~(60)Co-γ射线辐射对木薯组培苗的诱变效应
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摘要
为探究~(60)Co-γ射线辐射对不同木薯品种组培苗的诱变效应,以2个木薯品种新选048和SC205离体单芽茎段为试验材料,以不同~(60)Co-γ射线辐射剂量(0、5、10、20、40 Gy)进行处理,观察比较不同辐射处理条件下组培苗的生长情况以及移栽后植株的生长状况。结果表明,随着~(60)Co-γ辐射剂量的增加,2个品种组培苗的生根率和萌发率均显著降低;新选048和SC205的半致死剂量分别为15. 3 Gy和17. 2 Gy。随着辐射剂量的增加,对继代组培苗及其生根苗的生长伤害程度逐渐加强,株高、生根率、移栽成活率等指标均显著降低。不同品种对~(60)Co-γ射线辐射的敏感程度存在差异,新选048的辐射敏感性更强。经~(60)Co-γ射线辐射后,2个木薯品种组培苗均出现叶柄颜色变异、叶形变异、黄化苗、矮化苗等不同程度的形态变异。经大田移栽后统计,新选048变异率为3. 3%,SC205变异率为6. 6%。对突变材料进行SSR分子标记检测发现,其中有3条引物可检测SC205对照与突变体材料在分子水平上的差异; 5条引物可检测新选048对照与突变体材料在分子水平上的差异。本研究结果为利用~(60)Co-γ射线辐射诱变创造木薯新种质以及木薯诱变育种奠定了一定的理论基础。
To explore the mutagenic effects of ~(60)Co-γ irradiation,stem cutting of two cassava varieties,xinxuan 048 and SC205,were irradiated with 0,5,10,20 and 40 Gy of ~(60)Co-γ,respectively. The growth of tissue culture seedlings with different radiation treatment and the growth of plants after transplanting were observed and compared. The results showed that the rooting rate and germination rate of two varieties were decreased significantly with the increment of the irradiation dose. The median lethal dose( LD_(50)) for two varieties was defined as 15. 3 Gy,17. 2 Gy,respectively. With the increasement of the irradiation dose,the damage severity of tissue culture seedlings was reinforced,the seedling height,rooting rate and transplanting survival rate were decreased significantly. There were different vadiation sensitivity between varieties,xinxuan 048 are more radiation susception than SC205. After irradiation,different morphological variations such as petiole color,leaf shape,etiolated seedling and dwarf seeding were observed in both of two tissue culture seedlings. The variation rate of xinxuan048 and SC 205 was 4. 4% and 6. 6%,respectively. Three pairs of microsatellite( simple sequence repeat,SSR) primers were found to be polymorphism between genomic of SC205 wide type and mutants plants,five SSR primers were shown to be polymorphism between Xinxuan 048 wide type and it's corresponding mutants. The results laid a foundation for creating a new cassava germplasms by ~(60) Co-gamma rays radiation and opening a new method for cassava breeding.
To explore the mutagenic effects of ~(60)Co-γ irradiation,stem cutting of two cassava varieties,xinxuan 048 and SC205,were irradiated with 0,5,10,20 and 40 Gy of ~(60)Co-γ,respectively. The growth of tissue culture seedlings with different radiation treatment and the growth of plants after transplanting were observed and compared. The results showed that the rooting rate and germination rate of two varieties were decreased significantly with the increment of the irradiation dose. The median lethal dose( LD_(50)) for two varieties was defined as 15. 3 Gy,17. 2 Gy,respectively. With the increasement of the irradiation dose,the damage severity of tissue culture seedlings was reinforced,the seedling height,rooting rate and transplanting survival rate were decreased significantly. There were different vadiation sensitivity between varieties,xinxuan 048 are more radiation susception than SC205. After irradiation,different morphological variations such as petiole color,leaf shape,etiolated seedling and dwarf seeding were observed in both of two tissue culture seedlings. The variation rate of xinxuan048 and SC 205 was 4. 4% and 6. 6%,respectively. Three pairs of microsatellite( simple sequence repeat,SSR) primers were found to be polymorphism between genomic of SC205 wide type and mutants plants,five SSR primers were shown to be polymorphism between Xinxuan 048 wide type and it's corresponding mutants. The results laid a foundation for creating a new cassava germplasms by ~(60) Co-gamma rays radiation and opening a new method for cassava breeding.
引文
[1]Olkhov Y A,Allayarov S R,Tashmetov M Y,Dixon D A.Effect ofγ-irradiation on the molecular-topological structure of a copolymer of tetrafluoroethylene and perfluoro(propyl vinyl ether)[J].High Energy Chemistry,2017,51:101-107
[2]陈军.体外诱变与组织培养在菊芋种质创新中的应用研究[D].武汉:华中农业大学,2009
[3]孔滢,白锦荣,尚宏忠,王乃彦.重离子束辐射技术在花卉育种中的应用[J].园艺学报,2013,40(9):1837-1845
[4]张慧琴.辐射诱变结合组织培养在草莓育种中的应用[D].杭州:浙江大学,2007
[5]吴潇,齐开杰,殷豪,张绍铃.诱变技术在落叶果树育种中的应用[J].园艺学报,2016,43(9):1633-1652
[6]蒋彧,何俊蓉,刘菲,王海娥,卓碧萍.60Coγ辐射兰花春剑隆昌素根状茎分化苗的ISSR分析[J].核农学报,2013,27(9):1247-1252
[7]任羽,潘丙成,陆顺教,张志群,孔霞.60Co-γ辐射石斛兰组培苗的SRAP分析[J].中国农学通报,2016,32(31):85-89
[8]华婧.马铃薯高频再生体系建立及离体茎尖诱变筛选耐盐突变体的研究[D].大连:辽宁师范大学,2008
[9]杨锦芬,刘卉,彭苑霞,刘艳,邹伟权,詹若挺,徐晖,陈蔚文.阳春砂辐射诱变及基于ISSR的变异分析[J].植物遗传资源学报,2012,13(5):748-753
[10]Mandal A K,Chakrabarty D,Datta S K.In vitro isolation of solid novel flower colour mutants from induced chimeric ray florets of chrysan-themum[J].Euphytica,2000,114:9-12
[11]Das A,Gosal S S,Sidhu J S.Induction of mutations for heat tolerance with induced mutations for rice improvement[J].Euphytica,2001,120:401-408
[12]Mshara P J,Ganapath T R,Suprasannap P,BapatⅤ.Effect of single and recurrent gamma irradiation of in vitro shoot cultures of banana[J].International Journal of Fruit Science,2007,7(1):47-57
[13]严华兵,叶剑秋,李开绵.中国木薯育种研究进展[J].中国农学通报,2015,31(15):63-70
[14]曾文丹,谢向誉,严华兵.60Co-γ辐射对木薯体细胞胚子叶再生的诱变效应[J].热带作物学报,2017,38(10):1895-1900
[15]Nwachukwu E C,Mbanaso E N A,Ene L S O.Improvement of cassava for high dry matter,starch and low cyanogenic glucoside content by mutation induction[J].IAEA-tecdoc,1997,1997(951):93-98
[16]Sanchez T,Roserob A,Tofino A P.Induction and identification of useful mutations for root quality traits in cassava[C]//Shu Q Y.Induced plant mutations in the genomics era Rome:Food and Agriculture Organization of the United Nations,2009:266-269
[17]Ahiabu R K,Lokko Y,Danso K,Klu G Y P.Mutagenesis for ACMV resistance in a ghanian cassava cultivar‘Bosom Nsia’[J].IAEA-tecdoc,1997,1997(951):9-18
[18]王磊.丰花月季组培苗辐射诱变研究及遗传变异研究[D].哈尔滨:东北农业大学,2007
[19]闫庆祥,黄东益,李开绵,叶剑秋.利用改良CTAB法提取木薯基因组DNA[J].中国农学通报,2010,26(4):30-32
[20]王明,肖鑫辉,安飞飞,万仲卿,应东山,王琴飞,张如莲,李开绵,叶剑秋.利用SSR标记分析木薯遗传多样性[J].热带农业科学,2015,35(11):38-44
[21]Swartz H J.Post Culture Behavior:Genetic and Epigenetic Effects and Related Problems[M].Netherlands:Kluwer Academic Publishers,1991:95-121
[22]陆艳婷,陈金跃,张小明,叶胜海,王俊敏,金庆生.浙江省水稻辐射育种研究进展[J].核农学报,2017,31(8):1500-1508
[23]孙光祖,唐凤兰,王广金,李忠杰,张月学,阎文义,孙德全,孙岩.辐射诱变与生物技术结合创造小麦新种质的研究Ⅰ.小麦新种质的创造[J].麦类作物学报,1998,18(6):4-7
[24]彭振英,王兴军,田海莹,郑玲,单雷,范仲学,边斐,郭峰,王莹莹,万书波.花生60Co-γ辐射诱变和突变体库的构建[J].核农学报,2016,30(3):422-429
[25]宋华伟,刘颖,曹荣祥,张巨明.3种结缕草及其60Co-γ辐射诱变新品系抗旱性比较[J].华南农业大学学报,2015,36(6):55-61
[26]曲龙,卢夏茹,李阳生,韦双双,袁潜华,马启林.水稻辐射诱变突变体的种子蛋白质成分分析[J].核农学报,2015,29(2):209-214
[27]徐宏,胡小红,陈军顺,黄志勇,朱培林,汪江.辐射诱变选育栀子新品系[J].核农学报,2010,24(5):937-940,986
[28]郭泰,刘忠堂,胡喜平,王志新,吴秀红,郑伟,陈德祥,王雷.辐射诱变培育高油大豆新品种及其应用[J].核农学报,2005(3):163-167
[29]刘丽强,刘军丽,张杰,耿慧,张玉萍,沈红香,高遐红,姚允聪.60Co-γ辐射对观赏海棠组培苗的诱变效应[J].中国农业科学,2010,43(20):4255-4264
[30]邢莉莉,陈发棣,陈素梅.60Co-γ辐射对切花菊试管苗的诱变效应[J].园艺学报,2010,37(7):1117-1124
[31]范明霞.木薯再生体系的建立和HNL24b基因的克隆及辐照诱变育种[D].武汉:华中农业大学,2010
[32]陆柳英.木薯诱变处理的生物学效应以及抗寒突变体的初步筛选[D].海口:华南热带农业大学,2007
[33]方智振,叶新福,周丹蓉,潘少霖,姜翠翠.辐射三月李红肉迟熟突变体的ISSR分析[J].核农学报,2016,30(2):209-215
[34]陆波,郑玉红,陈默,彭峰,束晓春,高风.60Co-γ射线对彩色马蹄莲Parfait的辐照效应及其在高温高湿胁迫下的生理响应[J].核农学报,2014,28(8):1353-1357
[35]罗海燕.非洲菊60Co-γ辐射诱变的初步研究[D].长沙:湖南农业大学,2013
[36]郭安熙,范家霖,杨保友,王柏楠,张建伟,陈树国,王世荣,王然.菊花花色辐射诱变研究[J].核农学报,1997,11(2):65-73
[37]包建忠,李风童,孙叶,刘春贵,马辉,张甜,陈秀兰.60Co-γ射线辐照大花君子兰种子对其萌发特性及其性状的影响[J].核农学报,2013,27(11):1681-1685
[2]陈军.体外诱变与组织培养在菊芋种质创新中的应用研究[D].武汉:华中农业大学,2009
[3]孔滢,白锦荣,尚宏忠,王乃彦.重离子束辐射技术在花卉育种中的应用[J].园艺学报,2013,40(9):1837-1845
[4]张慧琴.辐射诱变结合组织培养在草莓育种中的应用[D].杭州:浙江大学,2007
[5]吴潇,齐开杰,殷豪,张绍铃.诱变技术在落叶果树育种中的应用[J].园艺学报,2016,43(9):1633-1652
[6]蒋彧,何俊蓉,刘菲,王海娥,卓碧萍.60Coγ辐射兰花春剑隆昌素根状茎分化苗的ISSR分析[J].核农学报,2013,27(9):1247-1252
[7]任羽,潘丙成,陆顺教,张志群,孔霞.60Co-γ辐射石斛兰组培苗的SRAP分析[J].中国农学通报,2016,32(31):85-89
[8]华婧.马铃薯高频再生体系建立及离体茎尖诱变筛选耐盐突变体的研究[D].大连:辽宁师范大学,2008
[9]杨锦芬,刘卉,彭苑霞,刘艳,邹伟权,詹若挺,徐晖,陈蔚文.阳春砂辐射诱变及基于ISSR的变异分析[J].植物遗传资源学报,2012,13(5):748-753
[10]Mandal A K,Chakrabarty D,Datta S K.In vitro isolation of solid novel flower colour mutants from induced chimeric ray florets of chrysan-themum[J].Euphytica,2000,114:9-12
[11]Das A,Gosal S S,Sidhu J S.Induction of mutations for heat tolerance with induced mutations for rice improvement[J].Euphytica,2001,120:401-408
[12]Mshara P J,Ganapath T R,Suprasannap P,BapatⅤ.Effect of single and recurrent gamma irradiation of in vitro shoot cultures of banana[J].International Journal of Fruit Science,2007,7(1):47-57
[13]严华兵,叶剑秋,李开绵.中国木薯育种研究进展[J].中国农学通报,2015,31(15):63-70
[14]曾文丹,谢向誉,严华兵.60Co-γ辐射对木薯体细胞胚子叶再生的诱变效应[J].热带作物学报,2017,38(10):1895-1900
[15]Nwachukwu E C,Mbanaso E N A,Ene L S O.Improvement of cassava for high dry matter,starch and low cyanogenic glucoside content by mutation induction[J].IAEA-tecdoc,1997,1997(951):93-98
[16]Sanchez T,Roserob A,Tofino A P.Induction and identification of useful mutations for root quality traits in cassava[C]//Shu Q Y.Induced plant mutations in the genomics era Rome:Food and Agriculture Organization of the United Nations,2009:266-269
[17]Ahiabu R K,Lokko Y,Danso K,Klu G Y P.Mutagenesis for ACMV resistance in a ghanian cassava cultivar‘Bosom Nsia’[J].IAEA-tecdoc,1997,1997(951):9-18
[18]王磊.丰花月季组培苗辐射诱变研究及遗传变异研究[D].哈尔滨:东北农业大学,2007
[19]闫庆祥,黄东益,李开绵,叶剑秋.利用改良CTAB法提取木薯基因组DNA[J].中国农学通报,2010,26(4):30-32
[20]王明,肖鑫辉,安飞飞,万仲卿,应东山,王琴飞,张如莲,李开绵,叶剑秋.利用SSR标记分析木薯遗传多样性[J].热带农业科学,2015,35(11):38-44
[21]Swartz H J.Post Culture Behavior:Genetic and Epigenetic Effects and Related Problems[M].Netherlands:Kluwer Academic Publishers,1991:95-121
[22]陆艳婷,陈金跃,张小明,叶胜海,王俊敏,金庆生.浙江省水稻辐射育种研究进展[J].核农学报,2017,31(8):1500-1508
[23]孙光祖,唐凤兰,王广金,李忠杰,张月学,阎文义,孙德全,孙岩.辐射诱变与生物技术结合创造小麦新种质的研究Ⅰ.小麦新种质的创造[J].麦类作物学报,1998,18(6):4-7
[24]彭振英,王兴军,田海莹,郑玲,单雷,范仲学,边斐,郭峰,王莹莹,万书波.花生60Co-γ辐射诱变和突变体库的构建[J].核农学报,2016,30(3):422-429
[25]宋华伟,刘颖,曹荣祥,张巨明.3种结缕草及其60Co-γ辐射诱变新品系抗旱性比较[J].华南农业大学学报,2015,36(6):55-61
[26]曲龙,卢夏茹,李阳生,韦双双,袁潜华,马启林.水稻辐射诱变突变体的种子蛋白质成分分析[J].核农学报,2015,29(2):209-214
[27]徐宏,胡小红,陈军顺,黄志勇,朱培林,汪江.辐射诱变选育栀子新品系[J].核农学报,2010,24(5):937-940,986
[28]郭泰,刘忠堂,胡喜平,王志新,吴秀红,郑伟,陈德祥,王雷.辐射诱变培育高油大豆新品种及其应用[J].核农学报,2005(3):163-167
[29]刘丽强,刘军丽,张杰,耿慧,张玉萍,沈红香,高遐红,姚允聪.60Co-γ辐射对观赏海棠组培苗的诱变效应[J].中国农业科学,2010,43(20):4255-4264
[30]邢莉莉,陈发棣,陈素梅.60Co-γ辐射对切花菊试管苗的诱变效应[J].园艺学报,2010,37(7):1117-1124
[31]范明霞.木薯再生体系的建立和HNL24b基因的克隆及辐照诱变育种[D].武汉:华中农业大学,2010
[32]陆柳英.木薯诱变处理的生物学效应以及抗寒突变体的初步筛选[D].海口:华南热带农业大学,2007
[33]方智振,叶新福,周丹蓉,潘少霖,姜翠翠.辐射三月李红肉迟熟突变体的ISSR分析[J].核农学报,2016,30(2):209-215
[34]陆波,郑玉红,陈默,彭峰,束晓春,高风.60Co-γ射线对彩色马蹄莲Parfait的辐照效应及其在高温高湿胁迫下的生理响应[J].核农学报,2014,28(8):1353-1357
[35]罗海燕.非洲菊60Co-γ辐射诱变的初步研究[D].长沙:湖南农业大学,2013
[36]郭安熙,范家霖,杨保友,王柏楠,张建伟,陈树国,王世荣,王然.菊花花色辐射诱变研究[J].核农学报,1997,11(2):65-73
[37]包建忠,李风童,孙叶,刘春贵,马辉,张甜,陈秀兰.60Co-γ射线辐照大花君子兰种子对其萌发特性及其性状的影响[J].核农学报,2013,27(11):1681-1685