类茶氨酸合成酶基因家族在番茄中的鉴定及表达研究

毛鹏, 王丽鸳, 白培贤, 韦康, 阮丽, 张亚真, 成浩

茶叶科学 ›› 2021, Vol. 41 ›› Issue (2) : 173-183.

PDF(606 KB)
PDF(606 KB)
茶叶科学 ›› 2021, Vol. 41 ›› Issue (2) : 173-183. DOI: 10.13305/j.cnki.jts.2021.02.003
研究报告

类茶氨酸合成酶基因家族在番茄中的鉴定及表达研究

  • 毛鹏1,2, 王丽鸳1,*, 白培贤1, 韦康1, 阮丽1, 张亚真1, 成浩1,*
作者信息 +

Identification and Analysis of Analogue Theanine Synthase Gene Family in Tomato

  • MAO Peng1,2, WANG Liyuan1,*, BAI Peixian1, WEI Kang1, RUAN Li1, ZHNAG Yazhen1, CHENG Hao1,*
Author information +
文章历史 +

摘要

茶树体内的茶氨酸合成酶(Theanine synthetase synthetase,TS)和部分谷氨酰胺合成酶(Glutamine synthetase,GS)家族成员都具有催化乙胺和谷氨酸合成茶氨酸的活性。我们将植物中这类与茶树TS序列高度相似,且具有茶氨酸合成活性的酶,统称为类茶氨酸合成酶(Analogue theanine synthase,ATS)。为了探究ATS在非茶植物中的分布,以及乙胺在茶氨酸合成中的作用,本研究以Mico-Tom番茄为试验材料,对水培番茄幼苗进行了外源添加乙胺处理。结果发现,仅在处理组叶片中检测到茶氨酸,并且处理组叶片中的谷氨酰胺(Glutamine,Gln)含量也极显著增加,而丙氨酸含量与对照相比无显著变化。利用生物信息学方法分别从茶树和番茄基因组中鉴定出12个和5个ATS家族成员,它们均含有谷氨酰胺合成酶催化功能的结构域Gln-synt_C;qRT-PCR检测发现,处理组叶片中有2个ATSSlGS1.1SlGS1)表达量极显著提高。上述结果表明,ATS广泛存在于植物中,而乙胺是合成茶氨酸的关键限制因子,能够提高ATS基因的表达,触发茶氨酸的合成。

Abstract

Theanine synthetase (TS) and some members of the glutamine synthetase (GS) family in tea plants show catalytic activity for the synthesis of theanine from ethylamine and glutamic acid. The enzymes in plants that show catalytic activity for the synthesis of theanine and are highly similar to TS sequence of tea plants are named as the analogue theanine synthase (ATS). To explore the existence of ATS in non-tea plants and effect of ethylamine on theanine synthesis, ‘Mico-Tom’ tomato was used as materials. As tomato seedlings was treated hydroponically with ethylamine, theanine was successfully detected in the leaves of the treated group. The content of glutamine (Gln) in the tomato leaves was also significantly increased, while the content of alanine did not change compared with the control group. Then, 12 and 5 members of the ATS family were identified respectively in tea and tomato through bioinformatics analysis and all of them contained the Gln-synt_C domain of glutamine synthase. Further qRT-PCR results show that the expressions of 2 ATS (SlGS1.1 and SlGS1) in the leaves were significantly increased under the treatment of ethylamine. In conclusion, the ATS may exist widely in plants, and ethylamine would be a key limiting factor in the synthesis of theanine as an important precursor. Meanwhile, ethylamine can greatly accelerate the production of theanine by promoting the transcripts of ATS.

关键词

茶氨酸 / 番茄 / 基因家族 / 类茶氨酸合成酶 / 乙胺

Key words

analogue theanine synthase / ethylamine / gene family / theanine / tomato

引用本文

导出引用
毛鹏, 王丽鸳, 白培贤, 韦康, 阮丽, 张亚真, 成浩. 类茶氨酸合成酶基因家族在番茄中的鉴定及表达研究[J]. 茶叶科学. 2021, 41(2): 173-183 https://doi.org/10.13305/j.cnki.jts.2021.02.003
MAO Peng, WANG Liyuan, BAI Peixian, WEI Kang, RUAN Li, ZHNAG Yazhen, CHENG Hao. Identification and Analysis of Analogue Theanine Synthase Gene Family in Tomato[J]. Journal of Tea Science. 2021, 41(2): 173-183 https://doi.org/10.13305/j.cnki.jts.2021.02.003
中图分类号: S571    Q52   

参考文献

[1] Chen Z M, Chen L.Delicious and healthy tea: an overview[M]//Chen L, Apostolides Z, Chen Z M. Global Tea Breeding. Berlin: Springer, 2012.
[2] Zhao J, Li P, Xia T, et al.Exploring plant metabolic genomics: chemical diversity, metabolic complexity in the biosynthesis and transport of specialized metabolites with the tea plant as a model[J]. Critical Reviews in Biotechnology, 2020, 40(5): 667-688.
[3] 毛雅琳, 汪芳, 尹军峰, 等. 不同茶树品种碾茶的品质分析[J]. 茶叶科学, 2020, 40(6): 782-794.
Mao Y L, Wang F, Yin J F, et al.Quality analysis of tencha made from different tea cultivars[J]. Journal of Tea Science, 2020, 40(6): 782-794.
[4] Yamaguchi S, Ninomiya K.Umami and food palatability[J]. The Journal of Nutrition, 2000, 130(4): 921-926.
[5] Yan Z M, Zhong Y Z, Duan Y H, et al.Antioxidant mechanism of tea polyphenols and its impact on health benefits[J]. Animal Nutrition, 2020, 6(2): 115-123.
[6] Yu Z, Yang Z.Understanding different regulatory mechanisms of proteinaceous and non-proteinaceous amino acid formation in tea (Camellia sinensis) provides new insights into the safe and effective alteration of tea flavor and function[J]. Critical Reviews in Food Science and Nutrition, 2020, 60(5): 844-858.
[7] Deng W W, Ogita S, Ashihara H.Distribution and biosynthesis of theanine in Theaceae plants[J]. Plant Physiology and Biochemistry, 2010, 48(1): 70-72.
[8] Cheng S H, Fu X M, Wang X Q, et al.Studies on the biochemical formation pathway of the amino acid L-theanine in tea (Camellia sinensis) and other plants[J]. Journal of Agricultural and Food Chemistry, 2017, 65(33): 7210-7216.
[9] Wei C L, Yang H, Wang S B, et al.Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality[J]. PNAS, 2018, 115(18): 4151-4158.
[10] Sasaoka K, Kito M, Onishi Y.Some properties of the theanine synthesizing enzyme in tea seedlings[J]. Agricultural and Biological Chemistry, 1965, 29(11): 984-988.
[11] Sasaoka K, Kito M, Inagaki H.Studies on the biosynthesis of theanine in tea seedlings[J]. Agricultural and Biological Chemistry, 1963, 27(6): 467-468.
[12] Okada Y, Koseki M, Chu M. Protein and cDNA sequences of two theanine synthetases from Camellia sinensis: JP2006254780 [P/OL].2006-09-28.
[13] Deng W W, Ogita S, Ashihara H.Biosynthesis of theanine (γ-ethylamino-L-glutamic acid) in seedlings of Camellia sinensis[J]. Phytochemistry Letters, 2008, 1(2): 115-119.
[14] Yamamoto S, Wakayama M, Tachiki T.Cloning and expression of Pseudomonas taetrolens Y-30 gene encoding glutamine synthetase: an enzyme available for theanine production by coupled fermentation with energy transfer[J]. Bioscience, Biotechnology and Biochemistry, 2006, 70(2): 500-507.
[15] Kei S, Makoto K.Synthesis of theanine by tea seedling homogenate[J]. Agricultural and Biological Chemistry, 1964, 28(5): 313-317.
[16] Fu X M, Liao Y Y, Cheng S H, et al.Nonaqueous fractionation and overexpression of fluorescent-tagged enzymes reveals the subcellular sites of L-theanine biosynthesis in tea[J]. Plant Biotechnology Journal, 2020, 19(1): 98-108.
[17] 朱文娴, 黎星辉, 王丽鸳, 等. 利用GS基因构建茶氨酸生物合成工程菌的研究[J]. 茶叶科学, 2008, 28(4): 242-248.
Zhu W X, Li X H, Wang L Y, et al.Construction of E. coli recombinant engineered strain for theanine biosynthesis with GS gene embedded[J]. Journal of Tea Science, 2008, 28(4): 242-248.
[18] Kimura T, Sugahara I, Hanai K, et al.Purification and characterization of γ-glutamylmethylamide synthetase from Methylophaga sp. AA-30[J]. Bioscience Biotechnology and Biochemistry, 1992, 56(5): 708-711.
[19] Chen X Y, Su L Q, Wu D, et al.Application of recombinant Bacillus subtilis γ-glutamyltranspeptidase to the production of L-theanine[J]. Process Biochemistry, 2014, 49(9): 1429-1439.
[20] 王丽鸳, 王贤波, 成浩, 等. 基因工程菌生物合成茶氨酸条件研究[J]. 茶叶科学, 2007, 27(2): 111-116.
Wang L Y, Wang X B, Cheng H, et al.Research on conditions of theanine biosynthesis by genetically engineered Escherichia coli[J]. Journal of Tea Science, 2007, 27(2): 111-116.
[21] Miyake K, Kakita S.A novel catalytic ability of γ-glutamylcysteine synthetase of Escherichia coli and its application in theanine production[J]. Bioscience Biotechnology and Biochemistry, 2009, 73(12): 2677-2683.
[22] Martí E, Gisbert C, Bishop G J, et al.Genetic and physiological characterization of tomato cv. Micro-Tom[J]. Journal of Experimental Botany, 2006, 57(9): 2037-2047.
[23] Zhang Y, Wang L, Wei K, et al.Differential regulatory mechanisms of secondary metabolites revealed at different leaf positions in two related tea cultivars[J]. Scientia Horticulturae, 2020, 272: 109579. doi: 10.1016/j.scienta.2020.109579.
[24] Xia E H, Li F D, Tong W, et al.Tea plant information archive: a comprehensive genomics and bioinformatics platform for tea plant[J]. Plant Biotechnology Journal, 2019, 17(10): 1938-1953.
[25] Chen C, Chen H, Zhang Y, et al.TBtools: an integrative toolkit developed for interactive analyses of big biological data[J]. Molecular Plant, 2020, 13(8): 1194-1202.
[26] Zouine M, Maza E, Djari A, et al.TomExpress, a unified tomato RNA-Seq platform for visualization of expression data, clustering and correlation networks[J]. The Plant Journal, 2017, 92(4): 727-735.
[27] Sasaoka K, Kito M, Onishi Y.Synthesis of theanine by pea seed acetone powder extract[J]. Agricultural and Biological Chemistry, 1964, 28(5): 318-324.
[28] 陈琪, 江雪梅, 孟祥宇, 等. 茶树茶氨酸合成酶基因的酶活性验证与蛋白三维结构分析[J]. 广西植物, 2015, 35(3): 384-392, 377.
Chen Q, Jiang X M, Meng X Y, et al.Enzymatic activity verification and protein three-dimensional structural analysis of theanine synthetase gene in Camellia sinensis[J]. Guihaia, 2015, 35(3): 384-392, 377.
[29] Li F, Dong C X, Yang T Y, et al.Seasonal theanine accumulation and related gene expression in the roots and leaf buds of tea plants (Camellia Sinensis L.)[J]. Frontiers in Plant Science, 2019, 10: 1397. doi: 10.3389/fpls.2019.01397.
[30] Crocomo O J, Fowden L.Amino acid decarboxylases of higher plants: the formation of ethylamine[J]. Phytochemistry, 1970, 9(3): 537-540.
[31] Dong C X, Li F, Yang T Y, et al.Theanine transporters identified in tea plants (Camellia sinensis L.)[J]. The Plant Journal, 2020, 101(1): 57-70.
[32] Fischer W N, Kwart M, Hummel S, et al.Substrate specificity and expression profile of amino acid transporters (AAPs) in Arabidopsis[J]. Journal of Biological Chemistry, 1995, 270(27): 16315-16320.
[33] Yao X H, Nie J, Bai R X, et al.Amino acid transporters in plants: identification and function[J]. Plants, 2020, 9(8): 972. doi: 10.3390/plants9080972.
[34] Zhang S H, Cheng Y, He X B, et al.Identification of MYB transcription factors regulating theanine biosynthesis in tea plant using omics-based gene coexpression analysis[J]. Journal of agricultural and food chemistry, 2020, 68(3): 918-926.
[35] Millard P S, Konrad W, Kragelund B B, et al.Specificity of MYB interactions relies on motifs in ordered and disordered contexts[J]. Nucleic Acids Research, 2019, 47(18): 9592-9608.
[36] Bai P X, Wei K, Wang L Y, et al.Identification of a novel gene encoding the specialized alanine decarboxylase in tea (Camellia sinensis) plants[J]. Molecules, 2019, 24(3): 540. doi: 10.3390/molecules24030540.

基金

现代农业产业技术体系(CARS-19)、浙江省农业新品种选育重大专项(2016C02053)

PDF(606 KB)

Accesses

Citation

Detail

段落导航
相关文章

/