Gene pool and connectivity patterns of Pinna nobilis in the Balearic Islands (Spain,Western Mediterranean Sea): Implications for its conservation through restocking

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Distribution and genetic diversity of Dothistroma septosporum in Pinus brutia forests of south-western Turkey

Dothistroma needle blight (DNB) is a serious disease of the Pinaceae, mainly Pinus species, caused by the fungi Dothistroma septosporum and D. pini. Both species are regarded as invasive forest pathogens worldwide, with rising incidence in central and northern Europe over the last three decades. In this work, 29 sites were investigated between 2013 and 2015 in south-western Turkey. Morphological examination of needles confirmed DNB infection (i.e., Dothistroma conidiospores observed) at 18 sites, and a total of 108 Dothistroma sp. isolates were obtained from 11 of the sites. Host age seemed to be an important factor in both occurrence and severity of DNB in Pinus brutia forests. Continuous rainy days, especially in December, may increase severity of disease; however, extreme rain events may reduce available conidiospores on plant tissues or in the air. Species-specific mating type primers showed that all isolates were D. septosporum; D. pini was not detected. The mating type ratio was close to 1:1, indicating sexual recombination was occurring. Eleven microsatellite markers revealed 59 unique multilocus haplotypes (MLHs) among the 73 isolates originating from different conidiomata. The majority of MLHs were represented by a single isolate (n = 52) and only one MLH was shared between two localities. Analyses showed high genetic diversity, isolation-by-distance, and clear population clusters. These findings suggest that D. septosporum is well established in south-western Turkey and is probably not a recent introduction.     

A highly accurate, single PCR reaction for parentage assignment in Senegal sole based on eight informative microsatellite loci

From a battery of microsatellite markers (100 loci), recently identified by our group, we have selected eight for parentage assignment in Senegal sole ( Solea senegalensis ). This tool is based on microsatellite loci obtained from four genomic DNA libraries and one cDNA library. Within the eight loci (six from anonymous genomic DNA sequences and two located in expressed sequence tags of known genes), we have found, in an analysis of a reproductive broodstock, between nine and 16 alleles. The expected heterozygosity was between 0.616 and 0.860. In addition, we have optimized the polymerase chain reaction (PCR) conditions to amplify all loci simultaneously in a single multiplex PCR reaction, and we have tested three lots of male and female (five to six individuals) and three offspring (50–60 larvae each). The use of the eight microsatellite loci, the possibility of amplifying them in a single PCR reaction and the high value of the exclusion probability (0.9992) make this multiplex PCR method a unique tool for parentage assignment.
Finally, analysing one meiotic gynogenetic progeny, we have determined the relative distance of six of these loci to the centromere, and we have also found that all of them are unlinked. All these characteristics confer this tool with a high accuracy for parentage studies and genetic population analyses of Senegal sole.     

鲢三、四核苷酸重复微卫星标记的筛选及其特征分析

通过磁珠富集法筛选鲢(Hypophthalmichtys molitrix)的三、四核苷酸重复微卫星分子标记。提取鲢血液大片段基因组DNA,用限制性内切酶Sau3AI进行酶切,蔗糖密度梯度离心收集400~900 bp长度的片段,构建鲢全基因组PCR文库,用生物素标记的微卫星探针(TGA)8、(CAG)8以及(AGAT)6进行筛选,磁珠富集含有微卫星序列的DNA片段,获得含有微卫星的单链序列;将这些序列通过PCR扩增,连接pMD 18-T载体,转入感受态大肠杆菌(Escherichia coli)DH5α,获得微卫星文库;再用γ-32P标记的放射性同位素探针进行二次杂交筛选,获得1 758个阳性克隆,选择测序348个克隆序列中,共含有280个微卫星座位,其中完美型167个(59.64%),非完美型28个(10%),混合型85个(30.36%)。Primer 3.0软件设计60对引物,并用珠江水系的野生鲢群体对引物进行多态性位点检测。统计软件分析显示其中22对具有较高的筛选效率和多态性,可作为鲢种质评价等遗传分析的工具。研究亮点:首次利用多碱基(三、四核苷酸重复)探针筛选出微卫星分子标记结合同位素探针进行两次筛选...     

The development and use of microsatellite markers for genetic analysis and plant breeding with emphasis on bread wheat

In recent years, a variety of molecular markers, based on microsatellites or simple sequence repeats (SSRs) have become the markers of choice, thus necessitating their development and use in a variety of plant systems. In this review, the basic principles underlying different hybridization-based (oligonucleotide fingerprinting) and PCR based approaches (STMS, MP-PCR, AMP-PCR/ ISSR/ ASSR, RAMPs/ dRAMPs, SAMPL), making use of microsatellites, have been outlined. Different methods for enrichment of genomic libraries for microsatellites have also been outlined. Relevant literature on the subject, giving a summary of results obtained using each approach, has been reviewed and critically discussed. The review also includes a discussion on literature, which deals with the use of microsatellites in genome mapping, gene tagging, DNA fingerprinting, characterization of germplasm and cytogenetics research. Special emphasis has been laid on the genome of bread wheat, where the work done in the authors' own laboratory has also been briefly reviewed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Two microsatellite markers flanking a dominant gene for resistance to soybean cust nematode race 3

The purpose of this work was to identifymicrosatellite markers linked to a gene forresistance to Heterodera glycinesIchinohe (Soybean Cyst Nematode – SCN) insoybean cultivar Hartwig. ABC₁F₂ mapping population derivedfrom a cross between Hartwig (resistant)and the Brazilian soybean line Y23(susceptible) was used. About 200microsatellite or simple sequence repeat(SSR) primer pairs were tested in a bulkedsegregant analysis (BSA). Those thatshowed clear polymorphisms were amplifiedin the BC₁F₂ population, whichhad been previously inoculated andevaluated for resistance/susceptibility toSCN Race 3. Three SSR markers linked toSCN resistance were detected in thepopulation. Two of them, Satt 038 and Satt163, flanking a dominant resistant gene(d/a = –0.90), explained 37% of thephenotypic variance. This gene was mappedat the edge of molecular linkage group G. Broad and narrow sense heritabilities wereestimated to be 50.54% and 37.73%,respectively. A selection efficiency of91.18% was obtained with the simultaneoususe of the two markers. The identified SSRmarkers will be useful tools for assistingthe selection of homozygous genotypes andfor expediting the introgression of the SCNresistance locus from cv. Hartwig tosoybean elite cultivars.     

Mapping QTLs for kernel oil content in a tropical maize population

Maize cultivars often have low kernel oil content. To increase the oil content, efficient maize breeding programs have to be developed, which require the knowledge of the inheritance of this trait. Thus, the objective of this research was to map quantitative trait locus (QTLs) and estimate their effects for kernel oil content in a tropical maize population. Two maize inbred lines, contrasting for kernel oil content, were used to develop an F₂ population. Four hundred and eight F₂ plants were self-pollinated, and their kernels (F_2:3 progenies) were used for kernel oil evaluation. A genetic map with 75 microsatellites was developed, and the QTLs were mapped using the composite interval map (CIM); also, estimates of genetic and phenotypic variances, and heritability coefficient were computed. The map presented 10 linkage groups, spanned 1,438.6 cM in length with an average interval of 19.18 cM between adjacent markers. The kernel oil content averaged 58.40 g kg^–1, and the broad-sense heritability was high (h²= 0.98). Thirteen QTLs were mapped, which were distributed into eight chromosomes, and explained 26.64% of the genetic variation. QTLs in chromosomes 1, 5, and 6 contributed the most for kernel oil content. Nine out of 13 QTLs with favorable alleles were from the parental inbred with the highest kernel oil content. The average level of dominance was partial, but gene action of the QTLs ranged from additive to overdominance. Eight out of 13 mapped QTLs were already reported for temperate maize populations.     

Comparative study of the discriminating capacity and effectiveness of AFLP, STMS and EST markers in assessing genetic relationships among evergreen azaleas

The application of amplified fragment length polymorphism (AFLP), sequence tagged microsatellite site (STMS) and expressed sequence tag (EST) markers to study the genetic relationships in an evergreen azalea gene pool was investigated. STMS and EST markers revealed a higher genetic distance detection capacity than AFLPs, which, nevertheless, were the most efficient marker system due to their highest polymorphism detection capacity. Similarity matrices showed weak, yet significant, correlations when Mantel's test was applied. To assess the usefulness of the overall information provided by these marker data for establishing phylogenetic relationships and horticultural classification, cluster analysis was performed. The joint AFLP, STMS and EST data were demonstrated to be remarkably effective for group discrimination and phylogenetic studies. The use of these polymerase chain reaction marker systems is discussed in terms of the choice of appropriate marker techniques for different aspects of evergreen azalea germplasm evaluation.     

Genetic mapping of the Acph1 locus in Aegilops tauschii

The Acph1 locus of Aegilops tauschii encodes a new electrophoretically ‘fast’ acid phosphatase, whose allelic variation could well be involved in intraspecies differentiation. Genetic mapping via microsatellite (simple sequence repeat) analysis revealed that Acph1 is tightly linked with the marker Xgwm157 near the centromere region of chromosome 2.     

Assessment of the uniformity of wheat and tomato varieties at DNA microsatellite loci

By analysing a number (20–38) of individuals from selected varieties of wheat and tomato, we have been able to assess intra-varietal uniformity at certain micro satellite (simple sequence repeat, SSR) loci. In total, 45 varieties of wheat were analysed at between 7–9 different SSR loci, and 10 varieties of tomato were analysed at six loci. The results showed that there was variation both between varieties and between microsatellites in the degree of non-uniformity observed, and it was possible to identify a number of different probable sources of non-uniformity. Twenty-four of the wheat varieties and nine of the tomato varieties were sufficiently uniform to meet the standards currently applied for distinctness, uniformity and stability (DUS) testing using phenotypic characteristics. The implications for the potential future use of SSRs in DUS testing are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.