逆境胁迫影响茶树生长发育及茶叶品质,ASR（abscisic acid, stress, ripening-induced）基因在植物抗逆响应中具有重要功能。本研究以龙井43品种茶树为材料,从中克隆了CsASR基因的全长cDNA序列、基因组序列及其启动子序列,分析了该基因的生物信息学特征及在组织间和不同胁迫处理下的表达模式。结果显示,CsASR的cDNA序列全长875βbp,含有546βbp的ORF序列,编码181个氨基酸,蛋白质分子量19.89βkD,理论等电点5.69;CsASR蛋白结构序列中74.5%的序列为无序结构,是一种无序蛋白;CsASR的C-端含有ABA/WDS功能结构域,主要定位于细胞质和细胞核中;茶树CsASR与海枣的ASR相似性最高,为87%,而在进化树中与枣的关系最近。CsASR基因含2个外显子,第1个外显子长363βbp,第2个外显子长183βbp,内含子较大为2β750βbp,内含子中含7种简单重复序列和2种DNA转座子序列。克隆获得起始密码子ATG上游2β554βbp的启动子区序列,该启动子上含有干旱、低温、高温以及ABA等相关的顺式作用元件。CsASR在根中的表达量最低;ABA抑制CsASR的表达,而干旱、NaCl和低温胁迫能够显著上调CsASR的表达。表明CsASR基因可能与茶树抗逆密切相关。
Abstract
Abiotic stress severely affects the growth and development of tea plant and the quality of tea. ASR (abscisic acid, stress, ripening) genes play crucial roles in the plant response to stresses. The full-length cDNA, genomic sequence and the promoter sequence of CsASR gene were cloned from tea cultivar Longjing 43 in this study, The bioinformatic and the expression analysis of CsASR in tissues under different stress treatments were performed. The results revealed that the full-length cDNA of CsASR was 875βbp, containing a 546βbp ORF encoding 181 amino acid. The predict protein molecular and theoretic isoelectric point of CsASR were 19.89βkD and 5.69. Most regions of the amino acid sequence (74.5%) were predicted as the non-ordered regions, indicating that CsASR is a disordered protein. C-terminal of CsASR contained an ABA/WDS functional domain which was primarily located in both the cytoplasm and the nucleus. The homologous alignment and phylogenetic tree analysis showed that CsASR had the highest similarity (87%) with Phoenix dactylifera, and had the closest genetic relationship with Ziziphus nummularia. The genomic sequence of CsASR gene was comprised by two exons with 363 bp and 183 bp in length, respectively, and had a 2 750 bp intron which contained seven simple repeats and two DNA transposons. The promoter sequence of CsASR was 2 554 bp in length and was predicted to contain several stress-responsive elements related to drought, cold, high temperature stresses and ABA-signaling. The expression analysis showed that CsASR had the lowest level in roots, and its expression was repressed by ABA treatment. While the drought, NaCl and cold stresses could significantly up-regulate the expression of CsASR. The results revealed that CsASR might be closely related to stress response in tea plant.
关键词
ASR基因 /
茶树 /
基因克隆 /
逆境胁迫
Key words
abiotic stress /
ASR gene /
gene cloning /
tea plant
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Gonzalez RM, Iusem ND.Twenty years of research on Asr (ABA-stress-ripening) genes and proteins[J]. Planta, 2014, 239(5): 941-949.
[2] Frankel N, Carrari F, Hasson E, et al.Evolutionary history of the Asr gene family[J]. Gene, 2006, 378: 74-83.
[3] Golan I, Dominguez PG, Konrad Z, et al.Tomato abscisic acid stress ripening (ASR) gene family revisited[J]. PLoS One, 2014, 9(10): e107117.
[4] Huang K, Zhong Y, Li Y, et al.Genome-wide identification and expression analysis of the apple ASR gene family in response to Alternaria alternata f. sp. Mali[J]. Genome, 2016, 59(10): 866-878.
[5] Virlouvet L, Jacquemot MP, Gerentes D, et al.The ZmASR1 protein influences branched-chain amino acid biosynthesis and maintains kernel yield in maize under water-limited conditions[J]. Plant Physiol, 2011, 157(2): 917-936.
[6] Jia H, Jiu S, Zhang C, et al.Abscisic acid and sucrose regulate tomato and strawberry fruit ripening through the abscisic acid-stress-ripening transcription factor[J]. Plant Biotechnology Journal, 2016, 14(10): 2045-2065.
[7] Tiwari V, Chaturvedi AK, Mishra A, et al.Introgression of the SbASR-1 gene cloned from a halophyte Salicornia brachiate enhances salinity and drought endurance in transgenic groundnut (arachis hypogaea) and acts as a transcription factor[J]. PLoS One, 2015, 10(7): e0131567.
[8] Hu W, Huang C, Deng X, et al.TaASR1, a transcription factor gene in wheat, confers drought stress tolerance in transgenic tobacco[J]. Plant, Cell & Environment, 2013, 36(8): 1449-1464.
[9] renhart RA, Bai Y, Valter de Oliveira LF, et al. New insights into aluminum tolerance in rice: The ASR5 protein binds the STAR1 promoter and other aluminum-responsive genes[J]. Molecular Plant, 2014, 7(4): 709-721.
[10] Kim IS, Kim YS, Yoon HS.Rice ASR1 protein with reactive oxygen species scavenging and chaperone-like activities enhances acquired tolerance to abiotic stresses in Saccharomyces cerevisiae [J]. Molecular and Cell, 2012, 33(3): 285-293.
[11] Konrad Z and Bar-Zvi D. Synergism between the chaperone-like activity of the stress regulated ASR1 protein and the osmolyte glycine-betaine[J]. Planta, 2008, 227(6): 1213-1219.
[12] Saumonneau A, Laloi M, Lallemand M, et al.Dissection of the transcriptional regulation of grape ASR and response to glucose and abscisic acid[J]. Journal of Experimental Botany, 2012, 63(3): 1495-1510.
[13] Arenhart RA, Lima JC, Pedron M, et al.Involvement of ASR genes in aluminium tolerance mechanisms in rice[J]. Plant, Cell & Environment, 2013, 36(1): 52-67.
[14] Arenhart RA, Schunemann M, Bucker Neto L, et al.Rice ASR1 and ASR5 are complementary transcription factors regulating aluminium responsive genes[J]. Plant, Cell & Environment, 2016, 39(3): 645-651.
[15] Sun P, Miao H, Yu X, et al.A Novel role for banana MaASR in the regulation of flowering time in transgenic Arabidopsis [J]. PLoS One, 2016, 11(8): e0160690.
[16] 岳川, 曹红利, 周艳华, 等. 茶树谷胱甘肽还原酶基因CsGRs的克隆与表达分析[J]. 中国农业科学, 2014, 47(16): 3277-3289.
[17] Yue C, Cao H, Wang L, et al.Molecular cloning and expression analysis of tea plant aquaporin (AQP) gene family[J]. Plant Physiology and Biochemistry, 2014, 83: 65-76.
[18] Hao X, Horvath D, Chao W, et al.Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant (Camellia sinensis (L.) O. Kuntze)[J]. International Journal of Molecular Sciences, 2014, 15(12): 22155-22172.
[19] Hu YX, Yang X, Li XL, et al.The SlASR gene cloned from the extreme halophyte Suaeda liaotungensis K. enhances abiotic stress tolerance in transgenic Arabidopsis thaliana [J]. Gene, 2014, 549(2): 243-251.
[20] Wang L, Hu W, Feng J, et al.Identification of the ASR gene family from Brachypodium distachyon and functional characterization of BdASR1 in response to drought stress[J]. Plant Cell Reports, 2016, 35(6): 1221-1234.
[21] Feng ZJ, Xu ZS, Sun J, et al.Investigation of the ASR family in foxtail millet and the role of ASR1 in drought/oxidative stress tolerance[J]. Plant Cell Reports, 2016, 35(1): 115-128.
[22] Henry IM, Carpentier SC, Pampurova S, et al.Structure and regulation of the Asr gene family in banana[J]. Planta, 2011, 234(4): 785-798.
[23] Cakir B, Agasse A, Gaillard C, et al.A grape ASR protein involved in sugar and abscisic acid signaling[J]. The Plant Cell, 2003, 15(9): 2165-2180.
[24] Li J, Li Y, Yin Z, et al.OsASR5 enhances drought tolerance through a stomatal closure pathway associated with ABA and H2O2 signaling in rice[J]. Plant Biotechnology Journal, 2017, 15(2): 183-196.
[25] Perez-Diaz J, Wu TM, Perez-Diaz R, et al.Organ- and stress-specific expression of the ASR genes in rice[J]. Plant Cell Reports, 2014, 33(1): 61-73.
[26] Zhang L, Hu W, Wang Y, et al.The MaASR gene as a crucial component in multiple drought stress response pathways in Arabidopsis [J]. Functional & Integrative Genomics, 2015, 15(2):247-260.
[27] Yang CY, Chen YC, Jauh GY, et al.A Lily ASR protein involves abscisic acid signaling and confers drought and salt resistance in Arabidopsis [J]. Plant Physiology, 2005. 139(2): 836-846.
[28] Liu HY, Dai JR, Feng DR, et al.Characterization of a novel plantain Asr gene, MpAsr, that is regulated in response to infection of Fusarium oxysporum f. sp. cubense and abiotic stresses[J]. Journal of Integrative Plant Biology, 2010, 52(3): 315-323.
[29] Shkolnik D, Bar-Zvi D.Tomato ASR1 abrogates the response to abscisic acid and glucose in Arabidopsis by competing with ABI4 for DNA binding[J]. Plant Biotechnology Journal, 2008, 6(4): 368-378.
基金
国家自然科学基金（31600555）、福建省自然科学基金项目（2017J01616）、国家茶叶产业技术体系（CARS-23）
PDF(1025 KB)
浙公网安备 33019902000101号