Integration of AFLP markers into a linkage map of sugar beet (Beta vulgaris L.)

The AFLP (amplified fragment length polymorphism) technique has been applied in establishing an extended linkage map of sugar beet. A total of 120 AFLPs were integrated into an existing linkage map based on RFLP markers. Four primer combinations yielded between 19 and 40 polymorphic bands in an F₂ population consisting of 94 plants. The AFLP loci were evenly distributed over the nine linkage groups, with the exception of linkage group V where the number of AFLPs was significantly low. The AFLPs were found to be reproducible even against the background of different combinations of Taq DNA polymerases and buffers. However, the quantity of higher molecular weight fragments (>400 bp) was reduced when using plant DNA of poor quality as a template. The results of these experiments are discussed, together with possible applications of AFLPs in sugar beet breeding.     

A molecular marker linked to the Prv1 gene that confers resistance to Papaya ringspot virus-type W in melon

Papaya ringspot virus‐type W (PRSV‐W) is the most prevalent and important viral pathogen of cucurbits in Brazil. It can be effectively controlled by the incorporation of genetic resistance into susceptible melon cultivars. The present study identified amplified fragment length polymorphic (AFLP) markers linked to the PRSV‐W resistance Prv¹ allele. The susceptible yellow‐fleshed melon‐breeding line AF426^prv1 and its nearly isogenic‐resistant line AF426^Prv1, which carries the Prv¹ allele resident in the Indian cantaloupe U.S. Plant Introduction (PI) 180280, were screened for AFLP marker polymorphisms. Of 30 251 AFLP loci, only three were polymorphic between the nearly isogenic lines. Segregation analyses for these three polymorphic markers and the Prv¹ allele using a BC₁ population of 197 plants indicated close linkage (0.5% recombination frequency) between marker EK190 (HindIII‐CGA and MseI‐GTG; 190 bp) and Prv¹. Thus, EK190 might be a useful marker in breeding programmes aiming to develop melon cultivars resistant to PRSV‐W. The other two markers are closely linked to each other, but distantly linked to Prv¹.     

Screening RAPD primers for molecular taxonomy and cultivar fingerprinting in the genus Actinidia

Summary Eighty ten-base long arbitrary primers were tested for PCR-based DNA amplification of three species of the genus Actinidia (A. deliciosa the kiwifruit, A. chinensis, and A. kolomikta), with the aim of screening species-specific and genotype-specific markers.Of the 80 primers tested, 30 gave an average of 3.5 bands which were monomorphic within one or two species and absent in the remaining one(s), thus resulting in useful markers for taxonomic and phylogenetic purposes. None of the primers tested produced bands linked to sex. Twenty primers out of the twenty-five selected from a preliminary screening showed high levels of polymorphism, producing two to eleven patterns each from the 13 kiwifruit cultivars examined.We found the Stoffel fragment and the Taq polymerase were both suitable for RAPD analysis, the most noticeable difference being the smaller size of fragments (0.4–1.2 kb) produced by the former in comparison to the latter (1.0–3.4 kb). We tested also three different annealing temperatures (35, 37, and 39° C) and found the intermediate one best for number of amplified bands and reproducibility of results.Abbreviations 2-BE 2-butoxyethanol - CTAB hexadecyltrimethylammonium bromide - MAS Marker-Assisted-Selection - PCR Polymerase Chain Reaction - RAPD Random Amplified Polymorphic DNA - RFLPs Restriction Fragment Length Polymorphisms     

Molecular mapping and characterization of traits controlling fiber quality in cotton

Cotton (Gossypium spp) is the world's leading natural fiber crop. Genetic manipulation continues to play a key role in the improvement of fiber quality properties. By use of DNA-based molecular markers and a polymorphic mapping population derived from an inter specific cross between TM-1 (G. hirsutum) and 3-79 (G. barbadense), thirteen quantitative trait loci (QTLs) controlling fiber quality properties were identified in 3-79, an extra long staple (ELS) cotton. Four QTLs influenced bundle fiber strength, three influenced fiber length, and six influenced fiber fineness. These QTLs were located on different chromosomes or linkage groups and collectively explained 30% to 60%of the total phenotypic variance for each fiber quality property in the F₂ population. The effects and modes of action for the individual QTLs were characterized with 3-79 alleles in TM-1 genetic background. The results indicated more recessive than dominant, with much less additive effect in the gene mode. Transgressive segregation was observed for fiber fineness that could be beneficial to improvement of this trait. Molecular markers linked to fiber quality QTLs would be most effective in marker-assisted selection (MAS) of these recessive alleles in cotton breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

适合RAPD分析的三种玉米基因组DNA提取方法比较

本文通过对3种玉米基因组DNA提取方法的比较得出，DNA提取时采用一次氯仿／异戊醇抽提方法，得到的DNA质和量都较为理想，并且能够达到RAPD实验的要求。     

Sex-linked SSR markers in hemp

Hemp is a dioecious plant with sex chromosomes X and Y, the male sex being heterogametic. The quality of the fibre depends on the sex type. The sex chromosomes can be characterized by molecular markers. In this report, sex‐linked simple sequence repeat (SSR) markers are described. One SSR marker was polymorphic in both the populations derived from single crosses, two other markers in but one of the two populations. Three alleles were detected for two SSR markers indicating polymorphism not only between X and Y, but also between different X chromosomes. In addition, several sex‐linked RAPD markers were detected in one population. Recombination within the sex chromosomes was observed for nearly all markers.     

Molecular mapping determines that Hessian fly resistance gene H9 is located on chromosome 1A of wheat

Hessian fly [Mayetiola destructor (Say)] is one of the major insect pests of wheat (Triticum aestivum L.) worldwide. Hessian fly resistance gene H9 was previously reported to condition resistance to Hessian fly biotype L that is prevalent in many wheat‐growing areas of eastern USA and an RAPD marker, OPO05₁₀₀₀, linked to H9 in wheat was developed using wheat near‐isogenic lines (NILs). However, marker‐assisted selection (MAS) with RAPD markers is not always feasible. One of the objectives in this study was to convert an RAPD marker linked to the gene H9 into a sequence characterized amplified region (SCAR) marker to facilitate MAS and to map H9 in the wheat genome. The RAPD fragment from OPO05₁₀₀₀ was cloned, sequenced, and converted into a SCAR marker SOPO05₉₀₉, whose linkage relationship with H9 was subsequently confirmed in two F₂ populations segregating for H9. Linkage analysis identified one sequence tagged site (STS) marker, STS‐Pm3, and the eight microsatellite markers Xbarc263, Xcfa2153, Xpsp2999, Xgwm136, Xgdm33, Xcnl76, Xcnl117 and Xwmc24 near the H9 locus on the distal region of the short arm of chromosome 1A, contrary to the previously reported location of H9 on chromosome 5A. Locus Xbarc263 was 1.2 cM distal to H9, which itself was 1.7 cM proximal to loci Xcfa2153, Xpsp2999 and Xgwm136. The loci Xgwm136, Xcfa2153 and SOPO05₉₀₉ were shown to be specific to H9 and not diagnostic to several other Hessian fly resistance genes, and therefore should be useful for pyramiding H9 with other Hessian fly resistance genes in a single genotype.     

Molecular marker analysis of genetic variation characterizing a grasspea (Lathyrus sativus) collection from central Italy

Levels of genetic similarity characterizing 20 grasspea (Lathyrus sativus L.) populations collected in central Italy (17 populations in the Marche region and three populations in the Abruzzo region) were analysed with amplified fragment length polymorphism (AFLP) molecular markers. Two main clusters were found: one included large‐seeded populations from farms that were not market‐oriented (named Household populations) and the second, small‐seeded populations, cultivated in market‐oriented farms (named Commercial populations). Relationships among populations collected in different regions were found, although one population of the Abruzzo region was placed between the two main clusters, suggesting a possible further genetic differentiation within this grasspea germplasm collection. Principal component analysis based on AFLP marker frequency was effective in identifying polymorphic markers showing high discriminating ability between clusters H and C. In particular, seven markers showing high positive and three markers with low negative PC1 scores showed an almost cluster‐specific distribution. These results will be useful for enhancing Italian grasspea germplasm use in plant‐breeding programmes and for extending grasspea cultivation within the sustainable agricultural systems of central Italy.     

两种聚丙烯酰胺凝胶银染方法的比较

比较了两种聚丙烯酰胺凝胶的DNA染色方法。对小麦SSR扩增产物的染色结果显示,改进的Bassam银染法颜色背景浅,灵敏度较高,能够看到副带染色。改进的Sanguinetti方法对条带的鉴别力稍差,但因省时省力,成本较低可用于大规模引物筛选时的染色。     

中国五大湖三角帆蚌群体遗传多样性的RAPD分析

用OPJ和OPM两组40条10碱基随机引物,对中国五大湖三角帆蚌地理群体及诸暨养殖蚌进行了随机扩增多态性DNA(RAPD)分析,其中12个引物的扩增结果具有丰富的群体多态性,多态率为55.6%～80%。群体内遗传相似度大小依次为:鄱阳湖(0.8889),太湖(0.8694),洞庭湖(0.8111),诸暨(0.7746),洪泽湖(0.7348),巢湖(0.7185)。依据群体间遗传距离指数及分子系统树,表明洞庭湖群体和洪泽湖群体亲缘关系最近,鄱阳湖群体则与巢湖群体的亲缘关系最近,并且诸暨人工养殖群体与鄱阳湖和巢湖的群体比较接近。