不同辣椒品种果实发育过程中DNA甲基化的MSAP分析
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摘要
【目的】分析辣椒GB14和GB38果实发育过程中酶切位点的甲基化,为在分子水平上研究辣椒红素调控规律、在辣椒遗传育种中奠定理论基础。【方法】对辣椒果实发育成熟过程中的DNA甲基化修饰进行分析。【结果】DNA半甲基化条带比率与全甲基化条带比率均随着籽粒的生长发育而逐渐升高,两种辣椒果实绿熟期和红熟期基因组总体甲基化比例分别为36. 51%、36. 38%、35. 57%和35. 45%。对甲基化表现呈多态性变化的8个片段回收、测序分析表明,DNA甲基化发生的位点既存在于基因组的编码区,也存在于非编码区。【结论】在辣椒果实发育过程中,DNA甲基化修饰发生了巨大的变化; CCGG位点的半甲基化水平与辣椒红素积累呈正比。
【Objective】This project aims to carry out methylation analysis of restriction endonuclease sites in GB14 and GB38 fruit development of capsicum in the hope of providing a new insight for exploring the molecular mechanism of capsicum fruit development.【Method】The modification of DNA methylation during the development and maturation of Capsicum annuumL. fruit was studied. 【Result】The data showed that the ratios of DNA methylation and total methylation bands both increased gradually with seeds growth and development. The total methylation ratio of the two varieties of Capsicum annuumL. in green maturity period and red ripening period were 36. 51%,36. 38%,35. 57% and 35. 45%,respectively. Eight polymorphic fragments of methylation showed that the DNA methylation sites existed both in the coding region of the genome and in the non-coding region by the recovery and sequencing.【Conclusion】During the development of capsicum fruit,the modification of DNA methylation was greatly changed and the level of semi-methylation of CCGG loci was positively related to the accumulation of Capsicum annuumL. These results suggest a molecular level method for further studying the regulation of Capsicum annuumL.,which provide a theoretical foundation for the breeding of Capsicum annuumL.
【Objective】This project aims to carry out methylation analysis of restriction endonuclease sites in GB14 and GB38 fruit development of capsicum in the hope of providing a new insight for exploring the molecular mechanism of capsicum fruit development.【Method】The modification of DNA methylation during the development and maturation of Capsicum annuumL. fruit was studied. 【Result】The data showed that the ratios of DNA methylation and total methylation bands both increased gradually with seeds growth and development. The total methylation ratio of the two varieties of Capsicum annuumL. in green maturity period and red ripening period were 36. 51%,36. 38%,35. 57% and 35. 45%,respectively. Eight polymorphic fragments of methylation showed that the DNA methylation sites existed both in the coding region of the genome and in the non-coding region by the recovery and sequencing.【Conclusion】During the development of capsicum fruit,the modification of DNA methylation was greatly changed and the level of semi-methylation of CCGG loci was positively related to the accumulation of Capsicum annuumL. These results suggest a molecular level method for further studying the regulation of Capsicum annuumL.,which provide a theoretical foundation for the breeding of Capsicum annuumL.
引文
[1]李艳,王亮,刘志刚.新疆绿洲干旱区制干辣椒生产技术现状与产业发展对策[J].北方园艺,2014,(13):189-192.LI Yan,WANG Liang,LIU Zhi-gang.(2014). The production status and development strategy of dried chili in Xinjiang oasis of arid area[J]. Northern Horticulture,(13):189-192.(in Chinese)
[2] Zhao,G.,Zhang,W.,&Zhang,G.(2010). Production of single cell protein using waste capsicum powder produced during capsanthin extraction. Letters in Applied Microbiology,50(2):187-191.
[3]Ha,S. H.,Kim,J. B.,Park,J. S.,Lee,S. W.,&Cho,K. J.(2007). A comparison of the carotenoid accumulation in capsicum varieties that show different ripening colors:deletion of the. Journal of Experimental Botany,58(12):3,135-3,144.
[4]Brand,A.,Borovsky,Y.,Meir,S.,Rogachev,I.,Aharoni,A.,&Paran,I.(2012). Pc8. 1,a major qtl for pigment content in pepper fruit,is associated with variation in plastid compartment size. Planta,235(3):579-588.
[5]Meng,D.,Dubin,M.,Zhang,P.,Osborne,E. J.,Stegle,O.,&Clark,R. M.,et al.(2016). Limited contribution of dna methylation variation to expression regulation in Arabidopsis thaliana. Plos Genetics,12(7):e1006141.
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[7]Zhang,X.,Li,Q.,Kong,L.,&Yu,H.(2018). Epigenetic variation of wild populations of the pacific oyster crassostrea gigas,determined by methylation-sensitive amplified polymorphism analysis. Fisheries Science,84(1):61-70.
[8]Bock,C.,Tomazou,E. M.,Brinkman,A.,Müller,F.,Simmer,F.,&Gu,H.,et al.(2010). Genome-wide mapping of dna methylation:a quantitative technology comparison. Nature Biotechnology,28(10):1,106-1,114.
[9]Peng,C.,Sui,Z.,Zhou,W.,Hu,Y.,Mi,P.,&Jiang,M.,et al.(2018). Analysis of dna methylation of gracilariopsis lemaneiformis,under temperature stress using the methylation sensitive amplification polymorphism(msap)technique. Journal of Ocean University of China,17(3):623-631.
[10]Seymour,D. K.,&Becker,C.(2017). The causes and consequences of dna methylome variation in plants. Current Opinion in Plant Biology,(36):56-63.
[11]Wang,W.,Zhao,X.,Pan,Y.,Zhu,L.,Fu,B.,&Li,Z.(2011). Dna methylation changes detected by methylationsensitive amplified polymorphism in two contrasting rice genotypes under salt stress. Journal of Genetics and Genomics,38(9):419-424.
[12]Allawati,A.,Albahry,S.,Victor,R.,Allawati,A. H.,&Yaish,M. W.(2016). Salt stress alters dna methylation levels in alfalfa(medicago spp). Genetics&Molecular Research Gmr,15(1):15018299.
[13]任茂,李博,徐延浩,等.高温胁迫诱导棉花甲基化变化分析[J].分子植物育种,2017,15(3):1 069-1 076.REN Mao,LI Bo,XU Yan-hao,et al.(2017). Methylationsensitive amplified polymorphism analysis of epigenetic changes in cotton(Gossypium hirsutum L.)under heat stress[J]. Molecular Plant Breeding,15(3):1,069-1,076.(in Chinese)
[14]李廷春,杨华应,张玮,等.紫玉米籽粒呈色过程中DNA甲基化变化的MSAP分析[J].核农学报,2014,28(11):1 978-1 984.LI Ting-chun,YANG Hua-ying,ZHANG Wei,et al.(2014).MSAP Analysis of DNA Methylation Variations during Coloring Process for Purple Corn Seed[J]. Journal of Nuclear Agricultural Sciences,28(11):1,978-1,984.(in Chinese)
[15]Bitonti,M. B.,Cozza,R.,Chiappetta,A.,Giannino,D.,Ruffini,C. M.,&Dewitte,W.,et al.(2002). Distinct nuclear organization,dna methylation pattern and cytokinin distribution mark juvenile,juvenile-like and adult vegetative apical meristems in peach(prunus persica(l.)batsch). Journal of Experimental Botany,53(371):1,047-1,054.
[16] Wang,S.,Luo,L.,Jin,Q.,Jin,L.,Tan,K.,&Liu,C.,et al.(2017). Analysis of DNA Methylation changes during the late fruit development stage in luotian persimmon. Agricultural Biotechnology,(4):6-9.
[17]Ausin,I.,Greenberg,M. V. C.,Simanshu,D. K.,Hale,C. J.,Vashisht,A. A.,&Simon,S. A.,et al.(2012). Involved in de novo 2-containing complex involved in RNA-directed dna methylation in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America,109(22):8,374-8,381.
[2] Zhao,G.,Zhang,W.,&Zhang,G.(2010). Production of single cell protein using waste capsicum powder produced during capsanthin extraction. Letters in Applied Microbiology,50(2):187-191.
[3]Ha,S. H.,Kim,J. B.,Park,J. S.,Lee,S. W.,&Cho,K. J.(2007). A comparison of the carotenoid accumulation in capsicum varieties that show different ripening colors:deletion of the. Journal of Experimental Botany,58(12):3,135-3,144.
[4]Brand,A.,Borovsky,Y.,Meir,S.,Rogachev,I.,Aharoni,A.,&Paran,I.(2012). Pc8. 1,a major qtl for pigment content in pepper fruit,is associated with variation in plastid compartment size. Planta,235(3):579-588.
[5]Meng,D.,Dubin,M.,Zhang,P.,Osborne,E. J.,Stegle,O.,&Clark,R. M.,et al.(2016). Limited contribution of dna methylation variation to expression regulation in Arabidopsis thaliana. Plos Genetics,12(7):e1006141.
[6]林国卫,王爱斌,石光禹,等.黄独冻后培养胚性愈伤组织DNA甲基化的MSAP分析[J].分子植物育种,2018,16(5):1 590-1 597.LIN Guo-wei,WANG Ai-bin,SHI Guang-yu,et al.(2018).MSAP analysis of DNA methylation of Dioscorea bulbifera L. embryogenic calli cultured after cryopreservation molecular[J].Plant Breeding,16(5):1,590-1,597.(in Chinese)
[7]Zhang,X.,Li,Q.,Kong,L.,&Yu,H.(2018). Epigenetic variation of wild populations of the pacific oyster crassostrea gigas,determined by methylation-sensitive amplified polymorphism analysis. Fisheries Science,84(1):61-70.
[8]Bock,C.,Tomazou,E. M.,Brinkman,A.,Müller,F.,Simmer,F.,&Gu,H.,et al.(2010). Genome-wide mapping of dna methylation:a quantitative technology comparison. Nature Biotechnology,28(10):1,106-1,114.
[9]Peng,C.,Sui,Z.,Zhou,W.,Hu,Y.,Mi,P.,&Jiang,M.,et al.(2018). Analysis of dna methylation of gracilariopsis lemaneiformis,under temperature stress using the methylation sensitive amplification polymorphism(msap)technique. Journal of Ocean University of China,17(3):623-631.
[10]Seymour,D. K.,&Becker,C.(2017). The causes and consequences of dna methylome variation in plants. Current Opinion in Plant Biology,(36):56-63.
[11]Wang,W.,Zhao,X.,Pan,Y.,Zhu,L.,Fu,B.,&Li,Z.(2011). Dna methylation changes detected by methylationsensitive amplified polymorphism in two contrasting rice genotypes under salt stress. Journal of Genetics and Genomics,38(9):419-424.
[12]Allawati,A.,Albahry,S.,Victor,R.,Allawati,A. H.,&Yaish,M. W.(2016). Salt stress alters dna methylation levels in alfalfa(medicago spp). Genetics&Molecular Research Gmr,15(1):15018299.
[13]任茂,李博,徐延浩,等.高温胁迫诱导棉花甲基化变化分析[J].分子植物育种,2017,15(3):1 069-1 076.REN Mao,LI Bo,XU Yan-hao,et al.(2017). Methylationsensitive amplified polymorphism analysis of epigenetic changes in cotton(Gossypium hirsutum L.)under heat stress[J]. Molecular Plant Breeding,15(3):1,069-1,076.(in Chinese)
[14]李廷春,杨华应,张玮,等.紫玉米籽粒呈色过程中DNA甲基化变化的MSAP分析[J].核农学报,2014,28(11):1 978-1 984.LI Ting-chun,YANG Hua-ying,ZHANG Wei,et al.(2014).MSAP Analysis of DNA Methylation Variations during Coloring Process for Purple Corn Seed[J]. Journal of Nuclear Agricultural Sciences,28(11):1,978-1,984.(in Chinese)
[15]Bitonti,M. B.,Cozza,R.,Chiappetta,A.,Giannino,D.,Ruffini,C. M.,&Dewitte,W.,et al.(2002). Distinct nuclear organization,dna methylation pattern and cytokinin distribution mark juvenile,juvenile-like and adult vegetative apical meristems in peach(prunus persica(l.)batsch). Journal of Experimental Botany,53(371):1,047-1,054.
[16] Wang,S.,Luo,L.,Jin,Q.,Jin,L.,Tan,K.,&Liu,C.,et al.(2017). Analysis of DNA Methylation changes during the late fruit development stage in luotian persimmon. Agricultural Biotechnology,(4):6-9.
[17]Ausin,I.,Greenberg,M. V. C.,Simanshu,D. K.,Hale,C. J.,Vashisht,A. A.,&Simon,S. A.,et al.(2012). Involved in de novo 2-containing complex involved in RNA-directed dna methylation in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America,109(22):8,374-8,381.