Cloning and Expression Analysis of Phosphoenolpyruvate Transporter Gene CsPPT2 in Tea Plant(Camellia sinensis)

JI Zhifang; GAN Yudi; CHEN Changsong; YANG Dingjun; SUN Kang; LI Xinghui; CHEN Xuan

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Journal of Tea Science ›› 2017, Vol. 37 ›› Issue (2) : 149-159.

Cloning and Expression Analysis of Phosphoenolpyruvate Transporter Gene CsPPT2 in Tea Plant(Camellia sinensis)

JI Zhifang¹, GAN Yudi¹, CHEN Changsong², YANG Dingjun¹, SUN Kang¹, LI Xinghui¹, CHEN Xuan^1,*

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Abstract

A full length cDNA sequence of phosphoenolpyruvate transporter gene (CsPPT2) was obtained from tea plant(Camellia sinensis) cultivar ‘Baiye 1’ by polymerase chain reaction (PCR) and rapid amplification of cDNA ends PCR(RACE-PCR). The nucleotide sequence length of this gene was 1469 bp, containing a complete open reading frame (1218 bp) encoding 406 amino acids. Bioinformatic analysis showed that the predicted molecular weight of the protein was 44.6 kD, and the theoretic isoelectric point was 9.90. CsPPT2 has 6 trans-membrane regions, which may belong to the hydrophobic chloroplast trans-membrane protein. Phylogenetic tree analysis showed the phosphoenolpyruvate transporter family in tea plant could be divided into two subgroups, with CsPPT1 and CsPPT2 belonged to different subgroups. The expression of CsPPT1 and CsPPT2 were compared in different periods and plant organs. It showed that CsPPT2 was expressed in all tested tissues, with higher expression level in roots and mature leaves than those of CsPPT1. But the expression level of CsPPT1 in young shoots was higher than CsPPT2. CsPPT2 was transiently elevated at the beginning of the albino, but inhibited during the development of albino, indicates that the inhibition of the expression of CsPPT2 may act as a key factor for low-catechins and high-amino acids in ‘Baiye 1’.

Key words

Baiye 1 / CsPPT2 / expression analysis / phosphoenolpyruvate transporter / Camellia sinensis

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JI Zhifang, GAN Yudi, CHEN Changsong, YANG Dingjun, SUN Kang, LI Xinghui, CHEN Xuan. Cloning and Expression Analysis of Phosphoenolpyruvate Transporter Gene CsPPT2 in Tea Plant(Camellia sinensis)[J]. Journal of Tea Science. 2017, 37(2): 149-159

References

[1] Fischer K, Kammerer B, Gutensohn M, et al.A new class of plastidic phosphate translocators: a putative link between primary and secondary metabolism by the phosphoenolpyruvate/phosphate antiporter[J]. Plant Cell, 1997, 9: 453-462.
[2] Voll L, Husler R E, Hecker R, et al.The phenotype of the Arabidopsis cue1 mutant is not simply caused by a general restriction of the shikimate pathway[J]. Plant Journal, 2003, 36: 301-317.
[3] Kerbarh O, Bulloch E M M, Payne R J, et al. Mechanistic and inhibition studies of chorismate-utilizing enzymes[J]. Biochemical Society Transactions, 2005, 33: 763-766.
[4] Li H, Culligan K, Dixon RA, et al.Expression of the functional mature chloroplast triose phosphate translocator in yeast internal membranes and purification of the histidine-tagged proteion by a single metalaffinity chromatography step[J]. Proc Natl Acad Sci, 1993, 90: 2155-2159.
[5] Streatfield SJ, Weber A, Kinsman EA, et al.The phosphoenolpyruvate/phosphater translocator is required for phenolic metabolism, plastid cell development and plastid-dependent nuclear gene expression[J]. Plant Cell, 1999, 11: 1609-1621.
[6] Knappe S, Flugge UI, Fischer K.Analysis of the plastidic phosphate translocator gene family in Arabidopsis and identification of new phosphate translocator-homologous transporters, classified by their putative substrate-binding site[J]. 2003, 131(3): 1178-1190.
[7] Knappe S, Löttgert T, Schneider A, et al.Characterization of two functional phosphoenolpyruvate/phosphate translocator (PPT) genes in Arabidopsis-AtPPT1 may be involved in the provision of signals for correct mesophyll development[J]. The Plant Journal, 2003, 36(3): 411-420.
[8] Staehr P, Löttgert T, Christmann A, et al.Reticulate leaves and stunted roots are independent phenotypes pointing at opposite roles of the phosphoenolpyruvate/phosphate translocator defective in cue1 in the plastids of both organs[J]. Front Plant Sci, 2014, 5: 126.
[9] 杨坤, 吴学龙, 郎春秀, 等. 甘蓝型油菜PEP转运子BnPPT1基因的克隆、序列分析和表达模式[J]. 浙江农业学报, 2011, 23(1): 1-7.
[10] 吴学龙. PPT1基因调控植物生长发育的研究及叶脉特异表达增强子的分离应用[D]. 杭州: 浙江大学, 2013: 12-131.
[11] 成浩, 李素芳, 陈明, 等. 安吉白茶特异性状的生理生化本质[J]. 茶叶科学, 1999, 19(2): 87-92.
[12] 李素芳, 成浩, 虞富莲, 等. 安吉白茶阶段性返白过程中氨基酸的变化[J]. 茶叶科学, 1996, 16(2): 153-154.
[13] 李素芳, 陈树尧, 成浩. 茶树阶段性返白现象的初步研究[J]. 中国茶叶, 1994, 16(2): 26-27.
[14] 赵真, 陈暄, 王明乐, 等. 茶树磷酸烯醇式丙酮酸转运子基因CsPPT的克隆与表达分析[J]. 茶叶科学, 2015, 35(5): 491-500.
[15] 孙美莲, 王云生, 杨冬青, 等. 茶树实时荧光定量PCR分析中内参基因的选择[J]. 植物学报, 2010, 45(5): 579-587.
[16] Bagge P, Larsson C.Biosynthesis of aromatic amino acids by highly purified spinach chloroplasts-Compartmentation and regulation of the reactions[J]. Physiol Plantarum, 1986, 68(4): 641-647.
[17] Van Der Straeten D, Rodrigues-Pousada RA, Goodman HM, et al. Plant enolase: Gene structure, expression and evolution[J]. Plant Cell, 1991, 3: 719-735.
[18] Journet EP, Douce R.Enzymic capacities of purified cauliflower bud plastids for lipidsynthesis and carbohydrate metabolism[J]. Plant Physiol, 1985, 79: 458-467.
[19] Schulze-Siebert D, Heineke D, Scharf H, et al.Pyruvate-derived amino acids in spinach chloroplasts: Synthesis and regulation during photosynthetic carbon metabolism[J]. Plant Physiol, 1984, 76: 465-471.
[20] Prabhaka V, Löttgert T, Gigolashvili T, et al.Molecular and functional characterization of the plastid-localized Phosphoenolpyruvate enolase (ENO1) from Arabidopsis thaliana[J]. FEBSLet, 2009, 583: 983-991.
[21] Li HM, Culligan K, Dixon RA, et al.CUE1: a mesophyll cell-specific positive regulator of light-controlled gene expression in Arabidopsis[J]. Plant Cell, 1995, 7: 1599-1610.