This communication reports detection of somaclonal variation among tissue culture-raised plants of Amorphophallus rivieri Durieu, an economically important crop in China, with high content of glucomannan in its corms. A population of regenerated plants was obtained from a single donor plant of A. rivieri via corm organogenesis, and 28 plants were randomly selected as a representative sample and subjected to analysis of somaclonal variation using inter-simple sequence repeat (ISSR) markers. Of the 26 ISSR primers screened, 13 gave distinct and reproducible band patterns, yielding 131 bands with an average of 10.1 bands per primer. Ten primers were polymorphic and generated 16 polymorphic bands with 12.2% mean polymorphism. Based on the ISSR data from the regenerated plants and the donor plant, Jaccard's similarity coefficients were calculated; they ranged from 0.961 to 1.000 with a mean of 0.982. A dendrogram was constructed using the unweighted pair group method with arithmetic mean (Upgma); it showed that a majority of regenerated plants (including the donor plant) clustered closely, with a mean similarity coefficient of 0.987. Low somaclonal variation observed in the regenerated plants indicates that rapid propagation of A. rivieri via corm organogenesis is a practicable method with a low risk of genetic instability.
This study used ISSR markers to assess the genetic diversity of a collection of 15 genotypes of Salix purpurea and 6 interspecific hybrids, employing 40 of 60 tested ISSR primers generating polymorphic amplification products. The PCR-ISSR method was adapted for S. purpurea by optimizing the annealing temperature for each primer. The polymorphism index of ISSR amplification products was 91.8% for all studied genotypes and 70.4% for S. purpurea genotypes. Nei's genetic identity statistics ranged from 0.538 to 0.958. Nei's genetic distance values were used to build a dendrogram (UPGMA) for the investigated genotypes. The dendrogram shows five clusters, and principal coordinate analysis yielded nearly the same genetic relationships among the studied genotypes. The results confirm the usefulness of ISSR markers for determining genetic diversity in S. purpurea.
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.