银杏CONSTANS-LIKE基因的克隆与序列分析
|
摘要
为阐明银杏开花调控机制,对不同时期的银杏叶片进行了转录组测序,通过分析开花前后表达量差异较大的基因,筛选出1个与开花调控相关的CONSTANS-LIKE基因。使用RT-PCR法从银杏叶片cDNA中克隆得到该基因的完整编码序列(CDS),命名为GbCOL。GbCOL基因的CDS序列全长为1 239 bp,编码产物含412个氨基酸。核苷酸和蛋白质序列多重比对结果显示,GbCOL与其他植物的CO基因的核苷酸与蛋白质序列具有一定的同源性,与裸子植物中的CO基因同源性较高。该基因的蛋白产物具有典型的CONSTANS蛋白保守结构,包含2个高度保守的B-box及CCT结构域。该基因可能在银杏开花调控过程中发挥重要作用。
In order to identify the genes involved in flowering regulation of Ginkgo biloba L., the differentially expressed genes(DGEs) among different developmental stages of inflorescence in G. biloba were screened by transcriptome analysis. A CONSTANS-LIKE gene with significant expression alteration between the stages before and after flowering was selected as target gene. The gene designated as GbCOL was cloned from the cDNA of the leaf of G. biloba by RT-PCR. GbCOL with a 1 239 bp CDS sequence encodes a 412-amino acid product. The nucleotide and amino acid sequences of GbCOL are homologous to the sequences of CO genes of other species, especially Gymnosperm plants. The protein sequence of GbCOL contained a B-box and a CCT domain, which are typical motifs of COgene family in plants. Target gene in this study might play a role in flowering regulation of G. biloba.
In order to identify the genes involved in flowering regulation of Ginkgo biloba L., the differentially expressed genes(DGEs) among different developmental stages of inflorescence in G. biloba were screened by transcriptome analysis. A CONSTANS-LIKE gene with significant expression alteration between the stages before and after flowering was selected as target gene. The gene designated as GbCOL was cloned from the cDNA of the leaf of G. biloba by RT-PCR. GbCOL with a 1 239 bp CDS sequence encodes a 412-amino acid product. The nucleotide and amino acid sequences of GbCOL are homologous to the sequences of CO genes of other species, especially Gymnosperm plants. The protein sequence of GbCOL contained a B-box and a CCT domain, which are typical motifs of COgene family in plants. Target gene in this study might play a role in flowering regulation of G. biloba.
引文
[1]国靖,汪贵斌,封超年,等.银杏林下经济模式分类及模式综合效益评价[J].中南林业科技大学学报,2017,37(1):118-122.
[2]Kubota Y, Tanaka N, Kagota S, et al. Effects of Ginkgo biloba extract on blood pressure and vascular endothelial response by acetylcholine in spontaneously hypertensive rats.[J]. Journal of Pharmacy&Pharmacology, 2006,58(2):243-249.
[3]Mahadevan S, Park Y, Park Y. Modulation of cholesterol metabolism by Ginkgo biloba L. nuts and their extract[J]. Food Research International, 2008,41(1):89-95.
[4]Tang Yijun,Xu Yongjian,XiongShengdao,elal. The effect of Ginkgo Biloba Extract on the expression of PKCαin the inflammatory cells and the level of IL-5 in induced sputum of asthmatic patients[J]. 2007(4):375-380.
[5]Amasino R M. Control of flowering time in plants.[J]. Current Opinion in Genetics&Devolopment, 1996,6(4):480-487.
[6]徐友,王欢利,汪贵斌,等.温度和光照强度对银杏叶黄酮合成的影响[J].中南林业科技大学学报, 2016,36(4):30-34.
[7]岑秀芬,黄春红,韦鹏霄.激素因子对铁皮石斛离体培养开花诱导的效应[J].安徽农业科学,2010(16):8308-8311.
[8]Amasino R. Seasonal and developmental timing of flowering[J].Plant Journal, 2010,61(6):1001-1013.
[9]Zhang S, Zuo J. Advance in the flowering time control of{\sl Arabidopsis}[J]. Progress in Biochemistry&Biophysics,2006,33(4):301-309.
[10]Yun Z, Xiao D S, Ni M. Timing of Photoperiodic Flowering:Light Perception and Circadian Clock[J].植物学报:英文版,2007, 49(1):28-34.
[11]Tamaki S. Adaptation of photoperiodic control pathways produces short-day flowering in rice.[J]. Nature, 2003,422(6933):719-22.
[12]吕佳斌,龙洪旭,李泽,等.油桐LACS4基因的克隆与原核及真核表达[J].经济林研究, 2017,35(2):65-71.
[13]朱云美,张玲,高用顺,等.温度对植物开花时间调控的研究进展[J].亚热带农业研究, 2016,12(2):130-135.
[14]Hayama R, Coupland G. The Molecular Basis of Diversity in the Photoperiodic Flowering Responses of Arabidopsis and Rice[J].Plant Physiology, 2004,135(2):677-684.
[15]张友润.氮素营养调控拟南芥开花诱导相关基因表达的研究[D].杭州:浙江大学, 2011.
[16]Bernier G, Périlleux C. A physiological overview of the genetics of flowering time control[J]. Plant Biotechnology Journal,2005,3(1):3-16.
[17]Simpson G G. The autonomous pathway:epigenetic and posttranscriptional gene regulation in the control of Arabidopsis flowering time[J]. Current Opinion in Plant Biology, 2004, 7(5):570-574.
[18]Blázquez M A, Ahn J H, Weigel D. A thermosensory pathway controlling flowering time in Arabidopsis thaliana.[J]. Nature Genetics, 2003,33(2):168-71.
[19]Komeda Y. GENETIC REGULATION OF TIME TO FLOWER IN ARABIDOPSIS THALIANA[J]. Annual Review of Plant Biology, 2004,55(1):521-535.
[20]SasaniS,HemmingMN,OliverSN,etal. The influence of vernalization and daylength on expression of flowering-time genes in the shoot apex and leaves of barley(Hordeum vulgare).[J].Journal of Experimental Botany, 2009, 60(7):2169-2178.
[21]Peng F Y, Hu Z, Yang R C. Bioinformatic prediction of transcription factor binding sites at promoter regions of genes for photoperiod and vernalization responses in model and temperate cereal plants[J]. Bmc Genomics, 2016, 17(1):573-576.
[22]龚付全,熊超,陈柳吉,等.银杏CONSTANS基因的克隆与序列分析[J].湖北农业科学, 2010(12):2945-2948.
[23]Martínezgarcía J F, Virgóssoler A, Prat S. Control of photoperiodregulated tuberization in potato by the Arabidopsis flowering-time gene CONSTANS[J]. Proceedings of the National Academy of Sciences of the United States of America, 2002,99(23):15211-15216.
[24]Florea L, Song L, Salzberg S L. Thousands of exon skipping events differentiate among splicing patterns in sixteen human tissues[J]. F1000research, 2013(2):188-191.
[25]RobsonF,CostaMM,HepworthSR,etal.Functional importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants.[J]. Plant Journal for Cell&Molecular Biology,2001, 28(6):619-631.
[26]Torok M, Etkin L D. Two B or not two B? Overview of the rapidly expanding B-box family of proteins.[J]. Differentiation,2001,67(3):63-71.
[27]艾育芳.早晚熟油菜成花机理的初步研究[D].福州:福建农林大学, 2011.
[28]赵志琴,安丽君,李芳,等.草莓CO基因克隆与表达分析[J].中国农业大学学报, 2010(1):19-24.
[29]Lemmetyinen J, Keinonen-Mett?l?K, L?nnenp??M, et al. Activity of the CaMV 35S promoter in various parts of transgenic early flowering birch clones[J]. Plant Cell Reports,1998,18(3-4):243-248.
[30]Pe?a L, Martín-Trillo M, Juárez J, et al. Constitutive expression of Arabidopsis LEAFY or APETALA1 genes in citrus reduces their generation time[J]. Nature Biotechnology, 2001, 19(3):263-267.
[31]刘永平,杨静,杨明峰.植物开花调控途径[J].生物工程学报,2015,31(11):1553-1566.
[32]ValverdeF.CONSTANSandtheevolutionaryoriginof photoperiodic timing of flowering.[J]. Journal of Experimental Botany, 2011,62(8):2453-2463.
[33]YangXQ,TangQL,SongM,etal.FloweringControl Mechanism of CO as a Key Factor in Photoperiod Pathway[J].Plant Physiology Journal, 2013,49(9):861-868.
[34]Fernández V, Takahashi Y, Gourrierec J L, et al. Photoperiodic andthermosensorypathwaysinteractthroughCONSTANSto promote flowering at high temperature under short days[J]. Plant Journal for Cell&Molecular Biology, 2016,86(5):426.
[35]Wenkel S. Analysis of the function of the CONSTANS protein and transcriptional regulation of FLOWERING LOCUS T[J].Journal of Sea Research, 2006, 62(2-3):93-105.
[36]Nakano Y, Higuchi Y, Sumitomo K, et al. Flowering retardation by high temperature in chrysanthemums:involvement of FLOWERING LOCUS T-like 3 gene repression[J]. Journal of Experimental Botany, 2013, 64(4):909-920.
[37]CastillejoC,PelazS. ThebalancebetweenCONSTANSand TEMPRANILLO activities determines FT expression to trigger flowering.[J]. Current Biology Cb, 2008,18(17):1338.
[38]陆莹.烟草CONSTANS同源基因的克隆与分析[D].北京:中国农业科学院, 2012.
[39]石玉波,张荻,申晓辉,等.百子莲CONSTANS同源基因的克隆及表达分析[J].北京林业大学学报,2014,36(4):113-120.
[40]MartinJ,StorgaardM, AndersenCH,etal.Photoperiodic regulation of flowering in perennial ryegrass involving a CONSTANS-like homolog[J]. Plant Molecular Biology, 2004,56(2):159-169.
[41]Ben-Naim O, Eshed R, Parnis A, et al. The CCAAT binding factor can mediate interactions between CONSTANS-like proteins and DNA[J]. Plant Journal, 2006,46(3):462-476.
[42]LiuJ, YuJ,McintoshL,etal.Isolation of a CONSTANS ortholog from Pharbitis nil and its role in flowering.[J]. Plant Physiology, 2001, 125(4):1821-1830.
[43]Griffiths S, Dunford R P, Coupland G, et al. The Evolution of CONSTANS-Like Gene Families in Barley, Rice, and Arabidopsis[J]. Plant Physiology, 2003,131(4):1855-1867.
[44]GuoJL,YangQ,LiangF,etal.Molecularcloningand expression analysis of a novel CONSTANS-like gene from potato[J]. Biochemistry Biokhimiia, 2007,72(11):1241-1246.
[45]LiuJ, YuJ,McintoshL,etal. Isolation of a CONSTANS ortholog from Pharbitis nil and its role in flowering.[J]. Plant Physiology, 2001, 125(4):1821-1830.
[46]Kim S K, Yun C H, Lee J H, et al. OsCO3, a CONSTANS-LIKE gene, controls flowering by negatively regulating the expression of FT-like genes under SD conditions in rice.[J]. Planta, 2008,228(2):355-365.
[47]吴连成,常丽丽,陈晓,等. CO基因的调控表达与植物光周期反应[J].中国农学通报, 2010,26(2):116-121.
[48]Kay S A. Molecular basis of seasonal time measurement in Arabidopsis.[J]. Nature, 2002,419(6904):308-312.
[49]ValverdeF,Mouradov A,SoppeW,etal.Photoreceptor regulation of CONSTANS protein in photoperiodic flowering.[J].Science, 2004, 303(5660):1003-1006.
[50]Wigge P A, Min C K, Jaeger K E, et al. Integration of spatial and temporal information during floral induction in Arabidopsis[J].Science, 2005,309(5737):1056-1059.
[51]Yano M, Katayose Y, Ashikari M, et al. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS[J]. Plant Cell,2000, 12(12):2473-2483.
[2]Kubota Y, Tanaka N, Kagota S, et al. Effects of Ginkgo biloba extract on blood pressure and vascular endothelial response by acetylcholine in spontaneously hypertensive rats.[J]. Journal of Pharmacy&Pharmacology, 2006,58(2):243-249.
[3]Mahadevan S, Park Y, Park Y. Modulation of cholesterol metabolism by Ginkgo biloba L. nuts and their extract[J]. Food Research International, 2008,41(1):89-95.
[4]Tang Yijun,Xu Yongjian,XiongShengdao,elal. The effect of Ginkgo Biloba Extract on the expression of PKCαin the inflammatory cells and the level of IL-5 in induced sputum of asthmatic patients[J]. 2007(4):375-380.
[5]Amasino R M. Control of flowering time in plants.[J]. Current Opinion in Genetics&Devolopment, 1996,6(4):480-487.
[6]徐友,王欢利,汪贵斌,等.温度和光照强度对银杏叶黄酮合成的影响[J].中南林业科技大学学报, 2016,36(4):30-34.
[7]岑秀芬,黄春红,韦鹏霄.激素因子对铁皮石斛离体培养开花诱导的效应[J].安徽农业科学,2010(16):8308-8311.
[8]Amasino R. Seasonal and developmental timing of flowering[J].Plant Journal, 2010,61(6):1001-1013.
[9]Zhang S, Zuo J. Advance in the flowering time control of{\sl Arabidopsis}[J]. Progress in Biochemistry&Biophysics,2006,33(4):301-309.
[10]Yun Z, Xiao D S, Ni M. Timing of Photoperiodic Flowering:Light Perception and Circadian Clock[J].植物学报:英文版,2007, 49(1):28-34.
[11]Tamaki S. Adaptation of photoperiodic control pathways produces short-day flowering in rice.[J]. Nature, 2003,422(6933):719-22.
[12]吕佳斌,龙洪旭,李泽,等.油桐LACS4基因的克隆与原核及真核表达[J].经济林研究, 2017,35(2):65-71.
[13]朱云美,张玲,高用顺,等.温度对植物开花时间调控的研究进展[J].亚热带农业研究, 2016,12(2):130-135.
[14]Hayama R, Coupland G. The Molecular Basis of Diversity in the Photoperiodic Flowering Responses of Arabidopsis and Rice[J].Plant Physiology, 2004,135(2):677-684.
[15]张友润.氮素营养调控拟南芥开花诱导相关基因表达的研究[D].杭州:浙江大学, 2011.
[16]Bernier G, Périlleux C. A physiological overview of the genetics of flowering time control[J]. Plant Biotechnology Journal,2005,3(1):3-16.
[17]Simpson G G. The autonomous pathway:epigenetic and posttranscriptional gene regulation in the control of Arabidopsis flowering time[J]. Current Opinion in Plant Biology, 2004, 7(5):570-574.
[18]Blázquez M A, Ahn J H, Weigel D. A thermosensory pathway controlling flowering time in Arabidopsis thaliana.[J]. Nature Genetics, 2003,33(2):168-71.
[19]Komeda Y. GENETIC REGULATION OF TIME TO FLOWER IN ARABIDOPSIS THALIANA[J]. Annual Review of Plant Biology, 2004,55(1):521-535.
[20]SasaniS,HemmingMN,OliverSN,etal. The influence of vernalization and daylength on expression of flowering-time genes in the shoot apex and leaves of barley(Hordeum vulgare).[J].Journal of Experimental Botany, 2009, 60(7):2169-2178.
[21]Peng F Y, Hu Z, Yang R C. Bioinformatic prediction of transcription factor binding sites at promoter regions of genes for photoperiod and vernalization responses in model and temperate cereal plants[J]. Bmc Genomics, 2016, 17(1):573-576.
[22]龚付全,熊超,陈柳吉,等.银杏CONSTANS基因的克隆与序列分析[J].湖北农业科学, 2010(12):2945-2948.
[23]Martínezgarcía J F, Virgóssoler A, Prat S. Control of photoperiodregulated tuberization in potato by the Arabidopsis flowering-time gene CONSTANS[J]. Proceedings of the National Academy of Sciences of the United States of America, 2002,99(23):15211-15216.
[24]Florea L, Song L, Salzberg S L. Thousands of exon skipping events differentiate among splicing patterns in sixteen human tissues[J]. F1000research, 2013(2):188-191.
[25]RobsonF,CostaMM,HepworthSR,etal.Functional importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants.[J]. Plant Journal for Cell&Molecular Biology,2001, 28(6):619-631.
[26]Torok M, Etkin L D. Two B or not two B? Overview of the rapidly expanding B-box family of proteins.[J]. Differentiation,2001,67(3):63-71.
[27]艾育芳.早晚熟油菜成花机理的初步研究[D].福州:福建农林大学, 2011.
[28]赵志琴,安丽君,李芳,等.草莓CO基因克隆与表达分析[J].中国农业大学学报, 2010(1):19-24.
[29]Lemmetyinen J, Keinonen-Mett?l?K, L?nnenp??M, et al. Activity of the CaMV 35S promoter in various parts of transgenic early flowering birch clones[J]. Plant Cell Reports,1998,18(3-4):243-248.
[30]Pe?a L, Martín-Trillo M, Juárez J, et al. Constitutive expression of Arabidopsis LEAFY or APETALA1 genes in citrus reduces their generation time[J]. Nature Biotechnology, 2001, 19(3):263-267.
[31]刘永平,杨静,杨明峰.植物开花调控途径[J].生物工程学报,2015,31(11):1553-1566.
[32]ValverdeF.CONSTANSandtheevolutionaryoriginof photoperiodic timing of flowering.[J]. Journal of Experimental Botany, 2011,62(8):2453-2463.
[33]YangXQ,TangQL,SongM,etal.FloweringControl Mechanism of CO as a Key Factor in Photoperiod Pathway[J].Plant Physiology Journal, 2013,49(9):861-868.
[34]Fernández V, Takahashi Y, Gourrierec J L, et al. Photoperiodic andthermosensorypathwaysinteractthroughCONSTANSto promote flowering at high temperature under short days[J]. Plant Journal for Cell&Molecular Biology, 2016,86(5):426.
[35]Wenkel S. Analysis of the function of the CONSTANS protein and transcriptional regulation of FLOWERING LOCUS T[J].Journal of Sea Research, 2006, 62(2-3):93-105.
[36]Nakano Y, Higuchi Y, Sumitomo K, et al. Flowering retardation by high temperature in chrysanthemums:involvement of FLOWERING LOCUS T-like 3 gene repression[J]. Journal of Experimental Botany, 2013, 64(4):909-920.
[37]CastillejoC,PelazS. ThebalancebetweenCONSTANSand TEMPRANILLO activities determines FT expression to trigger flowering.[J]. Current Biology Cb, 2008,18(17):1338.
[38]陆莹.烟草CONSTANS同源基因的克隆与分析[D].北京:中国农业科学院, 2012.
[39]石玉波,张荻,申晓辉,等.百子莲CONSTANS同源基因的克隆及表达分析[J].北京林业大学学报,2014,36(4):113-120.
[40]MartinJ,StorgaardM, AndersenCH,etal.Photoperiodic regulation of flowering in perennial ryegrass involving a CONSTANS-like homolog[J]. Plant Molecular Biology, 2004,56(2):159-169.
[41]Ben-Naim O, Eshed R, Parnis A, et al. The CCAAT binding factor can mediate interactions between CONSTANS-like proteins and DNA[J]. Plant Journal, 2006,46(3):462-476.
[42]LiuJ, YuJ,McintoshL,etal.Isolation of a CONSTANS ortholog from Pharbitis nil and its role in flowering.[J]. Plant Physiology, 2001, 125(4):1821-1830.
[43]Griffiths S, Dunford R P, Coupland G, et al. The Evolution of CONSTANS-Like Gene Families in Barley, Rice, and Arabidopsis[J]. Plant Physiology, 2003,131(4):1855-1867.
[44]GuoJL,YangQ,LiangF,etal.Molecularcloningand expression analysis of a novel CONSTANS-like gene from potato[J]. Biochemistry Biokhimiia, 2007,72(11):1241-1246.
[45]LiuJ, YuJ,McintoshL,etal. Isolation of a CONSTANS ortholog from Pharbitis nil and its role in flowering.[J]. Plant Physiology, 2001, 125(4):1821-1830.
[46]Kim S K, Yun C H, Lee J H, et al. OsCO3, a CONSTANS-LIKE gene, controls flowering by negatively regulating the expression of FT-like genes under SD conditions in rice.[J]. Planta, 2008,228(2):355-365.
[47]吴连成,常丽丽,陈晓,等. CO基因的调控表达与植物光周期反应[J].中国农学通报, 2010,26(2):116-121.
[48]Kay S A. Molecular basis of seasonal time measurement in Arabidopsis.[J]. Nature, 2002,419(6904):308-312.
[49]ValverdeF,Mouradov A,SoppeW,etal.Photoreceptor regulation of CONSTANS protein in photoperiodic flowering.[J].Science, 2004, 303(5660):1003-1006.
[50]Wigge P A, Min C K, Jaeger K E, et al. Integration of spatial and temporal information during floral induction in Arabidopsis[J].Science, 2005,309(5737):1056-1059.
[51]Yano M, Katayose Y, Ashikari M, et al. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS[J]. Plant Cell,2000, 12(12):2473-2483.