Random Amplified Polymorphic DNA Primers Research Articles

Determination of the Genetic Relationships Among Salvia Species by RAPD and ISSR Analyses.

Salvia L. is the largest genus of the family Lamiaceae, which includes approximately 1000 species. According to recent studies, 100 Salvia species in total grow in Turkey. At the same time, 53% of them are endemic. The purpose of this study was to investigate the genetic relationships among 15 Salvia species that grow in wild conditions in Turkey's Eastern Anatolia region. The genetic relationships among 15 Salvia species were investigated using inter-simple sequence repeat (ISSR) and random amplified polymorphic-DNA (RAPD) profiles in the present study. Thirteen ISSR primers and 11 RAPD primers were utilized. The ISSR and RAPD data were combined to construct the unweighted pair group method using arithmetic average cluster. Based on the RAPD and ISSR data, the Salvia species were classified into six groups. As a result of the combined analysis, it was shown that similarities between the species varied between 0.54 (S. rosifolia-S. sclarea, S. rosifolia-S. limbata, and S. staminea-S. verticillata) and 0.93 (S. sclera-S. divaricata). The findings show that the two markers represent powerful instruments for assessing the genetic diversity and relationships among Salvia species.

Induced in vitro adaptation for salt tolerance in date palm (Phoenix dactylifera L.) cultivar Khalas

BackgroundSoil salinity causes huge economic losses to agriculture productivity in arid and semiarid areas worldwide. The affected plants face disturbances in osmotic adjustment, nutrient transport, ionic toxicity and reduced photosynthesis. Conventional breeding approaches produce little success in combating various stresses in plants. However, non-conventional approaches, such as in vitro tissue culturing, produce genetic variability in the development of salt-tolerant plants, particularly in woody trees.ResultsEmbryogenic callus cultures of the date palm cultivar Khalas were subjected to various salt levels ranging from 0 to 300 mM in eight subcultures. The regenerants obtained from the salt-treated cultures were regenerated and evaluated using the same concentration of NaCl with which the calli were treated. All the salt-adapted (SA) regenerants showed improved growth characteristics, physiological performance, ion concentrations and K+/Na+ ratios than the salt non-adapted (SNA) regenerants and the control. Regression between the leaf Na+ concentration and net photosynthesis revealed an inverse nonlinear correlation in the SNA regenerants. Leaf K+ contents and stomatal conductance showed a strong linear relationship in SA regenerants compared with the inverse linear correlation, and a very poor coefficient of determination in SNA regenerants. The genetic fidelity of the selected SA regenerants was also tested using 36 random amplified polymorphic DNA (RAPD) primers, of which 26 produced scorable bands. The primers generated 1–10 bands, with an average of 5.4 bands per RAPD primer; there was no variation between SA regenerants and the negative control.ConclusionThis is the first report of the variants generated from salt-stressed cultures and their potential adaptation to salinity in date palm cv. Khalas. The massive production of salt stress-adapted date palm plants may be much easier using the salt adaptation approach. Such plants can perform better during exposure to salt stress compared to the non-treated date palm plants.

Genetic variability analysis of partially salt tolerant local and inbred rice (Oryza sativa L.) through molecular markers

Random Amplified Polymorphic DNA (RAPD) analysis was performed to assess the genetic variability in sixteen selected germplasms of rice, Oryza sativa L. using eight decamer RAPD primers. The data obtained from this investigation reveals a high level of polymorphism between cultivars. The primers produced a total of 255 bands of which all 255 bands were polymorphic indicating 100% polymorphism. The size of the amplified bands ranged from 220 bp to 2290 bp. The number of polymorphic fragments ranged from 24 to 49 with an average of 32 polymorphic fragments for each primer. The primer OPX04 produced the maximum number (49) of polymorphic bands while the OPB04 and OPB17 produced the minimum number (24) of polymorphic bands. The polymorphic information content (PIC) values ranged from 0.6616 to 0.8845 with an average of 0.832. The highest PIC value (0.8845) was obtained for primer OPL03. The RAPD data was analyzed to determine the pair-wise genetic similarity coefficients which ranged from 0.00 to 0.83. The BRRIdhan 23 and the BRRIdhan 41 varieties were the closest genotypes with the highest similarity index of 83%. This was followed by 77% similarity between a pair of cultivars Kalamona and Horkuch. On the other hand, 100% dissimilarity was seen between BRRIdhan 53, BRRIdhan 50, BRRIdhan 10, BRRIdhan 70, BRRIdhan 54, BRRIdhan 40, BRRIdhan 23, BRRIdhan 47, BRRIdhan 41 and Dadsail respectively and between BRRIdhan 53 and Horkuch; indicating a high level of variability between paired genotypes. Cluster analysis was performed using Unweighted Paired Group of Arithmetic Means (UPGMA). The UPGMA dendrogram resolved the selected rice cultivars into four clusters.

Genetic diversity of the Andean blackberry (Rubus glaucusBenth.) in Ecuador assessed by AFLP markers

AbstractAndean blackberry (Rubus glaucusBenth.) is an emerging fruit crop with significant commercial potential. Despite its growing popularity, basic research about its genetic resources and breeding remains insufficient. The aim of this study was to assess the genetic diversity of Andean blackberry cultivars and related berries species from the main production areas in Ecuador. We analysed a total of 106 samples and performed DNA screening with different molecular markers: random-amplified polymorphic DNAs (RAPDs), inter-simple sequence repeats (ISSRs) and a set of representative samples with amplified fragment length polymorphisms (AFLPs). The tested RAPD primers did not reveal any differentiation among accessions identified asR. glaucus, however one ISSR primer was useful to find polymorphisms allowing the selection of 29 accessions for the analysis with AFLP markers. AFLP-M13 technology was used for screen genetic variations among these accessions and eight wildRubusaccessions. We scored 203 bands using five primer combinations; out of these 152 were informative inR. glaucus. AFLP markers clearly distinguishR. glaucusfrom the screened wildRubusspecies, also an unexpected genetic structure was revealed amongR. glaucuscultivars. This genetic differentiation and detection of admixed genotypes suggest a possible introgression of wildRubusspecies inR. glaucus. Our findings are relevant for blackberry genetic breeding and use of these genetic resources.

Genetic diversity and pathogenic characteristics of Phomopsis asparagi obtained from Asparagus officinalis and A. kiusianus in Japan

Phomopsis asparagi has been reported as a serious disease, causing stem blight on asparagus cultivars (Asparagus officinalis), and this fungus was also recorded as a common spot disease on wild asparagus (A. kiusianus). In this study, isolates of P. asparagi obtained from A. officinalis and A. kiusianus in different locations of the Kyushu island were successfully assessed the genetic variation through mycelial compatibility grouping (MCG) and random amplified polymorphic DNA (RAPD). Thirty fungal isolates were identified into six MCGs; of these, fifteen isolates were representatively chosen for the RAPD analysis, pathogenicity and extracellular enzyme test. Eight RAPD primers, B62, B66, B68, B71, C83, D22, D32 and E10, generated 100 different patterns from the isolates with 91% of the polymorphic bands. Based on RAPD analysis, P. asparagi isolates were divided into 3 distinct genetic clusters, namely RAPD-I, RAPD-II and RAPD-III at a similarity of 73%. The result indicated that there was a great genetic variation within the isolates from A. kiusianus, presenting in the three clusters. While the isolates from A. officinalis showed high genetic homology and they belonged to the RAPD-II cluster. There is no conflict between genetic clusters distinguished by the RAPD markers and those of MCG. On the other hand, the pathogenicity test revealed that the isolates of RAPD-II and RAPD-III were highly virulent pathogens. In contrast, the isolates of RAPD-I were weak virulent pathogens when inoculated on A. officinalis. Screening of extracellular enzyme activities concluded that all isolates were positive for laccase, lipase and protease activity.

Polyamine elicited aristolochic acid production in in vitro clonally fidel Aristolochia indica L.: An ISSR and RAPD markers and HPTLC based study

The present investigation was conducted to find out the influence of different polyamines (PAs) viz. spermine (spm), spermidine (spd) and putrescine (put) and plant growth regulators (PGRs) on multiplication, rooting, callusing and regeneration in Aristolochia indica L. in vitro. Firstly, the A. indica plants were collected, surface sterilized and cultured in Schenk and Hildebrandt (SH) media fortified with various PGRs and PAs in different concentrations and combinations. SH media fortified with benzylaminopurine (BAP) (2.0 mg/l)+put (0.5 mM) and in another experiment, BAP (2.0 mg/l)+spd (1 mM) produced best results with 41.00 and 39.2 axillary shoots/ nodal explant respectively. Following 4-6 weeks of incubation, explants propagated in SH media augmented with various combinations and concentrations of BAP and kinetin (kin) (0.5, 1, 1.5 and 2 mg/l), spm, spd and put (0.5 and 1 mM) generated white friable callus. SH media supplemented with BAP (2.0 mg/l)+spd (0.5 mM) displayed best response with an average number of 47.5 base callus derived shoots . Following 6 weeks of incubation, SH+ indole-3-acetic acid (IAA) (1.0 mg/l)+spd (0.5 mM) resulted maximum average number (7) of roots per shoot. The genetic homogeneity of the mother plant (MP), hardened plant (HP) and in vitro regenerated plants was ascertained employing random-amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. In this present study, in randomly selected A. indica plants, by using 6 ISSR and 5 RAPD primers, 63 scorable bands were derived, 59 of which were recorded as monomorphic with a degree of monomorphism of 90%. High-performance thin layer chromatography (HPTLC) revealed aristolochic acid (AA) content in in vitro grown A. indica roots and the roots from the mother plant (MP) as 6074.54 µg/g and 5891.14 µg/g respectively.

Molecular, physiological, and biochemical characterization of extracellular lipase production by Aspergillus niger using submerged fermentation.

BackgroundExtracellular production of fungal lipases especially the lipases obtained from the Aspergilli has gained immense interest in recent years due to its diverse biotechnological applications. In this study, we focused on determining the fermentation parameters required for the optimal lipase production.MethodsA total of 256 fungal isolates were obtained from oil seeds. From each genus, one isolate was selected to evaluate lipase production using phenol red and tributyrin plate assays. Lipase activity was estimated using the spectrophotometric pNPP hydrolysis assay. The highest lipase producer isolates were identified using 18S ribosomal RNA gene sequencing. The genetic variability was determined by random amplified polymorphic DNA (RAPD) analysis and the dendrogram was constructed using the unweighted pair group method with arithmetic averages method. The isolates were examined in a submerged fermentation culture (Smf) to measure the effect of temperature, pH, incubation time, carbon source, nitrogen source, inoculum volume, and lipid source on lipase production.ResultsEleven isolates belonging to the genus Aspergillus were analyzed for lipase production where they were found to be the highest lipase producers among various fungal genera. All the tested isolates were identified as A. niger using 18s rRNA sequencing. Genetic diversity was evaluated among all of the studied A. niger isolates using RAPD primers. The RAPD primers were used to amplify 285 loci, of which five were polymorphic (1.75%) and seven were monomorphic (2.45%). Thus, a high level of genetic diversity was observed among all isolates. The tributyrin test and the lipase activity assay identified five strains of A. niger as high lipase producers, and their optimal enzyme activities were 709.74, 532.54, 735.64, 794.62, and 787.69 U/ml. The optimal conditions for lipase production were as follows: 40 °C, pH 7.5, 1% fructose as the carbon source, 1% yeast extract as the nitrogen source, 2% palm oil, 2.5 × 107 spores/ml suspension, and 3 days of incubation.ConclusionsThe current study provides a comprehensive characterization of the optimal conditions, which are essential to enhance lipase production in five A. niger isolates.

Genetic diversity of Sterculia quadrifida in Kupang, Indonesia based on RAPD (Random Amplified Polymorphic DNA) markers

Abstract. Uslan, Pharmawat M. 2020. Genetic diversity of Sterculia quadrifida in Kupang, Indonesia based on RAPD (Random Amplified Polymorphic DNA) markers. Biodiversitas 21: 3407-3414. This study aims to determine the genetic diversity of Sterculia quadrifida R.Br. in Kupang based on RAPD markers. Samples of S. quadrifida were collected from the yard and community forest in Kupang City (Sub-districts of Oebobo, Kelapa Lima, Maulafa, and Alak) and mixed forest in the Kupang District (Sub-districts of Kupang Barat, Nekamese, Taebenu, and Fatuleu). DNA was isolated by the CTAB method and amplified by six RAPD primers (OPD-11, OPF-11, UBC-106, UBC-127, UBC-250, and OPB-04). The data were analyzed in the MVSP software using UPGMA method and Nei & Li similarity coefficient. Total of 131 DNA bands ranging from 250-1400 bp was obtained. Populations of S. quadrifida in Kupang were divided into two main clusters and 12 sub-clusters with. The highest genetic diversity was found in Kelapa Lima of 0.1050, while the lowest genetic diversity was found in S. quadrifida population in Fatuleu of 0.0305. The population of S. quadrifida in Kupang has high genetic diversity and also clustered based on their geographical distribution

Efficiency and reliability of random DNA markers (RDMs) for evaluation of genetic variability and relationship in Ocimum accessions

The genus Ocimum is characterized by a great variability in both morphological features and chemical composition because the cross-pollination appears leads to various species, subspecies, varieties and forms. In addition, the taxonomy of Ocimum is complicated by the existence of numerous botanical varieties, cultivar names, and chemotypes within the species that may not differ significantly in morphology. In the present study, genetic diversity and relationships within and among 24 Ocimum accessions from five different agro-climatic regions of West Bengal were developed using PCR-based Random Amplified polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers. The RAPD primers developed 191 robust loci, which revealed 69.42% polymorphism and 83 ISSR markers generated 67.46% polymorphism. Genetic diversity parameters [average and effective number of alleles, percent polymorphism, average heterozygosity, intralocus gene diversity, polymorphic information content (PIC)] for RAPD, ISSR, and RAPD+ISSR along with UPGMA clustering based on Jaccard's coefficient were estimated with a view to assess efficiency of the marker system in Ocimum. The UPGMA dendrogram generated using cumulative data revealed significant relationships among the genotypes. All 24 genotypes studied grouped into three clusters irrespective of their geographical affiliations with 100% bootstrap value except few genotypes, suggesting considerable diversity among the genotypes. Further the results demonstrate the efficiency and reliability of RAPD and ISSR markers in determining the genetic diversity and relationships among the Ocimum germplasm. RAPD and ISSR profiling have identified diverse Ocimum genotypes, which could be further utilized in various genetic improvement programmes including conventional as well as marker assisted breeding towards development of new and desirable Ocimum genotypes.

Genetic diversity of grapevine (Vitis vinifera L.) cultivars in Al-Madinah Al-Munawara based on molecular markers and morphological traits

Grapevine is one of the major fruit trees in the Kingdom of Saudi Arabia. This study aims to discriminate and assess the genetic diversity in three grapevine cultivars in Al- Madinah through the combination of characterization using both classical ampelographic as well as ampelometric studies with molecular markers using Randomly Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Start Codon Targeted Polymorphism (SCoT). For the ampelographic analysis, twelve OIV descriptors were used, and for the ampelometric analysis, the fully expanded mature leaves area were automatedly measured. The genetic distance among the three grapevines cultivars, calculated using Jaccard's coefficient, ranged from 0.7577 (between AL Nakheel and Banati) to 0.4501 (between AL Nakheel and Ahmer). The molecular analysis was based on the use of thirty-one primers; ten RAPD primers, seven ISSR primers and fourteen SCoT primers. RAPD primers generated the highest polymorphism (67%), while the level of polymorphism with ISSR primers was 36% and with SCoT 44%. All of the three markers generated similar dendrograms, and the genetic distance generated with RAPD was higher compared with SCoT and ISSR. The three markers RAPD, ISSR and SCoT were combined and amalgamated with the morphological data, and combined dendrogram was generated and discussed. AL Nakheel and Ahmer cultivars were found to be more closely related to each other than Banati which was separated in a different cluster. Both methods were effectively efficient for complete identification of grapevine and for studying the genetic diversity between closely related cultivars.
 Bangladesh J. Plant Taxon. 27(1): 113-127, 2020 (June)

Molecular Phylogeny of Sunflower Cultivars of Teddy Bear, Skyscraper, Lemon Queen and Common Sunflower Using RAPD Markers

Helianthus or sunflower is a genus of plant comprising about 70 species. Common sunflower and other members of Helianthae are cultivated in temperate regions and some tropical regions as food crops for humans, cattle, poultry, and as ornamental plants. The common sunflower is valuable with respect of economic and ornamental point of view. There are many cultivars of sunflower including teddy bear, skyscraper, and lemon queen. Variation among these cultivars has been studied using molecular techniques and the result were used to develop the phylogeny among them. Random Amplified Polymorphic DNA (RAPD) is one of molecular techniques that were used for this purpose. The purpose of this study was to contruct the phylogeny of three sunflower cultivars and common sunflower based on RAPD markers. The RAPD primers used in this study were OPA-2, OPA-9, OPA-13, OPB-2, OPB-4, OPB5, OPB-7, and OPB-11. Data analysis based on molecular data showed that genetic relationship among Lemon Queen, Skyscraper, Teddy Bear and Common sunflower based on RAPD markers shows that the cultivars studied are grouped into three main groups, namely: Group I Lemon Queen and Skyscraper, Group II Teddy Bear, and Group III Common sunflower; the closest kinship is shown between Lemon Queen and Skyscraper.

Diversity analysis of Sweet Potato (Ipomoea batatas[L.] Lam) genotypes using morphological, biochemical and molecular markers

Sweet potato [Ipomeabatatas(L.) Lam.]is a nutritious food crop primarily grown by small and marginal farmers. successful breeding and germplasm conservation programs demands characterization of its germplasm. Here, we tried to determine genetic diversity among 21 sweet potato genotypes using morphological, biochemical and molecular markers. Ten morphological traits were studied and subjected to analysis of variance (ANOVA). Mean square due to germplasm were highly significant as well as wide mean range performance was observed for tuber number per plant, individual tuber weight, tuber fresh yield per plant, tuber dry yield per plant, tuber yield per plot and tuber length. UPGMA (Unweighted Pair Group Method Arithmetic Average) cluster analysis based on morphological traits separated the germplasm into three groups. The genotypes Gautam, Shree Arun, RS-92 and CO-3-4 appeared promising with regard to yield characters. Total phenol was maximum in in V-12 genotype (1.39 mg), while minimum was recorded in Samrat genotype (0.95 mg). The highest total antioxidant was observed in the genotype Samrat (0.30 mg), while minimum was recorded in the genotype Navsari Local (0.16 mg). Molecular diversity analysis was carried out using 25 RAPD (Random Amplified Polymorphic DNA) primers, out of which 13 primers produced 117 reproducible amplicons (106 polymorphic, 7 monomorphic and 4 unique amplicons). UPGMA dendogram based on RAPD data separated the genotypes into two major clusters having the similarity coefficient ranged from 0.56 to 0.76. The results can be used for sweet potato crop improvement through molecular breeding and marker assisted selection of for desired traits in future.

Anatomical profile and genetic variability of sweet potato (Ipomoea batatas) cultivars in Banyumas, Central Java, based on RAPD markers

Abstract. Samiyarsih S, Fitrianto N, Azizah E, Herawati W, Rochmatino. 2020. Anatomical profile and genetic variability of sweet potato (Ipomoea batatas) cultivars in Banyumas, Central Java, based on RAPD markers. Biodiversitas 21: 1755-1766. Indonesia has developed a food consumption diversification based on local sources. Sweet potato (Ipomoea batatas) is one of the tubers of carbohydrate sources in addition to wheat, rice, corn, potatoes, and cassava. It has a wide distribution, and high cultivars diversity produces a high variation on its morphology, anatomy, and genetic modification. The objectives of this research were to investigate the anatomical profile and genetic variability of eight sweet potatoes (Ipomoea batatas) cultivars found in Banyumas, Central Java, Indonesia. Anatomical profile was observed by the embedding method and subjected to analysis of variance. DNA amplification using Random Amplified Polymorphic DNA (RAPD) method with 10 primers, namely OPA-1, OPA-2, OPA-3, OPA-4, OPA-9, OPA-11, OPA-13, OPA-15, OPA-16, and OPA-18. The result of the anatomical study showed that eight sweet potato cultivars have significantly different (p<0.05) variation in leaf anatomical profile. RAPD primers produced 52 amplified fragments varying from 100 to 1300 bp in size, and 90.40% of the amplification bands were polymorphic. The genetic similarity level is ranged from 0.37 to 0.93. Two unspecific groups were forming at a coefficient of 55% from the dendrogram. The first group consisted of one cultivar, Cangkuang, and the second group consisted of seven cultivars Antin, Ungu Tua, Borobudur, Sukuh, Sari, Beta, and Kidal. The nearest relationships cultivars were Borobudur, and Ungu Tua had the highest similarity coefficient of 93%, and the lowest similarity found in Cangkuang and Antin with a coefficient of 37%. The range of genetic distance of eight cultivars was from 0.37-0.93. Implementation of the research would particularly useful for the identification and evaluation of the genetic improvement of sweet potato cultivars.

Genetic Diversity of Potato Based on Random Amplified Polymorphic DNA and Simple Sequence Repeat Marker

Various potato clones are cultivated by farmers in Banjarnegara and Wonosobo , Central Java, Indonesia such as MZ, NH1, NH2, Vega, Gareta, Granola, Bliss, Merah (Red Potato), Ungu (Purple Potato), Klon_17 (K17), Local Dieng, Margahayu, and X. This encourages the importance of genetic diversity and genetic similarities. This study aimed to analyze genetic diversity and genetic similarities between 13 accessions of potatoes based on seven Random Amplified Polymorphic DNA (RAPD) primer s and nine pairs of Simple Sequence Repeat Marker (SSR) primer s. The results showed that RAPD and SSR primers could be used to analyze genetic diversity and genetic similarities of 13 potatoes accessions from Banjarnegara and Wonosobo. The PLP value was 80.9% in the RAPD primer and 65.41% in the SSR primer. Four RAPD primer s were informative based on PIC value: OPG 08, OPM 19, OPP 13, and OPX 04. Three SSR primer were informative: STM 2005, RGH- SSR 8, and S tI 035. Genetic similarities presented by Phylogenetic tree analysis resulted in two main clusters. The first cluster consisted of Granola, MZ, Ungu (Purple potato), Merah (Red Potato), Local Dieng, Margahayu, Gareta, Vega, NH2, NH1, Klon_17 (K17), and Bliss. The second cluster consisted of X. Granola and MZ having a high genetic similarity with a genetic distance of 0.07 and 0.132. Meanwhile, K17 and X had a low genetic similarity with a genetic distance of 0.31 and 0.987.

Assessment of Genetic Variation in Soybean (<i>Glycine max</i>) Accessions from International Gene Pools Using RAPD Markers: Comparison with the ISSR System

Soybean (Glycine max) is one of the most important crops in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were to 1) assess the level of genetic variation among soybean (G. max) accessions from different countries using Random Amplified Polymorphic DNA (RAPD) markers and 2) compare Inter Simple Sequence Repeats (ISSR) and RAPD marker systems in detecting polymorphic loci in soybeans (G. max). Genomic DNAs from 108 soybeans (G. max) accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The average level of polymorphic loci detected with the RAPD primers was 35%. The soybean accessions from the China, Netherlands, and Canada gene pools were the least genetically variable with 25%, 26%, and 30% of polymorphic loci, respectively. Accessions from Hungary (43%) and France (48%) showed the highest level of polymorphism based on the RAPD analysis. Overall, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The levels of polymorphic loci detected with the RAPD and ISSR marker systems were in general moderate and similar even if they target different regions of the genome. A combination of different marker systems that include RAPD/ISSR, microsatellites (SSR), and SNPs should provide the most accurate information on genetic variation of soybean (G. max) accessions.

Genetic divergence of Anthurium affine germplasm using morphoagronomic and molecular descriptors

Anthurium affine is an ornamental Brazilian native plant very appreciated due to its long-lasting durability. To explore the genetic diversity available, 21 A. affine half-sibling progeny were evaluated using morphoagronomic and molecular descriptors . The morphoagronomic descriptors used were: plant height, plant diameter, growth habit, leaf number, leaf blade length, leaf blade width, petiole length, petiole diameter, leaf margin undulation, general appearance; soil cover rate and Random Amplified Polymorphic DNA (RAPD) markers the molecular one . All the descriptors within the progeny showed variability and general appearance, leaf margin undulation and soil cover rate are the ones that most contribute to the plants divergence. From 45 RAPD primers tested, 19 presented clear polymorphic bands (42.2%). Those generated 200 molecular markers, 81 (40.5%) of them being polymorphic. The number of polymorphic bands per RAPD primer ranged from one (OPB10) to eight (OPW11) and polymorphism ranged from 16.6% (OPB10) to 70% (OPW05). Grouping analysis of morphoagronomic and molecular descriptors formed five and four groups respectively . The dendrograms showed differentiation between accessions although there was no association between morphoagronomic and molecular groupings. There is genetic divergence within the half-sibling progeny of A. affine plants evaluated, which opens up possibilities for increase variability.

Polymorphic information and genetic diversity in Brassica species revealed by RAPD markers

Randomly amplified polymorphic DNA (RAPD) is a tremendously convenient approach used to discriminate between Brassica species owing to its accuracy and speed. RAPD primers generate adequate genetic information that can be used in the primer-marker system. In this work, twenty RAPD-PCR based markers were executed to generate polymorphic data, like polymorphic information content (PIC), mean resolving power (MRP), resolving power (RP), effective multiplex ratio (EMR), and marker index (MI) for the first time and genetic distance among and between six Brassica species were calculated. Our results indicated that 20 primers produced a total of 231 scored band and generated 87% polymorphic bands. Average PIC, MRP, RP, MI, and EMR values were 0.088, 0.65, 6.7, 0.78, and 8.9, respectively. PIC showed an overall negative correlation with MRP, RP, MI, and EMR, whereas MRP, RP, and EMR, were positively correlated with each other. Genetic identities ranged from 41.99% (between Brassica napus and Brassica oleracea) to 68.83% (between Brassica campestris and Brassica oleracea). Dendrogram results showed no clustering between species except between Brassica campestris and Brassica nigra. Nevertheless, these results will be helpful to acquire useful information about the markers and their use to determine the genomic structures of Brassica species. Further, based on genetic distance and polymorphic information, new hybrids can be developed for effective oilseed production.

Genetic Structure of Phlebotomus orientalis (Diptera: Psychodidae) in Leishmaniasis Endemic Foci of Sudan.

Visceral leishmaniasis (VL) remains a major concern in many parts of Sudan. The disease is transmitted by Phlebotomus orientalis. The objective of this study was to determine genetic structure of Phlebotomus orientalis population from 5 geographical regions in Sudan. A total of 194 individual sand flies were collected from 5 geographic regions in Sudan. The field collected sand flies were analyzed by Random Amplified Polymorphic DNA (RAPD) using 30 primers. Eight hundred and 65 bands from 4 RAPD primers were analyzed for genetic variation. A higher level of intrapopulational variability was detected in populations of P. orientalis from eastern Sudan compared to those populations from central and northern Sudan. Diagnostic bands were detected in populations of P. orientalis central Sudan. Hieratical clustering analysis showed clear clustering into 2 main populations with 1 population subdivided into 4 subpopulations. However, these populations did not show any correlation with their geographical origins. Furthermore, the low genetic differentiation among subpopulations was supported by fixation index (FST) estimated by analysis of molecular variance (AMOVA). It is concluded that the populations of P. orientalis from the selected areas in Sudan have a low genetic differentiation. However, assessment of genetic structure of P. orientalis populations is important for understanding the patterns of transmission of VL in different endemic areas.

Phylogenetic Analysis of Chrysochamela (Fenzl) Boiss. (Brassicaceae) Taxa Growing in Turkey

In this study, we performed a genetic diversity analysis using RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter simple sequence repeat) markers for some Chrysochamela species growing in Turkey. PCR was performed using RAPD and ISSR primers and genomic DNA samples from each specimen, and the resulting bands were scored. Phylogenetic relationships and genetic distances between the studied taxa were calculated using the PAUP analysis program. According to the RAPD analysis, the closest genetic distance was found to be 0.20000 between C. noeana populations, while the most distant values were found as 0.60000 between C. noeana and C. elliptica. The ISSR analysis also revealed the closest genetic distance as 0.34043 between C. noeana populations, while the most distant values were found to be 0.55319 between C. noeana and C. elliptica. UPGMA (unweighted pair group method with arithmetic mean) dendrogram generated for both RAPD and ISSR analyses consisted of two groups. As a result, it was found that RAPD data were more appropriate for comparison with the previous anatomical and morphological data of the studied taxa.

Selecting Primers for RAPD, Microsatellite and Mitochondrial Cytochrome Oxidase Subunit 1 for Genetic Variation Analysis of Asian Corn Borer (Ostrinia furnacalis Guenée) Population in Java, Indonesia

Primer plays an important role in studying genetic diversity of an insect species. This research was aimed to select the suitable primers to visualize the genetic diversity of Asian corn borer (Ostrinia furnacalis) using Random Amplified Polymorphic DNA (RAPD), microsatellite, and mitochondrial cytochrome oxidase subunit 1 gene (mtCO1). Twenty four RAPD primers (OPA1, OPA4, OPA7,OPA8, OPA10, OPA11, OPA12, OPA13, OPB7, OPB10, OPB11, OPB12, OPB15, OPC4, OPC5, OPC14, OPC16, OPC18, OPC20, OPD3, OPD8, OPD10, OPD13, OPD14) and five microsatellite primers (T3, T4, T5, T81, D25) resulted high polymorphic informations of the genetics of O. furnacalis in Java Indonesia. Universal primers, Lep and Heb were appropriateto do molecular identification of O. furnacalis based on BLAST system on GenBank and BOLD systems.