RAPD

About: RAPD is a research topic. Over the lifetime, 15960 publications have been published within this topic receiving 360391 citations.

RAPD variation within and among natural populations of outcrossing willow-leaved foxglove (Digitalis obscura L.)

Abstract: Random amplified polymorphic DNA (RAPD) markers were used to assess levels and patterns of genetic diversity in Digitalis obscura L. (Scrophulariaceae), an outcrossing cardenolide-producing medicinal plant species. A total of 50 plants from six natural populations on the Iberian Peninsula were analysed by six arbitrarily chosen decamer primers resulting in 96 highly reproducible RAPD bands. To avoid bias in parameter estimation, analyses of population genetic structure were restricted to bands (35 of 96) whose observed frequencies were less than 1–3/n in each population. The analysis of molecular variance (AMOVA) with distances among individuals corrected for the dominant nature of RAPDs (genotypic analysis) showed that most of the variation (84.8%) occurred among individuals within populations, which is expected for an outcrossing organism. Of the remaining variance, 9.7% was attributed to differences between regions, and 5.5% for differences among populations within regions. Estimates of the Wright, Weir and Cockerham and Lynch and Milligan FST from null-allele frequencies corroborated AMOVA partitioning and provided significant evidence for population differentiation in D. obscura. A non-parametric test for the homogeneity of molecular variance (HOMOVA) also showed significant differences in the amount of genetic variability present in the six populations. UPGMA cluster analyses, based on Apostol genetic distance, revealed grouping of some geographically proximate populations. Nevertheless, a Mantel test did not give a significant correlation between geographic and genetic distances. This is the first report of the partitioning of genetic variability within and between populations of D. obscura and provides important baseline data for optimising sampling strategies and for conserving the genetic resources of this medicinal species.

Natural Environmental Sources ofCryptococcus neoformansvar.gattii

Abstract: We sought evidence for new environmental sources ofCryptococcus neoformansvar.gattiiby random amplification of polymorphic DNA (RAPD) analysis of isolates from 29 animals with a restricted territorial range in five Australian states. Twenty-three of the 29 isolates and 45 of 45 eucalypt isolates tested previously exhibited one RAPD profile, VGI. RAPD profile VGII was identified in 6 of 17 isolates from domesticated species but in none of 12 native species. Four VGII isolates originated from an area of Western Australia with no natural stands of known eucalypt host, indicating the existence of at least one unrecognized natural source ofC. neoformansvar.gattii.

Comparison of three typing methods for clinical and environmental isolates of Aspergillus fumigatus.

Abstract: To evaluate procedures used for epidemiologic analysis of outbreaks of aspergillosis, we analyzed a collection of 35 Aspergillus fumigatus isolates using three typing methods: isoenzyme analysis (IEA), random amplified polymorphic DNA (RAPD) analysis, and restriction endonuclease analysis (REA). Twenty-one isolates were from a single hospital, with four isolates coming from different patients. Three clinical isolates came from a different hospital, and 11 clinical or environmental isolates were derived from a culture collection. With IEA, the patterns of alkaline phosphatase, esterase, and catalase discriminated nine types. In contrast, 22 types were obtained with five different RAPD primers, and 21 types could be detected with three of these (R108, R151, and UBC90). Restriction endonuclease analysis of genomic DNA, digested with either XbaI, XhoI, or SalI, detected 3, 17, and 13 different REA types, respectively, and 22 types were identified by combining the data from the XhoI and SalI REAs. Twenty-eight types were obtainable with a combination of REA, IEA, and RAPD patterns. Overall, the results pointed to substantial genetic variation among the isolates. Though two isolates had markedly distinct genotypes, their morphologic features and exoantigens were consistent with their being A. fumigatus. The analysis will help in planning epidemiologic studies of aspergillosis.

Variation in Fusarium graminearum isolates from Nepal associated with their host of origin

Abstract: A collection of group II Fusarium graminearum isolates obtained from maize, wheat and rice from different locations in Nepal were identified using a combination of morphological and molecular criteria. The variation within this collection was analysed using RAPD markers, intergenic spacer (IGS) RFLP and PCR polymorphisms. The isolates were divided into two groups, A and B, by RAPD analysis. Isolates in group A yielded four different PCR polymorphic markers, but all the isolates in group B yielded a single polymorphic marker. The IGS RFLP analysis was consistent with division of the isolates into two groups. Isolates from wheat and rice were more frequently placed in group A, with isolates from maize more evenly distributed between the groups. Results indicate that host preference might be a factor in the division of isolates, although the year of isolation may also have had an influence. No geographical factors or agricultural practices could be identified to account for the observed variation.

Rapid emergence of hybrids between the two subspecies of Ophiostoma novo‐ulmi with a high level of pathogenic fitness

Abstract: During the 1970s Europe was invaded by two subspecies of the Dutch elm disease pathogen Ophiostoma novo-ulmi: subsp. americana from the west and subsp. novo-ulmi from the east. As a result their geographic ranges began to overlap in several areas. Only a weak prezygotic barrier to hybridization exists between the subspecies and in 1980 two hybrids were detected in the Netherlands. A subset of 107 O. novo-ulmi isolates collected in a subspecies overlap zone in Limburg, Netherlands in 1983 was characterized for three phenotypic markers and seven RAPD PCR markers. By phenotype, 33% were shown to be hybrid whereas by RAPD markers 69% were shown to be hybrid. Some isolates shown to be hybrid by phenotype were not revealed to be hybrid by PCR and vice versa. Combining the phenotype and RAPD data the estimated hybrid frequency was ∼78%. The mean growth rate of Limburg hybrid isolates was significantly faster than that of the Limburg subsp. novo-ulmi isolates but not significantly different from Limburg subsp. americana isolates. The Limburg hybrid isolates were just as pathogenic as the parent subspecies on both clonal Ulmus procera and on U. × Commelin. A subset of 100 isolates collected in another subspecies overlap zone at Orvieto, Italy in 1986 was also assessed with RAPD markers and ∼72% were shown to be hybrids. When 20 isolates of a ‘pure’ subsp. novo-ulmi population in the Baltic Ports area of Poland collected in 1980 were assessed by RAPD markers three isolates exhibited early introgression of subsp. americana DNA. This study therefore demonstrates very rapid emergence of O. novo-ulmi subspecies hybrids and introgressants in Europe in the early 1980s. In terms of two major fitness characters, growth rate and pathogenicity, these early hybrids were as fit as their parent subspecies. It is likely that complex hybrid swarms are now expanding across the continent.