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Aim: The present study aimed to investigate the phosphate solubilization potential of agriculturally important fungi, i.e., Aspergillus sp. isolated from the rhizosphere of healthy plants in Abha city, Saudi Arabia.Methodology: Sixteen Aspergillus sp. isolated and tested for phosphate solubilization potential were identified by 5.8S-ITS region sequencing and characterized by 11 ISSR-PCR markers. Finally, the highest phosphate solubilization potential isolates were used in field experiments on cucumber and tomato plants. Results: All Aspergillus niger isolates showed 96–100% similarity to A. niger strains available at GenBank database, Isolate ASAB-5 was most efficient at solubilizing phosphate on Pikovskaya’s medium, with a solubilization index of 2.67, and 235.22 mg l-1 of solubilized phosphate. ISSR-PCR markers revealed is total 142 bands in all isolates, with about 32.3% showing monomorphism and 67.6% polymorphism. Based on genetic similarity and intraspecies variability, the Aspergillus isolates were grouped into two different clusters with about 67.9% genetic similarity. The results of field experiments showed no significant difference between seeds treated with culture filtrate or conidial suspension of ASAB-5; however, both differed remarkably from untreated seeds. Interpretation: The current study confirms the existence of several useful phosphate solubilizing fungi in plants, which may serve as potential biological fertilizers. They are safer than chemical fertilizers and increase the bioavailability of soil phosphates for plants
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Spathopsis rubens is freshwater bivalves distributed in the Nile River and its main canals all over Egypt.Four populations of S. rubens on Al-Mahmoudia irrigation canal at Damnhour, Egypt were collected andinvestigated for morphometric characters and genetic diversity by using inter simple sequence repeats (ISSR).The results declared that both the length and height measurements of the collected samples from the differentlocations showed slight significant differences; however, no significant differences between the total weightand the width of the samples have found. High degree of correlation between temperature and total weight,height and length was reported in samples collected from the third location. Genomic DNA from the selectedsamples of each population was extracted and amplified using 10 ISSR primers. The primers (M2, M3, M8,M12, M17, F2, F4, and F9) showed 100% polymorphism. Unweighted pair group method with arithmetic meandendrogram characterized the samples of S. rubens into two main definite clusters. The cluster of genotypes2 and 3 recorded the highest similarity and distance indices at a distance of 0.60859, while genotypes 1 and 3recorded the lowest similarity and distance indices at a distance of 0.1716.
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Aim: The present study was undertaken to analyze the extent of genetic variability existing among twenty accessions of Lawsonia inermis, collected from Rajasthan and Gujarat states of India, using gene targeted SCoT, arbitrarily amplified ISSR and nuclear rDNA markers. Methodology: Twenty henna accessions, vegetatively established at the Institute were collected from Rajasthan (7) and Gujarat (13). Twenty-six SCoT and twenty ISSR markers generating distinct, unambiguous and scorable fragments were selected, after preliminary screening for assessment of genetic diversity. Data analysis was performed using NTSYS-pc, GenAlEx 6 and POPGENE version 1.31 programs, and dendrograms were generated using unweighted pair group method for arithmetic mean (UPGMA). The internal transcribed spacer (ITS) region of ribosomal DNA was amplified using universal primers followed by sequencing and dendrogram generation. Results: SCoT markers revealed lower values of similarity coefficients ranging from 0.87 - 0.93 compared to 0.93 - 0.98 for ISSR. SCoT markers delineated the L. inermis cultivars into three distinct clusters while ISSR markers demarcated them into five clusters. Interpretation: The Gujarat population of L. inermis was richer in genetic diversity than that of Rajasthan. SCoT markers proved better than the ISSR markers for genetic diversity analysis. Substantial variation in ITS-1 region due to SNPs, INDELS and ITS length polymorphism the nucleotide sequences signified its phylogenetic utility in assessing genetic diversity in of L. inermis.
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In the present study, to investigate genetic diversity and phylogenetic relationships among honey bee populations of Iraq ISSR markers were used. Sampling was carried out during summer 2017 from 5 cities of Iraq (Dahuk, Arbil, Sulaymaniyah, Kirkuk, and Kafri). Total DNA was extracted from the head and thorax sections of each worker honey bee, using salting out method with minor modifications. PCR amplification of genomic DNA was performed using 10 ISSR marker primers (A1, A2, A3, A4, A5, A6, A7, A8, A9 and A10). The primers yielded 50 polymorphic bands and number of bands were variable from 8-12 (average 9.62), and percentage of polymorphic loci was 73.6. The estimated genetic diversity for the populations ranged from 0.39 in Kafri population to 0.47 in Arbil population, and total genetic diversity among loci was calculated as 0.47 while average within population genetic diversity was 0.44. GST value was 0.085. The Phylogenetic tree showed two main clusters; the first one comprised of three populations (Dahuk, Arbil, and Sulaymaniyah), and the second one included two communities (Kirkuk and Kafri). Heterozygosity values, Shannon index and the number of alleles of honey bee populations were minimal that could be caused by low definite geographic structure of honey bee populations. This research provided new information regarding genetic diversity in selected local honeybee in Kurdistan region of Iraq and will be useful for selection, future local biodiversity conservation and controlled breeding programs.
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Abstract Calvertius tuberosus (Curculionidae) lives exclusively on Araucaria araucana trees (commonly known as pehuen) in southern Chile. In this study, morphometric and molecular genetic analyses of Andean and coastal populations of C. tuberosus were performed to evaluate evolutionary divergence associated with the discontinuity of the Araucaria forest between the coastal and Andean regions. Specimens of C. tuberosus were collected in Nahuelbuta National Park, Villa Las Araucarias, and Malalcahuello National Reserve and were classified and stored at the Animal Biotechnology Researching Laboratory (LINBA), University of La Frontera, Temuco, Chile. Thirteen morphometric parameters and the expression patterns of ISSR (inter-simple sequence repeat) markers were analyzed. Morphometric data revealed high phenotypic similarity between coastal populations. The genetic analysis revealed a high similarity between coastal populations (genetic identity, 93%), which were differentiated from the Andean population (genetic identity, 84%). This study contributes new genotypic and phenotypic data for the C. tuberosus populations in forest ecosystems of A. araucana, and clarifies the associations between these characteristics and the geographic distributions of populations.
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Abstract Evolutionary analyses have been widely used for evaluation of genetic diversity of natural populations and correlate these data to the fitness of the species, especially in the case of threatened species. Calydorea crocoides occurs in a restricted area at altitudes from 800 to 1500 m in southern Brazil and is considered endangered. A study assessing genetic diversity, cytogenetic features and ecological niche was performed aiming to characterize C. crocoides by multidisciplinary approaches. Molecular data highlighted that most of the total variation (76%; p < 0.001) was found within populations and the parameters of genetic diversity were high at the species level (PPB = 98.97%; I = 0.4319; h = 0.2821). Gene flow (Nm) was estimated in 0.97 individuals per generation. Cytogenetically, C. crocoides presents a bimodal karyotype and low asymmetry. DAPI banding pattern was uniform, but the CMA-signal evidenced a pericentric inversion in the population ESC688. The species presents high pollen viability and two different morphologies of pollen grains. Our data showed high levels of polymorphism maintained in this species that could ensure conservationist practices in which the main goal is to preserve the evolutionary potential of the species through the maintenance of genetic diversity.
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Objective: To analyze the genetic diversities and phylogenetic relationships of Gynostemma pentaphyllun germplasm. Methods: Genetic diversities and phylogenetic relationships of 48 G. pentaphyllun germplasm from 12 provinces in different ecological environments were analyzed by inter simple sequence repeat (ISSR) molecular makers. Results: (1) Fifteen primers were selected from 100 primers which had clear and stable polymorphic bands. A total of 214 loci were obtained, including 206 loci, on average, amplification site of each primer 14.27 and polymorphic loci (PPL) was 96.26%. The averagely observed number of alleles (Na), effective number of alleles (Ne), Shannon information index (I) and Nei's genetic index (H) were 1.9626, 1.3358, 0.2211, and 0.3598, respectively. The enetic similarity coefficient (GS) ranged from 0.57 to 0.96. Cluster annlysis with UPGMA method showed that the 48 testing materials were divided into four groups at the level GS 0.72. Conclusion: The results of ISSR analysis reveal that G. pentaphyllun germplasm has a higher genetic diversity level and the genetic relationship is not entirely consistent with geographical distribution and ecological environment.
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Aims: Investigate the degree of polymorphism using 41 ISSR (Inter-simple sequence repeat) markers in 50 sorghum accessions from 11 different regions in Sudan and Republic of South Sudan. Study Design: UPGMA cluster analysis using STATISTCA- SPSS software Ver. 9. Place and Duration of Study: Department of Molecular Biology, Commission for Biotechnology and Genetic Engineering, National Center for Research, Khartoum, Sudan (2010-2012). Methodology: 50 sorghum accessions with important agronomic traits, representing 11 regions in Sudan and Republic of South Sudan were assayed for polymorphism using Inter-simple sequence repeats (ISSRs). Seven primers out of 41 tested (807, 808, 810, 814, 848, 872 and 879) showed high polymorphism among the Sorghum accessions. Results: The results indicated 75 polymorphic bands out of 78 bands with percentage of polymorphic bands of 97%. UPGMA analysis showed ISSR distance matrix ranged between (0.04-0.47) which reflected high genetic diversity. The ISSR UPGMA dendrogram showed high molecular variance within regions. Based on the results of this study ISSR technique showed differences among closely related accessions of sorghum. Also it proved to be useful technique to study genetic variation among the Sudanese Sorghum accessions. Conclusion: Sorghum accessions from Sudan exhibits high genetic variation within and among regions. ISSR marker technique used in this study proved that it is efficient and could be very useful for breeders and researchers community in various fields of sorghum improvement in Sudan.
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Objective: To study the morphological characteristics and genetic diversity of Atractylodes macrocephala from different varieties, in order to protect and select the germplasm resources and optimize the cultivated technology. Methods: The inter-simple sequence repeat (ISSR), morphological characteristics, and physiology were used to analyze the genetic diversity among 11 varieties of A. macrocephala in Dapan mountain, a national natural reserve in Zhejiang Province and the cluster analysis was performed by UPCMA method. Results: The varieties differed greatly in leaf-shape and leaf-color, stem and rhizomatous configuration, production and disease resistance, and no correlation was observed in these differences. However, in all samples, the photosynthesis rate was closely correlated to the biomass production, despite the great differences existing among different varieties. Simultaneously, fifteen screened ISSR primers were served as ISSR-PCR markers, and 129 sites were amplified, which included 107 polymorphic sites (82.95% of the total bands). At molecular level, the genetic differentiations occured among 11 varieties, some differentiations were quite distinctive according to cluster analysis. Conclusion: The germplasm resources of A. macrocephala. are abundant and the genetic diversity basis exists in all kinds of A. macrocephala in Dapan mountain.
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For a study of diversity and genetic structuring in Melipona quadrifasciata, 61 colonies were collected in eight locations in the state of Minas Gerais, Brazil. By means of PCR analysis, 119 ISSR bands were obtained, 80 (68 percent) being polymorphic. He and H B were 0.20 and 0.16, respectively. Two large groups were obtained by the UPGMA method, one formed by individuals from Januária, Urucuia, Rio Vermelho and Caeté and the other by individuals from São João Del Rei, Barbacena, Ressaquinha and Cristiano Otoni. The Φst and θB values were 0.65 and 0.58, respectively, thereby indicating high population structuring. UPGMA grouping did not reveal genetic structuring of M. quadrifasciata in function of the tergite stripe pattern. The significant correlation between dissimilarity values and geographic distances (r = 0.3998; p < 0.05) implies possible geographic isolation. The genetic differentiation in population grouping was probably the result of an interruption in gene flow, brought about by geographic barriers between mutually close geographical locations. Our results also demonstrate the potential of ISSR markers in the study of Melipona quadrifasciata population structuring, possibly applicable to the studies of other bee species.