AFLP analysis indicates no introgression of maize DNA in wheat x maize crosses

Amplified fragment length polymorphisms (AFLPs) were used to follow the possible introgression of maize DNA into haploids of wheat as a side‐effect of exploiting wheat x maize hybridization for haploid production. AFLPs were generated with 64 MseI/ EcoRI and 64 MseI/ PstI primer combinations, and the AFLP profiles of haploids were tested against those of maize and of the regular wheat varieties involved in the crosses. On average, 45.1 and 110.7 fragments were produced per assay with the MseI/EcoRI and MseI/PstI combinations, respectively. Different numbers of fragments were produced for wheat and maize: an average of 81 in the haploid, 80 in the wheat parent, and only 67.1 in maize. No evidence was found for introgression of maize into the wheat genome. Three unique AFLP fragments were detected in haploids, which were not present in the parental wheat genotypes. These ‘novel’ AFLP bands in the haploids could be caused by nucleo‐cytoplasmic interaction in the hybrid zygote. Such instability in the wheat genome is defined as temporal, as it was not detected in further generations when colchicine‐doubled progeny of the haploids was tested for the presence of polymorphic fragments.     

Phenotypic and genotypic analysis of a commercial cultivar and wild populations of Anemone coronaria

Seven wild populations of Anemonecoronaria were assessed for 11 phenotypic traits, most of them having economic value for the flower industry. The wild populations were sampled to represent the diversity in habitats, climates, rock and soil types, terrains, and elevations in Israel. AFLP analysis was carried out on 12 individuals from each of six out of the seven wild populations and for six individuals from the commercial cultivar ‘Mona-Lisa’. It was found that the Dorot population, which is located in the area bordering the semi-arid zone at the very end of the species distribution, exhibits extreme and different phenotypes with relatively low variability compared withthe other wild populations. The other six wild populations, that grow in more favorable geographic and climatic conditions exhibit phenotypes with larger plants, larger numbers of flowers and less dissected leaves. These populations were less uniform than that of Dorot. Genetic characterization by AFLP markers revealed a total of 165 bands. The wild populations exhibit wide variation within-population, with about 80% polymorphic bands and average gene diversity between pairs of about 30%. The Dorot population has the lowest genetic variation and the Megido population the highest. Thus, the phenotypic variation reflects the genetic variation. The cultivar ‘Mona-Lisa’, as expected, has much lower genetic variation. The Dorot population and the ‘Mona-Lisa’ cultivar were found to have the largest genetic distances from the other wild populations, and the highest genetic variation between themselves. Phenetic analysis yielded a dendrogram describing the genetic relatedness of these populations. This revised version was published online in August 2006 with corrections to the Cover Date.     

RFLP/AFLP mapping of a brown planthopper (Nilaparvata lugens Stål) resistance gene Bph1 in rice

We have constructed a linkage map of the rice brown planthopper (BPH)resistance gene, Bph1. RFLP and AFLP markers were selected by thebulked segregant analysis and used in the mapping study of 262 F₂sthat were derived from a cross of `Tsukushibare', a susceptible japonica cultivar, and `Norin-PL3', an authentic japonicaBph1-introgression line. Twenty markers were mapped within a 28.9-cMregion containing the Bph1 locus on the long arm of rice chromosome12. Combining the result of segregation analysis of BPH resistance by themass seedling test and that of the markers, the Bph1 locus wasmapped within a 5.8-cM region between two flanking markers. The closestAFLP markers, em5814N and em2802N, was at 2.7 cM proximal to theBph1 locus. Together with the previously constructed high-resolutionmap of bph2 locating the locus at ca. 10 cM proximal to the Bph1 locus, this improved version of the linkage map would facilitatepyramiding these two important BPH resistance genes.     

Genetic relationships among species and cultivars of Pistacia using RAPDs and AFLPs

The genus Pistacia (Anacardiaceae) includes 11 species divided into four sections, according to leaf characters and nut morphology. Recently two monophyletic groups have been proposed by using cpDNA,Lentiscus and Terebinthus,containing evergreen and deciduous species, respectively. In the present work molecular markers, derived from two different methods, RAPD and AFLP, are used to study the relationships of native and introducedPistacia species present in Greece. According to the cophenetic correlation coefficients best results for both methods were obtained by using the Jaccard algorithm and the UPGMA clustering method. However, phenograms were constructed using the NJ method (r_cs= 0.987 for RAPDs and r_cs= 0.982 for AFLP), since it is less sensitive against varying mutation rates. Correlation among the genetic similarity (GS) matrices for the two methods was high(r = 0.941). The AFLP and RAPD phenograms were comparable with minor clustering differences. According to our results, two main branches are obtained, one containing the evergreen species P. lentiscusand the resin producing P. lentiscuscv. Chia (cultivated only in the island of Chios), and the other branch containing the deciduous species P. terebinthus,P. palaestina and P. vera, P. chinensis was clustered either with the evergreen species (RAPD data) or with the deciduous species (AFLP data). P. palaestina is clustered to P. terebinthus and can be considered as a subspecies of P. terebinthus, since its GS values are close or smaller than GS values of entries within species (P. vera). The four female cultivars were found to have a narrow genetic basis, probably related to cultivar ‘Nazareth‘ with Syrian origin. This revised version was published online in July 2006 with corrections to the Cover Date.     

Bulked AFLP analysis for the assessment of genetic diversity in white clover (Trifolium repens L.)

The use of bulked leaf samples from individual plants for amplified fragment length polymorphism (AFLP) analysis was evaluated as a tool for assessment of genetic diversity in white clover (Trifolium repens L.). Bulking of leaf samples produced slightly simpler AFLP profiles compared to the combined profiles of individual plants from the same cultivar. Approximately 90% of bands which were present in individual plants were present in bulked samples of the same cultivar. The majority of those absent were rare bands, shared by less than 25% of individual plants. Replicate bulk samples gave almost identical banding patterns, demonstrating the robustness of the bulked AFLP technique. Cluster analysis of AFLP data derived from individual plants resulted in a phenogram similar to that produced from data derived from bulked samples of the same plants. AFLP analysis of bulked samples detected significant amounts of genetic variability among 52 cultivars and accessions with genetic similarity values ranging from 0.42 to 0.92. However, cluster analysis of AFLP data only partially reflected the geographic origin of cultivars and accessions and was not congruent with cluster analysis based on variation for morphophysiological characters. Bulked AFLP analysis provides a powerful tool for rapid assessment of genetic variability in white clover and may also be used for cultivar identification. This revised version was published online in August 2006 with corrections to the Cover Date.     

利用AFLP进行“甘蔗属复合体”系统演化和亲缘关系研究

采用AFLP分子标记技术，对“甘蔗属复合体”中4个属16个种的69份来自中国和澳大利亚的甘蔗种质资源材料进行系统演化和亲缘关系分析。2对AFLP引物共检测到173个标记，其中172个为多态标记，多态率达99.4%；通过计算Jaccard相似性系数，用UPGMA和PCA法构建了分子系统树和效应图。结果表明：(1)属间亲缘关系中，甘蔗属与芒属较     

Amplified fragment length polymorphism- and simple sequence repeat-based molecular tagging and mapping of greenbug resistance gene Gb3 in wheat

The greenbug, Schizaphis graminum (Rondani), is the most economically damaging aphid pest of wheat in the southern Great Plains of the USA. In this study, the single, dominant greenbug resistance gene, Gb3, was molecularly tagged and genetically mapped using amplified fragment length polymorphism (AFLP) and simple sequence repeat(SSR) markers. Three AFLP loci were associated with the Gb3 locus in linkage analysis with 75 F_2:3 families from the cross between two near‐isogenic lines (NILs) for Gb3,‘TXGBE273’ and ‘TXGBE281′. Two of these loci, XMgcc Pagg and Xmagg Patg cosegregate with Gb3 in the population analysed. Further analysis indicated that XMgcc Pagg and Xmagg Patg are specific for the Gb3 locus in diverse genetic backgrounds. Two SSR markers, Xgwm111 and Xgwm428 previously mapped in wheat chromosome 7D, were shown to be linked with Gb3, 22.5 cM and 33.1 cM from Gb3, respectively, in an F₂ population of ‘Largo’בTAM 107’, suggesting that Gb3 is located in the long arm of chromosome 7D. The two AFLP markers cosegregating with Gb3 are valuable tools in developing molecular markers for marker‐assisted selection of greenbug resistance in wheat breeding.     

Cultivar identification and genetic map of mango (Mangifera indica)

Amplified Fragment Length Polymorphism (AFLP) information was used for identification of mango (Mangifera indica L.) cultivars, for studying the genetic relationship among 16 mango cultivars and seven mango rootstocks and for the construction of a genetic linkage map. Six AFLP primer combinations produced 204 clear bands and on the average 34 bands for each combination. The average Band-Sharing between cultivars and rootstocks was 83% and 80%, respectively. The average Band-Sharing for mango is 81%. The probability of obtaining a similar pattern for two different mango cultivars and rootstocks is 6 × 10⁻³and 2 × 10⁻³, respectively. A preliminary genetic linkage map of the mango genome was constructed, based on the progeny of a cross between ‘Keitt’ and ‘Tommy-Atkins’. This linkage map consists of 13 linkage groups and covers 161.5 cm defined by 34 AFLP markers. This revised version was published online in August 2006 with corrections to the Cover Date.     

Amplified fragment length polymorphism as a tool for molecular characterization of almond germplasm: genetic diversity among cultivated genotypes and related wild species of almond,and its relationships with agronomic traits

Amplified fragment length polymorphism (AFLP) analysis is a rapid and efficient method for producing DNA fingerprints and molecular characterization. Our objectives were to: estimate genetic similarities (GS), marker indices, and polymorphic information contents (PICs) for AFLP markers in almond cultivars; assess the genetic diversity of almond cultivars and wild species, using GS estimated from AFLP fingerprints and molecular characterization; and facilitate the use of markers in inter-specific introgression and cultivar improvement. The genetic diversity of 45 almond cultivars from Iran, Europe, and America, were studied assaying 19 primer combinations. In addition, several agronomic traits were evaluated, including flowering and maturity times, self-incompatibility, and kernel and fruit properties. Out of the 813 polymerase chain reaction fragments that were scored, 781 (96.23%) were polymorphic. GS ranged from 0.5 to 0.96, marker indices ranged from 51.37 to 78.79, and PICs ranged from 0.56 to 0.86. Results allowed the unique molecular identification of all assayed genotypes. However, the correlation between genetic similarity clustering as based on AFLP and clustering for agronomic traits was low. Cluster analysis based on AFLP data clearly differentiated the genotypes and wild species according to their origin and pedigree, whereas, cluster analysis based on agronomic data differentiated according the pomological characterization. Our results showed the great genetic diversity of the almond cultivars and their interest for almond breeding.