Functional Analysis of Glutathione Peroxidase Encoding Gene CsGPX1 in Camellia sinensis

LIU Sai; LIU Shuoqian; LONG Jinhua; WU Dunchao; CHEN Yuhong; LIU Liping; LIU Zhonghua; TIAN Na

doi:10.13305/j.cnki.jts.2019.04.003

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Journal of Tea Science ›› 2019, Vol. 39 ›› Issue (4) : 382-391. DOI: 10.13305/j.cnki.jts.2019.04.003

Functional Analysis of Glutathione Peroxidase Encoding Gene CsGPX1 in Camellia sinensis

LIU Sai^1,2, LIU Shuoqian^1,2,3, LONG Jinhua^1,2, WU Dunchao^1,2, CHEN Yuhong^1,2, LIU Liping^1,2, LIU Zhonghua^1,2,3, TIAN Na^1,2,3,*

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Abstract

To clarify function of glutathione peroxidase-encoding gene CsGPX1 in Camellia sinensis (L.) O. Kuntze, the full-length sequence of the coding region of CsGPX1 was obtained by transcriptome data of tea plant, and sequence analysis and function prediction were performed. Then, the CsGPX1 was overexpressed in tobacco. Subsequently, the difference in drought tolerance between wild-type and transgenic tobacco was compared to verify the function of CsGPX1. Sequence analysis indicates that CsGPX1 was 723 bp in length, encoding 240 amino acids. The Blast alignment reveals that the GPX showed more than 85% of identity in amino acid with GPX from Prunus persica. The amino acid sequence encoded by CsGPX1 has a conserved characteristic sequence of the GPX protein and a phospholipid hydrogen glutathione peroxidase (PHGPX) which is unique to other family members. The results of drought stress treatment show that the drought resistance of the over-expressing of CsGPX1 in tobacco lines was stronger than that of wild-type. The results of GPX enzyme activity assay show that the GPX enzyme activity of the transgenic plants was higher than that of the wild type plants. All of above results verify that the over-expression of CsGPX1 improved the drought tolerance of transgenic tobacco, suggesting that CsGPX1 might be related to the drought resistance in tea plants. This study could provide a new strategy to improve the drought resistance of tea plants, which would significantly reduce the management cost of tea plantation.

Key words

abiotic stress / drought resistance mechanism / glutathione / peroxidase / Camellia sinensis

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LIU Sai, LIU Shuoqian, LONG Jinhua, WU Dunchao, CHEN Yuhong, LIU Liping, LIU Zhonghua, TIAN Na. Functional Analysis of Glutathione Peroxidase Encoding Gene CsGPX1 in Camellia sinensis[J]. Journal of Tea Science. 2019, 39(4): 382-391 https://doi.org/10.13305/j.cnki.jts.2019.04.003

References

[1] 肖蓉, 慧珍, 张小娟, 等. 干旱和盐胁迫条件下枣树谷胱甘肽过氧化物酶基因(ZjGPX)的差异表达及功能分析[J]. 中国农业科学, 2015, 48(14): 2806-2817.
[2] 乔新荣, 张继英. 植物谷胱甘肽过氧化物酶(GPX)研究进展[J]. 生物技术通报, 2016, 32(9): 7-13.
[3] 马亭亭, 周宜君, 高飞, 等. 盐芥谷胱甘肽过氧化物酶基因(ThGPX6)的克隆及表达分析[J]. 植物遗传资源学报, 2012, 13(2): 252-258.
[4] Sugimoto M, Sakamoto W.Putative phospholipid hydroperoxide glutathione peroxidase gene from Arabidopsis thaliana induced by oxidative stress[J]. Genes & Genetic Systems, 1997, 72(5): 311-316.
[5] Li W J, Feng H, Fan J H, et al.Molecular cloning and expression of a phospholipid hydroperoxide glutathione peroxidase homolog in Oryza sativa [J]. Biochim Biophys Acta, 2000, 1493(1/2): 225-230.
[6] Depege N, Drevet J, Boyer N.Molecular cloning and characterization of tomato cDNAs encoding glutathione peroxidase-like proteins[J]. European Journal of Biochemistry, 1998, 253(2): 445-451.
[7] Avsian-Kretchmer O, Eshdat Y, Gueta-Dahan Y, et al.Regulation of stress-induced phospholipid hydroperoxide glutathione peroxidase expression in citrus[J]. Planta, 1999, 209(4): 469-477.
[8] Sugimoto M, Furui S, Suzuki Y.Molecular cloning and characterization of a cDNA encoding putative phospholipid hydroperoxide glutathione peroxidase from spinach[J]. Biosciences Biotechnology and Biochemistry, 1997, 61(8): 1379-1381.
[9] 王菲菲, 丁明全, 邓澍荣, 等. 胡杨谷胱甘肽过氧化物酶PeGPX基因的克隆及转化植株耐盐性分析[J]. 基因组学与应用生物学, 2012, 31(3): 231-239.
[10] 陈义挺. 龙眼体胚发生过程中的CDC48和GPX基因克隆与表达[D]. 福州: 福建农林大学, 2009.
[11] 乔新荣, 张继英. 茶树CsGPX基因克隆及干旱胁迫下的表达分析[J]. 江苏农业科学, 2018, 46(8): 42-44.
[12] 谭和平, 周李华, 钱杉杉, 等. 茶树转基因技术研究进展[J]. 武汉植物学研究, 2009, 27(3): 323-326.
[13] Chen S, Songkumarn P, Liu J, et al.A Versatile zero background T-vector system for gene cloning and functional genomics[J]. Plant Physiology, 2009, 150(3): 1111-1121.
[14] Lazo G R, Stein P A, Ludwig R A.A DNA transformation-competent Arabidopsis genomic library in Agrobacterium[J]. Biotechnology. 1991, 9(10): 963-967.
[15] 赵东, 刘祖生, 陆建良, 等. 根癌农杆菌介导茶树转化研究[J]. 茶叶科学, 2001, 21(2): 108-111.
[16] 张金丽, 张罗霞, 杨坤梅, 等. 改良CTAB法对山茶属植物基因组DNA提取的比较研究[J]. 江西农业大学学报, 2017, 39(4): 785-791.
[17] 李合生. 现代植物生理学[M]. 北京: 高等教育出版社, 2011: 100.
[18] Faltin Z, Holland D, Velcheva M, et a1. Glutathione peroxidase regulation of reactive oxygen species level is crucial for in vitro plant differentiation[J]. Plant Cell Physiology, 2010, 51(7): 1151-1162
[19] Gaber A, Ogata T, Maruta T, et al.The Involvement of Arabidopsis glutathione peroxidase 8 in the suppression of oxidative damage in the nucleus and cytosol[J]. Plant and Cell Physiology, 2012, 53(9): 1596-1606.
[20] Milla M, Maurer A, Huete A R, et al.Glutathione peroxidase genes in Arabidopsis are ubiquitous and regulated by abiotic stresses though diverse signaling pathways[J]. The Plant Journal, 2003, 36(5): 602-615.
[21] Herbette S, Labrouhe D T, Drevet J R, et al.Transgenic tomatoes showing higher glutathione peroxydase antioxidant activity are more resistant to an abiotic stress but more susceptible to biotic stresses[J]. Plant Science, 2011, 180(3): 548-553.
[22] 齐增园, 陶鹏, 李必元, 等. 白菜谷胱甘肽过氧化物酶基因GPX的鉴定与分析[J]. 浙江农业学报, 2016, 28(01): 64-69.
[23] 徐泽, 胡翔, 邓敏. 干旱胁迫对茶树的几种抗旱性生理指标的影响[C]//中国茶叶学会, 台湾茶协会. 第四届海峡两岸茶业学术研讨会论文集. 2006: 113-118.
[24] 张进. 泛素基因在烟草耐盐性中的作用及其生理机制研究[D]. 泰安: 山东农业大学, 2009.
[25] 蔡永智. 转CBF1和KatG基因棉花抗旱性分析[D]. 石河子: 石河子大学, 2013.
杜世章, 代其林, 刘婷婷, 等. ⁶⁰Coγ射线辐照对转基因和非转基因烟草抗氧化酶活性的影响[J]. 核农学报, 2011, 25(3): 456-460, 497.