RAPD Markers Research Articles

Synergistic Effect of Plant Growth Regulators on Micropropagation of Eclipta alba: A Plant with Diverse Medicinal Properties

Synergism between various plant growth regulators is reported to be a key factor for the development of efficient in vitro propagation for any plant. Therefore, the current study examines the synergistic impact of plant growth regulators on in vitro propagation of Eclipta alba, an important medicinal plant possessing diverse medicinal properties. For the establishment of aseptic cultures, nodal segments were employed as explants on MS medium supplemented with 2.5 μM of 6-benzyle adenine (BA). Varying concentrations of BA and Kinetin (KIN)(0.0-5.0 μM), either alone or in combination with α- naphthalene acetic acid (NAA @ 0.0-5.0 μM) and indole 3-acetic acid (IAA@ 0.0-5.0 μM), were found to be effective for promoting shoot proliferation. Compared to KIN, BA was found to promote shoot proliferation and elongation more effectively. Further, the addition of 0.5μM NAA in the MS medium supplemented with 2.5 μM of BA increased shoot multiplication and elongation frequency from 58 and 17 percent to 65 and 21 percent respectively. The rooting frequency was found to be maximum on 1/2 strength MS medium supplemented with 5.0 μM of indole 3-butyric acid (IBA), which was found to be a superior auxin for inducing roots as compared to the NAA and IAA. With a 75% survival rate, in vitro raised plantlets were effectively acclimatized first in a poly house and later under greenhouse conditions. Molecular analysis was carried out using RAPD markers, with results indicating that the micropropagated plants were genetically identical to the mother plant. The developed micropropagation protocol for E. alba can be used at the commercial level for the mass multiplication of plants.

Use of molecular biology techniques for animal identification and traceability

Animal husbandry is considered a substantial part of agriculture which is important for the economic development of a country. The local potential for the livestock sector has remarkably developed in the past few decades with the introduction of technological advancements. Identification and traceability of livestock populations evolved as critical points in the production chain. Molecular marker-based genetic studies have addressed the preliminary complications of the livestock field. The intended objective of genetic studies is to select and maintain superior traits which have a direct impact on economic value. In that regard, the application of genetic markers has a significant role. RFLP, SCAR, AFLP, SSR, ISSR, EST, RAPD, STR, SCoT, and STS markers are prominently used despite animal identification and traceability, determination of genetic distance, sex determination, parentage determination, genetic conservation, gene mapping approaches in the livestock field. The present review is a critical study of current molecular biological techniques and their versatile applications in animal identificati on and traceability approaches and possible improvements of the available techniques to address the major concerns in the field.

Identification of Morphology and Molecular PCR-RAPD Bactrocera spp. in the Location of Red Guava Crops, Deli Serdang District

Various fruit flies are in Indonesia, but only a few species of fruit flies have been found at the red guava plant in Deli Serdang District. Knowing the species of fruit flies are needed to do the proper control. In addition to morphological identification, it is necessary to carry out molecular characterization to obtain accurate results in characterizing species differences. This study aims to identify the fruit fly based on morphology and molecularly PCR-RAPD for mapping the genetic closeness of the relationship between individual fruit flies. Bactrocera morphologically identified at LIPI, namely <em>B. carambolae, B. papayae, B. caudata, B. albistrigata, B. umbrosa, B. curcubitae, B. tau,</em><em> </em>and<em> </em><em>B. kinabalu</em>. For molecular identification PCR-RAPD, shows the dendrogram results from the Neighbor-Joining analysis based on RAPD markers of DNA band characters showing the genetic proximity of eight species divided into three groups of Bactrocera based on their location. The closeness of genetic relationships between individuals was analyzed using Pairwise Distance Calculation which describes the genetic distance between species. The results of Pairwise Distance Calculation ranged from 0.13-0.42. By knowing what species there fruit flies are in red guava plantations in the Deli Serdang district, it can make it easier for farmers to monitor and control fruit flies in red guava plantations.

Determination of Genetic Diversity in Selected Salt Tolerance and Sensitive Wheat by Random Amplified Polymorphic cDNA

The aim of this study was to investigate the genetic diversity and markers associated with salinity tolerance in three genotypes of wheat created for salt tolerance by plant breeding program, as well as two Iraqi varieties using random amplified polymorphic cDNA (RAPD-PCR) with eight primers were used. The results of RAPD marker revealed that there are genetic variations in several particular segments of various sizes between the selected genotypes and the local varieties with more genetic variation except for (OPG-09) did not appear any band with the selected genotypes and local cultivars. The results of the phylogenetic tree analysis (cluster) based on the presence or absence of DNA amplified for each primer were used to classify similar genotypes into two main groups (1H, Iraq) genotypes were clustered in first group with a genetic distance of 85% and (2H, Ibaa99) genotypes clustered in second group with a genetic distance of 84%.The results of genetic similarity based on RAPD data shows the relationship between the selected genotypes and local cultivars, the highest genetic distance was 0.428571 between the selected genotypes (1H and 2H) and the local cultivar of (I), and the lowest value was 0.066667 between the selected genotype (1H and 3H). The results confirmed the efficiency of RAPD based cDNA technique of those genotypes that are studied for determination of genetic diversity and distance among them.

Molecular characterization and diversity analysis of some local potato (Solanum tuberosum L.) genotypes of Bangladesh by RAPD markers

Genetic identification of cultivars and varieties is useful in preserving endangered potato germplasm and planning for new breeding programs. Molecular characterization and genetic diversity of 10 local potato genotypes of Bangladesh were investigated with five RAPD primers, namely OPA-18, OPB-06, OPC-01, OPD-02 and OPW. Genomic DNA was extracted from young leaves and PCR reactions were performed with selected primers. The selected primers generated 33 distinct and differential amplified bands (size ranged from 131-1188 bp), out of which 28 were polymorphic. The percentage of polymorphic loci was valued from 33.33% to 100%. Gene frequency was ranged from 0.300 to 1 and gene diversity was from 0 to 0.70. The inter-varietal similarity indices was ranged from 42.45% to 92.67%. Pair-wise comparisons of Nei's genetic identity value was from 0.4848 to 0.9394. The highest Nei's genetic identity (0.9394) was observed in Sheel Bilati and SAU Promising Genotype-5 genotype pair. Nei's (1972) genetic distance was from 0.0625 to 0.7239. Dendrogram based on Nei's genetic distance using Unweighted Pair Group Method with Arithmetic Means (UPGMA) indicated the segregation of studied potato genotypes into two main clusters. It was found that RAPD markers are sensitive enough to identify genetic variation in a variety of germplasm, which will be helpful for choosing genetically unique germplasm in potato breeding programs. SAARC J. Agric., 20(2): 43-55 (2022)

Determination of Some Morphological Traits and Genetic Factor of Two Species of Liriomyza spp (Diptera: Agromyzidae) on the Climbing Beans and Tomato Crops

This study was carried out to study some morphological parts of adult male of Liriomyza spp, including the antenna, the femur and the middle vein (Cu Al), as well as the shape of mines that larvae made them on the leaves. Results showed that there were two species of L.bryoinae on tomato crop and L.sativae on climbing beans crop, this was agreed with the molecular study that it was conducted using the RAPD Random Amplification of Polymorphic DNA based on the PCR technique to determine genetic variability of DNA that it was extraction from adults and pupae and obtained a quantity of it ranged from (35-556.9) mg and purity ranged between (1.7-2).
 The RAPD markers were studied using 19 random primers, 13 of which showed a bands and 6 of them didn’t show any bands, the total number of bands loci were 53 loci, 23 of which were main bands and 30 were polymorphism and genetic analysis was performed based on these results. The genetic distance was found and lowest was (0.125) among adults and pupae of bean crop and higher distance was (0.323) the among the adults and the pupae of the tomato crop and the genetic distance between the tomato pupae and bean pupae was (0.185), according to this values the genetic relationship was found which was the tomato adult's species was independent and the genetic relationship between Adults and pupae have closest. Results showed that the pupae which were taken from the crops, there are two species of Liriomyza spp .
 

Evaluation of the optimum threshold of gamma-ray for inducing mutation on Polianthes tuberosa cv. double and analysis of genetic variation with RAPD marker

Purpose This experiment aimed to investigate the effect of gamma irradiation on morpho-physiological characteristics and molecular-induced variations in Polianthes tuberosa L. Methods Experiments were designed according to a completely randomized design with eight different gamma-ray doses (0, 20, 30, 40, 50, 60, 70, and 80 Gy) via a source of cobalt-60 with three replications. Some morpho-physiological characteristics of tuberoses were screened and evaluated at the end of the flower growth and development phases. The RAPD-PCR molecular marker technique was further used to identify the mutants of phenotypic variation flowers. Results Results indicated that the effect of different levels of γ-rays on some morphological and physiological traits was significant as the gamma-ray level was increased up to 50 Gy. The doses higher than 50 Gy were found to cause stand or no growth. The 50 Gy gamma irradiation reduced germination by 70.59%, germination rate by 66.36%, dry weight by 88.15%, fresh weight by 87.41%, flowering stem height (cm) by 69.22%, leaf area (cm2) by 57.35%, leaf number by 34.41%, chlorophyll content (mg g−1 FW) by 44.79%, number of florets by 92.57%, spike height (cm) by 27.80%, bulblet number by 32.57%, and bulblet diameter (mm) by 30.21%. On the contrary, gamma radiation at 50 Gy increased relative water content (%) and electrolyte leakage (ds m−1) by 41.27 and 237.65%, respectively. The results also showed that bulbs treated with 20 Gy gamma ray had the highest germination percentage and dry weight. The RAPD analysis indicated that among 10 primers tested, nine primers showed clear bands as the highest number of amplified fragments (90) was related to the OPM13 primer and the lowest number (40) to the OPM10 primer. However, the DNA polymorphism was dose-dependent. Conclusion Overall results showed that although the plant morphology was changed with gamma-ray level, no changes occurred in tuberose color.

Genotoxic potentials of some selected heavy metals exposure on Clarias gariepinus (Burchell, 1822) and Oreochromis niloticus (Linnaeus, 1758) using RAPD-PCR technique

Fish health and the aquatic ecosystem are strongly interwoven and interrelated. The aquatic ecosystem receives a range of anthropogenic chemicals which have toxicological or lethal health effects on the aquatic animals. The aim of this study was to determine the level of induced mutation and genomic stability of different sub-lethal doses (25%, 50% and 75% LC50) of heavy metal [Nickel (Ni), Mercury (Hg), Lead (Pb)and Zinc (Zn)] in Clarias gariepinus (Burchell, 1822) andOreochromis niloticus (Linnaeus, 1758) using RAPD-PCR technology over a period of 21 days. Four highly polymorphic RAPD markers were evaluated on isolated DNA from both heavy metal exposed fishes and control fishes after exposure. Observation of RAPD profiles in C. gariepinus revealed more damaging effect as sub lethal doses increased with zinc (a + b = 44 bands) mercury (a + b = 41 bands) nickel (a + b = 37 bands) lead (a + b = 35 bands) than with nickel (a + b = 37 bands) zinc (a + b = 34 bands) lead (a + b = 32 bands) mercury (a + b = 31 bands) for O. niloticus when compare to the control groups. Although, the genomic stability template decreased sub lethal heavy metal doses, higher stability was observed in O. niloticus (GTSPb = 26.32% GTSHg = 18.42% GTSZn = 10.53% GTSNi = 2.63%) than in C. gariepinus (GTSPb = 5.41% GTSNi = 0.0% GTSHg = -10.81% GTSZn = -18.92%). The results obtained showed differential variation in heavy metal induced genetic mutation and genomic stabilty in C. gariepinus and O. niloticus. These observable differences might be due to the physiological structure of the fish species evaluated. This study also confirms that RAPD–PCR technology is a useful tools in detecting the genotoxic effect of heavy metals in aquatic organisms but due to low reproducibility of RAPD results, it is recommended that this technology should be used along with other molecular techniques in fish genotoxicity studies.Keywords: Heavy metals, Induced, Genotoxicity, Fish, RAPD-PCR Technique

Effect of gamma iradiated seeds of Jabon Merah (Neolamarckia macrophylla (Wall.) Bosser) to genetic diversity

Jabon merah (Neolamarckia macrophylla (Wall.) as one of the potential industrial forest plantations to meet the need for wood that has a moderate level of genetic diversity. This study aims to analyze the morphological and genetic characters and analyze the stability of genes in explants from red jabon seeds induced by gamma rays at doses of 0 gray, 5 gray, 10 gray, 15 gray, 30 gray, 45 gray and 60 gray, 75 gray, 90 gray, 120 gray and 240 gray. The induced seeds were cultured in vitro. The subculture was started with an initialization on MS0 medium and repeatedly cultured. Finally, the LD50 value was determined using curve fit analysis, explant morphological analysis with SPSS, genetic analysis of explants using RAPD markers and SSR/Microsatellite. The DNA analysis with 9 SSR/Microsatellite and RAPD marker primers showed that most of the induction doses produced more fragments than the control.

Morphological and molecular characterization of maize lines tolerance to drought stress

Abstract. Amzeri A, Badami K, Santoso SB, Sukma KPW. 2022. Morphological and molecular characterization of maize lines tolerance to drought stress. Biodiversitas 23: 5844-5853. The early step in assembling varieties with early maturity, high productivity, and resistance to drought stress is to carry out morphological and molecular characterization among lines resistant to drought stress. The research aimed to characterize the morphology and molecular characteristics of maize lines resistant to drought stress to develop breeding and cultivation programs. The research used a randomized complete block design with twenty genotypes (maize lines) as treatment and three replicates. A total of 57 morphological characters and seven RAPD primers were used to assess the relationship based on morphological characters and RAPD markers. The results showed that the similarity coefficient based on the morphological characters was 0.69-0.97, while the similarity coefficient based on the RAPD marker was 0.64-0.95. The phenotypic diversity coefficient (PDC) was greater than the Genotypic Diversity Coefficient (GDC) in all observed quantitative characters. The broad sense heritability values of the tested maize lines ranged from 0.83 to 0.99. Production per hectare was significantly positively correlated with eight tested quantitative characters. G1 and G15 were the genotypes that can be used as parents to assemble hybrid maize varieties, while G10, G11, G16, and G17 could be used as maize for cultivation programs.

Genetic Diversity of Durian (Durio Zibethinus Murr.) in Nias Island, North Sumatera Use Random Amplified Polymorphic Dna (Rapd) Markers

Durian (Durio zibethinus Murr.) is one of tropic fruits that has varian taste depending on the places of their production. Nias Island, well known as durian production place in North Sumatera. The information about it’s varian was less studied and now day we need that information to analyze variant of durian especially in Nias Island. The aim of this research was to analyze genetic diversity of durian in Nias Island with Random Amplified Polymorphic DNA (RAPD). Ten shoots of durian was collected from each 5 regencies, which were: Nias Selatan, Nias Barat, Nias Utara, Nias Kota, dan Nias Induk. Durian’s DNA has been isolated by using Cetyl Trimethyl Ammonium Bromide (CTAB) method and amplified with six RAPD primers namely: OPA 01, OPA 03, OPA 07, OPA 10, RAPD 05, and OPN 06. The amplified band were scored and translated to biner data, then analyzed using Numerical Taxonomy and Multivariate System (NTSys) and clustered by Unweighted Pair Group Method With Arithme Average (UPGMA) method. Isolated DNA showed a clear and unsmeared band. The size of amplified band was around 221-2855bp with 100% of polymorphism. The dendogram was constructed by 50 accession of Nias’s durian with 0.51-0.59 of similarity value in each population. All of Nias’s durian accession has similiar value of 67% and B2 (Nias Barat) as a farthest genetic distance from all accession.

Field Screening and Molecular Characterization of Urdbean Genotypes for Yellow Mosaic Disease

Background: Urdbean [Vigna mungo (L.) Hepper] is an important pulse crop in India as well as in South East Asia. It is an excellent source of easily digestible, good quality protein. Mungbean Yellow Mosaic Virus causing Yellow mosaic disease (YMD) is one of the most devastating diseases affecting urdbean production and productivity to a large extent. Identification of resistant genotypes is one of the most important aspects in the management of MYMV diseases. The current study aimed to identify the MYMV resistant urdbean genotypes in order to manage and control the MYMV disease. Methods: In present investigation, a total 100 urdbean genotypes were screened against MYMV under natural field conditions in the augmented design by planting 2 rows of two test entries each alternated with one row of susceptible check. Further, selected genotypes which showed disease free, highly resistant, resistant, moderately resistance and susceptible reaction in field screening were used to tag RAPD markers associated with MYMV resistance in urdbean. Result: Out of 100 genotypes tested, only one genotype, NDU 12-1 was found to be disease free, twenty seven genotypes found to be highly resistance and four genotypes showed resistance consistently for four consecutive seasons (two summer and two Kharif) during 2017-2018. However, four genotypes were found to be superior for seed yield among all the genotypes tested as well as resistant to MYMV. The RAPD analysis clearly grouped all the MYMV resistant genotypes studied in cluster I whereas the susceptible genotypes were clustered together in the cluster II. The present work has successfully identified the MYMV resistant urdbean genotypes which can be used in MYMV resistance breeding programme.

Mitotic chromosomal abnormalities and DNA polymorphism in the Nile tilapia (Oreochromis niloticus, Linnaeus, 1758) as a biomarker for water pollution by heavy metals.

Mitotic chromosomal aberrations and DNA polymorphism (RAPD marker) were carried out on the Nile tilapia Oreochromis niloticus collected from five sites in Minia governorate, Egypt to test their applicability as biomonitors for heavy metal contaminants of water. The diploid chromosome number of O. niloticus population was 2 n=44. Different types of chromosomal aberrations were recorded (e.g., deletion, ring, centromeric attenuation, end-to-end association, dicentric chromosome, stickiness chromosomes, endomitosis, fragments and chromatid gap). The chromosomal aberrations varied between O. niloticus population collected from five sites, and the most common type was ring (R) chromosomes. Samples obtained from Bahr Yousef and Irrigation drain exhibited the highest aberration frequency. The frequency of chromosomal aberration was positively correlated with the concentration of heavy metals where their concentration in the surface water of Irrigation drain and Bahr Yousef exceeded the limits defined by WHO as well as the concentration of Pb in muscles. The RAPD marker was also used to identify genetic variation among Nile tilapia samples collected from five different water sources. It created polymorphic and unique bands that can be used as genetic markers to track DNA variations. The dendrogram also revealed that exposure to heavy metal pollution causes gradual accumulation of variance, whereas areas subjected to environmental stress showed higher genetic variation and clustered together.

Phenotypic Variation and RAPD Polymorphism of Pisang Kepok Local Cultivars (Musa acuminata x Musa balbisiana, ABB, Saba Subgroup)

Pisang Kepok is a major local banana cultivar in Indonesia with high economic, social and cultural value. Particularly on the island of Java, there are several variations of Pisang Kepok recognised with their own local names, which makes difficulties in taxonomic identification and grouping. Morphological features are used in conventional banana cultivar classification, but they are deemed less precise due to their subjectivity, thus, it is supposed to be complemented with a molecular approach. This study aims to identify the phenotypic variation of Pisang Kepok local cultivars also their genetic polymorphism using Random Amplified Polymorphic DNA marker. Phenotypic variation was observed using 35 morphological characters. Six RAPD primers were used, i.e. OPA2, OPA3, OPA4, OPA11, OPA12, and OPA18. Clustering analysis, both phenotypic and genetic were performed using PAST v4.02. The morphological characterisation identified four variants of Pisang Kepok i.e. Kepok Abang, Kepok Putih, Kepok Manurun, and Kepok Australi; which all confirmed as ABB genome group. Phenotypic clustering showed that Pisang Kepok cultivars were separated into 3 clusters based on their local name, with a high similarity value of >90%. PCA biplot showed that the fruit flesh colour was the most important character contributed to the cultivar variation. RAPD marker also showed that each specimen was grouped according to its local name and source, with a similarity value of >80%. Both morphology and molecular (RAPD) markers resulted in the branching of Pisang Kepok which was closer to Pisang Klutuk than Pisang Barlin.

Rapid In Vitro Propagation of Fig (Ficus carica L.) ‘Violette de Solliès’ Supported by Molecular and Microscopy Analyses

Ficus carica L. is a common fig that is an incredibly nutritional fruit, well-known for its medicinal and economic values. This study aims to establish an efficient protocol for the mass propagation of fig plantlets (Ficus carica L.) for the cultivar ‘Violette de Solliès’. Surface-sterilized shoot-tip explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins (6-benzylaminopurine, BAP; thidiazuron, TDZ; kinetin, Kn; and zeatin, Zea). Induced shoots were rooted on Woody Plant Medium (WPM) with different concentrations of auxins (naphthalene-acetic acid, NAA; indole-3-acetic acid, IAA; and indole-3-butyric acid, IBA). Rooted explants were acclimatized in eight different soil substrates prior to cultivation in a commercial plot. The propagated plantlets were analyzed for genetic stability and clonal fidelity using RAPD and SCoT molecular markers, whereas scanning electron microscopy (SEM) was performed to observe the stomata morphology of post-acclimatized plants. MS media supplemented with 5.0 mg/L BAP was the optimal treatment for multiple shoot induction (15.20 ± 1.03 shoots), whereas the highest percentage of rooting (93.33%) was achieved in WPM supplemented with 3.0 mg/L IBA. Plantlets were successfully acclimatized in biochar soil with a survival rate of 100%. RAPD and SCoT analysis showed no polymorphism occurrences across six subculture cycles, whereas observations via SEM indicated normal stomata structures on the leaves of acclimatized plantlets. This study documents an efficient micropropagation protocol for Ficus carica cv. ‘Violette de Solliès’ for the production of uniformed and true-to-type plant stocks suitable for commercial propagation.

Molecular and quantitative genetic variation within and between populations of the declining grassland species Saxifraga granulata.

Formerly common plant species are expected to be particularly susceptible to recent habitat fragmentation. We studied the population genetics of 19 recently fragmented Saxifraga granulata populations (max. distance 61 km) in Luxembourg and neighboring Germany using RAPD markers and a common garden experiment. We assessed (1) the relationships between plant fitness, quantitative genetic variation, molecular genetic variation, and population size; and (2) the relative importance of genetic drift and selection in shaping genetic variation. Molecular genetic diversity was high but did not correlate with population size, habitat conditions, or plant performance. Genetic differentiation was low (F ST=0.079 ± 0.135), and there was no isolation by distance. Longevity, clonality, and the long-lived seed bank of S.granulata may have prevented strong genetic erosion and genetic differentiation among populations. However, genetic distinctness increased with decreasing genetic diversity indicating that random genetic drift occurred in the studied populations. Quantitative and molecular genetic variations were correlated, and their differentiation (Q ST vs. F ST) among S.granulata populations was similar, suggesting that mainly random processes have shaped the quantitative genetic differentiation among populations. However, pairwise quantitative genetic distances increased with geographic and climatic distances, even when adjusted for molecular genetic distances, indicating diversifying selection. Our results indicate that long-lived clonal species may be buffered at least temporarily against the negative effects of fragmentation. The relationship between quantitative genetic and geographic distance may be a more sensitive indicator of selection than Q ST-F ST differences.

Analysis of Genetic Diversity through Leaf Trichome Characters and RAPD Markers in Two Species of Dendrocalamus

Traditionally, phenotypic characters or protein profiling were used to establish the genetic relationships or for identification purpose of species. In recent times, DNA profiling is done using various molecular techniques. Analysis of micro-morphological characters can be used as preliminary tool as compared to phenotypic characters. Present study is an approach to find out the intra-generic and intra-specific relationship based on micromorphological and RAPD analysis in Dendrocalamus spp. For micromorphological analysis, observations on trichomes were recorded on both the surfaces of leaves. Total five genotypes, Dendrocalamus asper (mother and micropropagated plant) and D. strictus (seed raised three cultivars from various locations) were included in the study. Both the samples of D. asper showed close similarity on the basis of presence and size of trichomes (371.07 - 1019.17 μm) on the lower surface (abaxial) and being present in the same subcluster for all the seven primers tested out of total nine. However, one primer (B17899) gave no amplification whereas, OPB4 divided the genotypes in to three clusters-cluster I (sample 2), cluster II (sample 1) and cluster III (sample 3, 4 and 5). Out of three samples of D. strictus, one sample showed characteristically very large trichomes (1189.3 - 2166.61 μm) and was in different cluster by the primers OPE1, OPD1, OPA12 and A17898.

The genetic diversity and variation in crude protein content of wheat (Triticum aestivum L.) promising cultivars for breeding in Albania

The genetic diversity and variation in crude protein content among eleven wheat genotypes, comprising three elite local genotypes and eight wheat genotypes of foreign origin were investigated in the present study. Variability was evidenced in grain protein content estimated by biuret test, it ranged from 9.5 to 13.9% with mean of 11.58%. Comparative analysis between local and introduced wheat genotypes revealed that the local genotypes had lower protein content than those of foreign origin. Fourteen polymorphic RAPD markers were used to assess genetic diversity among selected wheat varieties. The mean similarity among wheat genotypes was 67%. Genetic similarity among local wheat varieties was higher (83%) than among those of foreign origin (66%). The wheat genotypes were grouped into two main clusters on UPGMA dendrogram constructed based on Dice similarity coefficients. A clear clustering of genotypes according to the origin wasobserved. This clustering was also supported by principal coordinate analysis (PCoA) results. There was no observed clustering based on the protein content. The data revealed that local wheat genetic had narrow genetic diversity, however the wheat genotypes of foreign origin constitute a promising material to be employed in breeding programs aiming the increase of wheat protein content and genetic diversity.

Physio-biochemical analysis and molecular characterization of induced lentil mutant lines.

Lens culinaris is a proteinaceous food crop that is consumed worldwide for protein requirements. Mutation breeding has been used to improve protein content, yield, and related traits, as well as to select highly desirable mutants that are economically significant. An investigation of genotypic variation in lentil germplasm was carried out using induced mutagenesis, with caffeine, ethyl methane sulfonate (EMS), lead nitrate, and cadmium nitrate as mutagens that resulted in 18 mutant lines in the M3 generation. For the present study, we analyzed the genetic diversity of lentil mutant lines using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and random amplified polymorphic DNA markers (RAPD). The heterozygosity of RAPD markers per primer ranged from 50.00–90.90% with an average of 71.04%. The genetic divergent analysis was performed using hierarchical clustering (UPGMA), exhibited that these mutant lines were classified mainly into five subpopulation or clusters. A close resemblance with highest genetic coefficient similarity (1.00) were observed between control and mutant H; between mutant M and E; between mutant Q and J2, while more divergent mutants were N2 with mutant B; and mutant R with mutant J1with least genetic coefficient similarity (0.22). Protein and mineral content (Fe, Zn and Cu) were increased significantly in some high yielding mutant lines concerning to the control plant, and showed polymorphic variations in polypeptide chains in terms of banding pattern. Stomatal morphology in high yielding mutants were perceived utilizing scanning electron microscopy (SEM), exhibiting variations in stomatal size, stomatal opening and number of stomata. The present study’s promising mutant lines’ biological, physiological, and molecular profiles provide a foundation for forthcoming preservation and consumption strategies to broaden the genetic diversity of the breeding population of lentil.

RAPD markers in the discrimination of genetic variability in ornamental peppers

Genetic variability in pepper genotypes has been widely assessed using phenotypic traits and molecular markers, which are essential to estimate the genetic divergence of the species and thus recommend parental individuals for breeding programs. In this scenario, this study aimed to evaluate the genetic divergence between ornamental pepper accessions (C. annuum L.) using RAPD molecular markers. Seventeen ornamental pepper accessions and one plant of the commercial cultivar Calypso were used in the study. The accessions were subjected to RAPD analysis with 15 primers. The analysis revealed high variability between the accessions and evidenced that accessions UFPB-355 and UFPB-348 were the most dissimilar, whereas accessions UFPB- 443 and UFPB-77.3 were the most similar. The accessions were grouped into three different groups, evidencing their genetic divergence The accessions characterized in this study are promising and can be used in future pepper breeding programs due to their high genetic variability. Moreover, the results obtained in this studyallow recommending accessions UFPB-45, UFPB-77.3, UFPB-137, UFPB-356, and UFPB- 443 as parent individualssince they are the most divergent. Finally, the RAPD markers were highly sensitive in driscriminating pepperaccessions and should be used as an auxiliary tool for choosing parents.r a divergência genética na espécie e recomendar indivíduos parentais para programas de melhoramento. Assim, este trabalho teve como objetivo avaliar a divergência genética entre acessos de pimenta ornamental ( C. annuumL.) usando marcadores moleculares RAPD. Foram utilizados 17 acessos de pimenta ornamental e uma planta da cultivar comercial Calypso. Os acessos foram submetidos à análise de RAPD, com 15 primers. A análise revelou grande variabilidade entre os acessos. Os resultados evidenciaram que os acessos UFPB-355 e UFPB-348 são os mais dissimilares, enquanto os acessos UFPB-443 e UFPB-77.3 são os mais semelhantes. Os acessos foram agrupados em três grupos, evidenciando a divergência genética entre eles. Os acessos caracterizados neste estudo são promissores e podem ser utilizados em futuros programas de melhoramento de pimenta devido à alta variabilidade genética detectada. Os resultados obtidos neste estudo permitiram indicar, como indivíduos parentais, os acessos UFPB-45, UFPB-77.3, UFPB-137, UFPB-356 e UFPB-443,