Polymorphic Bands Research Articles

Calcium-Binding Protein and Polymorphism in Musa spp. Somaclones Resistant to Fusarium oxysporum

The fresh fruits of ‘Grande Naine’ (Cavendish AAA—Musa spp.) dominate the world market, especially in countries with a population in a situation of social vulnerability. However, Fusarium wilt, caused by the fungus Fusarium oxysporum f.sp. cubense race 4 Subtropical (Foc ST4), emerges as a serious threat to banana production, requiring the development of resistant cultivars based on biotechnological strategies, such as the induction of mutation in tissue culture. This study aimed to identify and characterize genetic variation in somaclones resistant to Fusarium oxysporum f.sp. cubense subtropical race 4 (Foc ST4), derived from ‘Grand Naine’ bananas, by molecular markers based on retrotransposons IRAP (Inter-retrotransposon Amplified Polymorphism) and REMAP (Retrotransposon-Microsatellite Amplified Polymorphism). Nine combinations of IRAP and six combinations of REMAP primers were used. The low number of polymorphic bands did not allow for genetic diversity studies; however, ten polymorphic bands between the somaclones and control were sequenced. Of these, three presented good base calling and were aligned, namely, 1AF, 2AF, and 3AF bands. Only the 1AF band presented function related to stress response with homology to a calcium-binding protein. These proteins act early in plant infection as secondary messengers activated by pathogen-associated molecular patterns (PAMPs), initiating the cascade of plant defense signals. The fact that this band is present in all somaclones reinforces previous assessments of their resistance to Foc ST4. The use of markers IRAP and REMAP produced polymorphic bands that can, through future primer design and field validations, accelerate the identification of resistant banana genotypes for use in banana genetic breeding programs.

Identification of morphological and variety diagnostic molecular markers for commercial varieties of nutmeg (Myristica fragrans Houtt.)

Abstract Nutmeg (Myristica fragrans Houtt; family: Myristicaceae) is an important tree spice of great export value due to the presence of secondary metabolites with scope in health, wellness and cosmetic industries. Its dioecious in nature due to which it is propagated by budding and grafting which requires specialised skill and as a result authentic planting material production is very expensive. Based on the constant demand from farmers and stakeholders for a suitable, fast and easy technique for determining the authenticity of nutmeg varieties, an attempt was made to develop morphological and molecular markers for the identification of three commercial varieties that are in high demand in India; viz. IISR Vishwashree, IISR Keralashree, Sindhushree and a monoecious nutmeg (MN) accession, along with their biochemical profiles. Among the various morphological characters as per distinctiveness, uniformity and stability guidelines, the varieties showed distinctness mainly in leaf shape, shape of female flowers, fruit shape and seed shape. Out of 35 ISSR primers screened, six primers viz., IS 02, ISSR 12, ISSR 05, ISSR 14, ISSR 01 and UBC 834 generated clear, unique reproducible polymorphic bands capable of distinguishing the varieties. Among the varieties analysed, MN was found to be superior in terms of seed butter and essential oil in nut, mace and leaves. IISR Vishwashree was on par with MN in nut essential oil content. IISR Keralashree was superior in terms of oleoresin in nuts and mace. The morphological and molecular markers identified may be used for effective checking of authenticity of planting materials of commercially grown nutmeg varieties.

Assessment of Genomic Integrity of Vitex negundoL., An Important Indian Medicinal Plant, Using RAPD Markers

Vitex negundo is an Indian medicinal plant containing steroids, flavonoids, lignans and terpenoids that can be used as a precursor for commercial production. An efficient marker system such as Random Amplified Polymorphic DNA (RAPD) was used to assess the genetic integrity of V. negundo. The straightforward method RAPD can be used to evaluate genomic integrity because it uses a small amount of DNA for PCR amplification. Six out of thirteen RAPD primers generated 150 distinct bands, of which 31 were polymorphic, with an average of 5.16 polymorphic bands per primer. A maximum of up to 32 fragments were amplified, and an average of 25 per primer, and the amplicons varied in size between 100 and 2000bp. The percentage of polymorphism ranges from 12.9 to 22.5, with an average of 16.6. The PIC values ranged from 0.11 to 0.63 for RAPD primers. The study pointed out that RAPD markers evaluate the genetic fidelity in Vitex negundo. The UPGMA cluster analysis grouped all in vitro raised plantlets treated with different growth regulators such as BAP, DPU, TDZ, and mT. The principal component analysis also substantiates this clustering pattern. Thus, the phylogenetic relationship and a high genetic variation revealed in the present study could provide baseline data for the conservation and improvement of this plant in future. Also, the molecular marker identified in this study will be helpful in the authentication of this species to prevent adulteration in herbal medicine.

Evaluation of germplasm resources of Sarcomyxa edulis in the Changbai Mountains of Northeast China

ABSTRACT In this study, we evaluated 16 strains of Sarcomyxa edulis from the Changbai Mountains, China through antagonism tests and ISSR molecular marker technique for cytological and molecular biological evaluation. Q-type and R-type cluster analyses were utilised to evaluate and classify agronomic traits according to the test guidelines for distinctness, uniformity, and stability. The tested strains showed antagonistic reactions. The differences in antagonistic reactions among different germplasm resources are helpful for evaluating genetic diversity and provide a basis for better development and utilisation of fungal germplasm resources. A total of 330 bands were amplified by 21 ISSR primers, including 321 polymorphic bands. The percentage of polymorphic bands observed was 97.07%. Using PopGen32 software, the effective number of alleles (Ne), Nei's gene diversity index (H), and Shannon's diversity index (I) were calculated, yielding average values of 1.6638, 0.3796, and 0.5480, respectively. In addition, the genetic similarity coefficient of 16 strains ranged from 0.4306–0.7778. According to the Q-type clustering, the tested strains were classified into two categories based on the Euclidean distance of 8.52. While R-type cluster analysis revealed a significant correlation between the traits. At the Euclidean distance of 6.298, the traits could be classified into three distinct categories. According to the findings, all 16 strains demonstrated high genetic diversity. The ISSR clustering results revealed that strains with shorter genetic distances were more similar in specific traits. However, there were notable differences between the results of comprehensive trait clustering (Q-type clustering) and ISSR clustering.

Assessment of Genetic Diversity in Alfalfa Using DNA Polymorphism Analysis and Statistical Tools

The cultivation of alfalfa is crucial for farmers as it is an excellent forage crop with a high nitrogen-fixing capacity, making it indispensable in crop rotations. Breeding programs face challenges in advancing more rapidly in genetic diversity to achieve a higher heterosis effect and, consequently, greater yield. In this study, we used 30 alfalfa varieties, which were used for molecular analyses by 5 ISSR primers and 13 RAPD primers. The results obtained highlighted the greater efficiency of ISSR primers in identifying genetic diversity. On the other hand, the simultaneous use of ISSR + RAPD allowed for clearer clustering of varieties that enabled more efficiently distinguishing the genetic diversity. The most efficient ISSR primer, A17, generated 31 polymorphic bands, while the most efficient RAPD primer, L-07, generated only 21 bands. Varieties such as “Pastoral” and “F1413-02” exhibited low similarity coefficients (0.39), suggesting their potential for enhancing genetic variability through crossbreeding, thereby increasing the potential of achieving a greater heterosis effect. Conversely, varieties with high similarity coefficients, such as ”Cristal” and “Viking” (0.81) are less suited for this purpose. The correlation between specific markers highlights that using both ISSR and RAPD markers together offers a clear understanding of genetic diversity in alfalfa, aiding in more effective selection for crossbreeding in breeding programs.

Evaluation of oxidative stress, biochemical parameters and in silico markers in different pea accessions in response to drought stress.

ARG6 and ARG10 pea accessions exhibited better tolerance to drought by keeping drought-associated attributes stable and higher, that is, stable chlorophyll content, high antioxidant activity, and the presence of polymorphic bands with stress-responsive EST-SSR markers. Each year, a significant portion of crops is lost due to various abiotic stresses, and even pea (Pisum sativum) crop growth and yield are severely affected by the challenges posed by drought stress. Drought is a critical factor that limits crop growth and development, and its impact is exacerbated by changes in the magnitude of climatic conditions. Drought induces oxidative stress in plants, leading to the accumulation of high concentrations of reactive oxygen species that damage cell structures and vital functioning of cells. The primary objective was to identify stress-tolerant plants by evaluating different morphological and biochemical attributes, such as biomass, chlorophyll content, relative water content, ascorbate peroxidase (APX), superoxide dismutase (SOD), and DPPH scavenging activity, as well as protein, proline, and phenolic content. Our study revealed that pea accessions (ARG6 and ARG10) were more resilient to drought stress as their chlorophyll, relative water, protein, and proline contents increased under drought conditions. Antioxidant enzymes, such as SOD, APX, and DPPH activities, also increased under drought stress in ARG10 and ARG6, suggesting that these accessions could bolster the antioxidant defense system in response to drought stress. Based on putative (cellular, biological, and metabolic) functions, ten EST-SSR primers were selected for the amplification study. Three EST-SSR primers, AUMP06_110, AUMP18_300, and AUMP31_250, were used for ARG6 and ARG10. Based on the correlation between the presence or absence of specific EST-SSR alleles, various physiological and morphological traits, and DPPH scavenging activity, both ARG10 and ARG6 demonstrated resistance to drought stress.

Genetic Diversity of Local Chili (Capsicum annum L.) from West Sumatra Indonesia Based on Inter Simple Sequences Repeat (ISSR) Markers

Capsicum annum L. is the most widely cultivated species due to its economic importance and health benefits. West Sumatra is one of the largest chili pepper producers in Indonesia, so it has a lot of local chili genetic resources. There is inadequate information on the genetic diversity of local chilies in West Sumatra. The purpose of this research was to analyze the genetic diversity of 23 genotypes of chili pepper from West Sumatra amplified with 10 markers of ISSR. The material used was leaves from 23 genotypes of West Sumatra’s local chili which are collections of the Agriculture Faculty, Andalas University, Padang, West Sumatra. The results showed that ISSR markers (ISSR primer 12, ISSR UBC, and ISSR 3M) had produced 84 polymorphic DNA bands with a total polymorphic percentage of 61.45%. The dendrogram based on 10 ISSR markers showed that the 23 chili genotypes were retrieved into three main groups (I, II, III, IV) with genetic similarity coefficient values ranging from 0.67 to 0.97 or a diversity of features of 0.03–0.33 (30%). Group I consisted of 1 genotype, group II consisted of 3 genotypes, and group III consisted of 14 genotypes. Kopay chili (G20) has a large genetic distance compared to the other genotypes, while Lolai and Gero chilies have a very high similarity. This genetic diversity information can be used as a basis for generating crosses between genotypes in a breeding program. The farther the genetic distance will produce a high population diversity, the greater the chance of getting superior hybrids. Keywords: Capsicum annum, ISSR, local chili, West Sumatra

Physiological alterations and genotoxic damage under combined aluminum and cadmium treatments in Bryophyllum daigremontianum clones.

Cadmium (Cd) is one of the most important stress factors in plants, with its high mobility in soils, ease of uptake by plants and toxicity at low concentrations. Aluminum (Al) is another phytotoxic metal, the accumulation of which is a crucial agricultural complication for plants, especially in acidic soils. In this study, Bryophyllum daigremontianum clone plantlets were obtained from bulbiferous spurs of a mother plant and separated into four different groups and watered with Hoagland solution and mixtures containing 0, 50, 100, and 200 µM of AlCl3 and CdCl2 each for 75 days. Control groups were maintained under the same conditions without Al and Cd treatment. To simulate acidic soil conditions typical of environments where Al toxicity is prevalent, the soil pH was adjusted to 4.5 by spraying the sulphuric acid (0.2%) with 2-day intervals after each irrigation day. After harvesting, growth parameters such as shoot length and thickness, root, shoot and leaf fresh and dry weights were measured, along with physiological parameters like mineral nutrient status, total protein, and photosynthetic pigment concentrations (chlorophyll a, b, a/b, total chlorophyll, and carotenoid) in both control and experimental groups of B. daigremontianum clones. In response to Al and Cd applications, the plant height, shoot thickness and carotenoid levels were declined, whereas the increments were found in leaf/shoot/root fresh weight, root dry weight, and total protein content. Moreover, differences in genomic alterations were investigated using 21 ISSR and 19 RAPD markers, which both have been used extensively as genetic markers to specify phylogenetic relationships among different cultivars as well as stress-dependent genetic alterations. RAPD primers were used due to their arbitrary sequences and the unknown genome sequence of the plant material used. In contrast, ISSR primers were preferred for a genome-wide genotoxic effect scan via non-arbitrary and more common genetic markers. Distinct types of band polymorphisms detected via RAPD and ISSR markers include band loss, and new band formation under a combination of Al and Cd stress. 17 ISSR and 14 RAPD primers generated clear electrophoretic bands. The study revealed that combined application of Al and Cd affect B. daigremontianum clones in terms of growth, physiology and genotoxicity related to the increasing concentrations.

Development of microsatellite markers and evaluation of the genetic diversity of the edible sea anemone Paracondylactissinensis (Cnidaria, Anthozoa) in China.

Paracondylactissinensis Carlgren, 1934 is a sea anemone with economic value in China. The wild population of P.sinensis has been shrinking due to overfishing and environmental pollution, which have caused price instability. In winter, the price of P.sinensis can reach 25 USD per kilogram. Up to now, there are no genetic markers developed for P.sinensis, preventing a further exploration of their population genetic diversity. In this study, the full-length transcriptome of P.sinensis was sequenced and microsatellite DNA markers (simple sequence repeats [SSRs]) were developed from those transcripts. A total of 52 primer pairs, which can amplify specific polymorphic bands in PCR experiments, were designed for the SSR markers. Genetic diversity and population genetics were analysed for P.sinensis populations collected from the coasts of Taizhou and Rizhao using six microsatellite DNA loci. While inbreeding was detected in both populations (Fis > 0), the overall number of alleles (Na = 11.3) and bottleneck analysis suggested that the genetic diversity of P.sinensis has not been greatly impacted. Clustering analyses using STRUCTURE, principal coordinate analysis and unweighted pair group method with arithmetic mean tree revealed that the Taizhou population diverged from the Rizhao population; however, the genetic differentiation between the populations was moderate. Human-mediated commercial activities may be the principal reasons for the gene flow between the populations. Our study provides the first evaluation of the genetic resources of wild P.sinensis populations in China, which can serve as a useful reference for future comparative studies on population genetics and may guide policy-makers in initiating strategies for germplasm conservation and artificial breeding.

Biosystematic studies of genus Withania Pauquy in Egypt

Withania (Solanaceae, Solanoideae) is a widespread genus. Comparative macro-, micro-morphological, anatomical, and molecular features of this genus in Egypt were examined using light and scanning electron microscopy to reassess the conflicted taxonomic relationships between the two studied species. The most significant morphological differences that have been found were: the shape of the lamina, apex, anther, and stigma, and the ratio of calyx tube/lobe; anatomical examination of taxonomic interest are as follows: number of vascular bundles, presence of ears and distribution of accessory vascular bundles in petiole and shape of spongy cells, and number of lower parenchyma in the midrib region of the leaf; trichomes of both species showed no significant differences; pollen, and seed characters are of taxonomic significance in differentiation and characterization between them. Protein profiling revealed that W. somnifera has only conserved proteins, while W. obtusifolia possessed both conserved and additional proteins in their SDS-PAGE banding patterns. Eleven starts codon-targeted (ScoT) primers were applied and produced 96 amplicons with an average of 70.83% polymorphism/primer. W. obtusifolia generated more polymorphic bands and maintained monomorphic ones. SDS-PAGE disclosed that both Withania species were 50% related. While Scot-Dendrogram revealed that both Withania species were poorly related. So, protein and molecular analyses showed considerable genetic variations between these two species.

Genetic Diversity of Common Bean (Phaseolus vulgaris L.) Landraces Based on Morphological Traits and Molecular Markers.

The objective of this study was to evaluate the genetic diversity among traditional common bean accessions through morphological descriptors and molecular markers. Sixty-seven common bean accessions from the Germplasm bank of the Instituto Federal of Espírito Santo-Campus de Alegre were evaluated. For this, 25 specific morphological descriptors were used, namely 12 quantitative and 13 qualitative ones. A diversity analysis based on morphological descriptors was carried out using the Gower algorithm. For molecular characterization, 23 ISSR primers were used to estimate dissimilarity using the Jaccard Index. Based on the dendrograms obtained by the UPGMA method, for morphological and molecular characterization, high genetic variability was observed between the common bean genotypes studied, evidenced by cophenetic correlation values in the order of 0.99, indicating an accurate representation of the dissimilarity matrix by the UPGMA clustering. In the morphological characterization, high phenotypic diversity was observed between the accessions, with grains of different shapes, colors, and sizes, and the accessions were grouped into nine distinct groups. Molecular characterization was efficient in separating the genotypes in the Andean and Mesoamerican groups, with the 23 ISSR primers studied generating an average of 6.35 polymorphic bands. The work identified divergent accessions that can serve different market niches, which can be indicated as parents to form breeding programs in order to obtain progenies with high genetic variability.

Assessment of Population Genetic Diversity of Medicinal Meconopsis integrifolia (Maxim.) Franch. Using Newly Developed SSR Markers.

Meconopsis integrifolia is an endangered Tibetan medicinal plant with significant medicinal and ornamental value. Understanding its genetic diversity and structure is crucial for its sustainable utilization and effective conservation. Here, we develop a set of SSR markers based on transcriptome data to analyze the genetic diversity and structure of 185 individuals from 16 populations of M. integrifolia. The results indicate that M. integrifolia exhibits relatively high genetic diversity at the species level (the percentage of polymorphic bands PPB = 91.67%, Nei's genetic diversity index He = 0.2989, Shannon's information index I = 0.4514) but limited genetic variation within populations (PPB = 12.08%, He = 0.0399, I = 0.0610). The genetic differentiation among populations is relatively high (the coefficient of gene differentiation GST = 0.6902), and AMOVA analysis indicates that 63.39% of the total variation occurs among populations. This suggests that maintaining a limited number of populations is insufficient to preserve the overall diversity of M. integrifolia. Different populations are categorized into four representative subclusters, but they do not cluster strictly according to geographical distribution. Limited gene flow (Nm = 0.2244) is likely the main reason for the high differentiation among these populations. Limited seed and pollen dispersal abilities, along with habitat fragmentation, may explain the restricted gene flow among populations, highlighting the necessity of conserving as many populations in the wild as possible.

Analysis of Fluorescently Labeled ISSR Markers to Reveal the Genetic Diversity of Cassava Varieties (Manihot esculenta Crantz) in the Kerala’s Tribal Area of India

The study Inter Simple Sequence Repeat (ISSR) molecular markers to genotype distinct cassava (Manihot esculenta Crantz) cultivars within tribal communities in Kerala, India. Fluorescent ISSR primers were labelled with 6-carboxyfluorescein and used for PCR amplification. The total number of bands produced was 72, of which 49 were found to be polymorphic markers; an average of 17 polymorphic bands was obtained per primer. The manuscript is a scientific validation of the traditional knowledge of tribal people regarding the diversity and wealth of cassava crops in the region, and this validation can therefore help in protecting the farmers right on a particular landrace that they cultivate. The technique used in the diversity analysis is a time-saving process as compared to conventional breeding programs for crop improvement. The data in the manuscript also supports similarity and differences of each variety to one another, which is analysed statistically with the molecular marker-based data generated. By examining genetic diversity at a high resolution, ISSR markers can provide valuable insights into the evolutionary dynamics of these cassava populations. This research contributes to the broader goal of enhancing agricultural sustainability and food security in tribal communities.

Molecular evaluation of several wheat varieties of Triticum aestivum L.

This study was carried out with the aim of determining the extent of genetic variation among 10 varieties of wheat. In this work, 5 primers were used to do the PCR to detect the band patterns of ISSR markers. Within the molecular weight range of 100-1500 bp, the 5 primers combined to generate 42 bands, at a rate of 8 bands per primer. Polymorphic bands accounted for 37 of the totals and the ratio of polymorphic bands was 87.97 %. The highest percentage of divergent bands reached 88.88 %, shown by primers HPS13, 842F and HBG20. The starters 1955A recorded a lower rate of efficiency and distinctive ability of the starter reaching 16.22 %. According to the UPGMA method, a phylogenetic tree of the genetic affinity between the varieties was obtained, varieties were separated into 2 major groups: There were 2 varieties in the first major group, namely S7 and S6, 2 minor groups were formed from the division of the second major group. The S3 variant made up the first secondary group, as opposed to the second secondary group. Additionally, it was split into 2 sub-secondary groups: classes S5, S10, S9, S8 and S1 made up the first sub-secondary group and classes S4 and S2 made up the second. Based on the Nei and Li coefficient, the values of genetic similarity between the varieties were estimated, because the S6 variety and the S3 variety had the largest genetic divergence (lowest genetic similarity), with a value of 0.7073 and with the S4 variety, its value was 0.6829, while the highest genetic similarity was (least genetic divergence) between variety S9 and variety S8, where its value reached 0.9268. The findings of this study showed the high efficiency of ISSR makers in detecting genetic variation among several wheat varieties.

Genetic polymorphism amongst moth bean (Vigna aconitifolia (Jacq.) Marechal) accessions evaluated using various molecular markers

This study presents genetic level diversity of 50 moth bean legume accessions (Vigna aconotifolia (Jacq.) Marechal). Despite seeds being a rich source of soluble nutritional proteins, it still remains an underutilized crop commercially. To assess the molecular diversity amongst accessions, RAPD, ISSR, DAMD, SCoT and SSR were employed as PCR markers. Of the total 5064 polymorphic bands shown with these makers, DAMD1 and SCoT1 presented higher levels of polymorphism with 407 and 262 polymorphic bands respectively. These genotypes revealed logical similarity and variation under UPGMA clustering. The PCA analysis of IC-258167, IC-415167, and IC-402286 present three highest eigen values, hence seem fit to be considered as parents for moth bean breeding programme. The polymorphic bands as produced by SCoT and DAMD primers can serve unique genotypic identifiers. This study seems to present a maiden report in labelling genetic marker based polymorphism amongst the accessions of moth bean.

Determining the Genetic Markers of Schizophrenia using Molecular Diagnosis

Schizophrenia poses a considerable disease burden, notwithstanding its relatively low incidence rate. The escalation in population size and mean age has led to a substantial rise in the incidence of illness linked to schizophrenia, particularly in countries with moderate levels of prosperity. Therefore, it is necessary to find a way to detect and recognize the illness as fast as possible to stop the consequences from occurring and stop symptoms from appearing before they even start. Within this study’s scope, fifteen distinct primers were selected randomly to determine whether or not they could recognize schizophrenia. To accomplish this, a Randomly Amplified Polymorphic DNA-polymerase chain reaction (RAPD-PCR) was conducted utilizing DNA samples obtained from individuals afflicted with the condition and those in good health. The outcomes of the gel electrophoresis and subsequent analysis revealed that solely three primers including C12, E02 and E03 exhibited monomorphic characteristics. However, it was observed that 80% of the primers, precisely eleven out of fifteen, exhibited polymorphism. The results indicated that among the 15 primers tested, the primer labeled as H01 failed to generate any detectable bands. Subsequent examination of the polymorphic primers revealed that the primer efficiency varied between 0 and 0.0755, while the discrimination power ranged from 0 to 20 percent. The study observed variations in the frequency of disease occurrence between patients and healthy individuals. H05 primer utilization yielded three distinct polymorphic bands exhibiting diverse lengths. The band exhibiting the greatest length measured 1190 bp, whereas the other two bands had 480 and 430 bp lengths respectively. The two polymorphic bands exhibited 100% prevalence in the patient population while being completely absent in the healthy control group. Based on the available data, it appears that utilizing this primer to identify schizophrenia is a valid and valuable claim. Overall, DNA-based diagnostics may overlook a number of difficulties as laboratories have the ability to extract DNA from various sources easily. In this scenario, molecular and genetic techniques may be crucial for diagnosing some diseases.

Diversity in the population of Fusarium oxysporum f. sp. ricini causing castor wilt disease in India

A major constraint on castor crop productivity is wilt disease caused by Fusarium oxysporum f. sp. ricini (For). In this study, morphological, pathogenic and molecular features of F. oxysporum f. sp. ricini isolates collected from different castor-growing agro-climatic zones of India were analyzed. Eighty-nine F. oxysporum f. sp. ricini isolates were obtained from wilt-infected root samples of castor plants from different states of India viz., Andhra Pradesh, Bihar, Gujarat, Haryana, Karnataka, Madhya Pradesh, Maharashtra, Odisha, Rajasthan, Telangana and Tamil Nadu. Morphology assessment in terms of variability in radial growth, size of micro and macroconidia and number of septa, grouped the isolates into 5 clusters in biplot analysis. A genotype-by-isolates bi-plot analysis ranked genotypes for pathogen susceptibility, with JI-35 being closest to the ideal and most susceptible, and DCS-9, and Kranthi being less susceptible. In contrast, 48–1 and JI-258 showed resistance, while Haritha and VP-1 exhibited intermediate susceptibility levels. K-means cluster analysis found 5 groups for pathogenic variability studies of isolates. The six isolates viz., For-12-5 (61.8 %), For-12-10 (50.5 %), For-13-47 (51.4 %), For-113 (86.1 %), For-13-33 (77.9 %) and For-53 (62.2 %) recorded as highly virulent isolates against wilt disease which showed >50 % average wilt incidence mean value across cultivars. Out of 40 ISSR primers tested, 11 primers gave polymorphic bands and analysis of genetic variability among 36 F. oxysporum f. sp. ricini isolates showed polymorphism with PIC, heterogygosity index, effective multiplex ratio, Marker index, and Shannon Diversity Index which ranged (in %) between 0.22 and 0.69, 0.19–0.44, 11.09–27.9, 6.14–8.98, and 0.78–3.04 respectively. The dendrogram analysis revealed 53 % similarity among the isolates studied and these isolates grouped broadly into 6 clusters. Variability among isolates was significant in morphological, pathogenic and molecular characters of F. oxysporum f. sp. ricini.

PHYLOGENETIC STUDY OF TEN SPECIES FROM CENTAUREA (ASTERACEAE) IN DUHOK CITY, KURDISTAN REGION-IRAQ

The current research aimed to estimate the evolutionary relationships of ten Centaurea L. species growing naturally in the Duhok City, Kurdistan region of Iraq. The combing Start Codon Targeted (SCoT) markers with Internal Transcribed Spacer (ITS) gene region barcode were performed. To detect the DNA sequence variations and phylogenetic tree reconstruction, the Dice similarity matrix, the unweighted pair group method with arithmetic mean (UPGMA) clustering and Maximum Likelihood (ML) methods were applied. 104 polymorphic bands were scored with an average of 10.4. The Polymorphic Information Content (PIC) and Resolving Power (Rp) values ranged between (0.24 to 0.36) and 3.4 in primer (SCoT1) to 12 in primer (SCoT53) with an average of 0.319 and 5.74 respectively. The lowest similarity value was 0.52 between C. behen L. and C. solstitialis L., while the highest was 0.82 between C. balsamita and C. rigida. The reconstructed polytomous dendrogram was as follows: clade one; C. solstitialis L.; clade 2, C. balsamita Lam. and C. virgata Lam.; clade three subdivide into two subclades: C. iberica Trev. ex Spreng., C. hayalolepis Boiss., C. brugueriana (DC) Hand. Mazz. and C. gigantea Sch. Bip. Ex Boiss., C. regia Boiss., C. rigida Banks & Sol., C. behen Lam. Furthermore, C. brugueriana (DC) Hand. Mazz., C. iberica Trev. ex Spreng, C. behen L., C. solstiotialis L. and C. balsamita Lam. were nested with National Center for Biotechnology Information (NCBI). In contrast, the remaining taxa were mixed with other closely related species. Thus, ScoT markers and ITS DNA barcode were considerably effective for investigating the evolutionary relationships of Centaurea taxa.

Agarose-Resolvable SSR Markers Based on ddRADSeq in Chickpea

Exploitation of genetic diversity is essential for sustainable crop production. Molecular markers have potential to reach these goals in more rapid and efficient manner. Simple sequence repeats (SSRs) are one of the most popular molecular markers in breeding studies due to their co-dominant inheritance, abundance in the genome, multi-allelic and reproducible nature. Here, we developed genomic SSR markers from chickpea with the use of ddRADSeq data. 1,396 SSR regions with an average of 530 SSR/Mb in the whole genome were successfully identified. Considering different types of repeats, dinucleotides were the most frequent type accounting for 62.03% of the total SSR regions identified, followed by trinucleotides (25.50%) and tetranucleotides (4.58%). The AT/TA motif was greatly characterized among dinucleotide repeats, and it was also the most common type in the chickpea genome accounting for 36.5% of the total SSR regions identified, followed by AG/GA (139) and TC/CT (135) among dinucleotide motifs. Considering their genomic distribution and simple visualization on agarose gels, we examined SSR regions of 10 bp and longer for identification of SSR markers. A total of 10 SSR markers were successfully designed and resulted in successful polymorphic bands among chickpea genotypes. Consequently, the results show that ddRADSeq is effective for marker development and these markers might be valuable for biodiversity studies, marker-assisted selection (MAS) and linkage map construction in chickpea.

Assessment of elite pepper breeding lines using molecular markers

In this study, 38 elite breeding pepper lines were genetically analyzed using SRAP markers and tested for resistance to PVY, TSWV, and PMMoV viruses using molecular markers. In the virus resistance tests, 1 line (37-H–D-6) from the Three-lobs population was found to be resistant to all 3 viruses tested. The 19 SRAP primer combinations used for genetic diversity yielded a total of 85 bands, 57 of which were polymorphic among pepper lines. While 2–8 bands per primer were obtained, the number of polymorphic bands ranged from 1 to 6. The average polymorphism rate of the primers was 66.44%. The PIC values ranged from 0.06 to 0.40 (with a mean of 0.18). In addition, the average gene diversity, effective allele number, and Shannon information index values of the primers were 0.21, 1.34, and 0.31, respectively. STRUCTURE analysis showed that the pepper lines were grouped into 4 clusters. PCoA and Q-matrix plots supported the cluster distribution. Some lines of the Sivri and Three-lobs pepper populations were observed as outliers in the plots. Kapia and Three-lobs were more similar to each other. This study showed that SRAP markers can be successfully used for genetic diversity of pepper breeding lines.