Cloning and Expression Analysis of &#x025B3;12-fatty Acid Desaturase in Tea Plants

CHEN Dan; YU Ying; YUE Chuan; WANG Pengjie; CHEN Jing; CHEN Guixin; YE Naixing

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Journal of Tea Science ›› 2017, Vol. 37 ›› Issue (6) : 541-550.

Cloning and Expression Analysis of △12-fatty Acid Desaturase in Tea Plants

CHEN Dan, YU Ying, YUE Chuan, WANG Pengjie, CHEN Jing, CHEN Guixin^*, YE Naixing^*

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Abstract

Based on the transcriptome database of tea plants, the cDNAs ( CsFAD2 and CsFAD6) including full ORFs of the key rate-limiting enzyme-FAD (△12-fatty acid desaturase) in the unsaturated fatty acid synthesis pathway were cloned from the buds and leaves of tea cultivar Tieguanyin by RT-PCR .The full length of cDNA of CsFAD2 was 1 184 bp, which contained a 1β149βbp ORF encoding 382 amino acids with 97% homologous to the CoFAD2. Subcellular localization prediction showed that CsFAD2 was localized to the endoplasmic reticulum. The full length of cDNA of CsFAD6 was 1β425βbp and contained a 1β311βbp ORF encoding 436 amino acids, which was 81% homologous to VvFAD6. It was predicted to be located on the chloroplast. Quantitative PCR analysis showed that the expression of CsFAD2 and CsFAD6 were induced by cold stress (4℃), which had the highest expression levels at 48βh. Similarly, CsFAD2 and CsFAD6 were induced by 100βg·L^-1 PEG treatment for 12βh. The expression of CsFAD2 was significantly up-regulated under the treatment of ABA(100βμmol·L^-1) for 6-24βh. While the expression of CsFAD6 was not affected by ABA during this period and dramatically down-regulated at 72βh. The expression of CsFAD2 was repressed under NaCl (250βmmol·L^-1) treatment. Inversely;|the expression of CsFAD6 was significantly induced by NaCl treatment from 24 to 72βh.

Key words

△12-fatty acid desaturase enzyme / abiotic stresses / gene expression / Camellia sinensis

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CHEN Dan, YU Ying, YUE Chuan, WANG Pengjie, CHEN Jing, CHEN Guixin, YE Naixing. Cloning and Expression Analysis of △12-fatty Acid Desaturase in Tea Plants[J]. Journal of Tea Science. 2017, 37(6): 541-550

References

[1] 年洪娟, 陈丽梅. 不饱和脂肪酸在逆境胁迫中的作用[J]. 中国微生态学杂志, 2012(8): 760-762.
[2] 于超. 温度胁迫下番茄内质网ω-3脂肪酸去饱和酶基因的表达和功能研究[D]. 泰安: 山东农业大学, 2009.
[3] 叶静, 胡海涛, 王长春, 等. 植物脂肪酸去饱和酶与抗冷性研究进展[J]. 广东农业科学, 2011(12): 131-134.
[4] 李金金, 张晶晶, 年洪娟. △12-脂肪酸去饱和酶FAD2的基本特性及其在胁迫中的功能[J]. 生命科学研究, 2013(2): 174-178.
[5] 刘永红, 张丽静, 张洪荣, 等. △12-脂肪酸脱氢酶及其编码基因研究进展[J]. 草业学报, 2011(3): 256-267.
[6] Yang L, Ye J, Guo W, et al.Differences in cold tolerance and expression of two fatty acid desaturase genes in the leaves between fingered citron and its dwarf mutant[J]. Trees, 2012, 26(4): 1193-1201.
[7] Kargiotidou A, Deli D, Galanopoulou D, et al.Low temperature and light regulate delta 12 fatty acid desaturases (FAD2) at a transcriptional level in cotton (Gossypium hirsutum)[J]. Journal of ExperiMental Botany, 2008, 59(8): 2043-2056.
[8] Matteucci M, D'Angeili S, Errico S, et al. Cold affects the transcription of fatty acid desaturases and oil quality in the fruit of Olea europaea L. genotypes with different cold hardiness[J]. Journal of Experimental Botany, 2011, 62(10): 3403-3420.
[9] Hernández ML, Padilla MN, Sicardo MD, et al.Effect of different environmental stresses on the expression of oleate desaturase genes and fatty acid composition in olive fruit[J]. Phytochemistry, 2011, 72(2): 178-187.
[10] Teixeira MC, Coelho N, Olsson ME, et al.Molecular cloning and expression analysis of three omega-6 desaturase genes from purslane (Portulaca oleracea.L)[J]. Biotechnology Letters, 2009, 31(9): 1089-1101.
[11] Zhou Z,Wang M,Zhao S,et al.Changes in freezing tolerance in hybrid poplar caused by up-and down-regulation of PtFAD2 gene expression[J]. Transgenic Research, 2010, 19(4): 647-654.
[12] Hernandez M L, Mancha M, Martinez-Rivas J M, et al. Molecular cloning and characterization of genes encoding two microsomal oleate desaturases (FAD2) from olive[J]. Phytochemistry, 2005, 66(12): 1417-1426.
[13] Zhang YM, Wang CC, Hu HH, et al.Cloning and expression of three fatty acid desaturase genes from cold-sensitive lima bean (Phaseolus lunatus.L)[J]. Biotechnology Letters, 2011, 33(2): 395-401.
[14] Ma QP,You E,Wang J,et al.Isolation and expression of CsFAD7 and CsFAD8, two genes encoding ω-3 fatty acid desaturase from Camellia sinensis[J]. Acta Physiologiae Plantarum, 2014, 36(9): 2345-2352.
[15] 姚雪倩, 岳川, 杨国一, 等. 茶树牻牛儿基牻牛儿基焦磷酸合成酶基因CsGGDPS的克隆及表达分析[J]. 茶叶科学, 2017, 37(1): 86-96.
[16] 俞滢, 陈丹, 孙君, 等. 茉莉花萜类合成酶基因JsTPS的克隆及其表达分析[J]. 园艺学报, 2016, 43(2): 356-364.
[17] 曹红利, 岳川, 郝心愿, 等. 茶树胆碱单加氧酶CsCMO的克隆及甜菜碱合成关键基因的表达分析[J]. 中国农业科学, 2013, 46(15): 3087-3096.
[18] 岳川, 曹红利, 周艳华, 等. 茶树谷胱甘肽还原酶基因CsGRs的克隆与表达分析[J]. 中国农业科学, 2014, 47(16): 3277-3289.
[19] Wu ZJ, Tian C, Jiang Q, et al.Selection of suitable reference genes for qRT-PCR normalization during leaf development and hormonal stimuli in tea plant (Camellia sinensis)[J]. Scientific Reports, 2016: 1-10. DOI: 1038/srep19748.
[20] 陈静, 俞滢, 张丹丹, 等. 白茶萎凋过程中儿茶素合成关键酶基因表达分析[J]. 南方农业学报,2016(8): 1364-1369.
[21] Wada H, Gombos Z, Murata N, et al.Enhancement of chilling tolerance of a cyanobacterium by genetic manipulation of fatty acid desaturation[J]. Nature, 1990, 347(9): 200-203.
[22] Matos AR, Hourton-Cabassa C, Ciçek D, et al.Dominique ciçek.alternative oxidase involvement in cold stress response of arabidopsis thaliana fad2 and FAD3+ cell suspensions altered in membrane lipid composition[J]. Plant Cell Physiology, 2007, 48(6): 856-865.
[23] Zhang JT, Liu H,Sun J, et al.Arabidopsis fatty acid desaturase FAD2 is required for salt tolerance during seed germination and early seedling growth[J]. Plos One, 2012, 7(1): e30355.
[24] Sonia RV, Alicia SG, Jose Manuel MR, et al.Fluidization of membrane lipids enhances the tolerance of saccharomyces cerevisiae to freezing and salt stress[J]. Applied & Environmental Microbiology, 2007, 73(1): 110-116.
[25] Paula FMD, Thi ATP, Zuily-Fodil Y, et al.Effects of water stress on the biosynthesis and degradation of polyunsaturated lipid molecular species in leaves of Vigna unguiculata[J]. Plant Physiology & Biochemistry, 1993, 31(5): 707-715.
[26] Zhang M, Barq R, Yin M, et al.Modulated fatty acid desaturation via overexpression of two distinct ω-3 desaturases differentially alters[J]. Plant Journal, 2005, 44(3): 361-371.
[27] RG Upchurch.Fatty acid unsaturation, mobilization, and regulation in the response of plants to stress[J]. Biotechnology Letters,2008, 30(6): 967-977.