Random Amplified Polymorphic DNA Bands Research Articles

Variation of Fruit Color in Cakra Hijau, G1/M8 and HV-149 Chilli Pepper Cultivar: Physiology and Molecular Approach

The fruit color of chili pepper is an important characteristic in identification and classification and is often used as the basis for determining consumer preferences. Information on the relationship between chili fruit color and its molecular profile is very important in supporting selection activities in plant breeding. This study aims to identify genetic diversity associated with the fruit color of three genotypes of chili (Capsicum frutescens L.): Cakra Hijau, HV-149 and G1/M8, using Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeat (SSR). Morphological confirmation was carried out according to Capsicum descriptors. Nineteen RAPD markers and six SSR markers were used for genetic variability assessment. Genetic variation was analyzed using the unweighted pair group method with the arithmetic mean and the Jaccard similarity index. The three chili genotypes had different fruit colors at each maturation stage. The immature Cakra Hijau fruit is dark green and turns dark red as it ripens. The immature fruits of the G1/M8 line are light green and turn red when ripe. Finally, immature HV-149 fruits are dark green and yellow when ripe. The SSR markers used in this study were unable to show polymorphism. On the other hand, the RAPD marker successfully detected genetic variation in the three chili genotypes and resulted in a total of 49 alleles. The average value of polymorphic information content of the RAPD primers used ranged from 0 to 0.296, with the highest index indicated by OPA-1. The dendrogram shows the separation of the three genotypes into two main clusters, with the first cluster consisting of the HV-149 variety and the second cluster consisting of Cakra Hijau and G1/M8 lines. This study revealed that there are genetic variations based on the morphological characteristics of fruit color at each ripening stage and RAPD band profile. The RAPD marker was more effective than the SSR marker for identifying the genetic diversity of fruit color in the three chilies studied.

Genetic Analysis of Prunus salicina L. by Random Amplified Polymorphic DNA (RAPD) and Intersimple Sequence Repeat (ISSR).

Background Prunus salicina L. is an important fruit tree species of great economic value which is mainly distributed in the northern hemisphere. Methods 25 samples of Prunus salicina L. were collected from 8 provinces in China, Japan, USA, and New Zealand. The genetic variations of these samples were characterized by the random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) technique, respectively, and in combination. Results Totally, 257 RAPD bands ranging 200∼2300 bp was found, and 81.59% of these bands were polymorphic. ISSR analysis identified 179 bands ranging 300∼2500 bp, and 87.74% of the bands were polymorphic. ISSR results showed that the similarity coefficient index between samples P10 (Maihuangli in Anhui, Chin) and P13 (Longyuanqiuli in Heilongjiang, China) was lowest, while that between samples P10 (Maihuangli in Anhui, Chin) and P15 (Baili in Japan) was highest. Combined analysis of RAPD and ISSR demonstrated that the similarity coefficient index between samples P4 (Qiepili in Ningbo, Zhejiang, China) and P13 (Longyuanqiuli in Heilongjiang, China) was lowest, while that between samples P19 (Laroda in USA) and P20 (Red heart in USA) was highest. Conclusion RAPD combined with ISSR analysis can be used for genetic characterization of Prunus L. species.

Molecular characterization of Canna indica L. based on random amplified polymorphic DNA markers

Random amplified polymorphic DNA (RAPD) markers were employed for characterization, assessment of genetic variation and inferring relationships among six variants of Canna indica L. A total of 198 RAPD bands ranging from 200 bp to 3 kbp were generated by all the six variants. Among them, most of the bands were found to be polymorphic, four band were unique of which two bands (OPA022000 and OPA043000) were observed in the variant 2 (small red) and the other two (OPA013000 and OPA053000) were noticed in the variant 4 (orange), and the remaining bands were found to be monomorphic. The pair-wise genetic distance was determined among the six variants that ranged from 0.1446 to 0.6554. A dendogram was constructed based on the RAPD profiling to infer the relationship among the six variants of C. indica that resulted in two major clusters: the first one contained two variants, viz. variant 1 (local red) and variant 2 (small red), while the second cluster composed of the remaining four variants. The results as revealed from the RAPD analysis were found congruent with those of morphological and anatomical investigation of the species.
 Bangladesh J. Plant Taxon. 28(1): 75-81, 2021 (June)

Molecular and Alkaloid Characterization of Claviceps purpurea Sensu Lato From Grass Seed Production Areas of the U.S. Pacific Northwest.

Ergot, caused by Claviceps purpurea sensu lato, is an economically important seed replacement disease of Kentucky bluegrass (Poa pratensis) and perennial ryegrass (Lolium perenne) seed crops. C. purpurea sensu stricto is considered the primary Claviceps species responsible, but genetic diversity and cryptic species within C. purpurea sensu lato have previously been reported. Fifty-six C. purpurea sensu lato isolates collected from P. pratensis (n = 21) and L. perenne (n = 35) in Oregon and Washington between 2010 and 2014 were characterized via random amplified polymorphic DNA (RAPD), partial internal transcribed spacer (ITS), β-tubulin and elongation factor-1α (EF-1α) sequences, conidial size, and ergot alkaloid chemotype. Based on RAPD analysis, seven isolates from P. pratensis and 33 isolates from L. perenne collected in Oregon corresponded to C. purpurea sensu stricto, and 13 isolates collected from P. pratensis in Washington and Oregon were identified as C. humidiphila. Partial ITS, β-tubulin, and EF-1α sequences identified 10 isolates from P. pratensis as C. humidiphila, and seven isolates from P. pratensis and 33 isolates from L. perenne were identified as C. purpurea sensu stricto. Several isolates generated ambiguous RAPD bands or sequences that prevented identification. Ergot alkaloid chemotype profiling found that ergocornine and its epimer were predominant in sclerotia from P. pratensis, whereas ergotamine and its epimer were most abundant in sclerotia from L. perenne. This study confirms the presence of the C. purpurea sensu lato species complex in the U.S. Pacific Northwest and suggests that more research is needed to characterize and mitigate Claviceps spp. infection of grass seed crops in North America.

Zinc oxide nanocatalyst mediates cadmium and lead toxicity tolerance mechanism by differential regulation of photosynthetic machinery and antioxidant enzymes level in cotton seedlings

Cadmium (Cd) and Lead (Pb) heavy metal pollution induced toxicity severely affects the plant growth and yield of various agriculutral crops worldwide. The present study discuss the prime role of phycomolecules coated zinc oxide nanoparticles (ZnONPs) application on development of heavy metal tolerance mechanism in cotton (Gossypium hirsutum L.) seedlings better than exposed to Cd and Pb treatments alone. Co-exposure of ZnONPs along with heavy metal treatments significantly promoted the shoot, and root growth as well as biomass compared to control, while it was down-regulated in Cd and Pb exposed seedlings. The intervention of ZnONPs had up-regulated the level of chlorophyll a, b and carotenoid contents in leaves grown under Cd and Pb treatments than the untreated control. Similarly, the level of total soluble protein and malondialdehyde (MDA-lipid peroxidation) contents was significantly increased in the co-presence of ZnONPs along with Cd and Pb treatments over their respective control. Accumulation of antioxidant defense enzymes viz., superoxide dismutase (SOD), catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APX) was up-regulated significantly in seedlings upon co-exposure of ZnONPs with Cd and Pb treatments. Random amplified polymorphic DNA (RAPD) fingerprinting analysis exhibited no genomic changes/alterations in seedlings by co-existence of ZnONPs with heavy metals. Overall, the present results indicate that the addition of ZnONPs with Cd and Pb ion exposure protects cotton seedlings by alleviating heavy metal induced phytotoxicity and promoted physiochemical characteristics via differential regulation of photosynthetic machinery as well as antioxidative defense mechanisms in cotton seedlings. Results strongly suggest that phycomolecule coated ZnO nanoparticles could be effectively used as nanofertilizer to cultivate agronomically important crops in heavy metal polluted soil in the future.

Assessment of Somaclonal Variations in Embryo-derived Axillary Shoots of Chickpea using Molecular Markers

Background: Somaclonal variation is considered as a source of genetic variation for crop improvement. It has been investigated using cytological, biochemical and molecular techniques.Methods: Genetic stability in the embryo-derived axillary shoots of 4 chickpea genotypes was assessed using eight inter-simple sequence repeat (ISSR) and 19 random amplified polymorphic DNA (RAPD) primers. Result: RAPD primers produced 94 and ISSR primers produced 38 distinct and scorable alleles, with an average of 4.9 alleles for RAPD and 4.75 for ISSR primers. The polymorphic information content (PIC) ranged from 0.36 to 0.90 for RAPD and from 0.50 to 0.87 for ISSR. ISSR recognized a 90%, but RAPD recognized 82% similarity value. No absolute similarity value was between the mother plant and the regenerated shoots for the overall genotypes. At a 90% similarity value, 15 out of the 20 regenerated shoots from ‘Giza 88’ group with their mother plant using ISSR markers; however, 11 regenerated shoots grouped with their mother plant in one central cluster for ‘Giza 4’ using RAPD markers. The observed variations in the total number of polymorphic RAPD and ISSR bands and the number of bands specific to the mother and regenerated shoots, detected intra-clonal variation and genetic instability seem to be genotype-dependent.

Genetic variation of maiden lily (Lilium rubellum Baker) within and among three natural populations in Fukushima Prefecture in Japan

Lily ( genus Lilium ) is one of the most important flowers for bulb and cut flower production in the world. Lilium rubellum Baker is a native species of Japan that exhibits early flowering, is pink coloured, and possesses a pleasant fragrance, which makes it an important genetic resource for breeding lilies. In this work, the genetic diversity of L. rubellum was studied among three natural populations located in Fukushima Prefecture, Japan, which have been designated as the Mt. Azuma, Nango and Atsushio-kano populations. A total of 31 accessions collected in 1997 were analysed using Random Amplified Polymorphic DNA (RAPD) markers. Eighteen decamer primers produced 98% polymorphic RAPD bands; furthermore, 11 of the 18 primers produced 10 or more polymorphic bands with a mean per primer Polymorphic Information Content of 0.382. Among the three populations, the mean Shannon index, Nei´s gene diversity and percentage of polymorphic loci were 0.2749, 0.4099 and 76.7%, respectively. These data revealed that there is a high genetic diversity within all the populations. Analysis of the three genetic diversity indices within populations showed that the Nango population had the highest genetic diversity, whereas the Atsushio-kano population had the lowest. An unweighted pair group method with arithmetic mean dendrogram based on Jaccard´s similarity coefficient was constructed, and the three populations of L. rubellum collected in the Fukushima prefecture were clearly differentiated at 0.61 similarity index. The Nango and Mt. Azuma populations were genetically closer than the Atsushio-kano population. The analysis of molecular variance showed a 29.53% variance among the populations. This study revealed that there is high genetic diversity within populations and moderate genetic diversity among the three natural populations of L. rubellum Baker in the Fukushima Prefecture of Japan.

Evaluation of RAPD markers for molecular identification of five bamboo genera from Indonesia

Abstract Conservation of bamboos for future exploitation as fuel, fibre and as an ingredient for cosmetics depends on knowledge of its natural genetic variation. The study of molecular genetic diversity in bamboos will provide important information for its conservation. This article reports on the genetic diversity in 25 species representing five genera of bamboos found in Indonesia using Random Amplified Polymorphic DNA (RAPD) molecular markers. Out of 40 primers, 24 primers produced 1107 total bands and 86.21% of polymorphic bands across the 25 species. Sixteen bands were uniquely found in one species only and their presence or absence helped to define nine bamboo species. RAPD band sizes ranged from 162 to 2247 base pairs. A dendrogram based on the similarity coefficient of Dice divided the bamboo species into three big clusters. In conclusion, RAPD can capture the diversity among five different bamboo genera and has a great potential to be used in the study of genetic diversity in Indonesian bamboos.

Genetic Diversity of Artemisia herba-alba in Libyan Green Mountain

The genetic variability of 18 individuals of Artemisia herba-alba from 3 different elevations in Green Mountain—Libya was examined using sequence-related amplified polymorphism (SRAP) and random amplified polymorphic DNA (RAPD) techniques. Results showed distinctive SRAP and RAPD banding pattern in this species scoring a total of 17 SRAP and 61 RAPD bands including 94.12% and 100% polymorphic with amplicon size ranging from 50 to 1600 bp and 150 to 1800 bp, respectively. Phylogenetic relationship between the genotypes was able to divide the samples into two main groups with genetic similarity value ranged from 41 to 84%, indicating that the genetic relationship is slightly related to the geographical location of the studied plant area.

Genetic polymorphism among seven entomopathogenic nematode species (Steinernematidae) revealed by RAPD and SRAP analyses

The strength of sequence-related amplified polymorphism (SRAP) in comparison to random amplified polymorphic DNA (RAPD) in determining the genetic polymorphism among seven species of entomopathogenic nematodes (Steinernematidae) was tested. Nine RAPD and 12 SRAP primer pair’s combinations were used. The number of polymorphic bands and the polymorphism percentages was high in SRAP analysis (97 out of 107 bands were polymorphic (90.6%)) compared to RAPD (65 out of 89 bands were polymorphic (73%)). The highest number of RAPD bands was recorded for OPE-A-07 primer (14 band) while OPE-D-20 was scored the lowest band number (7 bands). The SRAP primers Me3-em2 registered the highest number of bands (13 bands) while Me3-em3and Me2-em2 showed the lowest band number (6 bands). The genotype-specific SRAP and RAPD markers for the different ENP species were recorded. The highest number of SRAP specific markers (7 markers) was scored for Steinernema glaseri, then 4 markers for S. abbasi 2 followed by S. riobrave (3 markers), 1 marker for S. abbasi 1, S. riobrave recorded the highest number of specific RAPD marker (4 specific markers) followed by S. carpocapsae DD 136 and S. glaseri (two specific markers), then only 1 specific marker for S. abbasi 1. Based on the data obtained from RAPD and SRAP analysis, the dendrogram was created to clarify the genetic distances among different species of the studied entomopathogenic nematodes. The present study indicated that SRAP was more powerful than RAPD analysis for detecting the genetic polymorphism among closely related species of nematodes. The genotype-specific markers detected from both RAPD and SRAP can be used in future biological control programs.

Genetic Variation among Five Egyptian Clover Cultivars Using Random Amplified Polymorphic DNA (RAPD) and Sequence-Related Amplified Polymorphism (SRAP) Molecular Markers

To estimate the genetic variation among five Egyptian clover cultivars, Random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) analyses were employed. Sixteen RAPD and six SRAP primers were used in the present study, a total of 34 and 12 bands showed 24.8 and 60% polymorphism percentage. The highest number of RAPD bands was recorded for primers OPE-B-07 (13 bands), followed by OPE-K-04 (12 bands), while the lowest was scored for OPE-B-05 (5 bands). For the SRAP Primers Me1-em2, Me3-em2 and Me3-em4 recorded the highest number of bands (four bands), followed by three bands for Me1-em1and Me3-em3, primer Me3-em1 recorded the lowest band number (two bands only). The dendrogram built based on combined data from RAPD and SRAP analyses clarify the genetic distances among the five Egyptian clover cultivars.

Utilizing RAPD Analysis as Molecular Markers for comparison between Various Callus and Mother Plant of Jatropha curcas L.

Calli were induced from different explants of Jatropha curcas L. seedling on MS basal agar medium in presence of different plant growth regulators. at different levels for 4 weeks Random Amplified Polymorphic DNA (RAPD) markers were used for the assessment of genetic variation of callus grown in different hormonal combinations in comparison with the mother plant. A total of 8 arbitrary sequence primers were evaluated. Of 8 primers used for the RAPD analysis, primers showed consistent band patterns. In total, scorable bands were observed with the primers. The total number of amplicons produced per primer varied from 5 for OPA-07 and OPB-12 to as many as 8 bands for OPB-10, OPC- 09 and OPR-17. The average number of bands per primer was 6.625. Out of 53 bands, 36 were polymorphic (67.92%). The average number of polymorphic RAPD bands was 4.5 per primer. the highest similarity (0.71) with mother plant was recorded in leaf-derived callus (S4) grown on MS solid medium with 2.5 μM NAA plus 2.5 μM IBA, while endosperm callus maintained on MS with hormonal combination 12.5 μM IBA + BAP- 25.0 μM was found to show least similarity (0.54) with mother plant.In this concern, results have been suggested to be useful fast methods for comparing of geneticchanges and variation in plants.

Genetic and phenotypic variation among Turkish terrestrial orchid species as revealed by RAPD and morphological characteristics

Terrestrial orchid species are natural sources of salep and a closely related group of plant species widely distributed throughout Turkey. The phylogenetic relationship among fourteen different tuber-producing orchid species was investigated after analyzing phenotypic and genetic variation within and among the natural population through fifteen morphometric traits and ten random amplified polymorphic DNA (RAPD) primer combinations. Statistical analyses (principal component analysis (PCA), principal coordinate analysis (PCoA), and cluster analysis) using the generated data identified taxonomic and genetic distance within the studied plant samples. The results of PCA from morphological traits show that there are no major groupings within and among different species instead somehow overlapping with few distinctly characterized species. In addition, the UPGMA-based phenogram with Euclidean distance (0-1) produces five major clusters among the studied orchid species according to their taxonomic status with few exceptions. On the other hand, PCoA and the phylogenetic dendrogram with the coefficient (0.56-0.79) from RAPD band profiles determine the true genetic diversity of those species. Although both combinations of genetic and phenotypic characteristics reveal the phylogenetic relationship of some those studied species very effectively, they are not clear for others. These results suggest that in the natural population of terrestrial orchid species significant amounts of gene flow are ongoing at intra/interspecies level. Therefore, it is recommended that conservation studies of these groups of orchid species should be done as a geographical unit rather than according to taxonomic status.

RAPD markers reveal genetic variation between Cichorium spinosum L. and Taraxacum sp.; a substantial medicinal plants of Greece

Fifteen Random Amplified Polymorphic DNA (RAPD) markers were used to measure genetic diversity and genetic relationships between five endemic genera of Mediterranean basin (Greece). Three species of Cichorium spinosum were collected; two from island Crete (Greece) and one from island Kythnos, and other two species of Taraxacom sp. (Asteraceae); are from Orhomenos and Athens. Two hundred-forty amplified products and 163 RAPD bands were scored with an average of 67.91% of them revealing polymorphism across accessions. In this research OPD-05 primer with 24 bands showed the highest number of bands, while the OPM-18 and OPB-16, both of them with 8 bands showed the least number of bands. Also OPV-06 primer with 18 polimorphic bands showed the highest number of bands. The least number of polymorphic bandswere found in OPX-18. UBC-292, OPAN-01, OPB-16, OPM-18, OPD-05 primers. Subsequently, OPD-05 primer with 29.16% showed the least percentage of polymorphism degree, whereas OPM-18 and OPB-16 primers with 87.50% showed the highest percentage of polymorphism. UPGMA clustering based on data from polymorphic RAPD bands revealed two distinct group which joined to form one major cluster at 32% level of similarity. Also Cichorium spinosum, Crete and Cichorium spinosum, Kythnos, varieties with 100% similarity are synonyms. The similarity indices of the RAPD dendrogram ranged between 30% and 100% averagely high enough to suggest useful variability for genetic diversity and plant breeding.

Induced mutation analysis with biochemical and molecular characterization of high yielding lentil mutant lines

Induced mutagenesis generates macromolecular variations which ultimately alters the bio-physiological and morphological nature of the crop genotypes. In the present study, molecular characterization of six high yielding lentil mutant lines, developed from hydrazine hydrates (HZ) and gamma rays mutagenesis, was carried out with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and random amplified polymorphic DNA (RAPD) markers. Activity of nitrate reductase (NR) and content of chlorophyll and carotenoid were found to be significantly high in the mutant lines. Protein and mineral (Fe, Zn & Cu) contents were also increased considerably in the mutant lines compared to their respective parent genotypes. SDS-PAGE profile of seed storage proteins showed 35 unique bands with 97.14% polymorphism. Genetic divergence analysis generated total 41 reproducible RAPD bands with average calculated polymorphic percentage of 63.06%. Among the primers, OPA-10 showed the highest polymorphism with significant PIC value. Genetic divergent analysis revealed that genome of cultivar DPL 62 mutated relatively more than the cultivar Pant L 406 due to the mutagen treatments, while DPL 62-B and Pant L406-A were the most divergent mutants induced in the present study. Biochemical and molecular profile of the induced mutant lines facilitates a basis for future conservation and utilization strategies to widen the genetic base of the current lentil breeding population.

Aluminum-Induced Changes on DNA Damage, DNA Methylation and LTR Retrotransposon Polymorphism in Maize

Aluminum (Al) toxicity is a kind of abiotic stresses that causes several damages on plants. In this study, Al-induced DNA damages, DNA methylation alterations and long terminal repeat retrotransposons (LTR RTs) polymorphisms were evaluated in Zea mays seedlings, using random amplified polymorphic DNA (RAPD), coupled restriction enzyme digestion–random amplification (CRED–RA) and inter-retrotransposon amplified polymorphisms assays, respectively. Our results fairly proved that all doses of \(\hbox {AlCl}_{3}\) (200, 400, 600 and 800 \(\upmu \)M) caused a reduction in genomic template stability and an increase in RAPD band frequencies (DNA damage). Furthermore, there was an increase in DNA methylation ratio based on CRED–RA results. Five LTR RTs (Nikita-E2647, N57 (Nikita), Sukkula, WLTR 2105 and Stowaway) that were determined in barley genome were investigated in maize genome. Polymorphism was determined under Al stress based on the used LTR primers. It was suggested that this mobilization and methylation status under Al stress might be a part of the defense mechanism in stress.

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Evidence of persistence of Prevotella spp. in the cystic fibrosis lung.

Purpose. Prevotella spp. represent a diverse genus of bacteria, frequently identified by both culture and molecular methods in the lungs of patients with chronic respiratory infection. However, their role in the pathogenesis of chronic lung infection is unclear; therefore, a more complete understanding of their molecular epidemiology is required.Methodology. Pulsed Field Gel Electrophoresis (PFGE) and Random Amplified Polymorphic DNA (RAPD) assays were developed and used to determine the degree of similarity between sequential isolates (n=42) from cystic fibrosis (CF) patients during periods of clinical stability and exacerbation.Results. A wide diversity of PFGE and RAPD banding patterns were observed, demonstrating considerable within-genus heterogeneity. In 8/12 (66.7 %) cases, where the same species was identified at sequential time points, pre- and post-antibiotic treatment of an exacerbation, PFGE/RAPD profiles were highly similar or identical. Congruence was observed between PFGE and RAPD (adjusted Rand coefficient, 0.200; adjusted Wallace RAPD->PFGE 0.459, PFGE->RAPD 0.128). Furthermore, some isolates could not be adequately assigned a species name on the basis of 16S rRNA analysis: these isolates had identical PFGE/RAPD profiles to Prevotella histicola.Conclusion. The similarity in PFGE and RAPD banding patterns observed in sequential CF Prevotella isolates may be indicative of the persistence of this genus in the CF lung. Further work is required to determine the clinical significance of this finding, and to more accurately distinguish differences in pathogenicity between species.

Identification and Differentiation of Egyptian Isolates of Erwinia amylovora using 16S rRNA Gene and RAPD-PCR

Erwinia amylovora the causative of fire blight disease on pear in Egypt was identified with some classical tests and molecular methods. Random Amplified Polymorphic DNA (RAPD) technique was used to identify the genetic variability among 6 Egyptian isolates of E. amylovora isolated from different governorates (Behera, Gharbiya, Nubariya, Menufiya, Kaluobiya and Ismailiya) and compared with one reference isolate. Profiles were obtained by amplification of DNA from these isolates with six random primers.The RAPD band patterns resulting from these primers were analyzed using UPGMA method to construct a similarity matrix and generate a dendogram indicating the relationships between the examinedisolates. The results obtained indicated that primers P4d, P5e and P6f were chosen because they reliably and reproducibly detected polymorphism among the selected isolates. Also, no variation in bacterial genome was observed among the examined isolates. In addition, 16SrRNA gene sequence was used to identify E. amylovora isolates using universal primers (8F and U1492R). The results showed that sequence of the isolates collected from different locations was matched with the sequence found in the Genbank database. The sequence of the fragment (16S rRNA gene) showed percentage homology with strain DSM30165 of Genbank strains ranged from 89–97%. The selected isolates were found to have theclosest homology to E. amylovora.

Genetic Assessment of the Shrub <i>Syncepalum dulcificum</i> (SCHUMACH & THONN.) Daniell in Nigeria Using the Randomly Amplified Polymorphic DNA (RAPD)

In order to advocate for informed management decisions with regards to endangered species, we assessed genetic diversity and genetic structure in 40 individuals of six natural populations of the shrub Syncepalum dulcificum (SCHUMACH & THONN.) DANIELL growing in South Western Nigeria. Twelve (12) Random Amplified Polymorphic DNA (RAPD) primers were tested on total genomic DNA extracted from silica gel dried leaves. Bands were then scored for reproducibility and scoring error calculated. Several genetic diversity parameters were then tested using the POPGENE v1.32 software. Five (5) primers produced 227 reproducible and clear RAPD bands of which 47 were polymorphic (20.7%). The percentage of polymorphic loci (PPI) within populations ranged from 36% to 68%. Nei’s gene diversity among population (Hs) was 0.03, while at species level (Ht) was 0.18. The coefficient of gene differentiation (Gst) among populations was estimated to be 0.83 with a gene flow rate (Nm) of 2.49 showing high genetic diversity within and among populations. The results however indicate a high similarity between the populations as well as close genetic relationship among them. We infer that S. dulcificum in Nigeria does not represent a genetically diverse population and this may be accounted for due the plant’s its breeding system which is mainly autogamous. This study further suggests an in-situ form of conservation be set up as immediate rescue conservation procedure for the plant.