Welcome to Journal of Tea Science,Today is

Cloning and Expression Analysis of GDP-D-mannose Pyrophosphorylase cDNA in Camellia sinensis

  • XIAO Yao ,
  • ZHOU Tianshan ,
  • LI Jiao ,
  • ZHANG Jiaxin ,
  • YU Youben
Expand
  • 1. College of Horticulture, Northwest Agriculture & Forestry University, Yangling 712100, China;
    2. Hanzhong Agricultural Science Institute, Hanzhong 723000, China

Received date: 2014-08-05

  Revised date: 2014-10-13

  Online published: 2019-08-23

Abstract

Ascorbic acid (Vitamin C, Vc) is an important antioxidant in tea plant, playing important roles in metabolism and responses to abiotic stress, and tea quality is positive correlation with the content of Vc. GDP-D-mannose pyrophosphorylase (GMP) is an important enzyme in the synthesis of ascorbic acid. The full-length cDNA sequence of GMP gene was isolated from the shoots of Camellia sinensis by RT-PCR and RACE. The entire GMP cDNA was 1β510βbp, containing a 1β086βbp complete open reading frame which encoding a protein with 361 amino acids and a calculated molecular weight of 39.599βkDa. Blast analysis showed that GMP gene in Camellia sinensis was most closely to Actinidia with 96% amino acids similarity. Quantitative real-time PCR analysis showed that the expression levels of GMP gene in the third leaves were highest while the stem is the lowest, and the different varieties also existed an obvious differences. High temperature stress stimulate the expression of GMP gene and the accumulation of Vc in incipient stage, then reduced rapidly.

Cite this article

XIAO Yao , ZHOU Tianshan , LI Jiao , ZHANG Jiaxin , YU Youben . Cloning and Expression Analysis of GDP-D-mannose Pyrophosphorylase cDNA in Camellia sinensis[J]. Journal of Tea Science, 2015 , 35(1) : 55 -63 . DOI: 10.13305/j.cnki.jts.2015.01.010

References

[1] Iqbal Y, Ihsanullah I, Shaheen N, et al. Significance of vitamin C in plants[J]. Journal of the Chemical Society of Pakistan, 2009, 31(1): 169-170.
[2] Smirnoff N.Ascorbic acid: metabolism and functions of a multi-facetted molecule[J]. Current opinion in plant biology, 2000, 3(3): 229-235.
[3] Mark W Davey, Marc Van Montagu, Dirk Inze, et al. Plant L-ascorbic acid: chemistry, function, metabolism, bioavailability and effects of processing[J]. Journal of the Science of Food and Agriculture, 2000 (80): 825-860.
[4] Jaleel C A, Riadh K, Gopi R, et al. Antioxidant defense responses: physiological plasticity in higher plants under abiotic constraints[J]. Acta Physiologiae Plantarum, 2009, 31(3): 427-436.
[5] Barth C, De Tullio M, Conklin P L.The role of ascorbic acid in the control of flowering time and the onset of senescence[J]. Journal of Experimental Botany, 2006, 57(8): 1657-1665.
[6] Pastori G M, Kiddle G, Antoniw J, et al. Leaf vitamin C contents modulate plant defense transcripts and regulate genes that control development through hormone signaling[J]. The Plant Cell Online, 2003, 15(4): 939-951.
[7] Debolt S, Melino V, Ford C M.Ascorbate as a biosynthetic precursor in plants[J]. Annals of botany, 2007, 99(1): 3-8.
[8] Ishikawa T, Shigeoka S.Recent advances in ascorbate biosynthesis and the physiological significance of ascorbate peroxidase in photosynthesizing organisms[J]. Bioscience, biotechnology, and biochemistry, 2008, 72(5): 1143-1154.
[9] 孙云. 茶叶抗坏血酸过氧化物酶(APX)的生理学与分子生物学研究[D]. 福州: 福建农林大学, 2009.
[10] Peters C M, Green R J, Janle E M, et al. Formulation with ascorbic acid and sucrose modulates catechin bioavailability from green tea[J]. Food research international, 2010, 43(1): 95-102.
[11] Dai F, Chen W F, Zhou B.Erratum to “Antioxidant synergism of green tea polyphenols with α-tocopherol and l-ascorbic acid in SDS micelles”[J]. Biochimie, 2009, 91(11): 1535.
[12] 李佼, 余有本, 周天山, 等. 茶树GDP-甘露糖-3′, 5′-异构酶基因cDNA全长的克隆与表达[J]. 西北农林科技大学学报: 自然科学版, 2013, 41(12): 100-106.
[13] 童启庆. 茶树栽培学[M]. 北京: 中国农业出版社, 2006: 93, 344.
[14] 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.
[15] Kampfenkel K, Vanmontagu M, Inze D.Extraction and determination of ascorbate and dehydroascorbate from plant tissue[J]. Analytical biochemistry, 1995, 225(1): 165-167.
[16] 安华明, 陈力耕, 樊卫国, 等. 高等植物中维生素C的功能、合成及代谢研究进展[J]. 植物学通报, 2004, 21(5): 608-617.
[17] Conklin P L, Norris S R, Wheeler G L, et al. Genetic evidence for the role of GDP-mannose in plant ascorbic acid (vitamin C) biosynthesis[J]. Proceedings of the National Academy of Sciences, 1999, 96(7): 4198-4203.
[18] Badejo A A, Tanaka N, Esaka M.Analysis of GDP-D-mannose pyrophosphorylase gene promoter from acerola (Malpighia glabra) and increase in ascorbate content of transgenic tobacco expressing the acerola gene[J]. Plant and cell physiology, 2008, 49(1): 126-132.
[19] Keller R, Kossmann J.Antisense inhibition of the GDP-mannose pyrophosphorylase reduces the ascorbate content in transgenic plants leading to developmental changes during senescence[J]. The Plant Journal, 1999, 19(2): 131-141.
[20] Cronje C, George G M, Fernie A R, et al. Manipulation of L-ascorbic acid biosynthesis pathways in Solanum lycopersicum: elevated GDP-mannose pyrophosphorylase activity enhances L-ascorbate levels in red fruit[J]. Planta, 2012, 235(3): 553-564.
[21] Ma Yuhua, Ma Fengwang, Zhang Junke, et al. Effects of high temperature on activities and gene expression of enzymes involved in ascorbate-glutathione cycle in apple leaves[J]. Plant Science, 2008, 175(6): 761-766.
Outlines

/