茶树CsAIL的克隆及在不同休眠阶段的表达分析
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摘要
从茶树不同休眠状态腋芽转录组中筛选出一条差异表达基因,经全长克隆和同源性分析证实该基因为与多年生植物休眠的形成和解除密切相关的AINTEGUMEN-LIKE(AIL)的同源基因,命名为CsAIL。生物信息分析表明,该基因包含1个1 872 bp的开放阅读框,编码623个氨基酸。编码的蛋白质分子质量为69.2kD,理论等电点为6.6,是一种非分泌性蛋白;该蛋白有两个保守的AP2结构域,是植物中特有的广泛参与生长发育调节的AP2亚家族成员。亚细胞定位预测CsAIL蛋白不存在于叶绿体或者线粒体中,可能定位于其他细胞器;经同源性分析,在‘云抗十号’和‘舒茶早’全基因水平上分别鉴定了11个和12个AIL同源基因。系统进化树及序列保守性分析显示,Cs AIL与‘云抗十号’CSA001743.1和‘舒茶早’TEA027750.1的进化关系最近,与已知的拟南芥AIL蛋白处于不同分支,但具有典型的保守结构域和已知的重要氨基酸位点。启动子序列分析显示该基因可能受生理节律、光及多种激素等信号共同调控。CsAIL在茶树顶芽、腋芽和茎中的表达量较高,叶片中较低,花中几乎不表达。在茶树冬季类休眠和生理休眠阶段,该基因的表达量在叶和越冬芽中均逐渐下调并维持在较低水平;在生态休眠及萌动阶段显著上调。分析其可能的下游调控基因CsCYCD3.2和CsCYCD6.1的表达,二者与Cs AIL的表达模式高度一致,与越冬芽的休眠状态存在高度关联。本研究表明,CsAIL可能是参与茶树休眠调控的关键基因,与CsCYCDs共同在茶树年休眠—生长循环中发挥作用。
In tea plant,on the basis of previous transcriptome study on the axillary buds at different dormancy states,a differentially expressed gene related to dormancy regulation was identified,cloned and sequenced. This gene was highly homologous to AINTEGUMEN-LIKE(AIL),and named as CsAIL.Bioinformatic analysis showed that the CsAIL gene contains an open reading frame(ORF)in 1 872 bp,encoding 623 amino acid residues. CsAIL is predicted as a secretory protein with a putative molecular mass of 69.2 kD and theoretical isoelectric point of 6.6. CsAIL contained two conserved AP2 domains and should be grouped into AP2 subfamily,a superfamily closely involved in plant growth and development.Subcellular localization showed that CsAIL protein possibly locates the other organelles except chloroplasts and mitochondria. Homology analysis indicated that 11 and 12 AIL homologous genes were identified at the level of‘Yunkang 10'and‘Shuchazao'tea plant genome,respectively. Analysis of phylogenetic tree and sequence conservatism showed that CsAIL had the closest evolutionary relationship with‘Yunkang 10'CSA001743.1 and‘Shuchazao'TEA027750.1,and had different branches with known Arabidopsis AIL protein,but had typical conserved domain and known important amino acid sites.Promoter sequence cloning and analysis showed that CsAIL gene may be co-regulated by physiological rhythm,light and multiple hormone signals. Comprehensive expression analyses indicated that CsAIL had high expression levels in tea apical buds,axillary buds and stems,but relative low in leaves and extremely in flowers. At the stage of paradormancy,the expression of CsAIL was gradually down-regulated. A pretty low transcript level was maintained during endodormancy period. On the contrary,rapid expression increase was detected during ecodormancy and bud flush. Furthermore,the expression patterns of CsCYCD3.2 and CsCYCD6.1,putative downstream targets of CsAIL,were highly consistent with CsAIL and a high coefficient of association was observed between them. These results suggest that CsAIL may be a pivotal gene in bud dormancy regulation by modulating CsCYCDs among the dormancy-growth cycles in tea plant.
In tea plant,on the basis of previous transcriptome study on the axillary buds at different dormancy states,a differentially expressed gene related to dormancy regulation was identified,cloned and sequenced. This gene was highly homologous to AINTEGUMEN-LIKE(AIL),and named as CsAIL.Bioinformatic analysis showed that the CsAIL gene contains an open reading frame(ORF)in 1 872 bp,encoding 623 amino acid residues. CsAIL is predicted as a secretory protein with a putative molecular mass of 69.2 kD and theoretical isoelectric point of 6.6. CsAIL contained two conserved AP2 domains and should be grouped into AP2 subfamily,a superfamily closely involved in plant growth and development.Subcellular localization showed that CsAIL protein possibly locates the other organelles except chloroplasts and mitochondria. Homology analysis indicated that 11 and 12 AIL homologous genes were identified at the level of‘Yunkang 10'and‘Shuchazao'tea plant genome,respectively. Analysis of phylogenetic tree and sequence conservatism showed that CsAIL had the closest evolutionary relationship with‘Yunkang 10'CSA001743.1 and‘Shuchazao'TEA027750.1,and had different branches with known Arabidopsis AIL protein,but had typical conserved domain and known important amino acid sites.Promoter sequence cloning and analysis showed that CsAIL gene may be co-regulated by physiological rhythm,light and multiple hormone signals. Comprehensive expression analyses indicated that CsAIL had high expression levels in tea apical buds,axillary buds and stems,but relative low in leaves and extremely in flowers. At the stage of paradormancy,the expression of CsAIL was gradually down-regulated. A pretty low transcript level was maintained during endodormancy period. On the contrary,rapid expression increase was detected during ecodormancy and bud flush. Furthermore,the expression patterns of CsCYCD3.2 and CsCYCD6.1,putative downstream targets of CsAIL,were highly consistent with CsAIL and a high coefficient of association was observed between them. These results suggest that CsAIL may be a pivotal gene in bud dormancy regulation by modulating CsCYCDs among the dormancy-growth cycles in tea plant.
引文
Allen M D,Yamasaki K,Ohme-Takagi M,Tateno M,Suzuki M.1998.A novel mode of DNA recognition by a beta-sheet revealed by the solution structure of the GCC-box binding domain in complex with DNA.Embo Journal,17(18):5484-5496.
Anastasiou E,Lenhard M.2007.Growing up to one’s standard.Current Opinion in Plant Biology,10(1):63-69.
Artlip T S,Wisniewski M E,Bassett C L,Norelli J L.2013.CBF gene expression in peach leaf and bark tissues is gated by a circadian clock.Tree Physiology,33(8):866-877.
Azeez A,Miskolczi P,Tylewicz S,Bhalerao R P.2014.A tree ortholog of APETALA1 mediates photoperiodic control of seasonal growth.Current Biology,24(7):717-724.
Chang S J,Puryear J,Cairney J.1993.A simple and efficient method for isolating RNA from pine trees.Plant Molecular Biology Reporter,11(2):113-116.
Devitt M L,Stafstrom J P.1995.Cell cycle regulation during growth-dormancy cycles in pea axillary buds.Plant Molecular Biology,29(2):255-265.
Elliott R C,Betzner A S,Huttner E,Oakes M P,Tucker W Q,Gerentes D,Perez P,Smyth D R.1996.AINTEGUMENTA,an APETALA2-like gene of Arabidopsis with pleiotropic roles in ovule development and floral organ growth.Plant Cell,8(2):155-168.
Espinosa-Ruiz A,Saxena S,Schmidt J,Mellerowicz E,Miskolczi P,Bako L,Bhalerao R P.2004.Differential stage-specific regulation of cyclin-dependent kinases during cambial dormancy in hybrid aspen.Plant Journal,38(4):603-615.
Foley M E F,Anderson J V A,Horvath D P H.2009.The effects of temperature,photoperiod,and vernalization on regrowth and flowering competence in Euphorbia esula(Euphorbiaceae)crown buds.Botany-botanique,87(10):986-992.
Hao X,Horvath D P,Chao W S,Yang Y,Wang X,Xiao B.2014.Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant[Camellia sinensis(L.)O.Kuntze].International Journal of Molecular Sciences,15(12):22155.
Hao X,Yang Y,Yue C,Wang L,Horvath D P,Wang X.2017.Comprehensive transcriptome analyses reveal differential gene expression profiles of Camellia sinensis axillary buds at para-,endo-,ecodormancy,and bud flush stages.Frontiers in Plant Science,8(481):553.
Hao Xinyuan.2015.Study on the molecular mechanism of winter bud dormancy in tea plant[Ph.D.Dissertation].Yangling:Northwest A&FUniversity.(in Chinese)郝心愿.2015.茶树越冬芽休眠的分子机理研究[博士论文].杨凌:西北农林科技大学.
Horvath D P,Anderson J V,Chao W S,Foley M E.2003.Knowing when to grow:signals regulating bud dormancy.Trends in Plant Science,8(11):534-540.
Horvath D P,Chao W S,Anderson J V.2002.Molecular analysis of signals controlling dormancy and growth in underground adventitious buds of leafy spurge.Plant Physiology,128(4):1439-1446.
Karlberg A,Bako L,Bhalerao R P.2011.Short day-mediated cessation of growth requires the downregulation of AINTEGUMENTALIKE1transcription factor in hybrid aspen.PLoS Genetics,7(11):e1002361.
Klucher K M,Chow H,Reiser L,Fischer R L.1996.The AINTEGUMENTA gene of Arabidopsis required for ovule and female gametophyte development is related to the floral homeotic gene APETALA2.Plant Cell,8(2):137-153.
Krizek B A.2003.AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain.Nucleic Acids Research,31(7):1859-1868.
Livak K J,Schmittgen T D.2001.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method.Methods,25(4):402-408.
Menges M,Samland A K,Planchais S,Murray J A.2006.The D-type cyclin CYCD3:1 is limiting for the G1-to-S-phase transition in Arabidopsis.Plant Cell,18(4):893-906.
Mizukami Y,Fischer R L.2000.Plant organ size control:AINTEGUMENTA regulates growth and cell numbers during organogenesis.Proceedings of the National Academy of Sciences of the United States of America,97(2):942-947.
Mudunkothge J S,Krizek B A.2012.Three Arabidopsis AIL/PLT genes act in combination to regulate shoot apical meristem function.Plant Journal,71(1):108-121.
Nole-Wilson S,Tranby T L,Krizek B A.2005.AINTEGUMENTA-like(AIL)genes are expressed in young tissues and may specify meristematic or division-competent states.Plant Molecular Biology,57(5):613-628.
Qian Wenjun,Yue Chuan,Cao Hongli,Hao Xinyuan,Wang Lu,Wang Yuchun,Huang Yuting,Wang Bo,Wang Xinchao,Xiao Bin,Yang Yajun.2016.Cloning and expression analysis of a neutral/alkaline invertase gene(CsINV10)in tea plant[Camellia sinensis(L.)O.Kuntze].Acta Agronomica Sinica,42(3):376-388.(in Chinese)钱文俊,岳川,曹红利,郝心愿,王璐,王玉春,黄玉婷,王博,王新超,肖斌,杨亚军.2016.茶树中性/碱性转化酶基因CsINV10的克隆与表达分析.作物学报,42(3):376-388.
Randall R S,Sornay E,Dewitte W,Murray J A.2015.AINTEGUMENTA and the D-type cyclin CYCD3;1 independently contribute to petal size control in Arabidopsis:evidence for organ size compensation being an emergent rather than a determined property.Journal of Experimental Botany,66(13):3991-4000.
Ren Y R,Zhao Q,Zhao X Y,Hao Y J,You C X.2016.Expression analysis of the MdCIbHLH1 gene in apple flower buds and seeds in the process of dormancy.Horticultural Plant Journal,2(2):61-66.
Riechmann J L,Meyerowitz E M.1998.The AP2/EREBP family of plant transcription factors.Biological Chemistry,379(6):633-646.
Rigal A,Yordanov Y S,Perrone I,Karlberg A,Tisserant E,Bellini C,Busov V B,Martin F,Kohler A,Bhalerao R,Legue V.2012.The AINTEGUMENTA LIKE1 homeotic transcription factor PtAIL1 controls the formation of adventitious root primordia in poplar.Plant Physiology,160(4):1996-2006.
Rohde A,Bhalerao R P.2007.Plant dormancy in the perennial context.Trends in Plant Science,12(5):217-223.
Ruttink T,Arend M,Morreel K,Storme V,Rombauts S,Fromm J,Bhalerao R P,Boerjan W,Rohde A.2007.A molecular timetable for apical bud formation and dormancy induction in poplar.Plant Cell,19(8):2370-2390.
Sakuma Y,Liu Q,Dubouzet J G,Abe H,Shinozaki K,Yamaguchi-Shinozaki K.2002.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression.Biochemical&Biophysical Research Communications,290(3):998-1009.
Samach A,Onouchi H,Gold S E,Ditta G S,Schwarzsommer Z,Yanofsky M F,Coupland G.2000.Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis.Science,288(5471):1613-1616.
Singh R K,Svystun T,AlDahmash B,Jonsson A M,Bhalerao R P.2017.Photoperiod-and temperature-mediated control of phenology in treesa molecular perspective.New Phytologist,213(2):511-524.
Tang Hu,Hao Xinyuan,Wang Lu,Xiao Bin,Wang Xinchao,Yang Yajun.2017.Molecular regulation and substance exchange dynamics at dormancy and budbreak stages in overwintering buds of tea plant.Acta Agronomica Sinica,43(5):669-677.(in Chinese)唐湖,郝心愿,王璐,肖斌,王新超,杨亚军.2017.茶树越冬芽在休眠与萌发时期的物质交流变化及其分子调控.作物学报,43(5):669-677.
Vergara R,Noriega X,Parada F,Dantas D,Perez F J.2016.Relationship between endodormancy,FLOWERING LOCUS T and cell cycle genes in Vitis vinifera.Planta,243(2):411-419.
Wang X,Hao X,Ma C,Cao H,Yue C,Wang L,Zeng J,Yang Y.2014.Identification of differential gene expression profiles between winter dormant and sprouting axillary buds in tea plant(Camellia sinensis)by suppression subtractive hybridization.Tree Genetics&Genomes,10(5):1149-1159.
Wang Xinchao,Ma Chunlei,Yang Yajun,Jin Jiqiang,Ma Jianqiang,Cao Hongli.2012.cDNA cloning and expression analysis of cyclin-dependent kinase(CsCDK)gene in tea plant.Acta Horticulturae Sinica,39(2):333-342.(in Chinese)王新超,马春雷,杨亚军,金基强,马建强,曹红利.2012.茶树细胞周期蛋白依赖激酶(CsCDK)基因cDNA全长克隆与分析.园艺学报,39(2):333-342.
Wang Xinchao,Yang Yajun,Ma Chunlei,Jin Jiqiang,Ma Jianqiang,Cao Hongli.2011.Cloning and expression analysis of cyclin gene(CsCYC1)of tea plant.Acta Botanica Boreali-Occidentalia Sinica,31(12):2365-2372.(in Chinese)王新超,杨亚军,马春雷,金基强,马建强,曹红利.2011.茶树细胞周期蛋白基因的克隆与表达.西北植物学报,31(12):2365-2372.
Wei C,Yang H,Wang S,Zhao J,Liu C,Gao L,Xia E,Lu Y,Tai Y,She G,Sun J,Cao H,Tong W,Gao Q,Li Y,Deng W,Jiang X,Wang W,Chen Q,Zhang S,Li H,Wu J,Wang P,Li P,Shi C,Zheng F,Jian J,Huang B,Shan D,Shi M,Fang C,Yue Y,Li F,Li D,Wei S,Han B,Jiang C,Yin Y,Xia T,Zhang Z,Bennetzen J L,Zhao S,Wan X.2018.Draft genome sequence of Camellia sinensis var.sinensis provides insights into the evolution of the tea genome and tea quality.Proceedings of the National Academy of Sciences of the United States of America,115(18):E4151-E4158.
Wisniewski M,Norelli J,Bassett C,Artlip T,Macarisin D.2011.Ectopic expression of a novel peach(Prunus persica)CBF transcription factor in apple(Malus×domestica)results in short-day induced dormancy and increased cold hardiness.Planta,233(5):971-983.
Yanovsky M J,Kay S A.2002.Molecular basis of seasonal time measurement in Arabidopsis.Nature,419(6904):308-312.
Anastasiou E,Lenhard M.2007.Growing up to one’s standard.Current Opinion in Plant Biology,10(1):63-69.
Artlip T S,Wisniewski M E,Bassett C L,Norelli J L.2013.CBF gene expression in peach leaf and bark tissues is gated by a circadian clock.Tree Physiology,33(8):866-877.
Azeez A,Miskolczi P,Tylewicz S,Bhalerao R P.2014.A tree ortholog of APETALA1 mediates photoperiodic control of seasonal growth.Current Biology,24(7):717-724.
Chang S J,Puryear J,Cairney J.1993.A simple and efficient method for isolating RNA from pine trees.Plant Molecular Biology Reporter,11(2):113-116.
Devitt M L,Stafstrom J P.1995.Cell cycle regulation during growth-dormancy cycles in pea axillary buds.Plant Molecular Biology,29(2):255-265.
Elliott R C,Betzner A S,Huttner E,Oakes M P,Tucker W Q,Gerentes D,Perez P,Smyth D R.1996.AINTEGUMENTA,an APETALA2-like gene of Arabidopsis with pleiotropic roles in ovule development and floral organ growth.Plant Cell,8(2):155-168.
Espinosa-Ruiz A,Saxena S,Schmidt J,Mellerowicz E,Miskolczi P,Bako L,Bhalerao R P.2004.Differential stage-specific regulation of cyclin-dependent kinases during cambial dormancy in hybrid aspen.Plant Journal,38(4):603-615.
Foley M E F,Anderson J V A,Horvath D P H.2009.The effects of temperature,photoperiod,and vernalization on regrowth and flowering competence in Euphorbia esula(Euphorbiaceae)crown buds.Botany-botanique,87(10):986-992.
Hao X,Horvath D P,Chao W S,Yang Y,Wang X,Xiao B.2014.Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant[Camellia sinensis(L.)O.Kuntze].International Journal of Molecular Sciences,15(12):22155.
Hao X,Yang Y,Yue C,Wang L,Horvath D P,Wang X.2017.Comprehensive transcriptome analyses reveal differential gene expression profiles of Camellia sinensis axillary buds at para-,endo-,ecodormancy,and bud flush stages.Frontiers in Plant Science,8(481):553.
Hao Xinyuan.2015.Study on the molecular mechanism of winter bud dormancy in tea plant[Ph.D.Dissertation].Yangling:Northwest A&FUniversity.(in Chinese)郝心愿.2015.茶树越冬芽休眠的分子机理研究[博士论文].杨凌:西北农林科技大学.
Horvath D P,Anderson J V,Chao W S,Foley M E.2003.Knowing when to grow:signals regulating bud dormancy.Trends in Plant Science,8(11):534-540.
Horvath D P,Chao W S,Anderson J V.2002.Molecular analysis of signals controlling dormancy and growth in underground adventitious buds of leafy spurge.Plant Physiology,128(4):1439-1446.
Karlberg A,Bako L,Bhalerao R P.2011.Short day-mediated cessation of growth requires the downregulation of AINTEGUMENTALIKE1transcription factor in hybrid aspen.PLoS Genetics,7(11):e1002361.
Klucher K M,Chow H,Reiser L,Fischer R L.1996.The AINTEGUMENTA gene of Arabidopsis required for ovule and female gametophyte development is related to the floral homeotic gene APETALA2.Plant Cell,8(2):137-153.
Krizek B A.2003.AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain.Nucleic Acids Research,31(7):1859-1868.
Livak K J,Schmittgen T D.2001.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method.Methods,25(4):402-408.
Menges M,Samland A K,Planchais S,Murray J A.2006.The D-type cyclin CYCD3:1 is limiting for the G1-to-S-phase transition in Arabidopsis.Plant Cell,18(4):893-906.
Mizukami Y,Fischer R L.2000.Plant organ size control:AINTEGUMENTA regulates growth and cell numbers during organogenesis.Proceedings of the National Academy of Sciences of the United States of America,97(2):942-947.
Mudunkothge J S,Krizek B A.2012.Three Arabidopsis AIL/PLT genes act in combination to regulate shoot apical meristem function.Plant Journal,71(1):108-121.
Nole-Wilson S,Tranby T L,Krizek B A.2005.AINTEGUMENTA-like(AIL)genes are expressed in young tissues and may specify meristematic or division-competent states.Plant Molecular Biology,57(5):613-628.
Qian Wenjun,Yue Chuan,Cao Hongli,Hao Xinyuan,Wang Lu,Wang Yuchun,Huang Yuting,Wang Bo,Wang Xinchao,Xiao Bin,Yang Yajun.2016.Cloning and expression analysis of a neutral/alkaline invertase gene(CsINV10)in tea plant[Camellia sinensis(L.)O.Kuntze].Acta Agronomica Sinica,42(3):376-388.(in Chinese)钱文俊,岳川,曹红利,郝心愿,王璐,王玉春,黄玉婷,王博,王新超,肖斌,杨亚军.2016.茶树中性/碱性转化酶基因CsINV10的克隆与表达分析.作物学报,42(3):376-388.
Randall R S,Sornay E,Dewitte W,Murray J A.2015.AINTEGUMENTA and the D-type cyclin CYCD3;1 independently contribute to petal size control in Arabidopsis:evidence for organ size compensation being an emergent rather than a determined property.Journal of Experimental Botany,66(13):3991-4000.
Ren Y R,Zhao Q,Zhao X Y,Hao Y J,You C X.2016.Expression analysis of the MdCIbHLH1 gene in apple flower buds and seeds in the process of dormancy.Horticultural Plant Journal,2(2):61-66.
Riechmann J L,Meyerowitz E M.1998.The AP2/EREBP family of plant transcription factors.Biological Chemistry,379(6):633-646.
Rigal A,Yordanov Y S,Perrone I,Karlberg A,Tisserant E,Bellini C,Busov V B,Martin F,Kohler A,Bhalerao R,Legue V.2012.The AINTEGUMENTA LIKE1 homeotic transcription factor PtAIL1 controls the formation of adventitious root primordia in poplar.Plant Physiology,160(4):1996-2006.
Rohde A,Bhalerao R P.2007.Plant dormancy in the perennial context.Trends in Plant Science,12(5):217-223.
Ruttink T,Arend M,Morreel K,Storme V,Rombauts S,Fromm J,Bhalerao R P,Boerjan W,Rohde A.2007.A molecular timetable for apical bud formation and dormancy induction in poplar.Plant Cell,19(8):2370-2390.
Sakuma Y,Liu Q,Dubouzet J G,Abe H,Shinozaki K,Yamaguchi-Shinozaki K.2002.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression.Biochemical&Biophysical Research Communications,290(3):998-1009.
Samach A,Onouchi H,Gold S E,Ditta G S,Schwarzsommer Z,Yanofsky M F,Coupland G.2000.Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis.Science,288(5471):1613-1616.
Singh R K,Svystun T,AlDahmash B,Jonsson A M,Bhalerao R P.2017.Photoperiod-and temperature-mediated control of phenology in treesa molecular perspective.New Phytologist,213(2):511-524.
Tang Hu,Hao Xinyuan,Wang Lu,Xiao Bin,Wang Xinchao,Yang Yajun.2017.Molecular regulation and substance exchange dynamics at dormancy and budbreak stages in overwintering buds of tea plant.Acta Agronomica Sinica,43(5):669-677.(in Chinese)唐湖,郝心愿,王璐,肖斌,王新超,杨亚军.2017.茶树越冬芽在休眠与萌发时期的物质交流变化及其分子调控.作物学报,43(5):669-677.
Vergara R,Noriega X,Parada F,Dantas D,Perez F J.2016.Relationship between endodormancy,FLOWERING LOCUS T and cell cycle genes in Vitis vinifera.Planta,243(2):411-419.
Wang X,Hao X,Ma C,Cao H,Yue C,Wang L,Zeng J,Yang Y.2014.Identification of differential gene expression profiles between winter dormant and sprouting axillary buds in tea plant(Camellia sinensis)by suppression subtractive hybridization.Tree Genetics&Genomes,10(5):1149-1159.
Wang Xinchao,Ma Chunlei,Yang Yajun,Jin Jiqiang,Ma Jianqiang,Cao Hongli.2012.cDNA cloning and expression analysis of cyclin-dependent kinase(CsCDK)gene in tea plant.Acta Horticulturae Sinica,39(2):333-342.(in Chinese)王新超,马春雷,杨亚军,金基强,马建强,曹红利.2012.茶树细胞周期蛋白依赖激酶(CsCDK)基因cDNA全长克隆与分析.园艺学报,39(2):333-342.
Wang Xinchao,Yang Yajun,Ma Chunlei,Jin Jiqiang,Ma Jianqiang,Cao Hongli.2011.Cloning and expression analysis of cyclin gene(CsCYC1)of tea plant.Acta Botanica Boreali-Occidentalia Sinica,31(12):2365-2372.(in Chinese)王新超,杨亚军,马春雷,金基强,马建强,曹红利.2011.茶树细胞周期蛋白基因的克隆与表达.西北植物学报,31(12):2365-2372.
Wei C,Yang H,Wang S,Zhao J,Liu C,Gao L,Xia E,Lu Y,Tai Y,She G,Sun J,Cao H,Tong W,Gao Q,Li Y,Deng W,Jiang X,Wang W,Chen Q,Zhang S,Li H,Wu J,Wang P,Li P,Shi C,Zheng F,Jian J,Huang B,Shan D,Shi M,Fang C,Yue Y,Li F,Li D,Wei S,Han B,Jiang C,Yin Y,Xia T,Zhang Z,Bennetzen J L,Zhao S,Wan X.2018.Draft genome sequence of Camellia sinensis var.sinensis provides insights into the evolution of the tea genome and tea quality.Proceedings of the National Academy of Sciences of the United States of America,115(18):E4151-E4158.
Wisniewski M,Norelli J,Bassett C,Artlip T,Macarisin D.2011.Ectopic expression of a novel peach(Prunus persica)CBF transcription factor in apple(Malus×domestica)results in short-day induced dormancy and increased cold hardiness.Planta,233(5):971-983.
Yanovsky M J,Kay S A.2002.Molecular basis of seasonal time measurement in Arabidopsis.Nature,419(6904):308-312.