孵化湿度对胚胎期中华鳖裙边外部形态和胶原沉积的影响  

Effect of Incubation Humidity on Development of Calipash Related Traits and Collagen Deposition in Embryos of Chinese Soft-Shelled Turtle Pelodiscus sinensis

作　　者：傅凯琳 陈培恒 杨明康 吴俊涛 李彩燕[1] FU Kailin;CHEN Peiheng;YANG Mingkang;WU Juntao;LI Caiyan(College of Biological and Environmental Sciences,Zhejiang Wanli University,Ningbo 315000,China)

机构地区：[1]浙江万里学院生物与环境学院,浙江宁波315000

出　　处：《水产科学》2024年第3期445-452,共8页Fisheries Science

基　　金：国家自然科学基金资助项目(31802272);浙江省水产新品种选育项目(2021C02069-8-3);浙江省生物工程一流学科创新项目(CX2021026);宁波市科技创新2025重大专项(2021Z009).

摘　　要：为探究孵化湿度对中华鳖营养品质的影响,采集5龄中华鳖所产的同批次受精卵[(3.05±0.05)g],在相对湿度65%和85%下于实验室全裸孵化箱中进行无接触式孵化,分别取18、20、22、24、26期胚胎各20枚,迅速解剖出裙边组织,用于荧光定量PCR分析测定胶原蛋白含量,并用天狼星红染色法进行组织学分析,研究2种孵化湿度对胚胎期中华鳖外部形态和胶原物质积累的影响。试验结果显示:随着胚胎发育,背甲指数(背甲质量/体质量)、裙边指数(裙边质量/体质量)、背甲比值(背甲长/体长)和裙边比值(体长/裙边宽)等性状总体上呈增加趋势。相对湿度85%下孵育的中华鳖胚胎在大多数发育阶段的外部形态特征性指标显著优于相对湿度65%(P<0.05)。相对湿度85%下孵育的裙边组织呈现强橙黄色或亮红色偏振光,随着胚胎发育,裙边组织Ⅰ型胶原蛋白面积的占比不断增加,极显著高于相对湿度65%下的此数值(P<0.01)。发育至第22期,相对湿度85%下孵育的裙边组织Ⅰ型胶原蛋白基因mRNA表达水平迅速增加,推测潮湿环境可能有利于胚胎期中华鳖发育过程中胶原蛋白的沉积和相关优良性状的形成。试验结果可为中华鳖生产实践中改善孵化湿度调控裙边性状发育提供基础数据。The same batch of fertilized eggs[(3.05±0.05)g]laid by 5-year-old Chinese soft-shelled turtle Pelodiscus sinensis were incubated without contact in a laboratory naked incubator,incubated at 65%and 85%relative humidity,starting from the 18th stage,20 embryos of 18,20,22,24 and 26 stages were taken respectively,the calipash tissue was quickly dissected,and the collagen content was analyzed and determined by fluorescence quantitative PCR,and some samples were fixed in paraformaldehyde.The effects of two hatching humidity on the external morphology and accumulation of collagen matter in embryonic Chinese soft-shelled turtles were studied by histological analysis by Sirius red staining.It was found that the calipash related traits including embryo length,dorsal carapace index(dorsal carapace weight/body weight),calipash index(calipash weight/body weight),dorsal carapace ratio(dorsal carapace length/body length)and calipash ratio(body length/calipash width)showed an overall increasing tendency as embryos developed.The embryonic development at relative humidity of 85%had significant bigger embryo size and higher collagen content at most development stages compared with those embryos incubated at relative humidity of 65%.The results of picrosirius red staining showed that the calipash tissue of embryos incubated under relative humidity of 85%showed strong orange yellow or bright red polarized light,and the proportion of type I collagen in the calipash tissue was increased with embryo development,which was significantly higher than that in relative humidity of 65%(P<0.01).The observed rapid promoted mRNA level of type I collagen gene of Chinese soft-shelled turtle incubated at relative humidity of 85%in developmental stage 22 indicated that the biosynthesis of collagen in Chinese soft-shelled turtle embryos was induced.The findings indicated that relative incubation humidity of 85%was conducive to more collagen deposition and more preferable calipash traits during the embryonic development,which provided the possible man

关 键 词：中华鳖 裙边 孵化湿度 胶原蛋白 

分 类 号：S966.5[农业科学—水产养殖]