Journal of Tropical Oceanography >
Establishment of parentage assignment techniques in Acanthopagrus latus and its applications in stock enhancement assessment
Copy editor: LIN Qiang
Received date: 2024-09-11
Revised date: 2024-10-22
Online published: 2024-10-28
Supported by
Science and Technology Planning Project of Fujian Province, China(2023N0011)
Project of Fujian Institute of Fisheries(S23311)
Open Project of the Fujian Province Key Laboratory of Marine Fisheries Resources and Ecological Environment
To evaluate the stock enhancement effectiveness of Acanthopagrus latus, this study employed highly polymorphic microsatellite markers developed by our research group using the species' genome and established a parentage assignment technology. Based on this, we assessed the stock enhancement effectiveness of A. latus in Xiamen Bay sampled in 2023-2024. The specific findings are as follows: (1) Polymorphic primer screening: from 110 randomly selected microsatellite loci in the A. latus genome, we designed primers and preliminarily screened 9 loci with high polymorphism using a wild A. latus population. (2) Establishment of parentage assignment technology: using known pedigree families of A. latus, including 20 parental fish and 72 offspring, we performed parentage analysis with the 9 selected microsatellite loci. As shown by the results, a total of 145 alleles were detected across the loci, with an average allele number (Na) of 16.11, an average observed heterozygosity (Ho) of 0.788, an average expected heterozygosity (He) of 0.810, and an average polymorphic information content (PIC) of 0.785, indicating high polymorphism. The analysis using the Cervus 3.0.7 software showed that under a 95% confidence level, the parentage assignment success rate was 98.61% when the sex of both parents was known, and 97.22% when the sex was unknown. (3) Stock enhancement evaluation: the parentage assignment technology was applied to 114 released parental fish and 264 recaptured individuals. Among the 264 recaptured A. latus, 8 were identified as offspring of the released parents, indicating a resource contribution rate of 3.03% to the wild A. latus population. The results provide reliable technical methods for future genetic resource identification and stock enhancement evaluation of A. latus.
LU Youjun , CHEN Qiang , HUANG Liangmin , YOU Songyuan , LIU Dingyu , MA Chao , WU Shuiqing , LIU Xiande . Establishment of parentage assignment techniques in Acanthopagrus latus and its applications in stock enhancement assessment[J]. Journal of Tropical Oceanography, 2025 , 44(3) : 141 -147 . DOI: 10.11978/2024175
表1 黄鳍棘鲷9个微卫星位点的基本信息Tab. 1 Basic information of 9 microsatellite loci in A. latus |
位点 | 重复单元 | 荧光标记 | 引物序列 | 片段大小 | 退火温度/℃ |
---|---|---|---|---|---|
AL1 | (GT)10 | FAM | F: TGTGTCTGTGTGCTCGTGT R: TGGAGCAACAGAAAACACGC | 141 | 57℃ |
AL2 | (GTG)7 | FAM | F: GATCTGGAAATCTGCGGCGA R: TGGCCCGATCACCACTACTA | 162 | 58℃ |
AL3 | (CACG)5 | FAM | F: CTGAATTACACACGTGCACACA R: AGTGGAAGATGAGCACCGTC | 140 | 57.5℃ |
AL4 | (GGAC)5 | HEX | F: GAGGTGATGTGGATGGAGGG R: GCCCCCCCTCTTCTTTCAAA | 142 | 58℃ |
AL5 | (TAG)7 | FAM | F: AAATGTTGGCGTGGTTCACC R: GCAACAGGACACGAAGAAGC | 150 | 57℃ |
AL6 | (TGC)7 | HEX | F: GGTGGGTGGTTAAGTTCAATCC R: AGAATCTTCCAGCCCATTAAGC | 195 | 56℃ |
AL7 | (AGG)3 | FAM | F: GATACTGTCGCAGGCTGAGG R: AACCACACACACACAAAGCC | 200 | 58℃ |
AL8 | (ATC)3 | HEX | F: AGGTAGTACGCAGCACAACC R: CCTTCCAGAGAGTCTCTTTGAC | 188 | 57.5℃ |
AL9 | (AGAT)4 | HEX | F: TACAGTTCTGTGTCGGCTGC R: GATGCTTGAATTTCCTCCGGG | 186 | 58℃ |
表2 9个微卫星位点在黄鳍棘鲷亲本与子代中的遗传信息Tab. 2 Genetic information of 9 microsatellite loci in A. latus parents and offspring |
位点 | Na | Ho | He | PIC | HWE | FN |
---|---|---|---|---|---|---|
AL1 | 18 | 0.787 | 0.821 | 0.796 | NS | 0.0204 |
AL2 | 9 | 0.522 | 0.747 | 0.700 | ** | 0.1716 |
AL3 | 15 | 0.815 | 0.854 | 0.834 | NS | 0.0228 |
AL4 | 21 | 0.978 | 0.906 | 0.893 | ND | -0.0427 |
AL5 | 17 | 0.978 | 0.852 | 0.832 | * | -0.0779 |
AL6 | 11 | 0.457 | 0.589 | 0.557 | NS | 0.1333 |
AL7 | 23 | 0.780 | 0.829 | 0.808 | * | 0.0068 |
AL8 | 10 | 0.784 | 0.764 | 0.725 | NS | -0.0197 |
AL9 | 21 | 0.989 | 0.932 | 0.922 | ND | -0.0331 |
平均值 | 16.11 | 0.788 | 0.8103 | 0.785 | - | - |
注: NS表示不显著偏离(P>0.05); ND表示无结果; *表示偏离(P<0.05); **表示显著偏离(P<0.01); ***表示极显著偏离(P<0.001)。 |
表3 7个微卫星位点在回捕个体中的遗传信息Tab. 3 Genetic information of 7 microsatellite loci in recaptured individuals |
位点 | Na | Ho | He | PIC | HWE |
---|---|---|---|---|---|
AL1 | 29 | 0.883 | 0.904 | 0.895 | NS |
AL2 | 11 | 0.727 | 0.728 | 0.682 | NS |
AL3 | 24 | 0.818 | 0.912 | 0.904 | ** |
AL4 | 39 | 0.864 | 0.918 | 0.911 | * |
AL5 | 24 | 0.896 | 0.882 | 0.870 | *** |
AL6 | 40 | 0.463 | 0.797 | 0.786 | *** |
AL7 | 43 | 0.939 | 0.947 | 0.943 | ND |
平均值 | 30 | 0.798 | 0.8698 | 0.8559 |
注: NS表示不显著偏离(P>0.05); ND表示无结果; *表示偏离 (P<0.05); **表示显著偏离 (P<0.01); ***表示极显著偏离(P<0.001)。 |
表4 成功鉴定出亲本的回捕个体信息Tab. 4 Information of recaptured individuals with successfully assigned parents |
子代编号 | 匹配座位数 | 体长/mm | 体重/g | 体高/mm | 似然率值 | 候选亲本编号 |
---|---|---|---|---|---|---|
HB15 | 7 | 140 | 59.6 | 52 | 8.31 | A33/A42 |
HB33 | 6 | 150 | 89.29 | 56 | 8.58 | A51/A102 |
HB49 | 6 | 148 | 76.87 | 59 | 8.41 | A56/A64 |
HB51 | 7 | 148 | 62.31 | 50 | 7.69 | A11/A28 |
HB66 | 7 | 155 | 106.35 | 64 | 8.41 | A71/A86 |
HB74 | 6 | 145 | 108.37 | 65 | 7.71 | A43/A46 |
HB135 | 6 | 132 | 75.74 | 57 | 8.44 | A1/A48 |
HB167 | 6 | 177 | 181.74 | 79 | 10.7 | A7/A38 |
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