生物学杂志

生物学杂志

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微卫星技术筛选大黄鱼耐低温标记

  

  1. 宁波大学 应用海洋生物技术教育部重点实验室， 宁波 315211
  • 出版日期:2017-02-18 发布日期:2017-02-18
  • 通讯作者: 李明云，教授，研究方向为鱼类遗传育种与种质保护，E-mail: limingyun@nbu.edu.cn
  • 作者简介:穆方申，硕士，研究方向为生物化学与分子生物学，E-mail:mufangshen@126.com
  • 基金资助:
    国家高技术研究发展计划（2012AA10A403-4）；浙江省重大科技专项优先主题（2009C12077; 2012C12907-8）；宁波市重大项目（2008C3005）

Screening of microsatellite markers associated with cold tolerance of large yellow croaker (Pseudosciaena crocea

  1. Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo 315211, China
  • Online:2017-02-18 Published:2017-02-18

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摘要： 为筛选大黄鱼耐低温性状相关分子标记，对大黄鱼进行降温实验（15℃降至6℃）后用SSR标记对低温敏感个体和低温耐受个体进行扩增分析。结果显示：10对SSR引物共扩增到39个条带，仅标记KPC43的扩增条带在低温敏感和低温耐受个体间的出现频率存在显著差异。标记KPC43共扩增到3个等位基因，其中KPC43170bp在低温耐受组中出现的频率（80.0%）显著高于低温敏感组（23.33%），该条带与大黄鱼耐低温能力存在极显著正相关（P<0.01），相关系数为0.498；KPC43150bp在低温敏感组中出现的频率（76.67%）显著高于低温耐受组（30.0%），与大黄鱼耐低温能力存在极显著负相关（P<0.01），相关系数达到-0.417。序列比对显示KPC43170bp仅核心序列比KPC43150bp多了10个（TG）重复。推测SSR标记KPC43可能与大黄鱼耐低温性状相关基因连锁，其核心序列的重复次数可能与该基因的表达活性有关，从而影响大黄鱼个体的耐低温能力。

关键词: 大黄鱼, 耐低温, 微卫星标记

Abstract: In this study, the large yellow croakers Donghai No.1 was divided into cold-tolerant and cold-sensitive two groups by reducing water temperature from 15℃ to 6℃ gradually, and then the molecular marker which linked to the resistance to low temperature was screened by microsatellite markers (SSR) analysis. The results showed that 39 alleles were amplified using 10 SSR primers, and only the alleles frequence of locus KPC43 had significant difference between the cold-tolerant group and the cold-sensitive group. In the three alleles of KPC43, the occurrence frequency of KPC43170bp was 80.0% in the cold-tolerant group, while its occurrence frequency was only 23.33% in the cold-sensitive group. Correlative analysis showed that there was a significant positive correlation between KPC43170bp and cold-tolerant trait (P<0.01), and the correlation coefficient was 0.498. In the cold-sensitive group, the frequency of the KPC43150bp (76.67%) was significantly higher than that in the cold-tolerant group (30.0 %). And there was a significant negative association with the KPC43150bp and cold-tolerant related character (P<0.01), and the correlation coefficient was-0.417. Sequence alignment showed that the microsatellite flanking regions of KPC43170bp and KPC43150bp were the same, while the repeat sequence was (TG) 12 in KPC43170bp and (TG) 2 in KPC43150bp. It can be speculated that SSR marker KPC43 may be linked to some cold tolerant gene of large yellow croaker, and the repeat numbers of (TG) may affect these gene′s expressing activity, which is associated to the ability of cold tolerance.

Key words: Pseudosciaena crocea, low temperature tolerant, microsatellite marker