膨腹海马生长性状相关微卫星标记筛选

Screening of microsatellite markers related to growth traits in Hippocampus abdominalis

  • 摘要:
    目的 筛选膨腹海马生长性状相关分子标记,利用分子标记来提高育种效率,加速育种进程。
    方法 本研究联合分群分离分析法(BSA)和微卫星(SSR)技术对同池养殖的同龄膨腹海马群体生长差异性状标记进行筛选。首先根据表型选择生长快慢极端个体构建极端混池,进行BSA全基因组重测序,初定位到生长性状候选区域,利用MISA工具扫描SSR位点并设计引物,采用二分法挑选引物,对生长快慢极端群体混池进行PCR扩增和毛细管电泳分析,筛选差异位点,对初筛差异位点开展极端混池个体基因型验证,最终通过家系水平极端混池及个体样本验证标记可靠性。
    结果 BSA重测序初定位到4 Mb生长性状候选区域,MISA结合Primer Premier 6.00共设计了16 691个SSR引物,二分法挑选的273对引物经群体极端混池毛细管电泳分析,共鉴定出55个差异位点,个体基因型验证显示,位点HA81、HA88分别存在266、245 bp的等位基因片段与生长性状呈现显著正相关关系,相关系数分别为0.202、0.183,而位点HA223存在260 bp的等位基因片段与生长性状呈现显著负相关关系,相关系数为−0.420。家系极端混池和极端个体有效验证了3个SSR位点与生长性状的显著关联性。
    结论 本研究首次获得了3个与膨腹海马生长性状显著相关的分子标记,为膨腹海马生长性状分子标记辅助选育提供了参考依据。

     

    Abstract: Hippocampus abdominalis was the largest known species of seahorse, primarily distributed in the sea of Australian and New Zealand. It has been included in the Red List of the World's Threatened Species due to overfishing, deterioration of the marine environment and global warming. It has been bred in captivity and was introduced to China in 2016. Current research primarily focused on reproductive physiology, pharmacological effects and aquaculture ecology. In the actual production and breeding of H. abdominalis, there are generally significant differences in growth traits in the same batch of seahorse populations. The breeding enterprises need to be screened regularly according to the size of the specifications to facilitate breeding management. The screening process is not only easy to cause the stress reaction of H. abdominalis but also increase the burden of production and breeding. Growth traits are important economic traits in aquatic animal genetic breeding. Therefore, combining trait molecular markers with breeding methods can improve breeding efficiency, accelerate breeding process and improve uniformity. To screen molecular markers related to growth traits of H. abdominalis, this study combined BSA and SSR techniques to screen differential growth trait markers of the same age H. abdominalis population cultured in the same pond. Firstly, the 4 Mb growth trait candidate region was initially mapped using BSA resequencing. Then, MISA tool was used to search for microsatellite locus in this candidate region and 16691 primers were designed. 273 primer pairs were selected for amplification analysis of two gene pools constructed from 60 individuals with fast and slow growth in H. abdominalis. A total of 55 differential locus were screened, and the genotype differences of these 55-differential locus in these 120 individuals were analyzed. The results showed that there were significant positive correlation (P<0.05) between allele fragments of 266 bp, 245 bp, 303 bp and 338 bp at locus HA81, HA88 and HA223 and growth traits, with correlation coefficients of 0.202, 0.183, 0.219, and 0.235, respectively; There were significant negative correlations (P<0.05) between allele fragments of 263 bp and 260 bp at locus HA88 and HA223 and growth traits, with correlation coefficients of -0.181 and -0.420, respectively; Finally, pedigree individual verification was conducted on these 3 locus, and the results showed that locus HA81 and HA88 were significantly positively correlated with growth traits at the 266 bp and 245 bp allele fragments, respectively, while locus HA223 was significantly negatively correlated with growth traits at the 260 bp allele fragment. Therefore, this study obtained for the first time 3 molecular markers significantly correlated with the growth traits of H. abdominalis, providing a reference for molecular marker assisted breeding of H. abdominalis growth traits.

     

/

返回文章
返回