2 75 and the length of the alleles ranged from 196 bp to 528 bp the number of alleles per locus ranged from 6 to 23 with an average of For the

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1 35 2 Vol. 35 No PROGRESS IN FISHERY SCIENCES Apr * bp Nei's UPGMA S A Genetic structure analysis between two generations of cultured Marsupenaeus japonicas by microsatellite markers HU Long-yang 1 2 LUAN Sheng 1 MAO Yong 3 ZHONG Sheng-ping 3 KONG Jie 1* 1 Key Laboratory of Sustainable Development of Marine Fisheries Ministry of Agriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Qingdao College of Fisheries and Life Science Shanghai Ocean University Shanghai College of Ocean & Earth Sciences Xiamen University Xiamen ABSTRACT The genetic structure for two generations of cultured Marsupenaeus japonicas was investigated using eight pairs of microsatellite primers. In this study a total of 108 alleles were tested AA10A409 * kongjie@ ysfri. ac. cn Tel hu0004@ 163. com Tel

2 2 75 and the length of the alleles ranged from 196 bp to 528 bp the number of alleles per locus ranged from 6 to 23 with an average of For the parents and offspring populations the average number of alleles was 8. 5 and respectively the observed heterozygosity was and respectively the average expected heterozygosity was and respectively and the average polymorphism information content was and respectively. The pedigree structure for the parents and progeny populations was inferred by maximum likelihood method according to the microsatellite marker information. The parent population was divided into six groups and the genetic distance among them ranged from to with an average of The offspring population was divided into 17 groups and the genetic distance among them ranged from 0 to with an average of The genetic distance between parents and offspring sub-groups ranged from 0 to with an average of An evolutionary tree for the 23 sub-group populations was drawn by unweighted pair-group method with arithmetic means UPGMA according to Nei's genetic distance. Eighteen sub-groups aggregated into the main stretch after forming three sub-stretches. However five sub-groups O15 O12 O13 P21 and P18 were obviously away from the main stretch. In this study the heterozygosity and polymorphism of the offspring population were lower than their parents. The result showed that the heterozygosity of the parents have partially lost during the genetic inbreeding. KEY WORDS Marsupenaeus japonicas Generation Genetic structure Genetic diversity Microsatellite Marsupenaeus japonicus Penaeidae Marsupenaeus t O'Connell et al Ben Naceur et al Balloux et al DNA Fenneropenaeus chinensis Litopenaeus vanname Cruz et al Pérez et al Ben Naceur et al Penaeus monodon Xu et al Wuthisuthim et al Pan et al Moore et al Luan et al Luan 2006

3 G1 G d DNA Wang 2006 DNA 0. 8% GENEQUANT pro Pharma-cia Biotech Ltd RNA /DNA DNA ng /μl PCR ' HEX FAM ROX PCR ABI 3130xl 8 1 PCR 25 μl 2 μl DNA 2 μl μl 2 Taq PCR MasterMix Taq dntps Buffer ddh 2 O 94 5 min s 1 min 72 1 min min 4 1 Table 1 Information of primers and grouping of amplification products Group Primers Primer sequence Motif Tm Length bp Fluorescence 1 MpSSR012 5'-CCATCCTGGCGAACATAC-3' TTCTTCTTA FAM 5'- CTCCACTTGCTACCCCTCC-3' MpSSR031 5'-AAGACAGACACGCCCTAA-3' TGT 14 TCA HEX 5'-CGTATGCGTGCGTGTATGAG-3' 2 MpSSR006 5'-CTCTCCTCTACCACAGTTCCC-3' CACG ROX 5'-CACCTATCCTTCAAATAAGTTCGT-3' MpSSR034 5'-CGTATGCGTGCGTGTATGAG-3' AAAAAT FAM 5'-CTGACGCCGTGCCAACAT-3' MsSSR076 5'-CTAGCAATCAGAAGCAGAA-3' TTAG HEX 5'-GTGCAAACGGTGTTCATCTG-3' 3 MpSSR039 5'-TACCTCACGGTTTCCTTACG-3' AC HEX 5'-GACGGTTCCTGAGAAGTTGAT-3' MpSSR064 5'-GACAACTCCTCCTGGTATG-3' GA FAM 5'-CTCCATCTTTTCCTTCCTCTT-3' MpSSR084 5'-TGAGCAGTCGTCCTCGGTT-3' CA ROX 5'-GTTGGTTATACAGACATGGGTAGTG-3'

4 ABI 3730xl Peak scanner Cervus 3. 0 Observed Heterozygosity Ho Expected Heterozygosity He Polymorphism Information Content PIC Hardy-Weinberg Colony Popgene Nei's Mega 5. 1 UPGMA bp Locus Type Table Genotype information of the eight SSR loci in cultured M. japonicas Hardy-Weinberg Allele Valid individual Ho He PIC HW MpSSR064 All * Parent ND Offspring NS MpSSR084 MpSSR076 MpSSR031 MpSSR034 MpSSR039 MpSSR012 All *** Parent NS Offspring *** All *** Parent NS Offspring *** All *** Parent NS Offspring NS All *** Parent ND Offspring *** All NS Parent ND Offspring *** All *** Parent *** Offspring ***

5 Locus Type Hardy-Weinberg Allele Valid individual Ho He PIC HW MpSSR006 Average All *** Parent NS Offspring *** All Parent Offspring ND Hardy-Weinberg NS Hardy-Weinberg * Hardy-Weinberg P < *** Hardy-Weinberg P < Note ND indicates that Hardy-Weinberg analysis was not done NS indicates that Hardy-Weinberg analysis is not significant the asterisk indicates significant differences at P < and triple-asterisk indicate significant differences at P < Table Number of individuals in different populations of M. japonicas ID of offspring Nei' s UPGMA O15 O12 O13 Number of shrimp ID of parent Number of shrimp O1 5 P18 1 O2 9 P19 2 O3 21 P20 1 O4 2 P21 1 O5 4 P22 10 O6 2 P23 46 O O O O O O O O O O O P21 P Botstein 1980 PIC PIC > <PIC < PIC < PIC MpSSR % PCR

6 第2 期胡龙洋等日本对虾养殖群体两个世代遗传结构的微卫星分析 79

7 MpSSR034 MpSSR039 Ho He He Ho 2012 MpSSR006 MpSSR012 60% 72. 6% Pemberton PIC Ho 1 23 PIC Fig. 1 Evolutionary tree for the 23 populations of two Ho generations of M. japonicas 3. 2 Hardy-Weinberg Bonferroni Hardy-Weinberg Colony Hardy-Weinberg Hardy-Weinberg Hardy-Weinberg Hardy-Weinberg Colony 1% Naceur et al. 2012

8 O12 O13 O15 P O15 O12 O13 P21 P18 1 Lande P19 P23 1 O5 P O4 O7 P COI DNA DNA PCR DNA fssr Larimichthys crocea WSSV PC STAT SSR SSR Balloux F Goudet J Perrin N Breeding system and genetic variance in the monogamous semi-social shrew Crocidura russula. Evolution 52 4

9 Ben Naceur A Chaabane R El-Falen C and 5 others Genetic diversity analysis of Nroth Africa's barley using SSR markers. J Gen Eng Biol Botstein D White RL Skolnick M and 1 other Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Human Gen Cruz P Ibarra AM Mejia-Ruiz H and 2 others Genetic variability assessed by microsatellites in a breeding program of Pacific white shrimp Litopenaeus vannamei. Mar Biotechnol Jerry DR Preston NP Crocos PJ and 3 others Parentage determination of Kuruma shrimp Penaeus Marsupenaeus japonicus using microsatellite markers Bate. Aquaculture Lande R Natural selection and random genetic drift in phenotypic evolution. Evol Luan S Kong J Wang QY Genetic variation of wild and cultured populations of the Kuruma prawn Marsupenaeus japonicus Bate 1888 using microsatellites. Aquaculture Res Moore SS Whan V Davis GP and 3 others The development and application of genetic markers for the Kuruma prawn Penaeus japonicus. Aquaculture Naceur AB Chaabane R El-Faleh C and 5 others Genetic diversity analysis of North Africa's barley using SSR markers. J Gen Eng Biotechnol O'Connell M Wright JM Microsatellite DNA in fishes. Rev Fish Biol Fish Pan YW Chou HH You EM and 1 other Isolation and characterization of 23 polymorphic microsatellite markers for diversity and stock analysis in tiger shrimp Penaeus monodon. Mol Ecol Notes Pérez F Ortiz J Zhinaula M and 3 others Development of EST-SSR markers by data mining in three species of shrimp Litopenaeus vannamei Litopenaeus stylirostris and Trachypenaeus birdy. Mar Biotechnol Pemberton J Slate J Bancroft D and 1 other Nonamplifying alleles at microsatellite loci a caution for parentage and population studies. Mol Ecol Wang WJ Kong J Dong SR and 3 others Genetic mapping of the Chinese shrimp Fenneropenaeus chinensis using AFLP markers. Curr Zool formerly Acta Zoologica Sinica Wuthisuthime TS Lumubol P Vanavichit A Development of microsatellite markers in black tiger shrimp Penaeus monodon Fabricius. Aquaculture Xu ZK Primavera JH Pena LD and 3 others Genetic diversity of wild and cultured black tiger shrimp Penaeus monodon in the Philippines using microsatellites. Aquaculture

AFLP % 1. 1 DNA 5 RAPD DNA 0. 1g % ~ ph 6 10 DNA 9 40μl TE SSR STS 20ng /L

AFLP % 1. 1 DNA 5 RAPD DNA 0. 1g % ~ ph 6 10 DNA 9 40μl TE SSR STS 20ng /L 2012 13 2 201-206 Journal of Plant Genetic Resources / 071001 41 SSR 21 SSR 41 52 2 ~ 4 2. 5 Shannon 0. 21 ~ 1. 40 0. 73 5. 54 3 7 9 13. 16% ~ 186. 49% 5. 48 6 11 SSR Genetic Diversity of Peanut Arachis