海水鱼类多倍体遗传育种研究

项目名称： 海水鱼类多倍体遗传育种研究

项目编号： No.30871914

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 林浩然

作者单位： 中山大学

项目金额： 36万元

中文摘要： 本项目在建立淡水鱼类多倍体育种技术和海水鱼类人工繁育技术基础上，开创性的将细胞工程和染色体工程等技术运用到海水鱼类杂交育种中，通过远缘杂交的方法获得异源多倍体海水杂交鱼。根据鲈形目海水鱼类彼此亲缘关系的远近，选择染色体数目相同且都具有一定经济价值的海水鱼类进行杂交组合，分别进行科间、亚科间、属间和种间杂交实验。采用细胞生物学和分子生物学方法对几个组合杂交子代的生长和繁殖等生物学特性进行系统研究，比较了不同杂交子代的受精率、孵化率和仔鱼的畸形率，观察了杂交子代的胚胎发育过程和性腺发育状态，比较了亲本和杂交子代的形态学特征和生殖生长性状。使用倍性分析仪、流式细胞术和染色体制片技术检测远缘杂交子代染色体数目和倍性，发现在斜带石斑鱼♀#215;鞍带石斑鱼♂#26434;交子代中存在三倍体鱼3N=72，其三倍体比率为7%。利用线粒体DNA基因和微卫星标记分析研究了杂交子代与亲本的遗传关系，采用分子生物学技术克隆得到亲本和杂交子代的GH基因和IGF-I基因及其编码的蛋白质序列。本研究成果在海水鱼的遗传育种方面具有重要意义。

中文关键词： 海水鱼类；石斑鱼；远缘杂交；多倍体；育种

英文摘要： In this project, we firstly apply techniques of cell engineering and chromosome engineering to cross-breeding in marine fish and obtained the allopolyploid marine fish by distant crossing based on polyploidy breeding in freshwater fish and artificial reproduction in marine fish. According to genetic relationship of Perciform fishes, we made different cross combinations by choosing different marine fish with economic value, which have different genetic relationship and have the same chromosomal number. And we performed interspecific hybridization from different subfamilies, different genus and different species, respectively. At the cell biology and molecular biology levels, we made a series of researches on ploidy, gonadal development, growth, reproduction and other biological characteristics of these offspring of cross combinations, in which the formation of allopolyploid marine fish like allotetraploids will be expected. In this study, the comparative analysis of morphological characteristics and growth traits were carried out to clarify the morphological and growth differences between the larvae and juvenile of hybrids and their parents. The ploidy diversity of hybrids was monitored by Flow CytoMeter and further by chromosome count on kidney tissue. The results showed that the triploid individuals were found in the hybrids of Epinephelus coioides♀×pinephelus lanceolatus♂ triploid individual chromosome number in 3N=72, diploid individual chromosome number is 2N=48. By the analysis of mtDNA COI and SSR, the genetic relationship of hybrids and their parents were studied to show the hybrids hereditary characters. Comparative analysis of the ORF and amino acid of GH and IGF-I among the hybrids and their parents were carried out to explore the molecular genetic background of the difference in growth traits. This research plays an important role in marine fish genetic breeding.

英文关键词： marine fish;grouper;interspecific hybridization; polyploid; artificial breeding