扬子鳄<i>β</i>防御素基因的血液表达谱及饲养密度对防御素基因表达的影响

doi:10.3785/j.issn.1008-9209.2019.12.271

浙江大学学报（农业与生命科学版）

2020, Vol. 46

Issue (5): 604-610 DOI: 10.3785/j.issn.1008-9209.2019.12.271

动物科学与动物医学

扬子鳄β防御素基因的血液表达谱及饲养密度对防御素基因表达的影响

陆瑶(

),方盛国(

)

浙江大学生命科学学院，生命系统稳态与保护教育部重点实验室/国家濒危野生动植物种质基因保护中心，杭州 310058

Expression profile of β-defensin genes and the effect of stocking density on them in Chinese alligator blood

Yao LU(

),Shengguo FANG(

)

MOE Key Laboratory of Biosystems Homeostasis and Protection/State Conservation Center for Gene Resources of Endangered Wildlife, College of Life Sciences, Zhejiang University, Hangzhou 310058, China

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摘要：

为研究扬子鳄血液中β防御素基因家族的表达水平和饲养密度对群体防御素基因表达的影响，利用反转录荧光定量聚合酶链式反应（reverse transcription quantitative polymerase chain reaction, RT-qPCR）技术检测41头扬子鳄七龄鳄血液中β防御素基因的表达情况，并分析不同性别和不同饲养密度群体间防御素基因相对表达量的差异。结果表明：在扬子鳄群体血液中，表达的防御素基因有AsBD5和AsBD8，并且这2个基因的表达量之间存在显著的正相关性。AsBD5和AsBD8在雌性和雄性扬子鳄间的表达水平没有显著性差异；但是在不同饲养密度下，2个扬子鳄群体间的表达水平存在显著差异，其中饲养密度较高的扬子鳄群体，整体的防御素表达水平更高，表明饲养密度对扬子鳄圈养种群的免疫状态有一定的影响。

关键词： 扬子鳄; β防御素基因; 饲养密度; 反转录荧光定量聚合酶链式反应

Abstract:

In order to investigate the expression profile of β-defensin genes in the blood of Chinese alligator and the effect of stocking density on them, reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression levels of β-defensin genes in blood samples from 41 seven-year-old Chinese alligators and study the defensin gene expression level differences between sexes and populations with different stocking densities. The results showed that only the AsBD5 and AsBD8 genes expressed in the blood of Chinese alligator, and there was a significantly positive correlation between their expression levels. There was no significant sex-biased difference of the defensin gene expression levels. However, their expression levels were significantly different between the two populations with different stocking densities. The defensin gene expression levels were significantly higher in the Chinese alligator population with higher stocking density, which indicating that the stocking density has some effect on the immune status of the captive Chinese alligator.

Key words: Chinese alligator β-defensin gene stocking density reverse transcription quantitative polymerase chain reaction (RT-qPCR)

收稿日期: 2019-12-27 出版日期: 2020-11-19

CLC:

S 86

基金资助: 国家自然科学基金重点项目(31530087);国家重点研发计划(2016YFC0503200);国家林业和草原局专项经费;中央直属高校基本科研业务费

通讯作者: 方盛国 E-mail: luyao2017@zju.edu.cn;sgfanglab@zju.edu.cn

作者简介: 陆瑶（https://orcid.org/0000-0002-4151-0555），E-mail：luyao2017@zju.edu.cn

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引用本文:

陆瑶,方盛国. 扬子鳄β防御素基因的血液表达谱及饲养密度对防御素基因表达的影响[J]. 浙江大学学报（农业与生命科学版）, 2020, 46(5): 604-610.

Yao LU,Shengguo FANG. Expression profile of β-defensin genes and the effect of stocking density on them in Chinese alligator blood. Journal of Zhejiang University (Agriculture and Life Sciences), 2020, 46(5): 604-610.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2019.12.271 或 http://www.zjujournals.com/agr/CN/Y2020/V46/I5/604

表1 扬子鳄七龄鳄群体特征和样本信息

图1 扬子鳄血液（A）和组织（B）总RNA的电泳结果M：DL2000分子标志物。图A中编号1～8代表8个扬子鳄个体的血液RNA；图B中编号1～8代表扬子鳄心、肝、脑、肺、肾、胃、小肠、肌肉等8个组织的RNA。

表2 qPCR引物信息

图2 扬子鳄七龄鳄群体血液中β防御素基因AsBD5和AsBD8的相对表达量

图3 不同性别（A）和不同饲养密度（B）的扬子鳄群体β防御素基因的相对表达量***表示在P＜0.001水平差异有极高度统计学意义。

1	JIANG H, WU X B. The IUCN red list of threatened species 2018. Alligator sinensis. 2018:e.T867A3146005.
2	NICOLA? M P J, MATZKE N J. Trait-based range expansion aided in the global radiation of Crocodylidae. Global Ecology and Biogeography, 2019,28(9):1244-1258. DOI:10.1111/geb.12929 doi: 10.1111/geb.12929
3	THORBJARNARSON J, WANG X. The Chinese alligator: ecology, behavior, conservation, and culture. American Society of Ichthyologists and Herpetologists, 2011,4:608-609.
4	吴荣,周永康.安徽扬子鳄国家级自然保护区现状分析. 安徽林业科技,2015,41(5):37-41. WU R, ZHOU Y K. Analysis on the actualities of Anhui Chinese Alligator National Nature Reserve. Anhui Forestry Science and Technology, 2015,41(5):37-41. (in Chinese with English abstract)
5	陈壁辉.扬子鳄研究.上海:上海科技教育出版社,2003. CHEN B H. Research on the Chinese Alligator. Shanghai: Shanghai Scientific and Technological Education Press, 2003. (in Chinese)
6	MORGAN K N, TROMBORG C T. Sources of stress in captivity. Applied Animal Behaviour Science, 2007,102(3):262-302. DOI: 10.1016/j.applanim.2006.05.032 doi: 10.1016/j.applanim.2006.05.032
7	王朝林.二年生扬子鳄饲养密度研究. 安徽林业,2004(3):22. WANG C L. Study on rearing density of two-year-old Chinese alligator. Anhui Linye, 2004(3):22. (in Chinese)
8	POLETTA G L, LARRIERA A, SIROSKI P A, et al. Broad snouted caiman (Caiman latirostris) growth under different rearing densities. Aquaculture, 2008,280(1):264-266. DOI: 10.1016/j.aquaculture.2008.04.032 doi: 10.1016/j.aquaculture.2008.04.032
9	BRIEN M L, WEBB G J, MCGUINNESS K A, et al. Effect of housing density on growth, agonistic behaviour, and activity in hatchling saltwater crocodiles (Crocodylus porosus). Applied Animal Behaviour Science, 2016,184:141-149. DOI:10.1016/j.applanim.2016.08.007 doi: 10.1016/j.applanim.2016.08.007
10	ELSEY R M, JOANEN T, MCNEASE L, et al. Growth rate and plasma corticosterone levels in juvenile alligators maintained at different stocking densities. Journal of Experimental Zoology, 1990,255(1):30-36.
11	ELSEY R M, JOANEN T, MCNEASE L, et al. Stress and plasma corticosterone levels in the American alligator-relationships with stocking density and nesting success. Comparative Biochemistry and Physiology Part A: Physiology, 1990,95(1):55-63.
12	HECKERT R A, ESTEVEZ I, RUSSEK-COHEN E, et al. Effects of density and perch availability on the immune status of broilers. Poultry Science, 2002,81(4):451-457. DOI:10.1093/ps/81.4.451 doi: 10.1093/ps/81.4.451
13	MUSTAFA M Y, MUNEER M A, ANJUM A A, et al. Influence of stocking density on immune response of broilers against Newcastle disease virus. Pakistan Journal of Life and Social Sciences, 2010,8(1):7-10.
14	SALAS-LEITON E, ANGUIS V, MARTíN-ANTONIO B, et al. Effects of stocking density and feed ration on growth and gene expression in the Senegalese sole (Solea senegalensis): potential effects on the immune response. Fish and Shellfish Immunology, 2010,28(2):296-302.
15	VALENZUELA C A, ESCOBAR-AGUIRRE S, ZULOAGA R, et al. Stocking density induces differential expression of immune-related genes in skeletal muscle and head kidney of fine flounder (Paralichthys adspersus). Veterinary Immunology and Immunopathology, 2019,210:23-27. DOI:10.1016/j.vetimm.2019.03.004 doi: 10.1016/j.vetimm.2019.03.004
16	FINGER J W JR, GOGAL R M JR. Endocrine-disrupting chemical exposure and the American alligator: a review of the potential role of environmental estrogens on the immune system of a top trophic carnivore. Archives of Environmental Contamination and Toxicology, 2013,65(4):704-714. DOI:10.1007/s00244-013-9953-x doi: 10.1007/s00244-013-9953-x
17	SELSTED M E, OUELLETTE A J. Mammalian defensins in the antimicrobial immune response. Nature Immunology, 2005,6(6):551-557. DOI:10.1038/ni1206 doi: 10.1038/ni1206
18	HAZLETT L, WU M. Defensins in innate immunity. Cell and Tissue Research, 2011,343(1):175-188. DOI:10.1007/s00441-010-1022-4 doi: 10.1007/s00441-010-1022-4
19	CONTRERAS G, SHIRDEL I, BRAUN M S, et al. Defensins: transcriptional regulation and function beyond antimicrobial activity. Developmental and Comparative Immunology, 2020,104:103556. DOI:10.1016/j.dci.2019.103556 doi: 10.1016/j.dci.2019.103556
20	TANG K Y, WANG X, WAN Q H, et al. A crucial role of paralogous β-defensin genes in the Chinese alligator innate immune system revealed by the first determination of a Crocodilia defensin cluster. Developmental and Comparative Immunology, 2018,81:193-203. DOI:10.1016/j.dci.2017.11.018 doi: 10.1016/j.dci.2017.11.018
21	PAL C, HURST L D. Evidence for co-evolution of gene order and recombination rate. Nature Genetics, 2003,33(3):392-395. DOI:10.1038/ng1111 doi: 10.1038/ng1111
22	DIJK A VAN, VELDHUIZEN E J A, HAAGSMAN H P. Avian defensins. Veterinary Immunology and Immunopathology, 2008,124(1/2):1-18. DOI:10.1016/j.vetimm.2007.12.006 doi: 10.1016/j.vetimm.2007.12.006
23	FANG X M, SHU Q, CHEN Q X, et al. Differential expression of α- and β-defensins in human peripheral blood. European Journal of Clinical Investigation, 2003,33(1):82-87. DOI:10.1046/j.1365-2362.2003.01076.x doi: 10.1046/j.1365-2362.2003.01076.x
24	CUPERUS T, COORENS M, DIJK A VAN, et al. Avian host defense peptides. Developmental and Comparative Immunology, 2013,41(3):352-369. DOI:10.1016/j.dci.2013.04.019 doi: 10.1016/j.dci.2013.04.019
25	VALENZUELA C A, ZULOAGA R, POBLETE-MORALES M, et al. Fish skeletal muscle tissue is an important focus of immune reactions during pathogen infection. Developmental and Comparative Immunology, 2017,73:1-9. DOI:10.1016/j.dci.2017.03.004 doi: 10.1016/j.dci.2017.03.004
26	HUCHZERMEYER F W. Diseases of farmed crocodiles and ostriches. Revue Scientifique et Technique, 2002,21(2):265-276. DOI:10.20506/rst.21.2.1334 doi: 10.20506/rst.21.2.1334
27	RAMOS M C C, COUTINHO S D, MATUSHIMA E R, et al. Poxvirus dermatitis outbreak in farmed Brazilian caimans (Caiman crocodilus yacare). Australian Veterinary Journal, 2002,80(6):371-372. DOI:10.1111/j.1751-0813.2002.tb14792.x doi: 10.1111/j.1751-0813.2002.tb14792.x
28	ACOBSON E R, GINN P E, TROUTMAN J M, et al. West Nile virus infection in farmed American alligators (Alligator mississippiensis) in Florida. Journal of Wildlife Diseases, 2005,41(1):96-106.
29	LOTT M J, MOORE R L, MILIC N L, et al. Dermatological conditions of farmed Crocodilians: a review of pathogenic agents and their proposed impact on skin quality. Veterinary Microbiology, 2018,225:89-100. DOI:10.1016/j.vetmic.2018.09.022 doi: 10.1016/j.vetmic.2018
30	KLENK K, SNOW J, MORGAN K, et al. Alligators as West Nile virus amplifiers. Emerging Infectious Diseases, 2004,10(12):2150-2155. DOI:10.3201/eid1012.040264 doi: 10.3201/eid1012.040264
31	ASHITANI J, MATSUMOTO N, NAKAZATO M. Elevated alpha-defensin levels in plasma of patients with pulmonary sarcoidosis. Respirology, 2007,12(3):339-345. DOI:10.1111/j.1440-1843.2007.01061.x doi: 10.1111/j.1440-1843.2007.01061.x
32	YAMAGUCHI N, ISOMOTO H, MUKAE H, et al. Concentrations of α- and β-defensins in plasma of patients with inflammatory bowel disease. Inflammation Research, 2009,58(4):192-197. DOI:10.1007/s00011-008-8120-8 doi: 10.1007/s00011-008-8120-8

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