鱼类经济性状遗传解析及分子育种应用研究

人类需要不断改良养殖鱼类的生产性状以提供更多优质的动物蛋白.鱼类的经济性状由多基因的数量性状位点所控制,其中大部分基因是微效的,但是少数基因可能具有决定性的影响.基于近10年来遗传学和基因组学研究中的主要进展,本文简要介绍和评述一些重要养殖鱼类在生长、抗病(逆)、性别等经济性状开展分子育种基础和应用研究的状况,探讨组学和下一代测序技术在鱼类经济性状遗传解析和分子设计育种中的应用潜力.这些研究有助于最终揭示鱼类经济性状的遗传调控机制并将相关研究成果应用于养殖鱼类目标性状的遗传改良.     

鱼类遗传育种中生物学方法的应用及研究进展

鱼类遗传育种是指利用生物学方法对鱼类进行遗传选择或改造,从而获得新型改良鱼类的过程;它可以通过人为选择优势遗传性状或者通过整合或改变已有的遗传性状而达到遗传改良的目的.鱼类遗传育种是一个从稳定品系中筛选或研制变异品系,再从变异品系中培育稳定品系的循环过程.鱼类良种的培育成功将在很大程度上带动良种的饲养、加工、销售、休闲等后续产业的发展,这在渔业经济发展中具有重要意义.本文系统地总结了包括传统选择育种、分子标记辅助育种、全基因组选择育种和单性控制育种在内的选择育种技术,综述了杂交育种(近缘和远缘杂交)、细胞核移植、生殖干细胞和生殖细胞移植、人工雌核发育和雄核发育技术、多倍体育种在内的性状整合育种技术,以及以转基因育种为代表的性状改造育种技术,乃至这些育种技术在鱼类育种中的应用情况,同时结合本实验室在鱼类远缘杂交、雌核发育和雄核发育等染色体倍性育种方面的研究成果,对国内外鱼类育种的研究现状以及存在的问题进行了系统的概述.     

lncRNA调控畜禽抗病力性状研究进展

长链非编码RNA(longnon-coding RNA,lncRNA)是一类长度大于200个核苷酸的非编码RNA分子。lncRNA虽然不具备蛋白编码能力,但可通过转录调控、转录后调控及表观遗传修饰调控等方式影响基因的表达,进而影响性状的表型。在现代畜牧业生产中,除提高生长发育和产量性状外,研究免疫因子、细胞因子等抗病力相关指标及性状的调控机制,对提高和改善畜禽的健康、福利及公共卫生尤为重要。近年来,利用lncRNA研究鸡(Gallus gallus)、猪(Sus scrofa)、牛(Bos taurus)等重要畜禽的抗病力性状的调控机制取得了一定进展,为将表观遗传标记应用于动物抗病遗传育种打下了一定的基础。本文介绍了lncRNA的生物学功能和产生机制,着重阐述了lncRNA对畜禽抗病力性状的调控作用及研究进展,以期为lncRNA在畜禽抗病遗传育种方面的研究及应用提供科学依据。     

水稻卷叶细胞学与分子机制研究进展

卷叶性状是水稻(Oryza sativa)重要的育种指标之一。研究表明,水稻叶片适度卷曲对植株的光合作用、株型及增产等均具有重要作用。该文综述了水稻叶片卷曲的相关研究进展,对水稻叶片卷曲的细胞学形成机制和相关调控基因的分子机制进行了阐述,以期深入阐明水稻叶片卷曲的细胞学和分子机制,促进卷叶性状在水稻育种中的应用。     

赤霉素生物合成与信号传递对植物株高的调控

植物株高是影响作物产量和品质的重要农艺性状。赤霉素(Gibberellins,GAs)是调控植物株高的重要激素,GA相关株高基因的克隆与分子机制研究对于合理调控作物生长发育和农业生产具有极其重要的利用价值,在水稻、小麦等粮食作物育种中得到了广泛应用。为了促进GA在果树、花卉等园艺作物育种中的有效利用,文中在分子生物学水平上介绍GA生物合成和GA信号传递途径对植物株高的调控。     

鱼类生长的神经内分泌调控

个体生长是由调控体内包括能量代谢和肌肉生长等多种生理信号通路共同影响的多基因调控性状.在参与脊椎动物生长过程的主要调控信号通路中,生长的内分泌调控轴及其组成信号起主要的作用.鱼类生长的内分泌调控轴和其生长调控的研究深受重视,包括生长激素、上游下丘脑激素、胰岛素样生长因子和下游分子等.许多的信号分子在体内不仅具有促进组织生长的作用,还具有对许多营养物质的代谢调控作用.体内其他内分泌调控轴也会对生长调控轴产生不可忽视的影响.本文主要归纳了近期硬骨鱼类生长内分泌调控轴的最新研究进展,包括最近利用遗传操作技术获得的相关体内模式的研究结果,并通过对这些最新研究的归纳,展示当前鱼类生长内分泌调控轴研究领域的新认识和新挑战.     

鱼类基因组操作与定向育种

水产养殖已成为全球范围内发展最快的农业产业之一,可持续发展水产养殖的关键在于培育具有优良性状的养殖品种.基因组操作技术为快速、定向的鱼类遗传育种提供了一条重要的可行性途径.本文回顾了基于经典基因组操作技术的鱼类育种方法学历史,如多倍体育种及细胞核移植等.然后重点介绍并展望了基于转基因技术及新近发展的基因组编辑技术的鱼类定向育种方法.后两种技术的发展和应用将会为未来的鱼类种业带来更加高效和更具预见性的育种新方法.     

山女鳟(Oncorhynchus masou masou)体重和叉长的遗传力估算

体重和体长作为鱼类的生长性状,是鱼类育种工作中最重要的生产性状之一,而在冷水鱼生产实践中通常利用叉长代替体长。文章采用不平衡巢式设计方法和定向人工授精技术建立了山女鳟(Oncorhynchus masoumasou)29个全同胞家系和14个半同胞家系,并测定全同胞家系和半同胞家系12月龄和24月龄的体重和叉长数据资料,进而利用方差分析法评估了作为引进种的山女鳟第六代群体体重和叉长的遗传力。结果表明:(1)山女鳟整个生长阶段叉长的变异系数明显低于体重变异系数,24月龄时的变异系数明显低于12月龄时的变异系数;(2)山女鳟在12月龄和24月龄雄鱼间和雄鱼内雌鱼间体重和叉长性状差异极显著(P<0.01);(3)山女鳟雌性亲本12月龄和24月龄体重和叉长的方差均大于雄性亲本的方差;(4)山女鳟12月龄体重的遗传力估计值为0.41~0.51,叉长的遗传力的估计值为0.46~0.54;24月龄时体重的遗传力估计值为0.55~0.60,叉长遗传力的估计值为0.53~0.59;(5)山女鳟体重和叉长性状属于中高遗传力,对山女鳟引进群体生长性状进行选择育种潜力巨大,预期能取得较好的遗传进展。研究结果为山女鳟选择育种相关工作提供了必要的参数依据。     

杉木育种亲本生长和结实性状早期测评与选择

为推进杉木(Cunninghamia lanceolata)育种进程,对广东乐昌市龙山林场和韶关市曲江区国有小坑林场杉木第3代育种园亲本群体的早期(3 a生)生长和结实性状进行分析.结果表明,同一育种园亲本无性系在生长性状(树高、胸径、单株材积、冠幅)和结实能力上存在显著差异,生长性状的差异甚至达到极显著水平(P<0....     

鱼类生长激素生物活性和定量免疫测定技术的研究进展

生长激素(growth hormone, GH)是调节鱼类生长、发育和代谢的一种重要激素.为了弄清鱼体内GH水平与鱼体生长的关系, 以及研究外源GH基因在受体鱼体内的表达部位、表达效率和表达调控等问题, 首先必须建立鱼类GH灵敏、特异的微量检测技术.此外, 在进行鱼类GH分离纯化中, 很重要的一点就是鉴定所获得的GH制品是否具有生物活性, 同样也需要建立鱼类GH灵敏、特异的生物活性检测技术.     

Fish genome manipulation and directional breeding

Aquaculture is one of the fastest developing agricultural industries worldwide.One of the most important factors for sustainable aquaculture is the development of high performing culture strains.Genome manipulation offers a powerful method to achieve rapid and directional breeding in fish.We review the history of fish breeding methods based on classical genome manipulation,including polyploidy breeding and nuclear transfer.Then,we discuss the advances and applications of fish directional breeding based on transgenic technology and recently developed genome editing technologies.These methods offer increased efficiency,precision and predictability in genetic improvement over traditional methods.     

Genome-wide association study reveals the genetic basis of growth trait in yellow catfish with sexual size dimorphism

Sexual size dimorphism has been widely observed in a large number of animals including fish species. Genome-wide association study (GWAS) is a powerful tool to dissect the genetic basis of complex traits, whereas the sex-differences in the genomics of animal complex traits have been ignored in the GWAS analysis. Yellow catfish (Pelteobagrus fulvidraco) is an important aquaculture fish in China with significant sexual size dimorphism. In this study, GWAS was conducted to identify candidate SNPs and genes related to body length (BL) and body weight (BW) in 125 female yellow catfish from a breeding population. In total, one BL-related SNP and three BW-related SNPs were identified to be significantly associated with the traits. Besides, one of these SNPs (Chr15:19195072) was shared in both the BW and BL traits in female yellow catfish, which was further validated in 185 male individuals and located on the exon of stat5b gene. Transgenic yellow catfish and zebrafish that expressed yellow catfish stat5b showed increased growth rate and reduction of sexual size dimorphism. These results not only reveal the genetic basis of growth trait and sexual size dimorphism in fish species, but also provide useful information for the marker-assisted breeding in yellow catfish.     

Quantifying the genetic parameters of feed efficiency in juvenile Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

Background

Improving feed efficiency in fish is crucial at the economic, social and environmental levels with respect to developing a more sustainable aquaculture. The important contribution of genetic improvement to achieve this goal has been hampered by the lack of accurate basic information on the genetic parameters of feed efficiency in fish. We used video assessment of feed intake on individual fish reared in groups to estimate the genetic parameters of six growth traits, feed intake, feed conversion ratio (FCR) and residual feed intake in 40 pedigreed families of the GIFT strain of Nile tilapia, Oreochromis niloticus. Feed intake and growth were measured on juvenile fish (22.4 g mean body weight) during 13 consecutive meals, representing 7 days of measurements. We used these data to estimate the FCR response to different selection criteria to assess the potential of genetics as a means of increasing FCR in tilapia.

Results

Our results demonstrate genetic control for FCR in tilapia, with a heritability estimate of 0.32?±?0.11. Response to selection estimates showed FCR could be efficiently improved by selective breeding. Due to low genetic correlations, selection for growth traits would not improve FCR. However, weight loss at fasting has a high genetic correlation with FCR (0.80?±?0.25) and a moderate heritability (0.23), and could be an easy to measure and efficient criterion to improve FCR by selective breeding in tilapia.

Conclusion

At this age, FCR is genetically determined in Nile tilapia. A selective breeding program could be possible and could help enabling the development of a more sustainable aquaculture production.

     

Fish Growth in Marine Culture Systems: A Challenge for Biotechnology

Aquaculture production is constrained largely by the growth efficiency of the species being produced. Nutritional approaches have played an important part in improving this situation, but, it is argued, the room for further improvement using such established techniques is limited. Alternative ways of improving fish production by utilizing recent biotechnological advances are explored and assessed as to their potential for commercialization in the near future. Transgenic technologies promise a revolution in aquaculture, but it is considered that consumer resistance may delay the use of transgenic fish for food production. An alternative approach could be the breeding of transgenic fodder plants without the amino acid deficiencies of existing alternatives to fish meal in aquaculture diets. The use of probiotics could reduce antibiotic use on fish farms while they might also provide the basis for ``smart' diets, tailored to specific purposes by the inclusion of microorganisms. The selection and genetic engineering of nitrifying and denitrifying bacteria could also pave the way for fully enclosed, recirculating marine culture systems, which would allow control of the environmental variables that currently restrain marine fish culture. Received August 10, 1998; accepted October 8, 1998.     

Development and application of biological technologies in fish genetic breeding

Fish genetic breeding is a process that remolds heritable traits to obtain neotype and improved varieties.For the purpose of genetic improvement,researchers can select for desirable genetic traits,integrate a suite of traits from different donors,or alter the innate genetic traits of a species.These improved varieties have,in many cases,facilitated the development of the aquaculture industry by lowering costs and increasing both quality and yield.In this review,we present the pertinent literatures and summarize the biological bases and application of selection breeding technologies(containing traditional selective breeding,molecular marker-assisted breeding,genome-wide selective breeding and breeding by controlling single-sex groups),integration breeding technologies(containing cross breeding,nuclear transplantation,germline stem cells and germ cells transplantation,artificial gynogenesis,artificial androgenesis and polyploid breeding)and modification breeding technologies(represented by transgenic breeding)in fish genetic breeding.Additionally,we discuss the progress our laboratory has made in the field of chromosomal ploidy breeding of fish,including distant hybridization,gynogenesis,and androgenesis.Finally,we systematically summarize the research status and known problems associated with each technology.     

Genetic basis and biotechnological manipulation of sexual dimorphism and sex determination in fish

Aquaculture has made an enormous contribution to the world food production,especially to the sustainable supply of animal proteins.The utility of diverse reproduction strategies in fish,such as the exploiting use of unisexual gynogenesis,has created a typical case of fish genetic breeding.A number of fish species show substantial sexual dimorphism that is closely linked to multiple economic traits including growth rate and body size,and the efficient development of sex-linked genetic markers and sex control biotechnologies has provided significant approaches to increase the production and value for commercial purposes.Along with the rapid development of genomics and molecular genetic techniques,the genetic basis of sexual dimorphism has been gradually deciphered,and great progress has been made in the mechanisms of fish sex determination and identification of sex-determining genes.This review summarizes the progress to provide some directive and objective thinking for further research in this field.     

鲢EST-SSR标记的开发及其与生长性状关联性分析

研究采用Illumina高通量测序技术对鲢(Hypophthalmichthys molitrix)卵组织进行转录组测序分析, 筛选多态EST-SSR标记并与生长性状进行关联性分析。研究结果显示: (1)测序组装总Unigene 77129个, SSR共24458个, 占所测EST数据的31.7%。鲢包含SSR的EST序列主要有二碱基、三碱基、四碱基和五碱基组成。二碱基中, AC含量最丰富; (2)随机挑选175条EST序列设计SSR引物, 筛选出多态EST-SSR标记36个, Blast结果显示有10条已知基因功能; (3)全同胞家系检测结果显示有两个标记与鲢的生长性状相关。SCE26与体长和体重显著相关(P<0.05), SCE65与肥满度显著相关(P<0.05)。研究结果将为鲢的分子标记辅助育种的应用提供基础理论指导。     

Development and testing of a combined species SNP array for the European seabass (Dicentrarchus labrax) and gilthead seabream (Sparus aurata)

SNP arrays are powerful tools for high-resolution studies of the genetic basis of complex traits, facilitating both selective breeding and population genomic research. The European seabass (Dicentrarchus labrax) and the gilthead seabream (Sparus aurata) are the two most important fish species for Mediterranean aquaculture. While selective breeding programmes increasingly underpin stock supply for this industry, genomic selection is not yet widespread. Genomic selection has major potential to expedite genetic gain, particularly for traits practically impossible to measure on selection candidates, such as disease resistance and fillet characteristics. The aim of our study was to design a combined-species 60 K SNP array for European seabass and gilthead seabream, and to test its performance on farmed and wild populations from numerous locations throughout the species range. To achieve this, high coverage Illumina whole-genome sequencing of pooled samples was performed for 24 populations of European seabass and 27 populations of gilthead seabream. This resulted in a database of ~20 million SNPs per species, which were then filtered to identify high-quality variants and create the final set for the development of the ‘MedFish’ SNP array. The array was then tested by genotyping a subset of the discovery populations, highlighting a high conversion rate to functioning polymorphic assays on the array (92% in seabass; 89% in seabream) and repeatability (99.4–99.7%). The platform interrogates ~30 K markers in each species, includes features such as SNPs previously shown to be associated with performance traits, and is enriched for SNPs predicted to have high functional effects on proteins. The array was demonstrated to be effective at detecting population structure across a wide range of fish populations from diverse geographical origins, and to examine the extent of haplotype sharing among Mediterranean farmed fish populations. In conclusion, the new MedFish array enables efficient and accurate high-throughput genotyping for genome-wide distributed SNPs for each fish species, and will facilitate stock management, population genomics approaches, and acceleration of selective breeding through genomic selection.     

Progress in the evaluation of transgenic fish for possible ecological risk and its containment strategies

Genetically improved transgenic fish possess many beneficial economic traits; however, the commercial aquaculture of transgenic fish has not been performed till date. One of the major reasons for this is the possible ecological risk associated with the escape or release of the transgenic fish. Using a growth hormone transgenic fish with rapid growth characteristics as a subject, this paper analyzes the following: the essence of the potential ecological risks posed by transgenic fish; ecological risk in the current situation due to transgenic fish via one-factor phenotypic and fitness analysis, and mathematical model deduction. Then, it expounds new ideas and the latest findings using an artificially simulated ecosystem for the evaluation of the ecological risks posed by transgenic fish. Further, the study comments on the strategies and principles of controlling these ecological risks by using a triploid approach. Based on these results, we propose that ecological risk evaluation and prevention strategies are indispensable important components and should be accompanied with breeding research in order to provide enlightments for transgenic fish breeding, evaluation of the ecological risks posed by transgenic fish, and development of containment strategies against the risks.