Effect of Low Temperature Stress on Function of Thymosins Gene in Pacific White Shrimp Litopenaeus vannamei
ZHANG Yi1, JIN Kuifeng1, LI Wenhui1, YANG Yingcan1, LAI Hui1,2, LIU Wenguang3
1. Meteorological Public Service Center of Guangdong Province, Guangzhou 510640, China; 2. Guangdong HuayunTechnology Development Company, Guangzhou 510640, China; 3. South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
Abstract:In order to investigate the molecular mechanism of thymosinsgene under low temperature stress in Pacific white shrimp Litopenaeus vannamei, the Lv-thymosins gene was cloned by rapid amplification of cDNA ends (RACE), and then the tissue distribution and the expression profiles of Lvthy gene under low temperature stress was detected by real-time quantitative PCR. Subsequently, RNA interference (RNAi) experiment was conducted to be knocked down the expression of Lvthy gene to verify the gene function under low temperature stress. It was found that the full length of the Lvthy gene cDNA sequences consisted of 1765 base pairs (bp), including a 729 bp open reading frame (ORF) encoding a 242-amino acid peptide with six thymosins beta actin-binding motif. The qRT-PCR showed that Lvthy gene was expressed in all tissues of the Pacific white shrimp, the maximal expression level in stomach, followed by intestine, heart and brain. Low temperature challenge showed that the expression level of the Lvthy gene was shown to be increased 1 h post low temperature stimulation. The maximal level (2.7 times compared with the control group) was found at 3 h in the shrimp challenged and then to be declined. RNA interference (RNAi) technology verified the gene function. The survival rate of the Pacific white shrimp was 31.1% under low temperature stress, significantly lower than that in the control group (83.3%). The findings indicate that the Lvthy gene plays critical role in the process of accumulating to cold stress and that provide a foundation for the study of adaptation mechanism of crustaceans under low temperature stress.
张毅, 靳奎峰, 李文慧, 杨颖璨, 来辉, 刘文广. 低温胁迫对凡纳滨对虾胸腺肽基因功能的影响[J]. 水产科学, 2024, 43(1): 104-110.
ZHANG Yi, JIN Kuifeng, LI Wenhui, YANG Yingcan, LAI Hui, LIU Wenguang. Effect of Low Temperature Stress on Function of Thymosins Gene in Pacific White Shrimp Litopenaeus vannamei. Fisheries Science, 2024, 43(1): 104-110.
[1] 彭自然,臧维玲,高杨,等.氨和亚硝酸盐对凡纳滨对虾幼虾的毒性影响[J].上海水产大学学报,2004,13(3):274-278. [2] 农业农村部渔业渔政局,全国水产推广总站,中国水产学会.2021年中国渔业统计年鉴[M].北京:中国农业出版社,2021:22. [3] LONG Y, SONG G L, YAN J J, et al. Transcriptomic characterization of cold acclimation in larval zebrafish[J].BMC Genomics,2013,14:612. [4] HUANG W, REN C H, LI H M, et al. Transcriptomic analyses on muscle tissues of Litopenaeus vannamei provide the first profile insight into the response to low temperature stress[J].PLoS One,2017,12(6):e0178604. [5] VORNANEN M, HASSINEN M, KOSKINEN H, et al. Steady-state effects of temperature acclimation on the transcriptome of the rainbow trout heart[J].American Journal of Physiology.Regulatory, Integrative and Comparative Physiology,2005,289(4):R1177-R1184. [6] JU Z, DUNHAM R, LIU Z. Differential gene expression in the brain of channel catfish (Ictalurus punctatus) in response to cold acclimation[J].Molecular Genetics and Genomics,2002,268(1):87-95. [7] GRACEY A Y, FRASER E J, LI W Z, et al. Coping with cold:an integrative, multitissue analysis of the transcriptome of a poikilothermic vertebrate[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(48):16970-16975. [8] 范永祥,霍秀英,王锋.对虾养殖的气象服务[J].气象,1992,18(5):49-54. [9] 李金枝,刘非.黄河三角洲地区南美白对虾养殖气象条件及智慧服务研究[J].中国水产,2020(1):60-62. [10] 宋丽莉.影响对虾养殖的主要气象因子初探[J].气象,1988,14(7):46-48. [11] 宋萍.雨季养虾管理[J].科学养鱼,2011(7):82. [12] 徐德峰,王雅玲,孙力军,等.降温及生态冰温条件对凡纳滨对虾无水存活时间的影响[J].广东海洋大学学报,2019,39(6):101-107. [13] 李色东,何建国,张存善,等.寒流对南美白对虾养殖的影响及防范措施探讨[J].海洋与渔业,2008(11):18-19. [14] 何有根,彭勇鳌,黄志斌.低温季节南美白对虾的处理方案[J].内陆水产,2009,34(12):40. [15] ZHONG P, XU Z N, CHEN W, et al. Molecular characterization of a c-Jun NH2-terminal kinase (JNK)-interacting protein 4 (Lv-JIP4) in Litopenaeus vannamei and its potential role in the Lv-p38MAPK pathway in response to low temperature[J].Aquaculture Reports,2021,21:100951. [16] 秋虎.高温天气谨防南美白对虾泛塘发病[J].渔业致富指南,2003(20):49. [17] 熊大林,段亚飞,陈成勋,等.高温与氨氮复合胁迫对凡纳滨对虾渗透调节的影响[J].水产科学,2021,40(4):475-482. [18] BOQUET I, BOUJEMAA R, CARLIER M F, et al. Ciboulot regulates actin assembly during Drosophila brain metamorphosis[J].Cell,2000,102(6):797-808. [19] VAN TROYS M, ONO K, DEWITTE D, et al. TetraThymosinbeta is required for actin dynamics in Caenorhabditis elegans and acts via functionally different actin-binding repeats[J].Molecular Biology of the Cell,2004,15(10):4735-4748. [20] LEELATANAWIT R, SITTIKANKEAW K, YOCAWIBUN P, et al. Identification, characterization and expression of sex-related genes in testes of the giant tiger shrimp Penaeus monodon[J].Comparative Biochemistry and Physiology Part A:Molecular & Integrative Physiology,2009,152(1):66-76. [21] GAI Y C, ZHAO J M, SONG L S, et al. Two thymosin-repeated molecules with structural and functional diversity coexist in Chinese mitten crab Eriocheir sinensis[J].Developmental & Comparative Immunology,2009,33(7):867-876. [22] LI D L, CHANG X J, XIE X L, et al. A thymosin repeated protein1 reduces white spot syndrome virus replication in red claw crayfish Cherax quadricarinatus[J].Developmental & Comparative Immunology,2018,84:109-116. [23] SHI X Z, SHI L J, ZHAO Y R, et al. β-Thymosins participate in antiviral immunity of red swamp crayfish (Procambarus clarkii)[J].Developmental & Comparative Immunology,2015,51(2):213-225. [24] ZHANG X, YANG H, LI H M, et al. Molecular identification of anion exchange protein 3 in Pacific white shrimp (Litopenaeus vannamei):mRNA profiles for tissues, ontogeny, molting, and ovarian development and its potential role in stress-induced gill damage[J].Frontiers in Physiology,2021,12:726600. [25] 陈清潮.中国海洋生物多样性的现状和展望[J].生物多样性,1997,5(2):142-146. [26] 农成万,张幸,李斌喜,等.钦州对虾养殖灾害性天气分析和气象服务要领[J].气象研究与应用,2013,34(1):60-63. [27] 曾琮,谢炯光.广东省寒潮、强冷空气的气候特征及与ENSO的关系[J].广东气象,2003,25(1):7-9. [28] 彭昌迪,郑建民,彭文国,等.南美白对虾的胚胎发育以及温度与盐度对胚胎发育的影响[J].上海水产大学学报,2002,11(4):310-316. [29] 陈昌生,叶兆弘,纪德华,等.南美白对虾摄食、生长及存活与温度的关系[J].集美大学学报(自然科学版),2001,6(4):296-300. [30] 沈文英,胡洪国,潘雅娟.温度和pH值对南美白对虾(Penaeus vannmei)消化酶活性的影响[J].海洋与湖沼,2004,35(6):543-548. [31] 黄燕华,王国霞,刘襄河,等.温度和pH对南美白对虾主要消化酶活性的影响[J].华南农业大学学报,2008,29(4):87-90. [32] HOCH K, VOLK D E. Structures of thymosin proteins[J].Vitamins and Hormones,2016,102:1-24. [33] 彭金霞,房振峰,韦嫔媛,等.低温处理凡纳滨对虾仔虾全长cDNA文库及troponin Ⅰ基因三种拷贝的比较分析[J].水产科学,2013,32(2):73-79. [34] 孙金辉,徐赟霞,季延滨,等.温度骤降对南美白对虾仔虾抗氧化机能的影响[J].天津农学院学报,2008,15(3):7-10. [35] SAELEE N, NOONIN C, NUPAN B, et al. β-thymosins and hemocyte homeostasis in a crustacean[J].PLoS One,2013,8(4):e60974. [36] 程光平,桑明远,黄国强,等.不同低温持续胁迫对凡纳滨对虾存活的影响[J].南方农业学报,2013,44(2):333-337. [37] MORISHITA S, NISHI Y, SATO E F, et al. Cold-stress induces thymocyte apoptosis in the rat[J].Pathophysiology,1997,4(3):213-219. [38] CHENG C H, GUO Z X, WANG A L. The protective effects of taurine on oxidative stress, cytoplasmic free-Ca2+ and apoptosis of pufferfish (Takifugu obscurus) under low temperature stress[J].Fish & Shellfish Immunology,2018,77:457-464. [39] ANVARIFAR H, AMIRKOLAIE A K, MIANDARE H K, et al. Apoptosis in fish:environmental factors and programmed cell death[J].Cell and Tissue Research,2017,368(3):425-439. [40] SOSNE G, SIDDIQI A, KURPAKUS-WHEATER M. Thymosin-beta4 inhibits corneal epithelial cell apoptosis after ethanol exposure in vitro[J].Investigative Ophthalmology & Visual Science,2004,45(4):1095-1100.