The full cDNA sequences of cytochrome P450 genes, CYP71A26 and CYP71B34 were cloned from Camellia sinensis Tieguanyin leaves based on NCBI transcriptome database. The complete cDNA lengths of CYP71A26 and CYP71B34 were 1β879βbp and 1β764βbp, containing the open reading frames (ORF) of 1β539βbp and 1β533βbp, which encoded 513 and 511 amino acids, respectively. The alignment of amino acid sequences showed 42.47% of conservation between CYP71A26 and CYP71B34, which indicated that the two genes belonged to different subfamilies. The structure analysis showed that both CYP71A26 and CYP71B34 contained the classic P450 structure domains, such as helix C, helix I, helix K, Meander and heme binding domain. Phylogenetic tree analysis illustrated that there were two branches of molecular evolution for the two genes with different genetic relationship. Via predicting of their tertiary structures and functional domains, the results indicated that both genes were constituted by β-pleated structure in N-terminal, α-helix structure in C-terminal, and had a P450 protein structure domain (Pfam domain). qRT-PCR results showed that the expression of two genes in leaves showed opposite trends under pest damage treatments. Moreover, the decrease of CYP71A26 and increase of CYP71B34 in expression levels were detected under cold or thermal stresses. In total, CYP71A26 and CYP71B34 genes were respond to biotic (like pest) and abiotic (like temperature) stresses, but the changing trends of the two genes were opposite, which might be correlated with opposite regulation systems.
SHAN Ruiyang
,
LIN Zhenghe
,
CHEN Zhihui
,
ZHONG Qiusheng
,
YOU Xiaomei
,
CHEN Changsong
. Molecular Cloning and Expression Analysis of Cytochrome P450 CYP71A26 and CYP71B34 Genes in Tea Plants (Camellia sinensis)[J]. Journal of Tea Science, 2018
, 38(5)
: 450
-460
.
DOI: 10.13305/j.cnki.jts.2018.05.002
[1] Axelrod J.The enzymatic demethylation of ephedrine[J]. Journal of Pharmacology and Experimental Therapeutics, 1955, 114(4): 430-438.
[2] Garfinkel D.Studies on pig liver microsomes I Enzymic and pigment composition of different microsomal fractions[J]. Archives of Biochemistry and Biophysics, 1958, 77(2): 493-509.
[3] Omura T, Sato R.A new cytochrome in liver microsomes[J]. Journal of Biological Chemistry, 1962, 237(4): 1375-1376.
[4] Schuler M A.Plant cytochrome P450 monooxygenases[J]. Critical Reviews in Plant Sciences, 1996, 15(3): 235-284.
[5] Zhao Y J, Cheng Q Q, Su P, et al.Research progress relating to the role of cytochrome P450 in the biosynthesis of terpenoids in medicinal plants[J]. Applied Microbiology & Biotechnology, 2014, 98(6): 2371-2383.
[6] 解敏敏. 烟草重要基因篇: 8. 烟草P450基因[J]. 中国烟草科学, 2015, 36(2): 118-120.
[7] Moore M T, Kröger R.Effect of three insecticides and two herbicides on rice (Oryza sativa) seedling germination and growth[J]. Archives of Environmental Contamination & Toxicology, 2010, 59(4): 574-581.
[8] Deng F, Hatzios K K.Characterization of cytochrome P450-mediated bensulfuron-methyl O-demethylation in rice[J]. Pesticide Biochemistry & Physiology, 2002, 74(2): 102-115.
[9] Zhou N, Tootle T L, Glazebrook J.Arabidopsis PAD3, a gene required for camalex in biosynthesis, encodes a putative cytochrome P450 monooxygenase[J]. The Plant Cell, 1999, 11(12): 2419-2428.
[10] 曹红利, 王璐, 钱文俊, 等. 茶树CsbZIP4转录因子正调控拟南芥对盐胁迫响应[J]. 作物学报, 2017, 43(7): 1012-1020.
[11] Ruan J, Gerendás J, Härdter R, et al.Effect of nitrogen form and root-zone pH on growth and nitrogen uptake of tea (Camellia sinensis) plants[J]. Annals of Botany, 2007, 99(2): 301-310.
[12] 曹运鹏, 程曦, 程俊, 等. 砀山酥梨细胞色素P450的基因组学分析[J]. 核农学报, 2016, 30(2): 259-266.
[13] Gilardi G, Di Nardo G.Heme iron centers in cytochrome P450: structure and catalytic activity[J]. Rendiconti Lincei, 2017, 28(1): 159-167.
[14] 轩丹丹, 孙廉平, 张沛沛, 等. 水稻无花粉型核雄性不育突变体whf41的鉴定与基因定位[J]. 中国水稻科学, 2017, 31(3): 247-256.
[15] 曾秀红. 水稻簇生穗基因cl-4的精细定位[D]. 扬州: 扬州大学, 2009: 1-2.
[16] Wang E, Wang R, DeParasis J, et al. Suppression of a P450 hydroxylase gene in plant trichome glands enhances natural-product-based aphid resistance[J]. Nature Biotechnology, 2001, 19(4): 371-374.
[17] 戴素明, 周程爱, 谢丙炎, 等. 细胞色素P450表达在植物防御反应中的作用[J]. 石河子大学学报, 2004, 22(7): 184-187.
[18] 代娅. 温度胁迫下多年生黑麦草P450家族基因表达及LpCYP72A161甲基化差异分析[D]. 北京: 中国科学院大学, 2016: 1-2.