利用CottonSNP63K芯片构建棉花品种的指纹图谱

【目的】利用SNP位点的单拷贝特性,结合陆地棉TM-1参考基因组序列信息,筛选基因组特异的SNP。【方法】以719份遗传背景来源广泛的陆地棉种质资源为材料,采用Illumina公司开发的Cotton SNP63K芯片,利用Genome Studio软件对芯片扫描所获得原始数据进行基因型数据质量控制,获得待测样品SNP位点的基因型数据。根据已公布的陆地棉TM-1基因组的两个版本——中国农业科学院棉花研究所版本Gossypium hirsutum(AD1)genome BGI v1.0与南京农业大学版本G.hirsutum(AD1)genome NBI v1.1为参考序列,对Cotton SNP63K芯片(63 058个SNP)各位点的侧翼序列分别进行全基因组Blast比对分析,以筛选具有单拷贝特性的特异SNP位点并用于样品指纹图谱的构建。【结果】利用Cotton SNP63K芯片对719份材料进行SNP位点基因分型,主要表现为无检出信号的SNP位点、无多态性的SNP位点、具有多态性的SNP位点,而具有多态性的SNP位点的分型结果又可分为单位点SNP(基因组特异SNP)、双位点SNP和多位点SNP。通过对两个已公布的陆地棉TM-1参考基因组序列Blast比对结果表明,中国农业科学院棉花研究所TM-1基因组版本比对获得基因组特异SNP标记为5 474个,而南京农业大学TM-1基因组版本比对获得基因组特异SNP标记仅为1 850个,两者共有的特异SNP为1 594个,进一步通过分型效果、检出率及多态性3个评价指标,筛选score值≥0.7,call frequency值≥0.95,且MAF值≥0.2的SNP位点,获得471个分型效果理想,检出率高且多态性较高的特异SNP位点。在471个SNP位点中,430个位于染色体上,41个位于scaffold片段上。考虑到标记间的连锁程度,剔除连锁标记37个,最终获得393个核心SNP位点。利用393个核心SNP构建了719份品种资源的特征DNA指纹图谱,除个别材料之间遗传背景高度相似、基因型完全一致外,97%的材料均能实现准确有效的鉴别。【结论】筛选出393个基因组特异的SNP,并利用这些核心SNP构建了719份资源材料的特征DNA指纹图谱,为SNP分子标记应用于棉花重要性状遗传改良提供了参考。     

A molecular tool for parentage analysis in the Mediterranean mussel (Mytilus galloprovincialis)

Estimation of selection response and genealogical tracing in family mixtures require an appropriate tool for parentage analysis. In this study, we tested 19 marker loci for parentage analysis allocation in Mediterranean mussel (Mytilus galloprovincialis). To this aim, we reared families in tanks isolated from wild mussel seed, analysed them using the 19 marker loci and characterized their performances based on Mendelian rules. Probabilities of exclusion of a false parent were estimated for different groups of loci and contrasted to the real paternity assignment. Based on this, we chose nine microsatellites with the highest exclusion probabilities and a real paternity assignment of 99.6%. Next, we analysed 600 individuals reared as in the usual production process, where contamination from wild seed is likely. We obtained a real assignment of 94.7% and were able to identify individuals from the wild as the most likely hypothesis to explain the observed incompatibilities with candidate parents. This information was used to evaluate parental contribution in offspring obtained from gamete mixtures of several parents, which bestowed results of interest for future breeding programs of Mediterranean mussel.     

229份水稻品种及重要育种材料抗稻瘟病Pik位点基因型鉴定

Pik位点包含Pi-k、Pi-km、Pi-kp、Pi1、Pi-kh等抗稻瘟病等位基因,在我国水稻抗病育种中具有重要应用价值。本研究对229份水稻品种及重要育种材料的Pik位点进行基因型鉴定。利用K/N-单元型分子标记对水稻材料Pik位点进行多态性分析,鉴定了80份材料为K1K2功能性基因型。进一步通过Pi-k、Pi-kp和Pi1功能分子标记检测,结合功能多态位点序列分析,从80份K1K2型材料中,鉴定了黑稻、京虎B为Pi-k基因型;软米谷为Pi-kp基因型;恩恢99-64为Pi1基因型。研究还发现,高配合力强优恢复系闽恢3301的Pik位点为功能性K1K2基因型,其K1K2片段的序列与已克隆Pi-k、Pi-km、Pi-kp、Pi1、Pi-kh等位基因的序列存在差异,表明闽恢3301的Pik位点可能存在一个新等位基因。本研究明确了229份水稻材料Pik位点的K/N-基因型,结果可为水稻抗稻瘟病育种和品种合理布局提供参考。     

Single nucleotide polymorphism detection at the Hypothenemus hampeiRdl gene by allele-specific PCR amplification with Tm-shift primers

The resistant Rdl allele for dieldrin insecticide was detected on the Hypothenemus hampei populations from Colombia using conventional PCR methods. Based on this sequence, a melting temperature (T_m) shift genotyping method that relies on allele-specific PCR is described for insecticide resistance-associated single nucleotide polymorphism (SNP) at the H. hampeiRdl gene. The method reported here uses GC-rich tails of unequal length attached to allele-specific primers containing 3′ terminal bases that correspond to SNP allelic variants. Specific PCR products are identified by inspection of a melting curve on a real-time PCR thermocycler using SYBR Green DNA binding dye. Resistant and susceptible alleles resulted in specific PCR products with T_m of 83.3 ± 0.1 °C and 86.0 ± 0.2 °C, respectively. The RdlT_m-shift genotyping method is a new method to identify the Rdl gene in the coffee berry borer H. hampei, the principal pest of coffee that in general show low genetic diversity and very few genetic strategies for control of this pest have been developed. The method supplies a high-throughput tool for dieldrin resistance-associated SNP diagnostic in the coffee berry borer which will be useful for resistance-management strategies and as genetic marker in the colombian insect populations for genetics research.     

Groundnut (Arachis hypogaea L.) improvement in sub-Saharan Africa: a review

Groundnut (Arachis hypogaea L.) is a multi-purpose legume crop widely cultivated in sub-Saharan Africa (SSA). However, yield levels of the crop has remained relatively low in SSA owing to a range of biotic, abiotic and socio-economic constraints. A dedicated groundnut improvement programme integrating new tools and methodologies to breed varieties suitable for current and emerging agro-ecologies and market needs is essential for enhanced and sustainable groundnut production in SSA. The objective of this review is to highlight breeding progress, opportunities and challenges on groundnut improvement with regard to cultivar development and deployment in SSA in order to guide future improvement of the crop. The review analysed the role of new tools in breeding such as, high-throughput and automated phenotyping techniques, rapid generation advancement, single seed descent approach, marker-assisted selection, genomic selection, next-generation sequencing, genetic engineering and genome editing for accelerated breeding and cultivar development of groundnut.     

基因组选择及其在羊育种中的应用研究进展

数量性状是羊育种中的重要性状,受微效多基因控制、遗传力低,而传统育种方法难以提高羊的育种效率。提高动物育种效率对于选种选配工作和经济生产效益至关重要。随着育种新技术的不断革新与发展,基因组选择(genomic selection, GS)方法已成为育种技术中强大的工具,且已成功运用于个体经济价值较大的物种中,其具有缩短世代间隔、提高育种准确性、减少生产成本、提高畜禽经济效益等优势。近年来,由于基因组技术的不断成熟和各个统计模型的升级优化,以及高密度SNP芯片价格的下调,报告有关于基因组选择育种的实证和模拟研究层出不穷,且基因组选择技术已在羊育种中逐步开展,特别是在羊的重要性状中已有不少报道。由于羊的品种较多,地方性状差异化较大,个体经济价值略低,尽管基因组育种的新技术已经非常成熟,但目前仍没有在羊育种中大范围普及。为了更全面地了解该技术在羊育种中的研究现状,且基于选种选配的重要地位,作者就基因组选择在羊育种中的研究进展展开综述,主要从表型测定、基因分型、不同模型方面介绍了基因组选择在羊的重要性状中的应用和现状,讨论了其优势与挑战,并展望了基因组选择的未来发展方向。     

Identification of potential gene‐associated major traits using GBS‐GWAS for Korean apple germplasm collections

Genomewide association study (GWAS), which queries the association between loci and a particular trait by examining single nucleotide polymorphisms (SNPs) of the entire genome, is used in many fields of study. The development of next‐generation sequencing techniques has facilitated GWASs by decreasing the sequencing costs and time. In particular, genotyping by sequencing (GBS) is useful for sequencing many samples simultaneously and at a moderate price. Herein, we describe a potential GWAS using GBS, focused on the apple germplasm, with the goal of developing an effective apple breeding strategy through the identification of useful markers. From 308 Korean apple germplasm, SNPs were selected after GBS, and major traits were investigated. Proprietary individuals were confirmed and grouped by association through genetic diversity and population structure analyses of the selected SNPs. Genes highly associated with the target traits were identified, respectively. As the first GWAS report on the apple germplasm, these results will be useful as base data for GWASs on other apple populations and traits.     

Genetic diversity analysis of common bean (Phaseolus vulgaris L.) genotypes for resistance to Mexican bean weevil (Zabrotes subfasciatus), using single nucleotide polymorphism and phenotypic markers

ABSTRACT

The objective of this study was to examine the genetic diversity present among 297 common bean genotypes using 2554 SNPs and 12 insects and seed-related traits. The phenotyping was done under laboratory condition while the genotyping was conducted by using the Illumina SNP BeadChip. High phenotypic diversity among traits were recorded, ranging from 0.87 to 0.96, with a mean of 0.92. Principal component and discriminant analyses identi?ed four PCs and three discriminant functions, which explained 82% and 100% of the total phenotypic variations among genotypes, respectively. Polymorphic Information Content ranged from 0.21 to 0.38, with a mean of 0.34. The mean gene diversity among genotypes ranged from 0.24 to 0.50, with a mean of 0.44. Genetic distance ranged from 0.19 to 0.82, with a mean of 0.62, while the phenotypic distance ranged from 0.00 to 1.00, with a mean of 0.64 were observed aamong genotypes. The analysis of molecular variance revealed highly signi?cant differences (p<0.001) among and within individuals and among populations. Both the SNP and the phenotypic markers grouped the 297 genotypes into two major distinct clusters and three sub-clusters. This information is useful for identi?cation and development of common bean germplasm with economically valuable traits and the conservation and utilization of genotypes.