JWSS - Journal of Water and Soil Science

---

The poor information available on variation of Ascochyta blight fungus is the most important factor limiting chickpea breeding programs for resistance to blight disease. In this study, efforts were made to detect genetic variation of the pathogen in Iran. The RAPD marker was employed to evaluate 26 isolates collected from 16 provinces. Twelve random primers were used to analyze genomic DNA of the isolates. Only ten primers showed polymorphism among isolates. Primer OPK-01 defined the highest number of polymorphism and primer OPK-09 confirmed relatively low degree of polymorphism. On the basis of this molecular marker, the estimated genetic diversity index was 98% and the pair-wise genetic distance of the isolates varied from 0.16 to 0.61. The least genetic distance belonged to isolates 20 and 22 from Qazvin and Golestan while the highest distance belonged to isolates 26 and 12 from Mazandaran and Markazi. The phylogenic tree was constructed by cluster analysis and all the isolates grouped to 22 genetic clusters at the 90% similarity level. The genetic groups were named from A to V and their distribution in Iran was determined. The results revealed that genetic variation among Iranian population of the pathogen is very high, and further that RAPD is a vigorous tool for genomic analysis of Ascochyta rabiei.

---

Genetic male sterility is controlled by one pair of ressesive allele (aa) in sugar beet. This trait is used in most breeding programes. The exsistance of the character in a line or population facilitates transfer of important trait to the breeding material (for example resistance to plant disease). Also, it is possible to increase genetic diversity of monogerm populations by using genetic male sterility. The time and cost of transferring of this gene will be decreased, if the character is tagged with a molecular marker. Bulked segregant analysis using 302 RAPD primers in two F2 populations (231 and 261 population) was performed for the the identification of RAPD markers linked to the genetic male sterility gene. DNA preparation from 8 male fertile and male sterile plants were separately mixed. At first, the primers were tested on bulks. The primers with polymorphic bands were tested on individual plants of the bulks. Only if the polymorphism of the primers was confirmed, they were tested on the other individual plants. Finally, 10 and 6 markers were identified in 231 and 261 populations, respectively, which their distances to male sterility gene were lower than 50 cM. AB-8-18-600r marker was the nearest marker to male sterility gene. This marker showed only 3 and 1 recombination in 231 and 261 populations, respectively. The distance of this marker and genetic male sterility locus was estimated as 5.3 cM in combined F₂ populations.

---

In order to detect genetic variation of native fowls in Mazandaran native Fowls breeding station, blood samples were collected from 100 male and female of birds (1:11). The DNA of the blood samples was extracted according to an optimized salting out protocol. The extracted DNA was amplified through polymerase chain reaction (PCR). Of the twenty random primers (10 mer) were used in this study, fourteen yielded satisfactory PCR. The total 63 polymorphic and 77 monomorphic bands were detected for the 14 primers. The number of bands displayed for each primer ranged from 4 to 16 with 200-2100 base pairs. The highest and lowest percentages of polymorphism band were observed for primer 9 (72%) and primer 14 (16%) respectively. The band sharing frequency was calculated for each primer, which ranged from 79 to 96. The genetic similarity within population and genetic variation were estimated as 89 and 11 percentage respectively. In conclusion, the existence of high level of polymorphism after ten generation of selection may indicate the accuracy of genetic evaluation program, suitable selection strategies and also large enough effective population size in this breeding flock.

---

Germplasm collection is the base of plant breeding. Iran is one of the most important centers of genetic diversity due to different climates and the old civilization.In this study we decided to collect melon accessions. The north and center of Iran were selected for this purpose. Fifteen qualitative and six quantitative traits were measured on thirty eight accessions. The cluster analysis by the use of UPGMA method and Jaccard coefficient helped separate the horticultural groups of Cucumis melo L. (Cantaloupensis, Inodorus, Flexousous, Reticulatus). The relationship between 30 of these accessions was assessed using 10 RAPD primers. The polymorphism was determined to be19%. The cluster analysis could not separate the horticultural groups of Cucumis melo L., showing that these groups are closely related. However, VB84 primer separated the tow Snakemelon.

---

Grapevine (Vitis vinifera L.) is a clonally propagated major fruit crop. In grapevine, identification of genotypes with amplographical features is often based on mature plant characteristics that may be affected by environmental conditions. This approach lacks objectivity and reliability. Recently, molecular markers have proved to be supplementary techniques to analyze genetic diversity and examine genetic relationships existing between cultivars in a range of horticultural crops. In this study, twenty genotypes from grapevine (V.vinifera species) grown in Isfahan province were characterized by RAPD technique to understand the extent of diversity and relatedness. Fifty random primers were used for the RAPD study. Of those, twenty four informative primers which generated reproducible polymorphic bands were used for grouping the genotypes. PCR products of the genotypes’genome revealed a total of 315 bands, out of which 282 were found to be polymorphic. Average number of 13 bands was obtained per primer and the amplification produced ranged in size from 300 bp to 3000 bp. The dendrogram constructed using UPGMA cluster analysis differentiated the genotypes into two major clusters, nineteen in one group and Madar-o-Bache genotype has been placed in a separate one, indicating its high genetic diversity compared to the rest of the genotypes. Intra-clustering within cluster A grouped the genotypes in four sub-clusters as expected from their genetic background. The results of the study revealed that the RAPD technique is a relevant technique to determine genetic diversity, genomic analysis and to examine genetic relationship in grapevines.

---

In this study, 45 Pleurotus isolates with a wide range of geographical origins (in five provinces of Iran: Isfahan, Chaharmahal Bakhtiary, Kohkiloye boyrahmad, Lorestan and Fars) were collected. A random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to assess the genetic diversity in ten populations of the Pleurotus eryngii complex. The use of ten primers allowed the production of 150 polymorphic bands. In addition, the evalution of the principal ecological and morphological characters provided further evidence for discriminating between populations. The results show a higher level of diversity of RAPD polymorphism in the populations. 28.67% of the RAPD diversity was among populations and 71.33% was within populations. In the RAPD patterns, Pleurotus strains growing on Ferula assa-foetida, Ferula ovina and Prangus ferulacea formed distinct groups. In this study, the resolution of the RAPD-PCR analysis was better than the morphological analysis.