摘要
研究旨在建立一套准确、经济的团头鲂(Megalobrama amblycephala)家系鉴定体系,以期为团头鲂的遗传参数评估及基因组和分子标记育种奠定基础。精选8个高多态性的微卫星标记对二段法筛选后的228尾耐低氧快生长团头鲂“浦江2号”子代进行亲子鉴定分析,利用CERVUS软件分析实验结果。等位基因频率分析结果显示8个微卫星标记的平均等位基因数(N_(a))为8个,平均观测杂合度(H_(o))为0.813,平均期望杂合度(H_(e))为0.775,平均多态信息含量(PIC)为0.743。亲子鉴定结果显示双亲性别已知时8个微卫星标记的累积排除概率为0.9999,说明筛选出的8个微卫星标记适用于团头鲂家系的亲子鉴定分析。当使用筛选的8个微卫星标记进行亲子鉴定时,228尾团头鲂子代中有221尾子代能找到其父母本,实际鉴定率为96.93%,这与实际记录的系谱信息一致。研究成功地为筛选后的耐低氧团头鲂个体找到父母本并划分家系,为进一步开展遗传参数的评估工作奠定了基础。
The objective of this study was to establish an accurate and economical family identification system for Megalobrama amblycephala,aimed at providing a foundation for genetic parameter evaluation and genome and molecular marker breeding of the species.Firstly,9 pairs of"Pujiang No.2"parents with high hypoxic tolerance and good growth performance were screened to establish 17 full-sib families.Subsequently,utilizing a"two-stage screening"process,228 progeny exhibiting hypoxia tolerance and rapid growth were obtained.The parentage analysis of these 228 progeny was conducted by using 8 highly polymorphic microsatellite loci specific to Megalobrama amblycephala Pujiang No.2".Parental and progeny DNA was amplified by a simple PCR reaction,and the resulting PCR products were separated by non-denatured polyacrylamide gel electrophoresis.With PBR322 DNA/MSPI as the marker,the size of the band alleles on the gel was determined by using Quantity One V4.6.2 software.Subsequently,the genotypes of both offspring and parents were formatted into text and input into the CERVUS software for parentage analysis.This analysis primarily involved allele frequency analysis,simulation analysis and paternity analysis in CERVUS software,with paternity relationships determined based on LOD values.Allele frequency analysis showed that the average number of alleles(N_(a))of the 8 microsatellite markers was 8,with an average observed heterozygosity(H_(o))of 0.813,average expected heterozygosity(H_(e))of 0.775,and average polymorphism information content(PIC)of 0.743.Hardy-Weinberg equilibrium testing showed that Mam-EST841,Mam-EST12,and Mam-EST158 were consistent with equilibrium,while other loci exhibited deviations.Exclusion probability analysis showed that when parental genotypes were unknown,the probability of exclusion at a single locus ranged from 0.339 to 0.540,with an average of 0.413.When one single parent genotype was known,the exclusion probability for the other of parents at a single locus ranged from 0.470 to 0.704,averaging 0.578.With both parental genotypes were known,the parent exclusion probability at a single locus ranged from 0.654 to 0.871,averaging 0.763.The cumulative exclusion probability were 98.47%(CE-1P)when both parents were unknown,and 99.99%(CE-2P)when one parent was known,and 99.99%(CE-PP)when both parents were known.Parentage analysis showed that the theoretical identification rate of 8 microsatellite loci was 99.44%when simulating 10000 offspring and 18 parents with known parental sex.However,the actual identification rate of parents for 221 offspring was 96.93%.This study successfully utilized eight highly polymorphic microsatellite loci were used to identify the offspring of hypoxia-tolerant and fast-growth Megalobrama amblycephala screened by the two-stage method,thereby identifying parents and corresponding families for 221(96.93%)offspring,mitigating the influence of common environmental effects in single family breeding.This accomplishment lays a foundation for further genetic parameter evaluation of hypoxic-tolerant and fast-growing families of Megalobrama amblycephala.
作者
王会虎
任超
郑国栋
邹曙明
WANG Hu-Hu;RE Chao;HENG Guo-Dong;ZOU Shu-Ming(Genetics and Breeding Center for Blunt Snout Bream,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;National Demonstration Center for Experimental es Science Education,Shanghai Ocean University,Shanghai 201306,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2024年第8期1378-1384,共7页
Acta Hydrobiologica Sinica
基金
国家重点研发计划(2022YFD2400600-2)
国家自然科学基金(32273100)
上海市农委科技兴农重点项目(2022-02-08-00-12-F01179)资助。
关键词
微卫星
亲子鉴定
累积排除概率
Microsatellites
Parentage analysis
Cumulative probability of exclusion
作者简介
王会虎(1999-),男,硕士研究生,研究方向为水产动物遗传育种。E-mail:1980954318@qq.com;通信作者:邹曙明,教授,研究方向为鱼类遗传育种工程。E-mail:smzoushou.edu.cn。
