durum wheat for reverse-and forward-genetic analyses[J].Crop&Pasture Science,2014,65(1):112-124.
[16]杨建胜,孙万仓,刘自刚,等.EMS诱变对北方白菜型冬油菜农艺性状与品质的影响[J].干旱地区农业研究,2018,36(1):213-220.
[17]弭宝彬,谢玲玲,王鑫,等.冬瓜EMS诱变条件优化及突变体筛选[J].中国农学通报,2018,34(21):35-41.
[18]白鸿雁,杨伟,武擘.高粱种子EMS诱变处理的研究[J].中国种业,2018(3):57-59.
[19]杨秀丽,杨丽萍,宁东贤,等.EMS诱变及其在构建花生突变体库中的应用[J].山西农业科学,2018,46(9):1577-1580.
[20]袁志平,李学军,张焕丽,等.不同温度下叠氮化钠浓度对辣椒种子萌发的影响[J].安徽农学通报,2018(6):17-18+31.
[21]苑平,吴娟娟,李先信,等.叠氮化钠对纽荷尔脐橙腋芽的半致死浓度、生理影响和诱变效率研究[J].湖南师范大学自然科学学报,2018,41(1):30-35.
[22]温日宇,刘建霞,宋亚静,等.叠氮化钠对绿豆种子和幼苗生长的诱变效应[J].山西农业科学,2017,45(12):1933-1936.
[23]全妙华,陈东明,蒋向辉,等.四棱豆新品种‘湘棱豆3号’[J].园艺学报,2011,38(9):1827-1828.
[24]许有嫔,熊文斌,钟程.凯里地区野生百合资源农艺性状研究[J].凯里学院学报,2013,31(6):56-60.
[25]肖雯,贾恢先,蒲陆梅,等.几种盐生植物抗盐生理指标的研究[J].西北植物学报,2000,32(7):818-825.
[26]陈立松,刘星辉.植物抗热性鉴定指标的种类[J].干旱地区农业研究,1997,23(6):34-37.
[27]路文静,李奕松.植物生理学实验教程[M].北京:中国林业出版社,2012.
[28]安学丽,蔡一林,王久光,等.化学诱变及其在农作物育种上应用[J].核农学报,2003(3):21.
[29]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.
[30]任少雄,王丹.植物理化复合诱变育种技术研究进展[J].安徽农业科学,2009,37(20):9345-9349.
[31]彭选明,易卫平,杨震,等.湖南水稻种质资源创制及辐射诱变育种应用成效[J].湖南农业科学,2015(4):10-13.
[32]卜婷,范小峰,刘丽琼,等.紫外辐射对矮牵牛愈伤组织增殖的影响[J].陇东学院学报,2013,24(5):49-52.
[33]NAGY Z,NMETH E,GUTH A,et al.Metabolic indicators of drought stress tolerance in wheat:Glutamine synthetase isoenzymes and Rubisco[J].Plant Physiology&Biochemistry Ppb,2013,67(3):48-54.
[34]蒋向辉,陈东明,全妙华.四棱豆的化学诱变及矮化突变性状初步分析[J].中国蔬菜,2010(6):219.
[35]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.
[36]何慧怡,樊丽娜,齐永文,等.EMS诱变甘蔗愈伤组织的初步研究[J].甘蔗糖业,2017(2):1-8.