生长素相关基因在茶树腋芽冬季休眠不同生长阶段的表达分析

doi:10.13305/j.cnki.jts.2012.06.012

摘要/Abstract

摘要： 利用实时定量PCR技术研究了从本实验室构建的茶树休眠芽与萌动芽抑制消减杂交文库中分离出来的与生长素相关的基因片段在茶树腋芽冬季休眠不同阶段的表达模式。研究结果表明：（1）与生长素响应因子同源的6条EST片段中,有5条的表达量休眠期高于萌发期,另外1条则相反。（2）与生长素原初反应基因同源的3条EST片段的表达模式存在差异,其中与SAURs同源的EST在休眠阶段上调表达,休眠解除以后下调表达。而与GH3和Aux/IAAs同源的2条EST在萌发阶段有一个表达高峰,而在休眠期和萌发以后,表达量都下降。（3）与生长素结合类蛋白同源的茶树CsGLP1表达量在萌发期高于休眠期。（4）与生长素内向运输载体基因同源的EST片段在深休眠期的表达量最高。（5）2条受生长素调控的EST表现出不同的表达模式,生长素诱导表达的EST在休眠期的表达量高于萌发期及随后阶段,而另外1条受生长素抑制的EST片段在萌发期的表达远远高于休眠期。这些结果初步说明生长素促进茶树腋芽的冬季休眠。

关键词: 茶树（Camellia sinensis）, 芽休眠与萌发, 生长素相关基因, 表达分析

Abstract: The expression patterns of some auxin-related genes, including auxin response factors (ARFs), primary auxin responsive genes, auxin-regulated genes, auxin influx carrier gene and auxin-binding protein (ABP) gene, were studied in different stages of tea plant axillary bud dormancy and its release. The results showed that (1) the expression levels of five in six ARF homologous EST fragments were up-regulated in the deep dormancy and down-regulated after sprouted, the level of the last one showed the opposite expression levels. (2) There were significant differences among the expression patterns of 3 ESTs which are homologous with primary auxin responsive genes. EST which is homologous with SAURs was up-regulated in the dormancy stage but down-regulated after dormancy release. On the contrary, the 2 ESTs which are homologous with GH3 and Aux/IAAs were up-regulated in sprouting stage while down-regulated in dormant stages and after sprouting stages. (3) The CsGLP1 gene which was homogenous with ABPs has a lower expression level in the dormancy stage but maintain rising trend after germination.(4) The EST which is homogenous with auxin influx carrier gene showed the highest expression levels during the deep dormant stage but maintain lower levels in other stages. (5) Two auxin-regulated ESTs showed different expression patterns. The expression levels of auxin-induced EST were higher in the dormant stages than those in the sprouting stages, while the levels of the other auxin-repressed EST were higher in the sprouting stage than that in the dormant stages. These results indicated that auxin promote the dormancy of tea axillary buds in winter.

Key words: tea plant (Camellia sinensis), bud dormancy and budbreak, auxin-related gene, expression analysis

中图分类号:

S571.1
Q52

王新超, 马春雷, 杨亚军, 曹红利, 郝心愿, 金基强. 生长素相关基因在茶树腋芽冬季休眠不同生长阶段的表达分析[J]. 茶叶科学, 2012, 32(6): 509-516. doi: 10.13305/j.cnki.jts.2012.06.012.

WANG Xin-chao, MA Chun-lei, YANG Ya-jun, CAO Hong-li, HAO Xin-yuan, JIN Ji-qiang. Expression Analysis of Auxin-related Genes at Different Winter Dormant Stages of Axillary Buds in Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2012, 32(6): 509-516. doi: 10.13305/j.cnki.jts.2012.06.012.

参考文献

[1] Arora R, Rowland L J, Tanino K.Induction and release of bud dormancy in woody perennials: a science comes of age[J]. HortScience, 2003, 38: 911-921.
[2] Vanneste S, Friml J.Auxin: a trigger for change in plant development[J]. Cell, 2009, 136: 1005-1016.
[3] Zhao Y.Auxin biosynthesis and its role in plant development[J]. Annual Review of Plant Biology, 2010, 61: 49-64.
[4] Horvath D P, Anderson J V, Chao W S, et al.Knowing when to grow: signals regulating bud dormancy[J]. Trends in Plant Science, 2003, 8: 534-540.
[5] Shimizu-Sato S, Tanaka M, Mori H.Auxin-cytokinin interactions in the control of shoot branching[J]. Plant Molecular Biology, 2009, 69: 429-435.
[6] 王新超, 杨亚军, 陈亮, 等. 茶树休眠芽与萌动芽抑制消减杂交文库的构建与初步分析[J]. 茶叶科学, 2010, 30(2): 129-135.
[7] 赵丽萍, 陈亮, 王新超, 等. 茶树新梢不同叶片中β-葡萄糖苷酶和β-樱草糖苷酶基因表达的实时定量PCR分析[J]. 茶叶科学, 2006, 26(1): 11-16.
[8] 王新超, 马春雷, 杨亚军, 等. 茶树生长素抑制蛋白基因CsARP1的克隆与表达分析[J]. 核农学报, 2011, 25(5): 910-915.
[9] Guilfoyle T J, Hagen G.Auxin response factors[J]. Current Opinion in Plant Biology, 2007, 10: 453-460.
[10] Kieffer M, Neve J, Kepinski S.Defining auxin response contexts in plant development[J]. Current Opinion in Plant Biology, 2010, 13:12-20.
[11] Hagen G, Guilfoyle T.Auxin-responsive gene expression: genes, promoters and regulatory factors[J]. Plant Molecular Biology, 2002, 49: 373-385.
[12] 倪迪安, 许智宏. 生长素的生物合成、代谢、受体和极性运输[J]. 植物生理学通讯, 2001, 37: 346-352.
[13] Dunwell J M, Gibbings J G, Mahmood T, et al. Germin and germin-like proteins: evolution, structure, and function[J]. Critical Reviews in Plant Sciences, 2008, 27: 342-375.
[14] Timpte C.Auxin binding protein: curiouser and curiouser[J]. Trends in Plant Science, 2001, 6: 586-590.
[15] Ulmasov T, Hagen G, Guilfoyle T J.Dimerization and DNA binding of auxin response factors[J]. The Plant Journal, 1999, 19: 309-319.
[16] Ulmasov T, Hagen G, Guilfoyle T J.Activation and repression of transcription by auxin-response factors[J]. Proceedings of National Academy of Sciences of the United States of America, 1999, 96: 5844-5849.
[17] Faivre-Rampant O, Cardle L, Marshall D, et al. Changes in gene expression during meristem activation processes in Solanum tuberosum with a focus on the regulation of an auxin response factor gene[J]. Journal of Experimental Botany, 2004, 55: 613-622.
[18] Schrader J, Moyle R, Bhalerao R, et al. Cambial meristem dormancy in trees involves extensive remodelling of the transcriptome[J]. The Plant Journal, 2004, 40: 173-187.
[19] Tiwari S B, Wang X J, Hagen G, et al. Aux/IAA proteins are active repressors and their stability and activity are modulated by auxin[J]. The Plant Cell, 2001, 13: 2809-2822.
[20] Staswick P E, Serban B, Rowe M, et al. Characterization of an Arabidopsis enzyme family that conjugates amino acids to indole-3-acetic acid[J]. The Plant Cell, 2005, 17: 616-627.
[21] Jain M, Tyagi A K, Khurana J P.Genome-wide analysis, evolutionary expansion, and expression of early auxin-responsive SAUR gene family in rice (Oryza sativa)[J]. Genomics, 2006, 88: 360-371.
[22] Grzelczak Z F, Lane B G.Signal resistance of a soluble protein to enzyme hydrolysis. An unorthodox approach to the isolation and purification of germin, a rare growth-related protein[J].Canadian Journal of Biochemistry and Cell Biology, 1984, 62: 1351-1352.
[23] Dunwell J M, Gibbings J G, Mahmood T, et al. Germin and germin-like proteins: evolution, structure, and function[J]. Critical Reviews in Plant Sciences, 2008, 27: 342-375.
[24] Ohmiya M, Sakai K S, Hayashi T.Purification and properties of an auxin-binding protein from the shoot apex of peach tree[J]. Plant Cell Physiology, 1993, 34: 177-183.
[25] Ohmiya A, Tanaka Y, Kadowaki K, et al. Cloning of genes encoding auxin-binding proteins (ABP19/20) from peach: significant peptide sequence similarity with germin-like proteins[J]. Plant Cell Physiology, 1998, 39: 492-499.
[26] Aubry C, Paven M C M L, Chateigner-Boutin A L, et al. A gene encoding a germin-like protein, identified by a cDNA-AFLP approach, is specifically expressed during germination of Phaseolus vulgaris[J]. Planta, 2003, 217: 466-475.
[27] Friml J, Vieten A, Sauer M, et al. Efflux-dependent auxin gradients establish the apical-basal axis of Arabidopsis[J]. Nature, 2003, 426: 147-153.
[28] Vieten A, Sauer M, Philip B, et al. Molecular and cellular aspects of auxin-transport-mediated development[J]. Trends in Plant Science, 2007, 12: 160-168.
[29] Bainbridge K, Guyomarc’h S, Bayer E, et al. Auxin influx carriers stabilize phyllotactic patterning[J]. Genes & Development, 2008, 22: 810-823.
[30] Ugartechea-Chirino Y, Swarup R, Swarup K, et al. The AUX1 LAX family of auxin influx carriers is required for the establishment of embryonic root cell organization in Arabidopsis thaliana[J]. Annals of Botany, 2010, 105: 277-289.
[31] Schrader J, Baba K, May S T, et al. Polar auxin transport in the wood-forming tissues of hybrid aspen is under simultaneous control of developmental and environmental signals[J]. Proceedings of National Academy of Sciences of the United States of America, 2003, 100: 10096-10101.
[32] Baba K, Karlberg A, Schmidt J, et al. Activity-dormancy transition in the cambial meristem involves stage-specific modulation of auxin response in hybrid aspen[J]. Proceedings of National Academy of Sciences of the United States of America, 2011, 108: 2418-2423.
[33] He D, Mathiason K, Fennell A.Auxin and cytokinin related gene expression during active shoot growth and latent bud paradormancy in Vitis riparia grapevine[J]. Journal of Plant Physiology, 2012, 169: 643-648
[34] John P C L. Hormonal regulation of cell cycle progression and its role in development [M]. //Inzè D (eds). Cell Cycle Control and Plant Development. Oxford: Blackwell Publishing, 2007: 311-334.
[35] McSteen P, Leyser O. Shoot branching[J]. Annual Review of Plant Biology, 2005, 56: 353-374.
[36] Napoli C A, Beveridge C A, Snowden K C.Reevaluating concepts of apical dominance and the control of axillary bud outgrowth[J]. Current Topics in Development Biology, 1999, 44: 127-169.
[37] Santner A, Calderon-Villalobos L I A, Estelle M. Plant hormones are versatile chemical regulators of plant growth[J]. Nature Chemical Biology, 2009, 5: 301-307.
[38] Shimizu S, Mori H.Control of ourgrowth and dormancy in axillary buds[J]. Plant Physiology, 2001, 127: 1405-1413.
[39] Aloni R, Peterson C A.Auxin promotes dormancy callose removal from the phloem of Magnolia kobus and callose accumulation and early wood vessel differentiation in Quercus robur[J]. Journal of Plant Research, 1997, 110: 37-44.
[40] Aloni R, Raviv A, Peterson C A.The role of auxin in the removal of dormancy callose and resumption of phloem activity in Vitis vinifera[J]. Canadian Journal of Botany, 1991, 69: 1825-1832.
[41] Nagar P K, Shweta S.Changes in endogenous auxins during winter dormancy in tea (Camellia sinensis (L.) O. Kuntze)[J]. Acta Physiologiae Plantarum, 2006, 28:165-169.
[42] 潘根生, 钱利生, 沈生荣, 等. 茶树休眠与内源激素的关系[J]. 茶叶, 2000, 26(4): 200-204.