| 382 | 6 | 90 |
| 下载次数 | 被引频次 | 阅读次数 |
作为新兴的能源植物,小桐子所具备的耐贫瘠、种子含油量高、油质优良等特性使其受到广泛关注。本研究对小桐子幼苗进行10%PEG模拟干旱锻炼与空气干旱胁迫处理,以叶片样品进行RNA-Seq文库构建并基于Illumina Hiseq2000平台进行高通量测序,对差异表达基因进行鉴定和通路富集分析,发现糖代谢通路得到了显著富集。其中,肌醇半乳糖苷合成酶(inositol galactoside synthase,GS)和棉子糖合成酶(raffinose synthase,RS)作为棉子糖系列寡糖合成途径的2类关键酶,其家族成员的不同编码序列在干旱胁迫下呈现不同的表达模式。实时荧光定量(RT-q PCR)验证了两个酶基因家族关键成员对干旱处理的诱导响应,暗示了棉子糖合成途径参与了干旱锻炼和胁迫耐受性的形成,为进一步揭示小桐子抗旱性形成的分子机制及其基因工程改良奠定了坚实的理论基础。
Abstract：Jatropha curcas L. has attracted wide attention as a new energy plant by the characteristics of its barren resistance, high seed oil content and good oil quality. In this study, Jatropha curcas seedlings were treated with10% PEG simulated drought hardening and air drought stress. RNA-Seq Libraries were constructed with leaf samples and high-throughput sequencing was carried out based on Illumina Hiseq2000 platform. Identification and enrichment analysis of the differentially expressed genes(DEG) indicated that the glucose metabolism pathways were significantly enriched. Among them, inositol galactoside synthase(GS) and rcarinose synthase(RS) acting as two kinds of key enzyme of raffinose oligosaccharide synthesis pathway, the different coding sequences of their family members showed different expression patterns during the drought treatment. Real-time PCR(RT-q PCR)results confirmed that the key isoforms of two enzyme gene families were induced by drought stress, and suggested that the raffinose synthesis pathway was involved in the response and formation of drought stress tolerance, which might establish a solid theoretical foundation for further revealing the molecular mechanism of drought-resistance formation and genetic engineering improvement of Jatropha curcas.
Chen K.,2011,Involvement of osmoregulation and their biochemical pathways in drought hardening enhanced drought resistance in the energy plant Jatropha curcas L.,Thesis for M.S.,Yunnan Normal University,Supervisor:Gong M.,pp.1-11(陈凯,2011,渗透调节在干旱锻炼提高能源植物小桐子抗旱性中的作用及其生化机制,硕士学位论文,云南师范大学,导师:龚明,pp.1-11)
Cheng Z.Y.,Li M.,Shi Y.,He P.,He L.L.,and Li F.S.,2015,Research of drought-response mechanism in sugarcane by high through-put sequencing-based digital gene expression profiling,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(9):2018-2028(程志远,李穆,石莹,何平,何丽莲,李富生,2015,利用高通量测序分析甘蔗的抗旱响应机制,分子植物育种,13(9):2018-2028)
Cui M.K.,Lin J.,Li C.H.,and Gong M.,2009,Studies on energy plant resources in Yunnan,Yunnan Shifan Daxue Xuebao(Journal of Yunnan Normal University(Natural Science)),29(4):39-46(崔明昆,林锦,李红朝,龚明,2009,云南的能源植物资源,云南师范大学学报(自然科学版),29(4):39-46)
Dai J.J.,Yu S.X.,Pang C.Y.,Song M.Z.,and Fan S.L.,2012,Application of next-generation sequencing technology and its effect on plant molecular breeding,Zhongguo Mianhua(China Cotton),39(4):1-5(代军杰,喻树迅,庞朝友,宋美珍,范术丽,2012,新一代测序技术的应用及其对作物分子育种的影响,中国棉花,39(4):1-5)
Dai X.,Li Z.G.,and Gong M.,2012,Effect of gibberellin,calcium and betaine on seed germination and resistance of Jat-ropha curcas L.seedlings to low temperature and drought stress,Zhiwu Kexue Xuebao(Plant Science Journal),30(2):204-212(代勋,李忠光,龚明,2012,赤霉素、钙和甜菜碱对小桐子种子萌发及幼苗抗低温和干旱的影响,植物科学学报,30(2):204-212)
Ding K.,and Wang X.F.,2012,Cloning and expression analysis of stachyose synthase gene in Brassica napus L.,Xibei Zhiwu Xuebao(Acta Bot.Boreal),32(3):439-446(丁可,汪晓峰,2012,甘蓝型油菜水苏糖合成酶基因克隆及表达分析,西北植物学报,32(3):439-446)
Eisen M.B.,Spellman P.T.,Brown P.O.,and Botstein D.,1998,Cluster analysis and display of genome-wide expression patterns,Proceedings of the National Academy of Sciences of the United States of America,95(25):14863-14868
Fini A.,Bellasio C.,Pollastri S.,Tattini M.,and Ferrini F.,2013,Water relations,growth,and leaf gas exchange as affected by water stress in Jatropha curcas,Journal of Arid Environments,89(1):21-29
Hoon M.J.L.,Imoto S.,Nolan J.,and Miyano S.,2004,Open source clustering software,Bioinformatics,20(9):1453-1454
Imanishi H.T.,Suzuki T.,Masuda K.,and Harada T.,1998,Accumulation of raffinose stachyose in shoot apices of Lonicera caerulea L.during cold acclimation,Scientia Horticulture,72(3-4):255-263
Langmead B.,Trapnell C.,Pop M.,and Salzberg S.L.,2009,Ultrafast and memory-efficient alignment of short DNA sequences to the human genome,Genome Biology,10(3):25-34
Li B.,and Dewey C.N.,2011,RSEM:accurate transcript quantification from RNA-Seq data with or without a reference genome,BMC Bioinformatics,12(1):2-16
Li F.,and Wang X.F.,2008,Advance in raffinose family oligosaccharides metabolism and the key enzymes in plants,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),28(4):852-859(李芳,汪晓峰,2008,植物中棉子糖系列寡糖代谢及其调控关键酶研究进展,西北植物学报,28(4):852-859)
Li H.,and Durbin R.,2009,Fast and accurate short read alignment with Burrows-Wheeler transfor m,Bioinformatics,25(14):1754-1760
Li Z.G.,and Gong M.,2010,Effects of different chemical disinfectant on seed germination and seedling growth of Jatropha curcas L.,Zhongzi(Seed),12(30):4-7(李忠光,龚明,2010,不同化学消毒剂对小桐子种子萌发和幼苗生长的影响,种子,12(30):4-7)
Liu Y.A.,Luo X.M.,Wei J.G.,Yang H.B.,Yuanteng L.M.,and Hu T.X.,2010,Photosynthetic physiological adaptation mechanism of Jatropha curcas seedling in different water conditions,Linye Kexue Yanjiu(Forest Research),23(1):108-113(刘永安,骆晓铭,魏建国,杨洪彬,远藤利明,胡庭兴,2010,不同水分条件下麻疯树幼苗的光合生理适应性研究,林业科学研究,23(1):108-113)
Luo F.Q.,Wang Z.Y.,Liang Y.,Pan J.R.,and Guo J.,2010,Physicochemical properties and viscosity-temperature characteristic of Jatropha curcas oil as fuel,Nongye Gongcheng Xuebao(Transactions of the CSAE),26(5):227-231(罗福强,王子玉,梁昱,潘军如,郭建,2010,作为燃油的小桐子油的物化性质及黏温特性,农业工程学报,26(5):227-231)
Maes W.H.,Trabucco A.,Achten W.M.J.,and Muys B.,2009,Climatic growing conditions of Jatropha curcas L.,Biomass and Bioenergy,33(10):1481-1485
Maghuly F.,and Laimer M.,2013,Jatropha curcas,a biofuel crop:functional genomics for understanding metabolic pathway and genetic improvement,Biotechnology Journal,8(10):1172-1182
Nishizawa A.,Yabuta Y.,and Shigeoka S.,2008,The contribution of carbohydrate sincluding raffinose family oligosaccharides and sugar alcohols to protection of plant cells from oxidative damage,Plant Signaling and Behavior,3(11):1016-1018
Osumi C.,Nozaki J.,and Kida T.,2005,Raffinose synthase gene,method for producing raffinose,and transgenic plant,United States Patent,US0009165A
Panikulangara T.J.,Eggers-Schumacher G.,Wunderlich M.,Stransky H.,and Schoffl F.,2004,Galactinol synthase1 a novel heat shock factor target gene responsible for heat-induced synthesis of raffinose family oligosaccharides in Arabidopsis,Plant Physiology,136(2):3148-3158
Peterbauer T.,Mach L.,Mucha J.,and Richter A.,2002,Functional expression of a c DNA encoding pea(Pisum sativum L.)raffinose synthase,partial purification of the enzyme from maturing seeds,and steady-state kinetic analysis of raffinose synthesis,Planta,215(5):839-846
Saldanha A.J.,2004,Java Treeview--extensible visualization of microarray data,Bioinformatics,20(17):3246-3248
Sapeta H.,Costa J.M.,Lourenco T.,Maroco J.,and Olivera M.M.,2013,Drought stress response in Jatropha curcas:Growth and physiology,Environmental and Experimental Botany,85:76-84
Sengupta S.,Mukherjee S.,Basak P.,and Majumder A.L.,2015,Significance of galactinol and raffinose family oligosaccharide synthesis in plants,Frontiers in Plant Science,6(656):1-11
Sonia T.,Garcia-Alcalde F.,Dopazo J.,Ferrer A.,and Conesa A.,2011,Differential expression in RNA-seq:a matter of depth,Genome Research,21(12):2213-2223
Sui X.L.,Meng X.L.,Wang H.Y.,Wei Y.X.,Li R.F.,Wang Z.Y.,Hu L.P.,Wang S.H.,Ren H.Z.,and Zhang Z.X.,2011,Response study of cloning,functional verification and low temperature of rape sugar synthase gene in cucumber,Yuanyi Xuebao(Acta Horticulturae Sinica),38(S):2591(眭晓蕾,孟凡珍,王虹云,韦玉霞,李瑞富,王振雨,胡丽萍,王绍辉,任华中,张振贤,2011,黄瓜棉子糖合成酶基因克隆,功能验证及其低温响应研究,园艺学报,38(S):2591)
Taji T.,Ohsumi C.,Iuchi S.,Seki M.,Kasuga M.,Kobayashi M.,Yamaguchi-Shinozaki K.,and Shinozaki K.,2002,Important roles of drought-and cold-inducible genes for galactinol synthase in stress to lerance in Arabidopsis thaliana,Plant,29(29):417-426
Wang H.B.,2015,RNA-Seq-based elucidation of the molecular mechanism,and cloning functional characterization of the key genes for cold resistance formation in Jatropha curcas L.,Thesis for D.S.,Yunnan Normal University,Supervisor:Gong M.,pp.33-52(王海波,2015,基于RNA-Seq解析小桐子抗冷性形成的分子机制及其关键基因的克隆与功能分析,博士学位论文,云南师范大学,导师:龚明,pp.33-52)
Wang H.B.,Zou Z.R.,and Gong M.,2014,Expression patterns of the cold tolerance-associated genes of Jatropha curcas screened based on RNA-Seq data and their cluster analysis,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),33(6):1196-1205(王海波,邹竹荣,龚明,2014,基于RNA-Seq数据筛选的小桐子抗冷相关基因的表达模式及其聚类分析,基因组学与应用生物学,33(6):1196-1205)
Wang Z.,Liu Y.X.,Wei J.H.,and Deng X.,2012,Cloning and expression of a gene encoding a raffinose synthase in the resurrection plant Boea hygrometrica,Zhiwu Xuebao(Chinese Bulletin of Botany),47(1):44-54(王智,刘永秀,魏建华,邓馨,2012,复苏植物漩涡苣苔棉子糖合酶家族基因的克隆和表达,植物学报,47(1):44-54)
Xu S.L.,Li Z.G.,and Gong M.,2011,Protective action of exogenous betaine on seed germination and seedling growth of Jatropha curcas under PEG stress,Zhongzi(Seed),30(9):29-33(许锁链,李忠光,龚明,2011,外源甜菜碱对PEG胁迫下小桐子种子萌发和幼苗生长的保护作用,种子,30(9):29-33)
Zhang Y.S.,Huang S.Z.,and Fu J.R.,2001,Progress in study on raffinose family oligosaccharides in seeds,Zhiwuxue Tongbao(Chinese Bulletin of Botany),18(1):16-22(张以顺,黄上志,傅家瑞,2001,种子中的棉子糖半乳糖苷系列寡糖研究进展,植物学通报,18(1):16-22)
Zhou Y.B.,Tian C.Y.,Zhu Y.M.,Zhang J.K.,and Sui Y.X.,2017,Fundamental research on reconstructed biosynthetic pathway of raffinose in saccharomyces cerevisiae,XiandaiShipin Keji(Modern Food Science and Technology),33(5):1-9(周英彪,田朝玉,朱玥明,张娟琨,孙媛霞,2017,在酿酒酵母中重构棉子糖生物合成途径的基本研究,现代食品科技,33(5):1-9)
基本信息:
DOI：10.13417/j.gab.037.003978
中图分类号:S794.9
引用信息:
[1]王君珂,王莎莎,张博,等.基于RNA-Seq解析小桐子棉子糖代谢途径在干旱适应中的作用[J].基因组学与应用生物学,2018,37(09):3978-3987.DOI:10.13417/j.gab.037.003978.
基金信息:
国家自然科学基金(31460059;31460182)项目资助
2018-09-20
2018-09-20