欢迎访问作物学报,今天是

作物学报 ›› 2017, Vol. 43 ›› Issue (03): 454-463.doi: 10.3724/SP.J.1006.2017.00454

• 研究简报 • 上一篇    下一篇

低温胁迫下马铃薯的数字基因表达谱分析

杨慧菊,郭华春*   

  1. 云南农业大学农学与生物技术学院薯类作物研究所,云南昆明 650201
  • 收稿日期:2016-07-06 修回日期:2016-11-02 出版日期:2017-03-12 网络出版日期:2016-11-18
  • 通讯作者: 郭华春,E-mail: ynghc@126.com;Tel: 0871-65227728
  • 基金资助:

    本研究由国家自然科学基金项目(C130407), 国家现代农业产业技术体系建设专项(GARS-10)和云南省重大科技专项(2013ZA007)项目资助。

Digital Gene Expression Profiling Analysis of Potato Under Low Temperature Stress

YANG Hui-Ju,GUO Hua-Chun*   

  1. Tuber-Root Crops Research Institute, College of Agronomy and Biotechnology, Yunnan Agriculture University, Kunming650201, China
  • Received:2016-07-06 Revised:2016-11-02 Published:2017-03-12 Published online:2016-11-18
  • Contact: 郭华春,E-mail: ynghc@126.com;Tel: 0871-65227728
  • Supported by:

    This study was supported by the National Natural Science Foundation of China(C130407), the China Agriculture Research System(GARS-10) and Specialized Research Fund for science and technology projects of Yunnan province(2013ZA007).

摘要:

以马铃薯品种合作88为材料,利用数字基因表达谱(DGE)技术,对–2℃低温胁迫处理后的马铃薯叶片cDNA文库进行差异基因表达谱分析。结果表明,有28505个基因受低温胁迫诱导差异表达,其中上调表达基因13703个,下调表达基因14802个。GO功能显著性富集分析表明,DEGs主要涉及信号生物代谢过程、氧化还原过程、能量代谢、次生代谢过程以及催化活性。KEGG富集分析表明,上调表达基因主要富集于苯丙烷、光合作用天线蛋白、类胡萝卜素的生物合成、苯丙氨酸代谢及淀粉与蔗糖代谢途径,而下调表达基因主要富集于植物激信号转导途径。利用实时荧光定量PCR(qRT-PCR)验证4DEGs在低温胁迫条件下的差异表达,其结果与DGE分析结果基本一致,证实了DGE测序结果的可靠性。

关键词: 马铃薯, 低温胁迫, 基因表达谱, 差异表达基因

Abstract:

Avariety Cooperation88was used to construct leaf cDNA library of potato treated with –2℃ low temperature stress by digital gene expression profiling (DGE) technology, and detect the differentially expressed genes(DEGs).The results of solexasequencing showed that a total of 28505 DEGs were screened out, 13703 DEGs of whichwere up-regulated and 14802 DEGs were down-regulated. GO enrichment analysis showed that these DEGs were involved insingle-organism metabolic process, oxidation-reduction process, metabolisms of carbohydrate, secondary metabolism and catalytic activity. The KEGGenrichment analysis showed that up-regulated genes were significantly associated withphenylpropanoid, photosynthesis-antennaproteins, carotenoidbiosynthesis, phenylalaninemetabolism, starch and sucrose metabolism pathways, while the down-regulated DEGs mainly enriched in plant hormone signal transduction pathway. The differential expressions of four genes were confirmed by Real-time quantitative PCR, showing that theexpression changes ofthese four genes weregenerally consistent with the results of DGE.

Key words: Potato, Low temperature stress, Gene expression profile, Differentially expressed genes

[1]Boyer J S. Plant productivity and environment. Science, 1982, 218: 443–448
[2]张珍. 马铃薯早熟防寒栽培技术. 河南农业, 2013, (3): 46
Zhang Z.Early winter cultivation techniques of potato. AgricHenan, 2013, (5): 46 (in Chinese with English abstract)
[3]ChenY K, Bamberg J B, Palta J P. Expression of freezing tolerance in the interspecific F1 and somatic hybrids of potatoes. Theor Appl Genet, 1999, 98: 995–1004
[4]Hijmans R J. Estimating frost risk in potato production on the Altiplano using interpolated climate data: CIP program report. Lima, International Potato Center, 1999.pp 373–380
[5]Van Swaaij A C, Jacobscn E, Fccnstra W J. Effect of cold hardening, wilting and exogenously applied proline on leaf proline content and frost tolerance of several genotypes of Solanum. Physiol Plant, 1985, 64: 230–236
[6]Chen T H H, Gusta L V. Abscisic acid-induced freezing resistance in cultured plant cells. Plant Physiol, 1983, 73, 71–75
[7]Wallis J G, Wang H, Guerra D J. Expression of a synthetic antifreeze protein in potato reduces electrolyte release at freezing temperatures. Plant Mol Biol, 1997, 35, 323–330
[8]Pino M T, Skinner J S, Park E J, Jeknic Z, Hayes P M., Thomashow M F, Chen T H H. Use of a stress inducible promoter to drive ectopic AtCBF expression improves potato freezing tolerance while minimizing negative effects on tuber yield. Plant Biotechnol J, 2007, 5, 591–604
[9]Pino M T, Skinner J S, Jeknic Z, Hayes P M, Soeldner A H, Thomashow M F, Chen T H H. Ectopic AtCBF1 over-expression enhances freezing tolerance and induces cold acclimation-associated physiological modifications in potato. Plant CellEnviron, 2008, 31: 393–406
[10]Li T, Kim M D, Yang K S, Kwon S Y, Kim S H, Kim J S, Yun D J, Kwak S S, Lee H S. Enhanced tolerance of transgenic potato plants overexpressing nucleoside diphosphate kinase 2 against multiple environmental stresses. Transgenic Res, 2008, 17, 705–715
[11]周华, 张新, 刘腾云, 余发新. 高通量转录组测序的数据分析与基因发掘. 江西科学, 2012, 30: 608–611
Zhou H, Zhang X, Liu T Y, Yu F X. Data processing and gene discovery of high-throughput transcriptome sequencing. Jiangxi Sci, 2012, 30, 608–611 (in Chinese with English abstract)
[12]Kakumanu A, Ambavaram M M R, Klumas C, Krishnan A, Batlang U, Myers E, Grene R, Pereira A. Effects of drought on gene expression in maize reproductive and leaf meristem tissue revealed by RNA-Seq.Plant Physiol, 2012, 160: 846–867
[13]Vidal R O, do Nascimento L C, Mondego J M C, Pereira G A G, Carazzolle M F. Identification of SNPs in RNA-seq data of two cultivars of Glycine max (soybean) differing in drought resistance. Genet Mol Biol, 2012, 35: 331–334
[14]卢坤, 张琳, 曲存民, 梁颖, 唐章林, 李加纳. 利用RNA-Seq鉴定甘蓝型油菜叶片干旱胁迫应答基因. 中国农业科学, 2015, 48: 630–645
Lu K, Zhang L, Qu C M, Liang Y, Tang Z L, Li J N. Identification of drought stress-responsive genes in leaves of Brassica napus by RNA sequencing.Sci Agric Sin, 2015, 48: 630–645 (in Chinese with English abstract)
[15]孙爱清, 张杰道, 万勇善, 刘风珍, 张昆, 孙利. 花生干旱胁迫响应基因的数字表达谱分析. 作物学报, 2013, 39: 1045–1053
Sun A Q, Zhang J D, Wan Y S, Liu F Z, Zhang K, Sun L. in silico expression profile of genes in response to drought in peanut. Acta Agron Sin, 2013, 39: 1045–1053 (in Chinese with English abstract)
[16]牙库甫江•阿西木, 关波, 张富春. 植物基因表达转录分析中内参基因的选择与应用. 生物技术通报, 2011, (7): 7–11
Yakupjan H X M, Guan B, Zhang F C. Research progress in plant reference genes. Biotechnol Bull, 2011, (7): 7–11 (in Chinese with English abstract)
[17]许英, 陈建华, 朱爱国, 奕明宝, 王晓飞, 孙志民. 低温胁迫下植物响应机理的研究进展. 中国麻业科学, 2015, 37(1): 40–49
Xu Y, Chen J H, Zhu A G, Luan M B, Wang X F, Sun Z M. Research progress on response mechanism of plant under low temperature stress. Plant Fiber Sci China, 2015, 37(1): 40–49 (in Chinese with English abstract)
[18]刘辉, 李德军, 邓治. 植物应答低温胁迫的转录调控网络研究进展. 中国农业科学, 2014, 47: 3523–3533
Liu H, Li D J, Deng Z. Advances in research of transcriptional regulatory network in response to cold stress in plants. Sci Agric Sin, 2014, 47: 3523–3533 (in Chinese with English abstract)
[19]祁云霞, 刘永斌, 荣威恒.  转录组研究新技术: RNA-Seq及其应用. 遗传, 2011: 1191–1202
Qi Y X, Liu Y B, R ong W H. RNA-Seq and its applications: a new technology for transcriptomics. Hereditas, 2011: 1191–1202 (in Chinese with English abstract)
[20]胡红柳, 侯晓明, 曲波, 高学军, 李庆章. 高通量基因表达谱的应用. 中国乳品工业, 2012, 40(12): 40–43
Hu H L, Hou X M, Qu B, Gao X J, Li Q Z. Application of high throughput gene expression profiling. China Dairy Ind, 2012, 40(12): 40–43 (in Chinese with English abstract)
[21]杨伟, 龚荣高, 石佳佳, 赖静, 廖明安, 梁国鲁. 低温胁迫下枇杷幼果转录组的De novo组装和功能注释. 西北农林科技大学学报(自然科学版), 2014, 42(8): 138–146
Yang W, Gong R G, Shi J J, Lai J, Liao M A, Liang G L. De novo assembly and functional annotation of the loquat young fruit transcriptome under chilling stress. J Northwest A&F Univ (Nat Sci Edn), 2014, 42(8): 138–146 (in Chinese with English abstract)
[22]Chen T, Li P H. Biochemical changes in tuber-bearing Solanmn species in relation to frost hardiness during cold acclimation. Plant Physiol, 1980, 66: 414–421
[23]Zheng M, Wang Y, Liu K, Shu H, Zhou Z. Protein expression changes during cotton fiber elongation in response to low temperature stress. J Plant Physiol, 2012, 169: 399–409
[24]Evers D, Legay S, Lamoureux D, Hausman J F, Hoffmann L, Renaut J. Towards a synthetic view of potato cold and salt stress response by transcriptomic and proteomic analyses. Plant Mol Biol, 2012, 78: 503–514
[25]秦玉芝, 陈珏, 邢铮, 何长征, 熊兴耀. 低温逆境对马铃薯叶片光合作用的影响. 湖南农业大学学报(自然科学版), 2013, 39(1): 26–30
Qin Y Z, Chen J, Xing Z, He C Z, Xiong X Y. Effects of low temperature stress on photosynthesis in potato leaves. J Hunan Agric Univ (NatSci), 2013, 39(1): 26–30 (in Chinese with English abstract)
[26]Niyogi K K. Photoprotection revisited: genetic and molecular approaches. Annu Rev Plant Physiol Plant Mol Biol, 1999, 50: 333–359
[27]Sun X, Tan Q, Nie Z, Hu C, An Y. Differential expression of proteins in response to molybdenum deficiency in winter wheat leaves under low-temperature stress.Plant Mol Biol Rep, 2014, 32:1057–1069
[28]Andreas T, Christophe C, Essaǐd A B. Physiological and molecular changes in plants grown at low temperatures. Planta, 2012, 235: 1091–1105
[29]Bhardwaj A R, Joshi G, Kukreja B, Malik V, Arora P, Pandey R, Shukla R N, Bankar K G, Agarwal S K, Goel S, Jagannath A, Kumar A, Agarwal M. Global insights into high temperature and drought stress regulated genes by RNA-Seq in economically important oilseed crop Brassica juncea. BMC Plant Biol, 2015, 15:1–15
[30]刘蕾, 杜海, 唐晓凤, 吴燕民,黄玉碧, 唐益雄. MYB转录因子在植物抗逆胁迫中的作用及其分子机理. 遗传, 2008, 30: 1265–1271
Liu L, Du H, Tang X F, Wu Y M, Huang Y B, Tang Y X. The roles of MYB transcription factors on plant defense responses and its molecular mechanism. Hereditas, 2008, 30: 1265–1271 (in Chinese with English abstract)
[31]李余良, 刘建华, 郑锦荣, 胡建广. 高温胁迫下甜玉米雌穗发育基因差异表达谱分析. 作物学报, 2013, 39: 269–279
Li Y L, Liu J H, Zheng J R, Hu J G. Gene expression profile of sweet corn ears under heat stress. Acta Agron Sin, 2013, 39: 269–279 (in Chinese with English abstract)
[32]Xu W, Li R, Zhang N, Ma F, Jiao Y T, Wang Z P. Transcriptome profiling of Vitis amurensis, an extremely cold-tolerant Chinese wild Vitis, species, reveals candidate genes and events that potentially connected to cold stress. Plant Mol Biol, 2014, 86:527–541
[33]唐寅, 张威威, 许锋, 程水源. 植物苯丙氨酸代谢相关酶基因启动子研究进展. 长江大学学报(自然科学版), 2010, 7(2): 68–71
Tang Y, Zhang W W, Xu F, Cheng S Y. The research progress of plant phenylalanine metabolism related enzyme gene promoter. J Yangtze Univ (NatSci Edn), 2010, 7(2): 68–71 (in Chinese with English abstract)

[1] 陈国欢, 张锐, 李艳迪, 赵佳琪, 任湧涛, 张天赐, 郭华春, 李俊, 杨芳. 外源硒叶面喷施对浅紫色马铃薯块茎花青素合成的影响[J]. 作物学报, 2026, 52(6): 1876-1890.
[2] 金昱何, 王雪菲, 徐张一娃, 缪怡宁, 蒋云杰, 伊莹, 缪德麟, 朱静仪, 钟一帆, 陈铭亨, 方芳, 刘鹏. 外源激素对低温胁迫下大豆叶片叶绿素荧光参数及抗氧化酶系统的影响[J]. 作物学报, 2026, 52(6): 1817-1829.
[3] 王文辕, 燕雪嘉, 刘玉霖, 孙晓彤, 李亚楠, 唐鑫华, 石瑛. 耐弱光马铃薯品种筛选及转录因子编码基因StPIF3的克隆与功能分析[J]. 作物学报, 2026, 52(6): 1631-1645.
[4] 徐苗苗, 邸太妹, 王洁, 吴叶蝶, 刘恩贝, 王玉春, 王新超, 王璐. 外源槲皮素增强茶树抗寒性的分子机制[J]. 作物学报, 2026, 52(5): 1418-1429.
[5] 田立涛, 丁宁, 王树林, 齐恩芳, 张荣, 王蕊蕊, 马利雯, 李建武, 杨江伟. 马铃薯Argonaute基因家族鉴定及晚疫病诱导表达分析[J]. 作物学报, 2026, 52(4): 1116-1126.
[6] 张宇, 刘芳, 蔡诚诚, 杨小华, 吉阿么石扎, 杨元军, 王西瑶. 溴乙烷与赤霉素协同处理破除马铃薯块茎休眠的机理初探[J]. 作物学报, 2026, 52(3): 825-838.
[7] 汪玲, 胡好, 宋家凤, 程洁蓝, 陈颖, 郑婷婷, 吕钊彦, 朱晓彪, 侯华兰. 马铃薯UDP-糖基转移酶基因StUGT52的克隆及功能验证[J]. 作物学报, 2026, 52(3): 665-676.
[8] 徐强, 谢奎忠, 胡新元, 岳云, 董博, 罗爱花. 连作对马铃薯根际土壤线虫群落结构与功能的影响[J]. 作物学报, 2026, 52(2): 527-538.
[9] 杨飚, 杜帅康, 张继旺, 石瑛, 张丽莉. 马铃薯III类POD基因家族的全基因组鉴定及其表达谱分析[J]. 作物学报, 2026, 52(2): 405-420.
[10] 姬炫彤, 卞春松, 金黎平, 李森, 秦军红, 李广存. 不同耐旱型马铃薯根际微生物对干旱的响应[J]. 作物学报, 2026, 52(1): 165-177.
[11] 田甲春, 葛霞, 李守强, 李梅, 田世龙, 张亚倩, 程建新, 李玉梅. 低O2高CO2贮藏环境延缓马铃薯块茎衰老的作用机制[J]. 作物学报, 2026, 52(1): 262-278.
[12] 王雅致, 杨飚, 季香林, 石瑛, 张丽莉. 二倍体马铃薯抗旱资源鉴定及抗旱基因初步筛选[J]. 作物学报, 2026, 52(1): 72-84.
[13] 刘迪, 黎瑞源, 石茂竹, 李洪有, 陈庆富, 石桃雄. 苦荞半矮秆突变体sd3的表型鉴定及转录组分析[J]. 作物学报, 2026, 52(1): 316-328.
[14] 卓峰琦, 唐振三, 雷雨俊, 程李香, 赵甜甜, 吕汰, 杨晨, 张峰. 基于烹饪方式及回生温度筛选低升糖马铃薯品种(系)[J]. 作物学报, 2025, 51(9): 2538-2546.
[15] 朱锦程, 杨秋华, 程李香, 李文丽, 石明明, 李惠霞, 张峰. 马铃薯抗南方根结线虫种质资源筛选及相关生理反应分析[J]. 作物学报, 2025, 51(9): 2307-2317.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!