宁夏固原致病疫霉群体结构特征

为揭示宁夏固原致病疫霉群体遗传结构,用对峙培养法和菌落直径法测定致病疫霉的交配型和甲霜灵抗性,利用限制性片段长度多态性聚合酶链反应（PCR-RFLP）技术检测致病疫霉线粒体DNA单倍型,并采用简单重复序列（SSR）和扩增片段长度多态性（AFLP）技术确定致病疫霉群体的基因型。在94株供试菌株中,发现了1株A1交配型、86株A2交配型及7株自育型菌株,全部为甲霜灵抗性菌株。菌株的线粒体单倍型均为Ⅰa型,共鉴定出2种SSR基因型,Ⅰ型占绝对优势。AFLP聚类分析显示,在相似系数0.68时分成α和β两组,宁夏固原菌株全部聚在β组。研究表明,宁夏固原致病疫霉群体结构比较单一、遗传多样性水平较低。     

连锁不平衡的研究与应用

早在20世纪初期，Jinnings就提出了连锁不平衡(Linkage disequilibrium，LD)的概念。近年来随着单核苷酸多态性(Single nucleotide polymorphisms，SNPs)及其单倍型(Haplotypes)研究的兴起，LD也日益成为人们研究的热点。本文借助SNPs和单倍型两种方法来论述LD的研究，并简要介绍了LD研究与关联分析的关系。     

The Northern Ireland Phytophthora infestans population 1998–2002 characterized by genotypic and phenotypic markers

A total of 204 isolates of Phytophthora infestans from Northern Ireland, almost all from commercial potato crops, were collected over 5 years (1998–2002). Phenotypic diversity was assessed using mating type and metalaxyl resistance; genotypic diversity was assessed using two allozyme loci (glucose-6-phosphate isomerase, Gpi, and peptidase, Pep ), mitochondrial DNA haplotype and the multilocus RFLP probe RG57. All isolates were A1 mating type and Gpi 100/100 . The majority were Pep 100/100 , but four Pep 83/100 and six Pep 96/100 isolates were identified. Three mtDNA haplotypes were detected; haplotype IIa was the most common, but each year up to 2001 its frequency declined, with a concomitant increase in Ia isolates. Three isolates had the rare haplotype IIb, but this was only detected in 1998. Metalaxyl resistance and mtDNA haplotype were markedly associated: most haplotype Ia isolates were metalaxyl-resistant, whereas haplotype IIa was more commonly associated with metalaxyl sensitivity. Analysis of a subsample of 91 isolates revealed nine RG57 genotypes, three associated exclusively with haplotype IIa and six exclusively with haplotype Ia. The most common RG57 genotype (51% of isolates) comprised both metalaxyl-resistant and -sensitive haplotype IIa isolates. However, of haplotype Ia isolates, all metalaxyl-resistant phenotypes belonged to one of four RG57 types, one of which was the second most frequent overall (29% of isolates), while all metalaxyl-sensitive isolates belonged to one of two other types. The P. infestans population in Northern Ireland appears to consist of a limited number of clones related to, but differentiated from, the populations in mainland Britain and elsewhere in Europe.     

Phenotypic and genetic variation in Finnish potato-late blight populations, 1997–2000

This study was conducted on potato late blight samples collected for monitoring early attacks in Finland from 1997 to 2000. Most of the 1726 Phytophthora infestans isolates were collected soon after the onset of the epidemics. Most of the isolates were tested for mating type as well as metalaxyl and propamocarb hydrochloride (HCl) sensitivity, while virulence on potato R-gene differentials and mitochondrial DNA (mtDNA) haplotype were determined for a subset of the isolates. In half of the fields from which more than one isolate was tested, both mating types were detected, indicating strong potential for sexual reproduction. The mating types coexisted more often in organic fields or gardens than in conventional fields. The proportion of A1 mating type decreased from approximately 80% (1997 and 1998) to 22% by the year 2000. Simultaneously, the proportion of isolates resistant to metalaxyl decreased from approximately 40% to 16%. Resistance to metalaxyl was confined to the IIa mtDNA haplotype and clearly associated with the A1 mating type, as resistance was 10 times more common among A1 isolates than among A2 isolates. Resistance to metalaxyl therefore probably derives from common descent from an isolate with the IIa haplotype. Most of the regional variation in metalaxyl sensitivity was also linked to mating type, as both metalaxyl resistance and the A1 mating type were most common in the north and south-west of Finland. In contrast to metalaxyl, only three propamocarb-HCl-insensitive isolates were found, but propamocarb-HCl sensitivity decreased significantly during the study.     

利用线粒体D-loop区分析家鸭品种遗传多态性与系统进化

通过线粒体DNA控制区的结构和多态性来研究我国家鸭的遗传多态性与系统进化。利用DNA测序技术测定了我国9个家鸭品种106个个体线粒体DNA控制区多变序列。序列分析结果显示:A、C、T、G碱基的平均含量分别为25.6%、33.3%、15.2%和25.9%。检测到34个变异位点,约占分析位点总数的5.1%,有转换、颠换、插入/缺失4种类型的变异。确定了31种单倍型,其中单倍型A7为家鸭的主体单倍型,品种之间有9种共享单倍型。9个家鸭品种单倍型多样度(Hd)平均为0.798,核苷酸多样度(Pi)平均为0.28%,单倍型多样度在荆江麻鸭中最高,其次是攸县麻鸭和恩施麻鸭,在文登黑鸭中最低。9个品种家鸭之间双参数距离范围为0.001 3~0.004 4。31个家鸭单倍型序列的系统发生分析表明,9个家鸭品种只有1个母系起源,没有发现东亚斑嘴鸭对9个家鸭品种起源有贡献的证据。     

Pinpointing the candidate region for muscular dystrophy in chickens with an abnormal muscle gene

Muscular dystrophies, a group of inherited diseases with the progressive weakness and degeneration of skeletal muscle, contain genetically variable diseases. Though chicken muscular dystrophy with abnormal muscle (AM) has long been known, the gene responsible has not yet been identified. In this study, a resource family for AM was established with 487 F2 individuals and 22 gene markers, including microsatellite and insertion–deletion markers, were developed. The haplotypes were analyzed with these markers for the candidate region of GGA2q described in a previous study. The candidate region was successfully narrowed down to approximately 1Mbp. The region included seven functional genes predicted as the most likely AM candidates.     

黑龙江腐烂茎线虫群体的分离和鉴定

 本研究对分离自黑龙江省马铃薯中的1个线虫群体Hp1进行形态观察与测量,采用线虫通用引物对其rDNA-ITS进行扩增与序列分析,利用PAUP软件以ML法构建系统发育树,并测定其致病性。结果显示,该线虫群体有6条侧线,头部略缢缩,口针明显,中食道球呈长纺锤形,食道峡部窄而细长;雌虫阴门稍突起,后阴子宫囊较长;雄虫交合刺略向腹面弯曲,引带短,具交合伞。其形态测量值与腐烂茎线虫(Ditylenchus destructor)基本一致。rDNA-ITS PCR扩增片段916 bp,经BLAST比对,该ITS序列与腐烂茎线虫序列相似度最高。系统发育树显示,该群体没有与腐烂茎线虫A、B基因型群体聚在一起,而是与C、D型群体聚为1个分支,且与C型亲缘关系更近。据此,将群体Hp1鉴定为腐烂茎线虫C基因型,这是首次在黑龙江省发现腐烂线虫为害马铃薯。     

鲁西牛4个Y-STR微卫星位点的遗传多态性分析

提取鲁西牛血液DNA样品,PCR扩增后用基因自动测序仪进行基因扫描分型,研究Y-STR微卫星位点UMN0929,UMN0108,UMN0920和INRA124在鲁西牛群体中的遗传多态性。结果发现,156头无亲缘关系的鲁西牛公牛在4个Y-STR微卫星位点均有遗传多态性;UMN0929检测到4种等位基因,UMN0108检测到5种等位基因,UMN0920和INRA124均检测到2种等位基因。UMN0929、UMN0108、UMN0920和INRA124位点各等位基因的频率分别为0.25,0.41,0.17,0.17;0.14,0.28,0.28,0.14,0.16;0.38,0.62和0.73,0.27;基因多样性(GD)分别为0.71,0.78,0.47,0.39。共发现了58种单倍型(Haplotype),单倍型多样性(Haplotype diversity,HD)为0.94。表明UMN0929、UMN0108、UMN0920和INR124 4个微卫星位点在鲁西牛群体中有遗传多态性,在鲁西牛的起源、个体识别和亲子鉴定的研究中有较高的应用价值。     

猪RXRB基因SNPs检测及其与生长育肥和繁殖性状的关联分析

旨在研究猪RXRB基因多态性位点,并筛选与猪生长育肥和繁殖性状显著相关的SNPs,为种猪的遗传改良提供新的遗传标记位点。本研究通过采集962头健康大白猪（美系大白459头,法系大白503头）的血液DNA,利用直接测序法和PCR-RFLP技术分别检测两个品系大白猪RXRB基因SNP位点,并进行遗传多态性分析。利用SAS 8.0软件中的混合线性模型（Mixed）对RXRB基因SNP位点与表型性状值进行关联分析。利用Haploview 4.2软件对RXRB基因SNP位点进行连锁不平衡分析。结果显示,在两个品系大白猪群中共检测到10个SNPs,其中,美系大白有5个SNPs,法系大白有5个SNPs。经X²适合性检验,该10个SNPs在大白猪群体中均处于Hardy-Weinberg平衡状态（P>0.05）。关联分析结果表明,rs340542491、rs330162688和rs326226767与达100 kg体重日龄达到显著相关（P<0.05）;rs340542491、rs330162688、rs336609453和rs332169586与活体背膘厚达到显著相关（P<0.05）;rs340542491、rs325538588、rs691889709和rs323107853与初生重达到显著相关（P<0.05）;在体长中,仅rs324141460的GA基因型个体显著大于GG基因型（P<0.05）;rs325538588和rs80789331与眼肌面积达到显著相关（P<0.05）;在乳头数中,rs340542491和rs330162688与左乳头数达到极显著相关（P<0.01）,rs324141460与左、右乳头数达到极显著相关（P<0.01）,rs336609453和rs332169586与右乳头数达到显著相关（P<0.05）。连锁不平衡分析结果显示,在美系大白中,rs326226767-rs325538588处于强连锁不平衡状态（D’=0.96,r²=0.91>0.33）,rs330162688-rs324141460处于完全连锁状态（D’=1）,rs324141460-rs340542491处于完全连锁状态（D’=1）,单倍型组合关联分析结果与单个SNP关联分析结果一致,单倍型组合与达100 kg体重日龄、活体背膘厚、初生重和乳头数等性状存在极显著（P<0.01）或显著（P<0.05）相关;在法系大白中,rs691889709-rs323107853处于完美连锁状态（D’=1,r²=1）,单倍型组合关联分析结果显示,单倍型组合与体长和乳头数达到极显著（P<0.01）或显著（P<0.05）相关。以上结果提示,RXRB基因内的10个SNPs可以作为猪育种改良的分子标记,同时对深入研究该基因功能具有一定的参考意义。