基于微卫星标记建立翘嘴鳜亲子鉴定技术

利用9个多态性较高的微卫星标记对翘嘴鳜(Siniperca chuatsi)进行亲子鉴定分析。结果显示,22个家系翘嘴鳜个体中共观测到95个等位基因,其观测杂合度和期望杂合度的平均值(范围)分别为0.578(0.219~0.800)和0.607(0.200~0.806),多态信息含量(范围)为0.552(0.187~0.787)。通过软件Cervus统计分析得到9个微卫星座位累计排除率为99.9658%,在27个可能的父母对条件下,混养养殖家系后代个体鉴定准确率为91%,研究结果表明筛选的这些多态微卫星分子标记可以用于翘嘴鳜优势家系的鉴定和分离,通过验证的微卫星标记建立的亲子鉴定技术为基础选育翘嘴鳜生长速度快的优势家系,为培育出生长速度快、抗病力强的翘嘴鳜新品种奠定基础。     

Marker‐assisted parentage analysis reveals high individual selfing rates in tetraploid red clover genotypes selected for seed yield

Seed yield is a major breeding target in tetraploid red clover. We investigated if marker‐assisted parentage analysis can identify progeny plants with two high seed‐yielding parents in tetraploid red clover and if this technique is more advantageous than traditional half‐sib selection. Parentage analysis was successfully performed on the progeny from the 10% highest seed‐yielding genotypes from a second‐cycle family selection trial: 16.0% of progeny were identified with a high seed‐yielding father. However, progeny plants with two high seed‐yielding parents did not produce more seeds than traditionally selected progeny (27.3 g vs. 30.7 g/plant, respectively). The 10% highest seed‐yielding genotypes displayed on average 2% self‐fertilization. Four genotypes were self‐fertile with individual selfing rates up to 20%. Our results discourage the use of marker‐assisted parentage analysis to improve seed yield in tetraploid red clover when the material has been preselected for seed yield. Breeders should be aware that intensive selection for seed yield in tetraploid red clover may inadvertently lead to selection for increased self‐fertility, which may increase inbreeding in the long term.     

柏氏鲤与荷包红鲤抗寒品系杂交子二代的RAPD分析

采用随机扩增多态性DNA（RAPD）分子标记技术，对荷包红鲤抗寒品系与柏氏鲤进行杂交所获得的F2代个体进行亲子鉴定分析。从180个随机引物中筛选出5个多态性较丰富的引物，并对其产生的多态性DNA片段进行统计分析，结果显示：双亲的RAPD谱带全部在F2代中表现出来，而子代中产生的所有谱带在双亲中的表现率在AB02、AB05、AC20、AI10、AI13几个引物中分别为87．0％、100．0％、92．1％、82．8％、92．6％。这也表明BAPD技术在对鲤的亲子鉴定中具有很强的可行性。     

中国育成黄麻主要品种间的亲缘关系分析

【目的】探讨新中国成立以来中国育成黄麻主要品种间的亲缘关系和遗传多样性。【方法】选用1949年以来,在中国有一定的推广面积和具有清晰系谱关系资料记录,以引进或育成的20个长果种和40个圆果种黄麻品种为供试材料,采用亲缘系数与聚类分析的方法,对中国60多年来黄麻生产上历经6次品种更替的主要品种间进行亲缘关系和遗传多样性分析。【结果】在长果黄麻中,20份供试品种共组成190对组合,31.58%存在亲缘关系,COP总和为38.5625,平均为0.0964,表明2/3的品种在系谱上的遗传关系较少;在圆果黄麻中,40份供试品种共组成780对组合,其中50.26%存在亲缘关系,COP平均为0.1179,表明50%的圆果品种存在亲缘关系,有近一半的品种亲缘关系相对较远。圆果黄麻品种间的亲缘关系比长果品种间的遗传相似性更高。聚类分析较直观地反映了新中国成立以来中国育成黄麻主要品种间的亲缘关系和品种演变特点。20份供试长果黄麻品种在欧式距离值0.95处,聚为5个类群。其中,2个国外引进品种各自聚为一个类群,其余18个品种分别以翠绿、广丰长果、巴长4号、马里野生长果等优势亲本聚为不同的大类群与亚类群。40份供试圆果黄麻品种在欧式距离值0.90处,聚为6个类群,除了2个地方品种各自聚为一个类群外,其余38个品种分别以D154、琼山、粤圆1号、卢滨圆果、选46等优势亲本聚为不同的大类群与亚类群。共祖先度分析表明,翠绿、广丰长果、宽叶长果、巴长4号等品种在新中国成立以来中国长果黄麻育种中利用价值很高;D154、卢滨圆果、粤圆1号、JRC-212、选46等品种是对中国圆果黄麻培育贡献较大的骨干亲本。【结论】在黄麻育种中,利用较多的资源是广丰长果、宽叶长果、粤圆1和早期引进的D154、JRC-212等少数骨干亲本,而近年引进的品种资源相对较少,由此导致现有黄麻品种遗传基础狭窄。在以后的育种工作中需加强黄麻种质资源的挖掘和引进,以提高黄麻品种的遗传多样性。     

Dispersal and tributary immigration by juvenile coho salmon contribute to spatial expansion during colonisation

Anadromous fishes are frequently restricted by artificial barriers to movement such as dams and culverts, so measuring dispersal helps identify sites where improved connectivity could promote range expansion and population viability. We used a combination of DNA‐based parentage analysis and mark–recapture techniques to evaluate dispersal by juvenile coho salmon (Oncorhynchus kisutch) in a population in the initial stages of colonisation following installation of fish passage structures at a previously impassable dam on the Cedar River, WA, USA. The spatial distribution of individuals within maternal families revealed that dispersal was common. Among the offspring of radio‐tagged mothers, 28% were collected outside the spawning reach and dispersed up to 6.3 km (median = 1.5 km). Most juveniles captured in a tributary (Rock Creek, where few adults spawned) had immigrated from the Cedar River and represented many different families. Juvenile dispersal therefore provided a secondary phase of spatial expansion following initial colonisation by adults. Consistent with the condition‐dependent dispersal hypothesis, juveniles that dispersed farther upstream in the tributary were larger than fish collected near the tributary mouth. Overall, the results demonstrated widespread dispersal in a system with low coho salmon densities, and this might increase the rate of population growth if it reduces the effects of local density dependence. By implication, juveniles can take advantage of rearing habitats reconnected through barrier removal, even when such areas are located several kilometres from adult breeding grounds.     

Parentage testing of Thoroughbred horse in Korea using microsatellite DNA typing

The present study was to construct a parentage testing system for Thoroughbred (TB) horse. A total number of 1,285 TB horse samples including 962 foals for parentage testing, 9 sires and 314 dams for individual identification were genotyped. Genomic DNA was extracted from 5 hair roots and genotyped by using 14 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, HMS1, HMS3, HMS6, HMS7, HTG4, HTG10, LEX3 and VHL20). This method consisted of multiplexing PCR procedure and showed reasonable amplification of all PCR products. Genotypes were determined by genetic analyzer. The number of alleles per locus varied from 3 to 9 with a mean value of 6.36 in TB horse. The expected heterozygosity was ranged from 0.548 to 0.831 (mean 0.699), and the total exclusion probability of 14 microstellite loci was 0.9998. Of the 14 markers, ASB2, ASB17, ASB23, HMS7 and HTG10 loci have relatively high PIC value (> 0.7). Of the 962 foals, 960 foals were qualified by compatibility according to the Mendelism. These results suggest that the DNA typing method has high potential for parentage verification and individual identification of TB horses.     

Multiple paternity,reproductive skew and correlates of male reproductive success in a wild population of the Trinidadian guppy

Abstract – In most species, males have a higher reproductive potential than females, leading to skewed reproductive success, particularly in mating systems where pre‐ or postcopulatory sexual selection reinforces inequality in male mating success. We investigated multiple paternity, reproductive skew and correlates of male reproductive success in a wild population of the Trinidadian guppy (Poecilia reticulata). We used nine microsatellite loci to assess the frequency of multiple paternity, number of sires per brood and reproductive skew. Across broods, the frequency of multiple paternity was high with 94% of broods having multiple sires (range: 1–5), resulting in a reproductive skew of 0.14. Variation in male reproductive success was high (range: 0–14 offspring per male), suggesting that there is considerable opportunity for sexual selection. Next, we examined correlations between male reproductive success and sexual coloration, sperm velocity and gonopodium length. Relative area of orange, black, iridescent and total coloration, and sperm velocity were not correlated with reproductive success. However, gonopodium length explained 14% of the variation in reproductive success, suggesting that gonopodium length is likely a sexually selected trait. We discuss these findings in the light of other studies that genetically dissect joint‐sex parentage and examine correlates of male reproductive success in wild populations.     

Parentage assignment of stocked black sea bream <Emphasis Type="Italic">Acanthopagrus schlegelii</Emphasis> in Hiroshima Bay using microsatellite DNA markers

The genetic contribution of 51 broodstock, comprising 29 females and 22 males, reared at Hiroshima City Marine Products Promotion Center for the production of stocked black sea bream was monitored during two consecutive years using seven microsatellite DNA loci. The high discrimination ability of these markers was reflected in the polymorphic identification content (PIC=0.831), the exclusion probability (Q≈1), and the low probability of identity index (I=3.635⁻¹⁰). The total number of breeders contributing to the mating process was estimated at 32 (62.7%) in 2000 and 30 (58.8%) in 2001. On pedigree reconstruction, 69.3% of the offspring were successfully assigned to a single broodstock pair. Loss of alleles accounted for 16.9% during seed production; nevertheless, 90.9% of males and 69.0% of females participated in the mating process. Based on microsatellite genetic tagging, 58.9% of the fish sampled during the two months after release were identified as hatchery stock, presenting no significant differences from wild conspecifics in either fork length or body weight.     

Microsatellite variability in natural populations of the blackspot seabream Pagellus bogaraveo (Brünnick, 1768): a database to access parentage assignment in aquaculture

Eight microsatellite loci previously reported were assessed for their utility in parentage assignment in 96 individuals belonging to natural populations of the blackspot seabream Pagellus bogaraveo (Brünnick, 1768) from the Mediterranean and Northeast Atlantic regions. At the mtDNA level, no differentiation was found between these two regions but based on microsatellite data an overall discrete genetic differentiation is perceivable between the two regions separated by the Strait of Gibraltar. The number of alleles per locus ranged from 8 to 30. A database constructed with allele frequency data from six populations was used in a simulation parentage assignment test using the software cervus . The test showed that the number of markers used is enough to perform parentage assignments with real data. The polymorphic information content for each locus was very high (mean value of 0.849), with a total exclusionary power of 0.9995. In summary, seven of the eight microsatellites analysed proved to be sufficient and powerful tools for parentage assignment in hatcheries and the allele frequency data given here can be used to perform pedigree analysis against which real data may be tested.