RAPD

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

Genetic variability and relatedness in the complex group of black Aspergilli based on random amplification of polymorphic DNA

Abstract: A PCR-based technique, involving the random amplification of polymorphic DNA (RAPD), was used for assessing genomic variability among a wide range of culture collection strains of black Aspergilli and related species. The performance of this technique is compared with that of the two other genetic techniques most commonly used, namely restriction fragment length polymorphisms on rDNA and isozyme analysis. The eight main groups as assigned by RFLP were also distinguished by RAPD patterns. On the basis of 122 polymorphic RAPD products using six random primers, the 17 collection strains examined could be subdivided into 15 distinct sub-groups. We suggest that the RAPD method is a quick and reliable tool for establishing the amount of genetic variability in closely related species. Our study indicates that the complex group of black Aspergilli is characterized by a high degree of genetic differentiation. This is also apparent from the considerable karyotype variation present in the group.

Chinese melon (Cucumis melo L.) diversity analyses provide strategies for germplasm curation, genetic improvement, and evidentiary support of domestication patterns

Abstract: The genetic diversity of melon market types (Cucumis melo L., 2n = 2x = 24) in China, an important secondary center of diversity, has not been examined. Therefore, reference accessions from India and Africa, Crete/Greece, Japan, Europe, U.S.A., Spain, and 68 Chinese cultigens (fresh-market non-netted thin-skinned; non-netted thick-skinned; netted thick-skinned; and non-netted thin-skinned, and vegetable) were evaluated by using 17 10-mer RAPD primers (32 mapped loci), days to flower, sex expression, lateral-branch number, and fruit number and weight per plant. While Chinese thin-skinned melons differed from vegetable melon types only in sex expression, the U.S. Western Shipping market type reference accession “Top Mark” and Chinese thick-skinned melons were similar for all of the morphological traits examined. The average similarity (Jaccard Coefficient) between any two pairs of accessions examined as estimated by RAPD variation was 0.47 ± 0.14. Within-group genetic similarities ranged between 0.94 (thin-skinned type) and 0.08 (non-netted thick-skinned type). The average/standard deviation, maximum, and minimum similarity between any two Chinese reference accessions was 0.41 ± 0.13, 0.75, and 0.12, respectively. Cluster analysis partitioned accessions into two main branches consisting of Group Cantalupensis and Inodorus reference accessions (clade 1) and Chinese accessions (clade 2). A second cluster analysis partitioned China, India, and Africa accessions into one major group, and accessions from Japan, Europe, and U.S.A. into another. Results indicate that Chinese accessions are a rich source of genetic diversity for plant improvement, and that molecular assessments support previously described theoretical melon domestication patterns constructed from historical and archeological evidence.

DNA-based marker systems to determine genetic diversity of weedy species and their application to biocontrol.

Abstract: DNA-based molecular markers may provide infor- mation about introduced weedy species that would be useful in biological weed control efforts Chloroplast DNA restric- tion fragment length polymorphisms (cpDNA RFLP) and random amplified polymorphic DNA (RAPD) analysis are two DNA-based marker techniques that can provide esti- mates of genetic variation in native and introduced popula- tions of weedy species Profiles provided by these techniques could furnish the necessary information to determine the geographic origins of introduced species and provide evi- dence for multiple introductions Although DNA-based markers would not necessarily identify the genetic basis for host-pest compatibility, they would enable identification of specific host genotypes Current criteria for selecting a weedy species as a target for biological control are primarily politi- cal and economic The importance of genetic diversity and population structure in determining the vulnerability of plant populations to insects or diseases has not been fully appreciated Estimates of genetic diversity based on DNA marker analysis could be used as one criteria for determining which plants are targeted for biological control The success of biological weed control efforts has been limited by the high levels of genetic diversity occurring in target weed species and the lack of biocontrol agent and target weed compatibili- ties DNA-based markers may be used to increase our under- standing of these factors and contribute to the success of biological weed control by helping to target the most vulner- able species and provide more realistic expectations of the potential for success given available resources Additional index words Chloroplast DNA, restriction frag- ment length polymorphisms, polymerase chain reaction, ran- dom amplified polymorphic DNA, biocontrol

Development of sex-linked PCR markers for gender identification in Actinidia

Abstract: Two sex-linked random amplified polymorphic DNA (RAPD) markers identified from Actinidia chinensis were converted into sequence-characterised amplified regions (SCARs) for the large-scale screening of Actinidia breeding populations. Initial SCAR primers converted one RAPD (SmX) into a dominant marker, but the other (SmY), which was potentially more useful because of its linkage to the male determining ‘Y’ locus, failed to retain polymorphism. This difficulty was overcome by cloning and sequencing the alternate ‘allele’ from female plants, and then designing ‘allele’-specific primers that utilised nucleotide differences between the sexes. Using a quick squash-blot method of DNA extraction, the SCAR primers were tested in 120 A. chinensis plants to determine their gender. The system is now in use for large-scale screening of seedling populations in the Actinidia breeding programme. The sex-linked SCAR primers also functioned with plants from some other geographically separate accessions of A. chinensis and with plants in the closely related polyploid species A. deliciosa, but did not amplify a sex-linked band in more distantly related species of Actinidia.

Genomic instability in phenotypically normal regenerants of medicinal plant Codonopsis lanceolata Benth. et Hook. f., as revealed by ISSR and RAPD markers

Abstract: Codonopsis lanceolata Benth. et Hook. f., commonly known as bonnet bellflower, is a high-valued herb medicine and vegetable. In this study, a large number of plants were regenerated via organogenesis from immature seed-derived calli in C. lanceolata by a simple and efficient method. Compared with the mother donor plant, the regenerated plants did not exhibit visible phenotypic variations in six major morphological traits examined at the stage of one-season-maturity under field conditions. To gain insight into the genomic stability of these regenerated plants, 63 individuals were randomly tagged among a population of more than 2,000 regenerants, and were compared with the single mother donor plant by two molecular markers, the inter-simple sequence repeats (ISSR) and randomly amplified polymorphic DNA (RAPD). Apparent genomic variation was detected in the 63 regenerants, whereas preexisting heterozygosiy in the donor plant was deemed minimal by testing 30 seedlings germinated from selfed seeds of the same donor plant. The percentages of polymorphic bands (PPB) in the ISSR and RAPD analysis were respectively 15.7 and 24.9% for the 63 regenerated plants. Cluster analysis indicates that the genetic similarity values calculated on the basis of RAPD and ISSR data among the 64 plants (63 regenerated and one donor) were respectively 0.894 and 0.933, which allow classification of the plants into distinct groups. Nineteen randomly isolated bands underlying the changed RAPD or ISSR patterns were sequenced, and three of them showed significant homology to known-function genes. Detailed pairwise sequence comparison at one locus between the donor plant and a regenerant revealed that insertion of two short (24 and 19 bp) stretches of nucleotides in the regenerated plant relative to the donor plant occurred in an apparently stochastic manner.