Bacterial speck of tomato caused by Pseudomonas syringae pv. tomato appeared to be recently the most important disease on tomato in Poland. The genetic relationships among four Polish strains of race 0 P. syringae pv. tomato of different origin, isolated from tomato plants, were examined by RAPD and PCR-RFLP techniques. Amplification of bacterial DNA using 33 primers with RAPD technique showed, that similarity of strains expressed by the Nei-Li coefficient was very high (above 0.8). Next, the restriction analysis of amplified region ITS with the use of 5 endonucleases revealed, that profiles obtained from electrophoretic separation of DNA fragments were also very similar. On the basis of those analyses it was concluded that all strains tested constituted a closely related group. However, they showed various level of virulence as was demonstrated on the inoculated leaves of tomato plants growing in the greenhouse.
Stem canker and black scurf of potato (Solanum tuberosum L.) caused by Rhizoctonia solani Kühn are important and epidemic diseases in potato-growing regions worldwide, including Iran. In this study, 120 isolates were retrieved from infected stem canker from six potato- growing regions in Iran (Isfahan, Ardebil, Fars, Hamedan, Kurdestan and Kerman). Out of these, 30 isolates were selected as representatives for genetic and virulence analysis. The isolates were analyzed by one sequence analyzes of the ITS-rDNA region, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies. Based on sequence analysis of the ITS-rDNA region, all 30 isolates were assigned to the anastomosis group (AG) and all were assigned to AG-3 PT. Cluster analysis using the unweighted pair group method with the arithmetic averages (UPGMA) method for both RAPD and ISSR markers revealed that they were divided into three main groups, with no correlation to geographical regions of the isolates. Pathogenicity tests showed that all isolates were pathogenic on potato cv. Agria; however, virulence variability was observed among the isolates. The grouping based on RAPD analysis and virulence variability was not correlated.
Gaeumannomyces graminis is an etiologic agent of take-all, economically important disease of cereals worldwide. A polymerase chain reaction with variety-specific primers was successfully used for detection of G. graminis var. tritici in plant tissue. Obtained results showed that this diagnostic method is a very sensitive and useful tool for detection of the pathogen even before disease symptoms arise. DNA polymorphism revealed by RAPD-PCR with three arbitrary primers was suitable for assessing genetic variation among Ggt isolates originating from wheat and rye.
Potato leaf blight disease caused by Ulocladium atrum (Syn. Stemphylium atrum) is an important and epidemic disease in potato-growing regions of Iran. In this study, 30 isolates of the disease were collected from the main potato-growing regions of Iran and were analyzed on the basis of morphological characterization and pathogenicity. Based on morphological characteristics, all isolates were identified as U. atrum. Pathogenicity studies indicated that all 30 isolates were pathogenic on potato “Agria” to varying degrees. Five U. atrum isolates causing potato leaf blight disease, obtained from the Plant Pathology Laboratory, Isfahan Research Center for Agriculture and Natural Resources, Isfahan, Iran, were also examined in this study. A total of 35 isolates were genetically analyzed using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers. Cluster analysis using the un-weighted pair group method with the arithmetic average (UPGMA) method for RAPD marker revealed no clear grouping of the isolates obtained from different geographical regions. The groupings, based on morphological characteristics, virulence variability and RAPD analysis, were not correlated. Cluster analysis using Jaccard’s coefficient for ISSR divided the U. atrum isolates into four main groups, in which there was no significant correlation between the isolate groupings regarding their geographic location and pathogenicity. Using molecular techniques genetic variability was detected among the accessions, with cophenetic correlation coefficients (CCC) of 0.80 for RAPDs and 0.89 for ISSRs. The RAPD and ISSR marker results corresponded well, with a correlation of 0.55.
Three problems in the taxonomy of Pancratium in Egypt are the lack of publications, a lack of clarity about the relationships between recently distinguished species, and the lack of markers for examining the levels and patterns of variation in rare and endemic species; the latter hinders work in plant conservation genetics. In this study we reassessed the taxonomic status of the Pancratium species of Egypt, and examined morphological and genetic variation within and between species, using specimens from different populations collected throughout its distribution range in the country. Our assessment was based on 38 macromorphological characters mainly representing vegetative parts, flowers, fruits and seeds, in addition to RAPD data. The results revealed five morphologically distinguished Pancratium species in Egypt, of which P. trianthum Herb. is newly recorded. Species identification was confirmed by two phenetic dendrograms generated with 26 quantitative morphological characters and RAPD data, while species delimitation was verified by principal component analysis. The diagnostic floral characters are those of the perianth, corona teeth, pistil, stamens, aerial scape, spathe, and number of flowers. The retrieved RAPD polymorphic bands show better resolution of the morphologically and ecologically closely allied Pancratium species (P. arabicum and P. maritimum), and also confirm the morphological and ecological divergence of P. tortuosum from the other studied species. These results are supported by the constructed UPGMA dendrogram.
Root associated bacteria were isolated from Suaeda nudiflora and two isolates were selected for this study: rhizospheric Bacillus megaterium and endophytic Pseudomonas aeruginosa. These isolates were inoculated into maize variety Narmada Moti during its germination. TTC (2, 3, 5-triphenyl tetrazolium chloride) staining was used to confirm the association of the isolates with the maize root. The effects of these root associated bacteria were tested alone and in combinations for cell wall reinforcement and the induction of defense enzymes such as phenylalanine ammonia lyase (PAL) and β-1,3-glucanase in the presence of fungal pathogen Aspergillus niger in maize. The results indicated that the rhizospheric bacteria had a greater fight response to fungal infection than the endophhytic bacteria due to cell wall lignification as well as the rapid induction of higher concentrations of defense related enzymes.
Genotypic differentiation among 10 isolates of Phytophthora cinnamomi Rands and 24 isolates of Phytophthora citricola Sawada from 12 different plant species grown in Polish ornamental nurseries was determined. DNA was extracted from pure pathogen cultures and amplified by the PCR technique using ISSR and RAPD primers. 9 primers were used to amplify P. cinnamomi and 8 to amplify P. citricola DNA. The analyzed amplification products were between 300 and 2300 bp. The genotypical differentiation was from 17 to 35% in P. cinnamomi and from 10 to 60% in P. citricola. Isolates from host plants of the same family showed, with some exceptions, similar levels of differentiation.
Hladnikia pastinacifolia RCHB., a narrow endemic, has an extremely restricted distribution in Trnovski gozd (Slovenia), despite the presence of many sites with suitable habitats. We compared the morphological traits of plants from different populations and habitats. The overall pattern showed that the smallest plants, with low fruit number, are found on Èaven (locus classicus or type locality); the largest individuals, with high fruit number, grow in the Golobnica gorge. As judged by plant size and seed set, the optimal habitats are screes. We used RAPD markers to estimate genetic variation between and within populations, as well as between and within the northern and the southern parts of the distribution area. Hladnikia showed only a low level of RAPD variability. AMOVA partitioned the majority of genetic diversity within selected populations. The low genetic differentiation between populations and their genetic depauperation indicates survival in situ, since the Trnovski gozd plateau most likely was a nunatak region in the southern Prealps during Pleistocene glaciations. Later range expansion of extant populations was limited by poor seed dispersal. We also analyzed the cpDNA trnL-F intergenic spacer to check whether the sequence is useful for studying the phylogenetic relationships of Hladnikia within the family Apiaceae (Umbelliferae). Our results support the assertion that H. pastinacifolia is an old taxon.