中国荷斯坦牛κ-酪蛋白基因第4和第5外显子多态性与泌乳性状的关联分析

 【目的】研究中国荷斯坦牛κ-酪蛋白基因第4和第5外显子多态性与泌乳性状的关联性,为培育高乳蛋白及高乳脂率奶牛提供参考依据。【方法】采用DNA测序、PCR-RFLP和CRS-PCR技术,对544头中国荷斯坦牛κ-酪蛋白基因的外显子4和5序列进行研究,通过SHEsis 软件和PHASE软件进行配对连锁不平衡分析和单倍型分析。【结果】发现6个新的SNPs,包括外显子5上A12907G、G12950A、C12989T、A13028G、T12980C共5个SNP位点,证实前4个位点紧密连锁;外显子4上A10996T共1个SNP位点。单位点基因型与泌乳性状关联分析表明,A10996T突变位点的TT和AT基因型个体乳脂率显著高于AA基因型个体(P＜0.05);T12980C突变位点的TC和CC基因型个体乳脂率、乳蛋白率分别极显著(P＜0.01)、显著(P＜0.05)高于TT基因型个体;紧密连锁的4个突变位点的DE基因型个体乳脂率极显著高于DD基因型个体(P＜0.01)。与泌乳性状的关联分析表明,共构建出7种单倍型,发现16种单倍型组合;其中H6H6单倍型组合个体乳脂率最高,H1H4次之;H1H4单倍型组合个体乳蛋白率最高;H5H6单倍型组合个体乳蛋白率及乳脂率最低。【结论】H1H4单倍型组合在乳蛋白率和乳脂率方面为有利单倍型组合,可作为选择高乳蛋白高乳脂率牛群的分子标记。     

Current situation and characterization of Pseudomonas syringae pv. actinidiae on kiwifruit in Galicia (northwest Spain)

Bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae (Psa), is a disease that is spreading rapidly in several kiwifruit‐producing countries, causing significant economic losses. In 2011, it was detected for the first time in Spain, in the south of Galicia (northwest Spain). Kiwifruit orchards were therefore inspected and sampled in 2011 and 2012 to determine the pathogen distribution, and the isolates obtained were characterized by morphology, fatty acids profile, biochemical tests and molecular techniques. Isolates were obtained from Actinidia deliciosa ‘Hayward’ (from leaves, canes, flower buds, fruits and roots), from A. deliciosa ‘Summer’, from Actinidia chinensis ‘Jin Tao’ (from canes and leaves) and from A. chinensis pollinator ‘Belén’ (from canes). Results of the analysis of the cfl gene (phytotoxin production‐related), the tox–argK gene cluster and phylogenetic analysis of the cts gene demonstrated that all Psa isolates from northwest Spain correspond to the Psa3 population, which includes strains of haplotype 2. This is the first record of Psa3 and haplotype 2 in Spain.     

Phylogenetic relationships and population structure of Ralstonia solanacearum isolated from diverse origins in Taiwan

Bacterial wilt caused by Ralstonia solanacearum is a destructive disease for many crops. The aim of this study was to investigate the phylogenetic relationships and genetic structure of an R. solanacearum population from diverse origins in Taiwan. All 58 tested isolates belonged to phylotype I, except the two biovar 2 isolates from potato. These belonged to phylotype IIB sequevar 1 and were identical to known potato brown rot strains, which were probably introduced. Phylotype I isolates were grouped into 10 sequevars. Sequevar 15 was predominant (34 out of 56 isolates). Its distribution covered the whole island and it was largely associated with solanaceous crops such as tomato, and with tomato field soil. A total of 14 haplotypes were identified based on a partial endoglucanase gene sequence. Parsimony network analysis revealed that haplotype A was the oldest haplotype in the local population. It encompassed all but one of the sequevar 15 isolates. Large variation in virulence on tomato was observed among the 58 isolates, and seven pathotypes were identified. Significant genetic differentiation was detected among pathotypes. Moreover, genetic differentiation was detected between biovar 3 and biovar 4 subgroups and between the strains associated with solanaceous and non‐solanaceous species, but none was detected between strains from different geographic origins. The results suggest that the phylotype I population in Taiwan is homogeneous, while mutation and local adaptation to specific ecological niches keep shaping the population.     

SCAR分子标记在甘蓝型油菜S单倍型分类中的应用

根据克隆到的ISLGa片段与ISLGb片段核苷酸序列之间的差异,发展了1个SCAR分子标记ISLGb(ISLGb-L+PS15),其引物只在具有ISLGb基因型的3个材料中扩增,并且均得到了大小为1265bp的单一亮带。通过与已有的相关分子标记整合得到了1套SCAR分子标记体系。包括2个多重PCR分子标记:①ISLG(PS5+PS15)和SP110(SP11-L+SP11-R')用于鉴别ISLG和IISP11b;②ISLGb(ISLGb-L+PS15)与SP11a(SP11-L+SP11-R)用于鉴别ISLGb和IISP11。还包括1个单PCR分子标记:SRKa(S40-5+S60-2)用于鉴别IISRKa。     

塔里木马鹿单倍型特性及其与产茸性状的关系

为了解塔里木马鹿单倍型特性及其与产茸性状的关系,以马鹿Y染色体上的ZFY基因为候选基因,以塔里木马鹿为试验群体,采用PCR直接测序法,利用Y染色体ZFY基因片段对塔里木马鹿的单倍型进行分析,共定义了4个单倍型,单倍型Z1、Z2为优势单倍型,利用SAS9.3软件分析单倍型与产茸性状之间的相关性。结果表明:ZFY基因对产茸性状有一定的影响(P0.2),其中单倍型2和单倍型3的影响尤为显著。     

中国荷斯坦奶牛与鲁西黄牛线粒体DNA多态性和系统进化分析

为阐明中国荷斯坦奶牛和鲁西黄牛的线粒体DNA多态性及其起源关系.提取了2个品种牛外周血液中的线粒体DNA(mtDNA),用PCR方法将牛mtDNA全长分为4段进行扩增,并用NlaⅢ、HpaⅡ、BglⅡ等16种限制性内切酶进行酶解消化,分析2个品种牛mtDNA的多态性,最终经7种限制性内切酶片段长度多态性分析,共归纳出4种主要的单倍型.通过对2个品种牛mtDNAD环全序列的聚类分析表明:鲁西黄牛和中国荷斯坦奶牛可能均为混合母系起源,源于欧洲普通牛和印度瘤牛.该研究结果对提高动物体外受精和体细胞核移植效率等生物技术平台有一定的理论指导意义.     

Nonrandom distribution of chum salmon stocks in the Bering Sea and the North Pacific Ocean estimated using mitochondrial DNA microarray

More than 1,000 age-identified chum salmon Oncorhynchus keta collected at 23 stations in the Bering Sea and the North Pacific Ocean in June to July 2003 were used to estimate their origin of stocks using a DNA microarray developed for analyzing the mitochondrial (mt)DNA haplotypes. The observed haplotype distribution was nearly the same as that reported previously for fish collected in September 2002 and 2003 in the present surveyed areas. A conditional maximum-likelihood method for estimation of stock compositions indicated that the Japanese stocks mainly distributed in north central Bering Sea, whereas the Russian stocks were mainly in western Bering Sea. The North American stocks were abundant in eastern Bering Sea and around the Aleutian Islands. Such an area-specific stock composition was not significantly different between mature and immature fish. Thus, the combined results of 2 years suggest that the distribution of chum salmon is nonrandom in the surveyed areas in summer and autumn, and that fish of the same origin migrate together to the same area irrespective of age.     

东亚4个黑山羊群体mtDNA D-环遗传多样性及其起源

为了研究地方山羊品种的起源与分化，了解其遗传背景，为山羊资源的合理利用提供基础资料，利用PCR产物直接测序法对3个中国黑山羊群体和1个韩国黑山羊群体共39个个体的mtDNA D-环部分序列进行了测定和分析。测定的序列经排列比对后选取441bp分析，发现了55个多态位点，单一多态位点11个，简约信息位点44个，确定了29种单倍型，单倍型多样度为0.984。构建系统发育树将29种单倍型分成了明显的3个分支，角猾羊聚入A分支。同时利用群体间的单倍型的错配分布和遗传距离分析了各群体的亲缘关系。结果表明：4个黑山羊群体遗传多样性丰富，至少拥有3个不同的母系起源，角猾羊是家山羊的一个母系祖先。     

杜洛克与二花脸杂交猪群体SNP偏分离分析

偏分离（TRD）是生物中普遍存在的现象,本研究拟在高密度基因芯片判断基因型的大规模群体中挖掘猪基因组标记位点的偏分离位点并探究其潜在的遗传机制。本研究用60K Illumina Porcine SNP芯片对1 020头F2资源家系（杜洛克与二花脸杂交F0-F2群体）进行基因型分型,用偏分离分析软件TRDscan （BF>100）和TDT （P<0.01）方法在单个位点上检测偏分离信号,并对二者共有的显著信号位点附近100 kb窗口的基因进行功能分析。此外,本研究还利用单倍型分型的软件包PHASEBOOK采用多位点连锁分析的策略分析了父源和母源配子中的偏分离区域,并在该区域内搜寻潜在的QTLs。结果表明：1）在单位点的偏分离分析中共鉴定到44个显著的偏分离位点,筛选到23个相关基因;对父本和母本特异性偏分离效应进行分析时,分别鉴定到27和35个显著偏分离位点,其中,分别有11和25个位点的100 kb附近有已注释的功能基因。2）基于单倍型多位点连锁的偏分离结果显示,父本显著偏分离位点位于5和13号染色体上,在该偏分离区域内搜寻功能相关的QTL,共搜寻到3个与繁殖性状（木乃伊数、产活仔数、黄体数）有关的QTLs;分析母本偏分离位点时,在4、6和12号染色体上定位到显著偏分离区域,结合已知的猪QTL数据库,共找到了5个与繁殖性状相关的QTLs。本研究利用大规模猪家系数据系统地鉴别了猪基因组的偏分离位点,为解析猪中的偏分离现象和进一步探究其生物学机制提供了一定的参考作用。     

Markers, old and new, for examining Phytophthora infestans diversity

Late blight, caused by Phytophthora infestans , is an ongoing threat to potato and tomato crop production worldwide and considerable fundamental and applied research is conducted with the long-term aim of improved disease control. Understanding the mechanisms, processes and rates of P. infestans evolution is an important factor in predicting the effectiveness and durability of new management practices. A range of phenotypic and genotypic tests has been applied to achieve this goal, but each has limitations and new methods are sought. Recent progress in P. infestans genomics is providing the raw data for such methods and new high-throughput codominant biomolecular markers are currently being developed that have tremendous potential in the study of P. infestans population biology, epidemiology, ecology, genetics and evolution. This paper reviews some key applications, recommends some changes in approach and reports on the status and potential of new and existing methods for probing P. infestans genetic diversity.