研究报告

茶树CsPDAT1基因克隆及抗旱性分析

  • 沈英姿 ,
  • 李朵姣 ,
  • 胡新荣 ,
  • 江丽 ,
  • 郑寨生 ,
  • 留惠康 ,
  • 袁名安
展开
  • 金华市农业科学研究院,浙江 金华 321017
沈英姿,女,农艺师,从事茶树遗传育种方面的研究。

收稿日期: 2025-07-23

  网络出版日期: 2025-12-10

基金资助

浙江省科技计划项目(2023C04010)、浙江省基础公益研究项目(LGN22C160010)

Cloning of the CsPDAT1 Gene from Camellia sinensis and Its Role in Drought Tolerance

  • SHEN Yingzi ,
  • LI Duojiao ,
  • HU Xingrong ,
  • JIANG Li ,
  • ZHENG Zhaisheng ,
  • LIU Huikang ,
  • YUAN Ming′an
Expand
  • Jinhua Academy of Agricultural Sciences, Jinhua 321017, China

Received date: 2025-07-23

  Online published: 2025-12-10

摘要

在茶树(Camellia sinensis)的生长发育过程中,干旱、盐害等非生物胁迫频繁发生,对茶叶的产量和品质造成了显著不利影响。聚焦于茶树磷脂:二酰甘油酰基转移酶1基因(CsPDAT1),通过生物信息学分析、基因表达模式检测以及转基因功能验证等手段,系统探究了该基因在非生物胁迫响应中的功能。生物信息学分析表明,CsPDAT1编码的蛋白具有典型的溶血磷脂酰基转移酶(lysoPLA)结构域。实时荧光定量PCR结果显示,CsPDAT1在干旱及高盐处理下表达量显著上调,尤其在干旱胁迫初期表现明显。为深入解析其功能,构建了CsPDAT1过表达载体并转化拟南芥,成功获得3个纯合的T3代转基因株系。表型分析发现,在干旱胁迫下,过表达CsPDAT1的拟南芥植株种子萌发率显著高于野生型,丙二醛(MDA)含量降低,脯氨酸含量增加,且抗氧化酶活性显著增强。本研究证实CsPDAT1通过调控细胞膜脂代谢和渗透调节物质积累,增强植株对非生物胁迫的耐受性,为茶树抗逆分子育种提供了理论基础和基因资源。

本文引用格式

沈英姿 , 李朵姣 , 胡新荣 , 江丽 , 郑寨生 , 留惠康 , 袁名安 . 茶树CsPDAT1基因克隆及抗旱性分析[J]. 茶叶科学, 2025 , 45(6) : 920 -930 . DOI: 10.13305/j.cnki.jts.2025.06.002

Abstract

During the growth and development of tea plants (Camellia sinensis), abiotic stresses such as drought and salinity frequently occur, causing significant adverse effects on tea yield and quality. This study focused on the tea phospholipid: diacylglycerol acyltransferase 1 (PDAT1) gene and systematically investigated its function in response to abiotic stresses through bioinformatics analysis, gene expression pattern detection, and transgenic functional verification. Bioinformatics analysis reveals that the protein encoded by the CsPDAT1 gene possesses a typical lysophospholipid acyltransferase (lysoPLA) domain. Real-time quantitative PCR (qRT-PCR) results demonstrats that the expression of the CsPDAT1 gene was significantly upregulated under drought and high-salt treatments, particularly during the early stages of drought stress. To further elucidate its function, researchers constructed an overexpression vector for the CsPDAT1 gene and transformed it into Arabidopsis thaliana. Finally three homozygous T3-generation transgenic lines were obtained. Phenotypic analysis reveals that under drought stress, Arabidopsis plants overexpressing the CsPDAT1 gene exhibited a significantly higher seed germination rate compared to that of wild-type plants, along with reduced malondialdehyde (MDA) content, increased proline content, and significantly enhanced antioxidant enzyme activity. This study confirmed that the CsPDAT1 gene in tea plants enhances plant tolerance to abiotic stress by regulating cell membrane lipid metabolism and the accumulation of osmoprotectants, providing a theoretical foundation and genetic resources for stress-resistant molecular breeding in tea plants.

参考文献

[1] 刘声传, 陈亮. 茶树耐旱机理及抗旱节水研究进展[J]. 茶叶科学, 2014, 34(2): 111-121.
Liu S C, Chen L.Research advances on the drought-resistance mechanism and strategy of tea plant[J]. Journal of Tea Science, 2014, 34(2): 111-121.
[2] Chaeikar S S, Marzvan S, Khiavi S J, et al.Changes in growth, biochemical, and chemical characteristics and alteration of the antioxidant defense system in the leaves of tea clones (Camellia sinensis L.) under drought stress[J]. Scientia Horticulturae, 2020, 265: 109257. doi: 10.1016/j.scienta.2020.109257.
[3] 牛素贞, 宋秦飞, 樊卫国, 等. 干旱胁迫对喀斯特地区野生茶树幼苗生理特性及根系生长的影响[J]. 生态学报, 2017, 37(21): 7333-7341.
Niu S Z, Song Q F, Fan W G, et al.Effects of drought stress on leaf physiological characteristics and root growth of the clone seedlings of wild tea plants[J]. Acta Ecologica Sinica, 2017, 37(21): 7333-7341.
[4] 王铭涵, 丁玎, 张晨禹, 等. 干旱胁迫对茶树幼苗生长及叶绿素荧光特性的影响[J]. 茶叶科学, 2020, 40(4): 478-491.
Wang M H, Ding D, Zhang C Y, et al.Effects of drought stress on the growth and chlorophyll fluorescence characteristics of tea seedlings[J]. Journal of Tea Science, 2020, 40(4): 478-491.
[5] 田永辉, 梁远发, 魏杰, 等. 灾害性气候对茶树的影响[J]. 贵州农业科学, 2003, 31(2): 20-23.
Tian Y H, Liang Y F, Wei J, et al.Effect of disastrous climate on tea trees[J]. Guizhou Agricultural Sciences, 2003, 31(2): 20-23.
[6] 柯玉琴, 庄重光, 何华勤, 等. 不同灌溉处理对铁观音茶树光合作用的影响[J]. 应用生态学报, 2008(10): 2132-2136.
Ke Y Q, Zhuang C G, He H Q, et al.Effects of different irrigation treatments on photosynthesis of Tieguanyin tea plants[J]. Journal of Applied Ecology, 2008(10): 2132-2136.
[7] Gasulla F, Vom Dorp K, Dombrink I, et al.The role of lipid metabolism in the acquisition of desiccation tolerance in Craterostigma plantagineum: a comparative approach[J]. The Plant Journal, 2013, 75(5): 726-741.
[8] Liu X Y, Ouyang L L, Zhou Z G.Phospholipid: diacylglycerol acyltransferase contributes to the conversion of membrane lipids into triacylglycerol in Myrmecia incisa during the nitrogen starvation stress[J]. Scientific Reports, 2016, 6(18): 26610. doi: 10.1038/srep26610.
[9] Fan J L, Yan C S, Zhang X B, et al.Dual role for phospholipid: diacylglycerol acyltransferase: enhancing fatty acid synthesis and diverting fatty acids from membrane lipids to triacylglycerol in Arabidopsis leaves[J]. Plant Cell, 2013, 25(9): 3506-3518.
[10] Mueller S P, Unger M, Guender L, et al.Phospholipid: diacylglycerol acyltransferase-mediated triacylglycerol synthesis augments basal thermotolerance[J]. Plant Physiology, 2017, 175(1): 486-497.
[11] Demski K, Łosiewska A, Jasieniecka-Gazarkiewicz K, et al.Phospholipid: diacylglycerol acyltransferase1 overexpression delays senescence and enhances post-heat and cold exposure fitness[J]. Front Plant Science, 2020, 11: 611897. doi: 10.3389/fpls.2020.611897.
[12] Yuan L X, Mao X, Zhao K, et al.Characterisation of phospholipid: diacylglycerol acyltransferases (PDATs) from Camelina sativa and their roles in stress responses[J]. Biology Open, 2017, 6(7): 1024-1034.
[13] Qiu S, Zhang J, He J Q, et al.Overexpression of GmGolS2-1, a soybean galactinol synthase gene, enhances transgenic tobacco drought tolerance[J]. Plant Cell Tissue Organ Culture PCTOC, 2020, 143(3): 507-516.
[14] Sun X D, Lian H F, Liu X C, et al.The garlic NF-YC gene, AsNFYC8, positively regulates non-ionic hyperosmotic stress tolerance in tobacco[J]. Protoplasma, 2017, 254(3): 1353-1366.
[15] Yoon K, Han D, Li Y, et al.Phospholipid: diacylglycerol acyltransferase is a multifunctional enzyme involved in membrane lipid turnover and degradation while synthesizing triacylglycerol in the unicellular green microalga Chlamydomonas reinhardtii[J]. Plant Cell, 2012, 24: 3708-3724.
[16] Higashi Y, Okazaki Y, Myouga F, et al.Landscape of the lipidome and transcriptome under heat stress in Arabidopsis thaliana[J]. Scientific Reports, 2015, 5: 10533. doi: 10.1038/srep10533.
[17] Hernández M L, Moretti S, Sicardo M D, et al.Distinct physiological roles of three phospholipid: diacylglycerol acyltransferase genes in olive fruit with respect to oil accumulation and the response to abiotic stress[J]. Frontiers in Plant Science, 2021, 12: 751959. doi: 10.3389/fpls.2021.751959.
[18] 徐赫, 潘丽娟, 陈娜, 等. 磷脂二酰甘油酰基转移酶(PDAT)基因的克隆与表达分析[J]. 花生学报, 2018, 47(4): 33-40, 54.
Xu H, Pan L J, Chen N, et al.Cloning and expression analysis of two phospholipids: diacylglycerol acyltransferase genes in peanut[J]. Journal of Peanut Science, 2018, 47(4): 33-40, 54.
[19] Liu H C, Zhang J Q, Zhou J H, et al.Cloning of PsGRP gene from paeonia suffruticosa and waterlogging tolerance analysis of transgenic Arabidopsis[J]. Plant Physiology Journal, 2021, 57(2): 373-384.
[20] Chi K W, Song Y, Li S L.Genome-wide identification and expressional analysis of the LIM gene family in Medicago truncatula[J]. Plant Physiology Journal, 2021, 57(5): 1074-1086.
[21] Niazian M, Sadat-Noori S A, Tohidfar M, et al. Betaine aldehyde dehydrogenase (BADH) vs. Flavodoxin (Fld): two important genes for enhancing plants stress tolerance and productivity[J]. Frontiers in Plant Science, 2021, 12: 650215. doi: 10.3389/fpls.2021.650215.
[22] Fan J, Yan C, Roston R, et al.Arabidopsis lipins, PDAT1 acyltransferase, and SDP1 triacylglycerol lipase synergistically direct fatty acids toward β-oxidation, thereby maintaining membrane lipid homeostasis[J]. Plant Cell, 2014, 26(10): 4119-4134.
[23] Demidchik V, Straltsova D, Medvedev S S, et al.Stress-induced electrolyte leakage: the role of K+-permeable channels and involvement in programmed cell death and metabolic adjustment[J]. Journal of Experimental Botany, 2014, 65(5): 1259-1270.
[24] Weber H, Chételat A, Reymond P, et al.Selective and powerful stress gene expression in Arabidopsis in response to malondialdehyde[J]. Plant Journal, 2004, 37(6): 877-888.
[25] Gill S S, Tuteja N.Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants[J]. Plant Physiology Biochemistry, 2010, 48(12): 909-930.
[26] Mittler R.Oxidative stress, antioxidants and stress tolerance[J]. Trends in Plant Science, 2002, 7(9): 405-410.
文章导航

/