欢迎访问作物学报,今天是

作物学报 ›› 2014, Vol. 40 ›› Issue (10): 1767-1775.doi: 10.3724/SP.J.1006.2014.01767

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

花生属人工异源多倍体进化早期基因表达变化的cDNA-SCoT分析

贺梁琼1,2,3,熊发前1,唐秀梅1,蒋菁1,韩柱强1,钟瑞春1,高忠奎4,李忠1,何新华2,*,唐荣华1,*   

  1. 1广西农业科学院经济作物研究所,广西南宁 530007;2广西大学农学院,广西南宁 530004;3广西作物遗传改良生物技术重点开放实验室,广西南宁 530007;4广西农业科学院,广西南宁 530007
  • 收稿日期:2014-04-15 修回日期:2014-07-06 出版日期:2014-10-12 网络出版日期:2014-07-25
  • 通讯作者: 何新华, E-mail: honest66222@163.com; 唐荣华, E-mail: tronghua@163.com
  • 作者简介:何新华, E-mail: honest66222@163.com; 唐荣华, E-mail: tronghua@163.com
  • 基金资助:

    本研究由广西自然科学基金项目(2012GXNSFBA053051),广西农业科学院科技发展基金项目(桂农科2012JM15),国家自然科学基金项目(31160294,31240059),广西科学研究与技术开发计划项目(桂科能14121008-1-4),广西农业科学院基本科研业务专项项目(桂农科2014YZ05)和广西重点实验室建设项目(12-071-09)资助。

Analysis of Gene Expression Variation by cDNA-SCoT Technique at the Early Period of Arachis Artificial Allopolypoidy Evolution

HE Liang-Qiong1,2,3,XIONG Fa-Qian1,TANG Xiu-Mei1,JIANG Jing1,HAN Zhu-Qiang1,ZHONG Rui-Chun1,GAO Zhong-Kui4,Li Zhong1,HE Xin-Hua2,*,TANG Rong-Hua1,*   

  1. 1 Cash Crops Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, China; 2 Agricultural College of Guangxi University, Nanning 530004, China; 3 Guangxi Crop Genetic Improvement and Biotechnology Laboratory, Nanning 530007, China; 4 Guangxi Academy of Agricultural Sciences, Nanning 530007, China?
  • Received:2014-04-15 Revised:2014-07-06 Published:2014-10-12 Published online:2014-07-25
  • Contact: 何新华, E-mail: honest66222@163.com; 唐荣华, E-mail: tronghua@163.com
  • About author:何新华, E-mail: honest66222@163.com; 唐荣华, E-mail: tronghua@163.com

摘要:

为了探索花生属异源多倍体进化理论和种间杂交过程所涉及的遗传机制,以四倍体栽培种花生与二倍体野生种A. doigoi及其种间杂种F1和早期多倍体世代(S0~S3)为材料,采用cDNA-SCoT技术研究花生属人工异源多倍体进化早期基因表达变化规律。12SCoT引物共扩增出108cDNA片段,获得差异片段80个,占扩增总条带数的74.07%对其中的35个差异片段进行克隆测序,有26个和GenBank数据库中已录入的基因具有较高的相似性,包括能量与代谢相关基因(8)、未知功能蛋白基因(3)、抗逆性相关基因(4)、信号传导相关基因(2)和反转录转座子相关基因(9)。这说明花生属种间杂交人工异源多倍化早期世代发生着快速、剧烈的基因表达变化;从中获得的一些差异基因片段可用于花生属异源多倍化的分子机制研究。

关键词: 花生, 异源多倍化, cDNA-SCoT技术, 基因表达

Abstract:

To explore the allopolyploidy evolutionism and the genetic mechanism of Arachis interspecific hybridization, study the gene expression variation by cDNA-SCoT technique in the early period of Arachis artificial allopolypoidy evolution, using the progenitors, F1 and early polyploidy generations (S0 to S3) of the hybridization between tetraploid cultivated peanut and diploid wild peanut A. doigoi. Among 108 cDNA fragments amplified by 12 SCoT primers 80 were differentially expressed with the polymorphism frequency of 74.07%. Among 80 TDFs (transcripts derived fragments) 35 were cloned and sequenced, and the sequences of 26 TDFs shared high similarity with the genes documented in the GenBank. These genes included energy and metabolism-related genes (8), resistance-related genes (4), unknown functional protein genes (3), signal transduction-related genes (2) and retrotransposon-related genes (9). The results indicated that gene expression changes happened rapidly and drastically in the early generations during artificial allopolyploidization of peanut interspecific hybridization, and some obtained TDFs probably could be used in the research of molecular mechanism of Arachis allopolyploidization.

Key words: Peanut, Allopolyploidization, cDNA-SCoT technique, Gene expression

[1]Otto S P. The evolutionary consequences of polyploidy. Cell, 2007, 131: 452–462



[2]宋灿, 刘少军, 肖军, 何伟国, 周毅, 覃钦博, 张纯, 刘筠. 多倍体生物研究进展. 中国科学C辑(生命科学), 2012, 42: 173–184



Song C, Liu S J, Xiao J, He W G, Zhou Y, Qin Q B, Zhang C, Liu J. Polyploidy organisms. Sci China (Ser C: Life Sci), 2012, 42: 173–184 (in Chinese with English abstract)



[3]赵旭博, 李爱丽, 毛龙. 植物多倍化过程中小分子RNA调控基因表达机制研究进展. 作物学报, 2013, 39: 1331–1338



Zhao X B, Li A L, Mao L. Progress on gene regulatory mechanisms by small RNAs during plant polyploidization. Acta Agron Sin, 2013, 39: 1331–1338 (in Chinese with English abstract)



[4]Jiao Y, Wickett N J, Ayyampalayam S, Chanderbali A S, Landherr L, Ralph P E, Tomsho L P, Hu Y, Liang H, Soltis P S, Soltis D E, Clifton S W, Schlarbaum S E, Schuster S C, Ma H, Leebens-Mack J, Pamphilis C W. Ancestral polyploidy in seed plants and angiosperms. Nature, 2011, 473: 97–100



[5]Feldman M, Levy A A. Genome evolution due to allopolyploidization in wheat. Genetics, 2012, 192: 763–774



[6]Adams K L, Wendel J F. Polyploidy and genome evolution in plants. Curr Opin Plant Biol, 2005, 8: 135–141



[7]Higgins J, Magusin A, Trick M, Fraser F, Bancroft I. Use of mRNA-seq to discriminate contributions to the transcriptome from the constituent genomes of the polyploid crop species Brassica napus. BMC Genomics, 2012, 13: 247



[8]彭海, 张静, 吴先军. 植物基因表达中的倍性效应: 研究进展、问题与展望. 中国科学C辑(生命科学), 2008, 38: 1–7



Peng H, Zhang J, Wu X J. The ploidy effect of plant gene expression: research progress, problem and expectation. Sci China (Ser C: Life Sci), 2008, 38: 1–7 (in Chinese with English abstract)



[9]蔡得田, 陈建国, 陈冬玲, 戴兵成, 张维, 宋兆建, 杨之帆, 杜超群, 唐志强, 何玉池, 张道生, 何光存, 朱英国. 两个具多倍体减数分裂稳定性的多倍体水稻品系的选育. 中国科学C辑(生命科学), 2007, 37: 217–226



Cai D T, Chen J G, Chen D L, Dai B C, Zhang W, Song Z J, Yang Z F, Du C Q, Tamg Z Q, He Y C, Zhang D S, He G C, Zhu Y G. Breeding of two polyploidy rice strains with meiotic stability. Sci China (Ser C: Life Sci), 2007, 37: 217–226 (in Chinese with English abstract)



[10]周汉群, 唐荣华, 周翠球, 钟瑞春, 韩柱强. 花生亲和种远缘杂交育种研究. 花生学报, 2003, 32(suppl): 155–161



Zhou H Q, Tang R H, Zhou C Q, Zhong R C, Han Z Q. Interspecific hybridization between cultivated peanut and compatible wild species in Arachis. J Peanut Sci, 2003, 32(suppl): 155–161 (in Chinese with English abstract)



[11]王纪娟, 杨淑娟, 任兰柱. 植物远缘杂交及其在小麦育种的应用. 粮食作物, 2010, (6): 109–113



Wang J J, Yang S J, Ren L Z. Wide hybridization of plant and its application in wheat breeding. Grain Crops, 2010, (6): 109–113 (in Chinese with English abstract)



[12]Lu J, Zhang C, Baulcombe D C, Chen Z J. Maternal siRNAs as regulators of parental genome imbalance and gene expression in endosperm of Arabidopsis seeds. Proc Nat Acad Sci USA, 2012, 109: 5529–5534



[13]Kenan-Eichler M, Leshkowitz D, Tal L, Noor E, Melamed-Bessudo C, Feldman M, Levy A A. Wheat hybridization and polyploidization results in deregulation of small RNAs. Genetics, 2011, 188: 263–272



[14]Miller M, Zhang C, Chen Z J. Ploidy and hybridity effects on growth vigor and gene expression in Arabidopsis thaliana hybrids and their parents. Genes Genomes Genet, 2012, 2: 505–513



[15]Pumphrey M, Bai J, Laudencia-Chingcuanco D, Anderson O, Gill B S. Nonadditive expression of homoeologous genes is established upon polyploidization in hexaploid wheat. Genetics, 2009, 181: 1147–1157



[16]Chen Z J. Genetic and epigenetic mechanisms for gene expression and phenotypic variation in plant polyploids. Annu Rev Plant Biol, 2007, 58: 377–406



[17]Wang J, Tian L, Lee H S, Wei N E, Jiang H, Watson B, Madlung A, Osborn T C, Doerge R W, Comai L, Chen Z J. Genome wide nonadditive gene regulation in Arabidopsis allotetraploids. Genetics, 2006, 172: 507–517



[18]Kashkush K, Feldman M, Levy A A. Gene loss, silencing and activation in a newly synthesized wheat allotetraploid. Genetics, 2002, 160: 1651–1659



[19]He P, Friebe B R, Gill B S, Zhou J M. Allopolyploidy alters gene expression in the highly stable hexaploid wheat. Plant Mol Biol, 2003, 52: 401–414



[20]Adams K L, Percifield R. Wendel J F. Organ-specific silencing of duplicated genes in a newly synthesized cotton allotetraploid. Genetics, 2004, 168: 2217–2226



[21]Albertin W, Balliau T, Brabant P, Chevre A M, Eber F, Malosse C, Thiellement H. Numerous and rapid nonstochastic modifications of gene products in newly synthesized Brassica napus allotetraploids. Genetics, 2006, 173: 1101–1113



[22]Hegarty M J, Jones J M, Wilson I D, Barker G L, Coghill J A, Sanchez-Baracaldo P, Liu G, Buggs R J, Abbott R J, Edwards K J, Hiscock S J. Development of anonymous cDNA microarrays to study changes to the Senecio floral transcriptome during hybrid speciation. Mol Ecol, 2005, 14: 2493–2510



[23]Zhuang Y, Chen J F. Changes of gene expression in early generations of the synthetic allotetraploid Cucumis×hytivus Chen et Kirkbride. Genet Resour Crop Evol, 2009, 56: 1071–1076



[24]Kochert G, Halward T, Branchi W D. RFLP variability in peanut (Arachis hypogaea L.) cultivars and wild species. Theor Appl Genet, 1991, 81: 565–570



[25]Gimenes M A, Lopes C R, Valls F M. Genetic relationships among Arachis species based on AFLP. Genet Mol Biol, 2002, 25: 349–353



[26]Gimenes M A, Lopes C R, Galgaro M L, Valls J, Kochert G. RFLP analysis of genetic variation in species of section Arachis, genus Arachis (Leguminosae). Euphytica, 2002, 123: 421-429



[27]任小平, 廖伯寿, 黄家权, 张晓杰, 姜慧芳. 利用SRAP标记分析花生属花生区组种质亲缘关系. 中国油料作物学报, 2009, 31: 449–454



Ren X P, Liao B S, Huang J Q, Zhang X J, Jiang H F. Genomic affinities of Arachis section Arachis revealed by SRAP markers. Chin J Oil Crop Sci, 2009, 31: 449–454 (in Chinese with English abstract)



[28]Garcia G M, Tallury S, Kochert G S. Molecular analysis of Arachis interspecific hybrids. Theor Appl Genet, 2006, 112: 1342–1348



[29]贺梁琼, 熊发前, 钟瑞春, 韩柱强, 李忠, 唐秀梅, 蒋菁, 唐荣华, 何新华. 利用SCoT标记分析花生栽培种×A. chacoensis组合异源多倍化的早期基因组变化. 中国农业科学, 2013, 46: 1555–1563



He L Q, Xiong F Q, Zhong R C, Han Z Q, Li Z, Tang X M, Jiang J, Tang R H, He X H. Study on genome variations by using SCoT markers during allopolyploidization of the cultivated peanut × A. chacoensis. Sci Agric Sin, 2013, 46: 1555–1563 (in Chinese with English abstract)



[30]贺梁琼, 熊发前, 韩柱强, 韩柱强, 蒋菁, 唐秀梅, 李忠, 何新华, 唐荣华. 花生种间杂种异源多倍化早期世代性状和微卫星变化研究. 中国油料作物学报, 2013, 35: 499–507



He L Q, Xiong F Q, Han Z Q, Zhong R C, Jiang J, Tang X M, Li Z, He X H, Tang R H. Traits and microsatellites variation research of early generations during allopolyploidization in Arachis interspecific hybridization. Chin J Oil Crop Sci, 2013, 35: 499–507 (in Chinese with English abstract)



[31]Collard B C Y, Mackill D J. Start codon targeted (SCoT) polymorphism: A simple, novel DNA marker technique for generating gene-targeted markers in plants. Plant Mol Biol Rep, 2009, 27: 86–93



[32]Xiong F Q, Zhong R C, Han Z Q, Jiang J, He L Q, Zhuang W J, Tang R H. Start codon targeted polymorphism for evaluation of functional genetic variation and relationships in cultivated peanut (Arachis hypogaea L.) genotypes. Mol Biol Rep, 2011, 38: 3487–3494



[33]熊发前, 唐荣华, 陈忠良, 潘玲华, 庄伟建. 目标起始密码子多态性(SCoT): 一种基于翻译起始位点的目的基因标记新技术. 分子植物育种, 2009, 7: 635–638



Xiong F Q, Tang R H, Chen Z L, Pan L H, Zhuang W J. Start codon target polymorphism (SCoT): a novel gene targeted marker technique based on the translation start codon. Mol Plant Breed, 2009, 7: 635–638 (in Chinese with English abstract)



[34]熊发前, 蒋菁, 钟瑞春, 韩柱强, 贺梁琼, 李忠, 庄伟建, 唐荣华. 目标起始密码子多态性(SCoT)分子标记技术在花生属中的应用. 作物学报, 2010, 36: 2055–2061



Xiong F Q, Jiang J, Zhong R C, Han Z Q, He L Q, Li Z, Zhuang W J, Tang R H. Application of SCoT molecular marker in genus Arachis. Acta Agron Sin, 2010, 36: 2055–2061 (in Chinese with English abstract)



[35]陈虎, 何新华, 罗聪, 高美萍. 龙眼24个品种的SCoT遗传多样性分析. 园艺学报, 2010, 37: 1651–1654



Chen H, He X H, Luo C, Gao M P. Analysis on the genetic diversity of 24 longan (Dimocarpus longan) accessions by SCoT markers. Acta Hortic Sin, 2010, 37: 1651–1654 (in Chinese with English abstract)



[36]Luo C, He X H, Chen H, Ou S J, Gao M P, Brown J S, Tondo C T, Schnell R J. Genetic diversity of mango cultivars estimated using SCoT and ISSR markers. Biochem Syst Ecol, 2011, 39: 676–684



[37]Luo C, He X H, Chen H, Hu Y, Ou S J. Genetic relationship and diversity of Mangifera indica L.rrevealed through SCoT analysis. Genet Resour Crop Evol, 2012, 59: 1505–1515



[38]陈香玲, 苏伟强, 刘业强, 任惠, 陆玉英. 36份菠萝种质的遗传多样性SCoT分析. 西南农业学报, 2012, 25: 625–629



Chen X L, Su W Q, Liu Y Q, Ren H, Lu Y Y. Analysis on genetic diversity of 36 pineapple collections by SCoT markers. Southwest China J Agric Sci, 2012, 25: 625–629 (in Chinese with English abstract)



[39]吴建明, 李杨瑞, 王爱琴, 杨柳, 杨丽涛. 赤霉素诱导甘蔗节间伸长基因的 cDNA-SCoT差异表达分析. 作物学报, 2010, 36: 1883–1890



Wu J M, Li Y R, Wang A Q, Yang L, Yang L T. Differential expression of genes in gibberellin-induced stalk elongation of sugarcane analyzed with cDNA-SCoT. Acta Agron Sin, 2010, 36: 1883–1890 (in Chinese with English abstract)



[40]陈香玲, 李杨瑞, 杨丽涛, 吴建明, 罗聪, 熊发前, 杨柳. 低温胁迫下甘蔗抗寒相关基因的cDNA-SCoT差异显示. 生物技术通报, 2010, (8): 120–124



Chen X L, Li Y R, Yang L T, Wu J M, Luo C, Xiong F Q, Yang L. cDNA-SCoT differential display of cold resistance related genes in sugarcane under low temperature stress. Biotechnol Bull, 2010, (8): 120–124 (in Chinese with English abstract)



[41]熊发前, 刘俊仙, 王丛丛, 蒋菁, 钟瑞春, 韩柱强, 贺梁琼, 李忠, 唐秀梅, 唐荣华. mCTAB-dLiCl法高效提取花生各组织部位RNA及其验证. 南方农业学报, 2013, 44: 1781–1784



Xiong F Q, Liu J X, Wang C C, He L Q, Han Z Q, Jiang J, Zhong R C, Li Z, Tang X M, Tang R H. mCTAB-dLiCl method for efficiently extracting RNA from various tissues of cultivated peanut and its verification. J Southern Agric, 2013, 44: 1781–1784 (in Chinese with English abstract)



[42]Comai L, Tyagi A P, Winter K, Holmes-Davis R, Reynolds S H, Stevens Y, Byers B. Phenotypic instability and rapid gene silencing in newly formed Arabidopsis allotetraploids. Plant Cell, 2000, 12: 1551–1567



[43]Hegarty M J, Barker G L, Wilson I D, Abbott R J, Edwards K J, Hiscock S J. Transcriptome shock after interspecific hybridization in Senecio is ameliorated by genome duplication. Curr Biol, 2006, 16: 1652–1659



[44]Auger D L, Gray A D, Ream T S, Kato A, Jr. E H C, Birchler J A. Nonadditive gene expression in diploid and triploid hybrids of maize. Genetics, 2005, 169: 389–397



[45]Bennetzen J L. Transposable element contributions to plant gene and genome evolution. Plant Mol Biol, 2000, 42: 251–269

[1] 郑玉珍, 齐飞艳, 孙子淇, 刘华, 秦利, 石磊, 王娟, 汪蒙蒙, 韩锁义, 徐静, 苗利娟, 黄冰艳, 董文召, 郑峥, 张新友. 花生籽仁总超长链脂肪酸和7种脂肪酸组分的QTL定位[J]. 作物学报, 2026, 52(6): 1646-1657.
[2] 陆一楚, 李振影, 买春海, 赵晓蕊, 王利祥. 夜间复合体关键基因AhLUX1正向调控花生结瘤的功能研究[J]. 作物学报, 2026, 52(6): 1658-1668.
[3] 谷春苗, 王润风, 黄璐, 刘浩, 鲁清, 李海芬, 李少雄, 何双呈, 洪彦彬, 陈小平, 谭斌, 余倩霞. 花生WOX基因家族的全基因组分析及不定芽再生候选基因的鉴定[J]. 作物学报, 2026, 52(5): 1326-1340.
[4] 于天一, 王春晓, 肖丽, 钟召迪, 王宣仓, 赵勇, 路亚, 吴月, 吴正锋. 不同结瘤特性花生品种氮素累积、产量及品质特性对氮肥用量的响应[J]. 作物学报, 2026, 52(3): 881-894.
[5] 詹戈锐, 余文, 李锋, 武明珠, 徐馨, 罗朝鹏, 巫升鑫, 杨军, 张智强, 王中. 烟草NtWRKY6基因响应ABA表达及其调控多酚合成的功能研究[J]. 作物学报, 2026, 52(2): 446-458.
[6] 张胜忠, 李国卫, 戈立江, 王菲菲, 胡晓辉, 苗华荣, 李燕, 钟文, 陈静. 花生机械脱壳损伤相关农艺指标筛选与QTL定位[J]. 作物学报, 2026, 52(2): 644-652.
[7] 王菲菲, 张胜忠, 杨贵华, 苗华荣, 胡晓辉, 张则林, 刘莎莎, 乔利仙, 单世华, 陈静. 331份花生种质苗期耐盐性综合评价和强耐盐种质鉴选[J]. 作物学报, 2026, 52(1): 279-294.
[8] 金欣欣, 宋亚辉, 苏俏, 杨永庆, 王瑾. 高产高油高油酸花生品种的生长发育及干物质生产特征[J]. 作物学报, 2026, 52(1): 191-201.
[9] 迟晓元, 刘庆, 张君, 赵旭红, 李美, 于天一, 潘丽娟, 许静, 姜骁, 殷祥贞, 马俊卿, 陈娜. 不同花生品种(系)耐盐碱性田间鉴定及各性状指标相关性研究[J]. 作物学报, 2026, 52(1): 85-98.
[10] 孙辰硕, 张月, 田泽锴, 晏立英, 康彦平, 陈玉宁, 王欣, 淮东欣, 王前前, 姜慧芳, 罗怀勇, 黄莉, 廖伯寿, 王志慧, 雷永. 花生种质果柄强度的遗传分化与主要影响因子分析[J]. 作物学报, 2026, 52(1): 118-130.
[11] 景秀清, 蔡永朵, 邓宁, 赵晓东, 翟飞红, 曾群. 藜麦RopGEF家族基因的鉴定及表达模式分析[J]. 作物学报, 2026, 52(1): 28-43.
[12] 万书波, 张佳蕾, 高华鑫, 王才斌. 中国花生高产栽培研究进展与展望[J]. 作物学报, 2025, 51(7): 1703-1711.
[13] 郭腾达, 崔梦杰, 陈琳杰, 韩锁义, 郭敬坤, 吴晨迪, 付留洋, 黄冰艳, 董文召, 张新友. 花生磷脂酰肌醇转运蛋白基因AhSFH的克隆及其响应黄曲霉菌侵染的表达特征分析[J]. 作物学报, 2025, 51(6): 1489-1500.
[14] 李文佳, 廖泳俊, 黄璐, 鲁清, 李少雄, 陈小平, 金晶炜, 王润风. 花生开花时间的全基因组关联分析及候选基因筛选[J]. 作物学报, 2025, 51(5): 1400-1408.
[15] 林伟津, 郭泽佳, 刘浩, 李海芬, 王润风, 黄璐, 余倩霞, 陈小平, 洪彦彬, 李少雄, 鲁清. 花生荚果产量相关性状QTL定位与候选基因分析[J]. 作物学报, 2025, 51(4): 969-981.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!