~(60)Co-γ射线辐照翠竹的生物学效应及RAPD分析
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摘要
用不同剂量(5、10、15、20和25 Gy)的~(60)Co-γ射线对翠竹幼苗进行辐照处理,结果显示,辐照导致翠竹矮化,植株的株高、节间长度、叶长及叶宽等性状较对照组显著变短;并且10 Gy辐照导致翠竹部分叶片白化。基因组RAPD分子标记检测显示,辐照组扩增的DNA片段图谱发生显著变化,表明经辐照处理的翠竹基因组发生了变异。其中,5 Gy辐照实验组的变异程度最小,25 Gy辐照实验组的变异程度最大。研究表明,~(60)Co-γ射线辐照可以引起翠竹植株矮化,并导致基因组变异。本研究为进一步进行翠竹辐照诱变育种打下了基础。
Various radiation dosages( 5 Gy、10 Gy、15 Gy、20 Gy and 25 Gy) of~(60)Co-γ-radiation are used to treat the seedlings of Sasapygmaea. The results show that irradiation can induce dwarf effect,and the plant height,internode length,leaf length and width of irradiation group are significantly shorter than the control group. The leaves of Sasapygmaea in 10 Gy radiation dosage group show albinism. By genomic RAPD assay,the amplification of DNA bands show significantly change,which suggests the genome of Sasapygmaea underwent mutation after irradiation treatment. Specifically,the minimum extent of genetic mutation is 5 Gy radiation dosage group and the maximum is 25 Gy radiation dosage group. Our finding indicates that~(60)Co-γ-radiation can induce dwarf plant accompanying with genomic mutation. This study lays a foundation for further breeding of Sasapygmaea via radiation.
Various radiation dosages( 5 Gy、10 Gy、15 Gy、20 Gy and 25 Gy) of~(60)Co-γ-radiation are used to treat the seedlings of Sasapygmaea. The results show that irradiation can induce dwarf effect,and the plant height,internode length,leaf length and width of irradiation group are significantly shorter than the control group. The leaves of Sasapygmaea in 10 Gy radiation dosage group show albinism. By genomic RAPD assay,the amplification of DNA bands show significantly change,which suggests the genome of Sasapygmaea underwent mutation after irradiation treatment. Specifically,the minimum extent of genetic mutation is 5 Gy radiation dosage group and the maximum is 25 Gy radiation dosage group. Our finding indicates that~(60)Co-γ-radiation can induce dwarf plant accompanying with genomic mutation. This study lays a foundation for further breeding of Sasapygmaea via radiation.
引文
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[4]张强.辐照技术在农业中的应用[J].农业工程,2012,2(S1):42-46.
[5]乔广军.诱变在我国亚麻育种及资源创新中的利用[J].中国麻业科学,2006,28(1):17-20.
[6]朱丽兵,周云清,丁捷,等.CANDU重水反应堆钴调节棒组件结构设计[J].原子能科学技术,2010,44(B09):418-422.
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[8]任少雄,王丹,李卫锋,等.60Coγ射线辐射唐菖蒲鳞茎诱变育种试验[J].福建林业科技,2006,33(2):34-36.
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[15]NEI M,LI W H.Mathematical Nodel for Studying Genetic Variation in Terms of Restriction Endonucleases[J].Proceedings of the National Academy of Sciences of the United States of America,1979,76(10):5269-5273.
[16]周延清.DNA分子标记技术在植物研究中的应用[M].北京:化学工业出版社,2005.
[17]DE MICCO V,ARENA C,PIGNALOSA D,et al.Effects of Sparsely and Densely Ionizing Radiation on Plants[J].Radiation and Environmental Biophysics,2011,50(1):1-19.
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[19]谭美莲,汪磊,严明芳,等.60Co伽马射线辐照对苏子生长及生理特性的影响[J].中国油料作物学报,2012,34(4):396-401.
[20]BIRKY C W,PERLMAN P S,BYERS T J.Genetics and Biogenesis of Mitochondria and Chloroplasts[M].Columbus:The Ohio State University Press,1975:268-308.
[21]CONNETT M B.Mechanisms of maternal inheritance of plastids and mitochondria:Developmental and ultrastructuralevidence[J].Plant Molecular Biology Report,1986,4(4):193-205.
[22]傅汀,王丽辉,林伟强,等.核基因突变引起的拟南芥菜叶片花斑及其机制[J].细胞生物学杂志,2005 27(5):505-508.
[23]YU Fei,FU Aigen,ALURU M,et al.Variegation Mutants and Mechanisms of Chloroplast Biogenesis[J].Plant Cell Environment,2007,30(3):350-365.
[24]GUHAMAJUMDAR M,BALDWIN S,SEARS BB.Chloroplast Mutations Induced by 9-aminoacridine Hydrochloride are Independent of the Plastome Mutator in Oenothera[J].Theoretical and Applied Genetics,2004,108(3),543-549.
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[27]KIRK J T O,TILNEY-BASSETT R A E.The Plastids[M].2nd ed.Amsterdam:Elsevier/North-Holland,1978.
[28]Naested H,Holm A,Jenkins T,et al.Arabidopsis VARIEGATED 3Encodes a Chloroplast-targeted,Zinc-finger Protein Required for Chloroplast and Palisade Cell Development[J].Journal of Cell Science,2004,117(Pt20):4807-4818.
[29]HSIEH M H,GOODMAN H M.The Arabidopsis Isp H Homolog is Involved in the Plastid Nonmevalonate Pathway of Isoprenoid Biosynthesis[J].Plant Physiology,2005,138(2):641-653.
[30]WALBOT V,COE E H.Nuclear Gene Iojap Conditions a Programmed Change to Ribosome-less Plastids in Zea Mays[J].Proceedings of the National Academy of Sciences of the United States of America,1979,76(6):2760-2764.
[31]AKOYUNOGLOU G.Regulation of Chloroplast Differentiation[M].New York:Alan R.Liss,1986:455-466.
[32]张兴芬,何忠俊,梁社往,等.60Co-γ辐照对三七叶片叶绿体和线粒体超微结构的影响[J].中国农学通报,2015,31(16):127-133.
[33]袁丽钗,李雪平,彭镇华,等.菲白竹组培苗白化、绿化突变体的超微结构及15个叶绿体编码基因的表达[J].植物学报,2010,45(4):451-459.
[34]李卫锋,王丹,黄海涛.电子束辐射羽衣甘蓝红鸥干种子对植物性状的影响[J].福建林业科技,2008,35(2):87-90.
[35]王新新,高健,周立人.60Co-γ射线辐照毛竹种子的过氧化物同工酶和RAPD分子检测的研究[J].激光生物学报,2010,19(6):758-763.
[36]吴立蓉,贺红,张燕玲,等.60Co-γ射线对广藿香的辐照诱变及再生植株的RAPD分析[J].激光生物学报,2010,19(2):174-178.
[37]张启明,周瑜,李佳,等.电子束辐照对睡莲植株的诱变效应及RAPD分析[J].世界科技研究与发展,2015,37(3):281-285.
[38]丁亮,陈睦传,沈明山,等.RAPD分析氮离子注入甜菊种子后的幼苗基因组DNA变异[J].生物物理学报,1999,15(4):798-803.
[39]李岩,董喜存,李文建,等.碳离子辐照对甜高梁幼苗保护酶同工酶表达的影响[J].安徽农业科学,2009,37(35):17440-17441,17444.
[40]王丹,张志伟,张冬雪,等.唐菖蒲电子束诱变作用同功酶及SDS-PAGE分析[J].北方园艺,2007,31(8):180-182.
[2]中国科学院中国植物志编辑委员会.中国植物志[M].北京:科学出版社,2004:669.
[3]方伟,邹云.竹子矮化与盆栽技艺[J].浙江林业科技,1989,9(2):32-39.
[4]张强.辐照技术在农业中的应用[J].农业工程,2012,2(S1):42-46.
[5]乔广军.诱变在我国亚麻育种及资源创新中的利用[J].中国麻业科学,2006,28(1):17-20.
[6]朱丽兵,周云清,丁捷,等.CANDU重水反应堆钴调节棒组件结构设计[J].原子能科学技术,2010,44(B09):418-422.
[7]黄训端,周立人,何家庆,等.60Coγ射线辐照花魔芋球茎的早期诱变效应研究[J].激光生物学报,2004,13(4):306-313.
[8]任少雄,王丹,李卫锋,等.60Coγ射线辐射唐菖蒲鳞茎诱变育种试验[J].福建林业科技,2006,33(2):34-36.
[9]胡瑞阳,吴博,纳静,等.60Co-γ射线辐照处理对杉木种子萌发及幼苗生长的影响[J].中国农业通报,2016,32(4):1-4.
[10]伏毅,黄敏,陈浩,等.高剂量60Co-γ射线辐照对紫花苜蓿种子的诱变及致死效应[J].湖北农业科学,2015,54(15):3708-3711.
[11]陈子元.从辐射育种的发展来展望航天育种的前景[J].核农学报,2002,16(5):261-263.
[12]WILLIAMS J G,KUBELIK A R,LIVAK K J,et al.DNA Polymorphisms Amplified by Arbitrary Primers are Useful as Genetic Markers[J].Nucleic Acids Research,1990,18(22):6531-6535.
[13]胡裕清,赵树进.RAPD技术及其在植物研究中的应用[J].生物技术通报,2010,26(5):74-77.
[14]赵莹,沈志敏,刘雄伟,等.斑马鱼分子遗传位标及多态性研究进展[J].实验动物与比较医学,2011,31(2):147-152.
[15]NEI M,LI W H.Mathematical Nodel for Studying Genetic Variation in Terms of Restriction Endonucleases[J].Proceedings of the National Academy of Sciences of the United States of America,1979,76(10):5269-5273.
[16]周延清.DNA分子标记技术在植物研究中的应用[M].北京:化学工业出版社,2005.
[17]DE MICCO V,ARENA C,PIGNALOSA D,et al.Effects of Sparsely and Densely Ionizing Radiation on Plants[J].Radiation and Environmental Biophysics,2011,50(1):1-19.
[18]翟敬宇.电子束辐照翠竹幼苗的诱变效应及RAPD分析[J].竹子研究汇刊,2013,32(3):22-27.
[19]谭美莲,汪磊,严明芳,等.60Co伽马射线辐照对苏子生长及生理特性的影响[J].中国油料作物学报,2012,34(4):396-401.
[20]BIRKY C W,PERLMAN P S,BYERS T J.Genetics and Biogenesis of Mitochondria and Chloroplasts[M].Columbus:The Ohio State University Press,1975:268-308.
[21]CONNETT M B.Mechanisms of maternal inheritance of plastids and mitochondria:Developmental and ultrastructuralevidence[J].Plant Molecular Biology Report,1986,4(4):193-205.
[22]傅汀,王丽辉,林伟强,等.核基因突变引起的拟南芥菜叶片花斑及其机制[J].细胞生物学杂志,2005 27(5):505-508.
[23]YU Fei,FU Aigen,ALURU M,et al.Variegation Mutants and Mechanisms of Chloroplast Biogenesis[J].Plant Cell Environment,2007,30(3):350-365.
[24]GUHAMAJUMDAR M,BALDWIN S,SEARS BB.Chloroplast Mutations Induced by 9-aminoacridine Hydrochloride are Independent of the Plastome Mutator in Oenothera[J].Theoretical and Applied Genetics,2004,108(3),543-549.
[25]NEWTON K J,COE E H.Mitochondrial DNA Changes in Abnormal Growth(Nonchromosomal Stripe)Mutants of Maize[J].Proceedings of the National Academy of Sciences of the United States of America,1986,83(19),7363-7366.
[26]NEWTON K J,KNUDSEN C,GABAY-LAUGHNAN S,et al.An Abnormal Growth Mutant in Maize has a Defective Mitochondrial Cytochrome Oxidase Gene[J].Plant Cell,1990,2(2):107-113.
[27]KIRK J T O,TILNEY-BASSETT R A E.The Plastids[M].2nd ed.Amsterdam:Elsevier/North-Holland,1978.
[28]Naested H,Holm A,Jenkins T,et al.Arabidopsis VARIEGATED 3Encodes a Chloroplast-targeted,Zinc-finger Protein Required for Chloroplast and Palisade Cell Development[J].Journal of Cell Science,2004,117(Pt20):4807-4818.
[29]HSIEH M H,GOODMAN H M.The Arabidopsis Isp H Homolog is Involved in the Plastid Nonmevalonate Pathway of Isoprenoid Biosynthesis[J].Plant Physiology,2005,138(2):641-653.
[30]WALBOT V,COE E H.Nuclear Gene Iojap Conditions a Programmed Change to Ribosome-less Plastids in Zea Mays[J].Proceedings of the National Academy of Sciences of the United States of America,1979,76(6):2760-2764.
[31]AKOYUNOGLOU G.Regulation of Chloroplast Differentiation[M].New York:Alan R.Liss,1986:455-466.
[32]张兴芬,何忠俊,梁社往,等.60Co-γ辐照对三七叶片叶绿体和线粒体超微结构的影响[J].中国农学通报,2015,31(16):127-133.
[33]袁丽钗,李雪平,彭镇华,等.菲白竹组培苗白化、绿化突变体的超微结构及15个叶绿体编码基因的表达[J].植物学报,2010,45(4):451-459.
[34]李卫锋,王丹,黄海涛.电子束辐射羽衣甘蓝红鸥干种子对植物性状的影响[J].福建林业科技,2008,35(2):87-90.
[35]王新新,高健,周立人.60Co-γ射线辐照毛竹种子的过氧化物同工酶和RAPD分子检测的研究[J].激光生物学报,2010,19(6):758-763.
[36]吴立蓉,贺红,张燕玲,等.60Co-γ射线对广藿香的辐照诱变及再生植株的RAPD分析[J].激光生物学报,2010,19(2):174-178.
[37]张启明,周瑜,李佳,等.电子束辐照对睡莲植株的诱变效应及RAPD分析[J].世界科技研究与发展,2015,37(3):281-285.
[38]丁亮,陈睦传,沈明山,等.RAPD分析氮离子注入甜菊种子后的幼苗基因组DNA变异[J].生物物理学报,1999,15(4):798-803.
[39]李岩,董喜存,李文建,等.碳离子辐照对甜高梁幼苗保护酶同工酶表达的影响[J].安徽农业科学,2009,37(35):17440-17441,17444.
[40]王丹,张志伟,张冬雪,等.唐菖蒲电子束诱变作用同功酶及SDS-PAGE分析[J].北方园艺,2007,31(8):180-182.