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The in vitro cultures of Rindera graeca, a rare endemic plant, were developed as a sustainable source of phenolic acids. Various shoot and root cultures were established and scaled up in a sprinkle bioreactor. A multiplication rate of 7.2 shoots per explant was achieved. HPLC–PDA–ESI–HRMS analysis revealed the presence of rosmarinic acid (RA) and lithospermic acid B (LAB) as the main secondary metabolites in both the shoot and root cultures. The maximum RA (30.0 ± 3.2 mg/g DW) and LAB (49.3 ± 15.5 mg/g DW) yields were determined in root-regenerated shoots. The strongest free radical scavenging activity (87.4 ± 1.1%), according to 2,2-diphenyl-1-picrylhydrazyl-hydrate assay, was noted for roots cultivated in a DCR medium. The highest reducing power (2.3 µM ± 0.4 TE/g DW), determined by the ferric-reducing antioxidant power assay, was noted for shoots cultivated on an SH medium containing 0.5 mg/L 6-benzylaminopurine. A genetic analysis performed using random amplified polymorphic DNA and start codon targeted markers revealed genetic variation of 62.8% to 96.5% among the investigated shoots and roots. This variability reflects the capacity of cultivated shoots and roots to produce phenolic compounds. [ABSTRACT FROM AUTHOR]

Whitefly, Bemisia tabaci (Gennadius) is a cryptic species complex that infests various plant species and act as a vector for many plant viruses all over the world. To understand the B. tabaci cryptic species diversity in Bihar more comprehensively, interhost and interlocation surveys were conducted during the year 2020-2021. The genetic variability among 29 populations (16 interhost and 13 interlocation) was explored using nuclear markers viz. Random Amplified Polymorphic DNA (RAPD), Simple Sequence Repeats (SSR) along with mitochondrial marker, mtCOI. The dissimilarity coefficients of 29 populations clustered in a dendrogram with RAPD and SSR primers showed that interlocation populations were less diverged than the interhost populations. Bayesian phylogenetic analysis of 657 bp mtCOI sequences identified the presence of four cryptic species viz. Asia I, Asia II 1, Asia II 7 and China 3 belonging to two genetic groups (Asia and China) with high variations in interhost unlike in interlocation. Among the four cryptic species, Asia I was the most prevalent in Bihar, establishing 86.20% of all the sequenced samples and Asia II 7 and China 3 were reported for the first time in Bihar region. We believe that the information generated in this study is important from the perspective of identifying cryptic species diversity and to develop long term pest management strategies. [ABSTRACT FROM AUTHOR]

Toxigenic and non-toxigenic V. cholerae strains can be monitored for changes in clones or serogroups, linkages between clinical and environmental isolates, genesis and clonal selection of epidemic strains, and population structure. Also, determining genetic relatedness among V. cholerae strains is critical for determining population genetic structure and evolutionary trends. In collaboration with the Sarawak Government Hospital, the present work was carried out on a total of 16 V. cholerae isolates in order to determine the genetic relatedness or heterogeneity of V. cholerae isolates from a foodborne outbreak among guests who attended a wedding ceremony in Limbang, Sarawak, Malaysia. The random amplified polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC) were conducted to compare and determine strains and trace disease-causing microorganism. The RAPD fingerprinting was conducted using a total of 10-mer oligos 100 nmole random primers (OPAE1 - OPAE20). The primers OPAE7, OPAE10, OPAE14, and OPAE17 were selected because they were the most stable and discriminatory for V. cholerae. The PCR fingerprinting of ERIC-PCR was carried out using primer set of ERIC, ERIC1R (5'-ATGTAAGCTCCTGGGGATTCAC-3'), and ERIC2F (5'-AAGTAAGTGACTGGGGTGAGCG-3'). As a result, the 1 confirmed V. cholerae O1 samples were successfully fingerprinted. Based on the profiling results, the genetic fingerprint of some of the isolates from the clinical and environmental samples had 100% similarity, as indicated by the dendrogram. This indicated that the strains shared the same genetic profile. The smaller the genetic distance, the more homogeneous the strains are. The clinical and environmental strains shared some genetic characteristics. As indicated by the dendrogram, some strains were found to be closely linked to one another, while others were heterogeneous. Therefore, RAPD-PCR and ERIC-PCR produced the highest discrimination index. By combining these typing methods, evaluation of isolates' genetic diversity may be improved. The findings of the present work demonstrated the need for continued surveillance of V. cholera in Sarawak, Malaysia. [ABSTRACT FROM AUTHOR]

The pomegranate (Punica granatum L.) is one of the fruit species with the oldest cultural history. There are many traits to determine the quality of pomegranate fruits. Among them, soft-seeded feature of pomegranate fruit is important trait for the market value of the fruit. For this reason, the demand for pomegranate varieties with soft seeds has been increasing, especially in recent years. In this study, molecular markers associated with seed hardness were developed to distinguish pomegranate cultivars with soft-seeded feature based on genomic DNA at the early stages of the pomegranate breeding process. For this purpose, pomegranate genotypes and/or cultivars from the population involved in reciprocal crosses of hard-seeded Ernar, medium-hard-seeded Hicaznar, and soft-seeded Fellahyemez cultivars were grouped as soft-seeded or hard-seeded. Further, leaf samples were collected from individuals belonging to each group. Then, the genomic DNA was isolated from each plant separately, and equal amount of genomic DNA from individuals with the similar seed hardness were mixed for bulked segregant analysis (BSA). The bulked genomic DNAs of opposite characters were analyzed by polymerase chain reaction (PCR) using random decamer primers to develop random amplified polymorphic DNA (RAPD) markers associated with soft-seeded or hard-seeded pomegranates. A total of three RAPD markers were determined to distinguish the individuals having soft- or hard-seeded pomegranate genotypes and/or cultivars. As a result of the comparison of the DNA sequences of these RAPD markers, insertion-deletions (inDels) primers were designed to developed and validate a PCR assay to distinguish the soft- and hard-seeded pomegranate genotypes/cultivars from each other. The molecular markers developed in this study will enable us to distinguish soft-seeded pomegranate types easily in a short time at the early stages of the pomegranate breeding programs. [ABSTRACT FROM AUTHOR]

Aims: This study was aimed to characterise nine clinical isolates in our culture collection that were categorized as Diutina species based on their molecular genetic profiles. D. rugosa is a species complex comprising four taxa., i.e., D. rugosa sensu stricto, D. pseudorugosa, D. neorugosa and D. mesorugosa. The most commonly used phenotypic identification methods for yeasts often lead to the misidentification of this species complex. Methodology and results: The Diutina isolates were received from two local referral hospitals as pure cultures. Species confirmation was performed using conventional phenotypic methods; CHROMagar and RapID Yeast Plus Kit. To study the inter- and intraspecific relationships among the clinical isolates, ITS region, D1/D2 domain and random amplified polymorphic DNA (RAPD) analyses were performed. The results were further validated using the housekeeping gene sequence similarity technique coupled with pairwise sequence alignment. The results from phenotypic methods results were ambiguous and inconclusive. The sequence analyses of ITS regions and D1/D2 domains revealed that the samples consisted of three yeast species; D. rugosa complex: D. rugosa (n=1), D. mesorugosa (n=6), Candida pararugosa (n=1) and Meyerozyma guilliermondii (n=1). The RAPD analysis with random primers, OPG4, OPG11 and OPA18, demonstrated good banding patterns that could distinguish between the Diutina isolates. The pairwise sequence alignment revealed that the Diutina isolates were genetically similar to D. rugosa ATCC 10571. Conclusion, significance and impact of study: The molecular methods, D1/D2 domain, ITS1 and ITS4 region, and RAPD analyses have proven helpful for accurately identifying the yeasts, especially closely related species; D. rugosa and D. mesorugosa. [ABSTRACT FROM AUTHOR]

Genetic improvement mainly depends on the level of genetic variability present in the population, and the degree of genetic diversity in a population largely determines the rate of genetic advancement. For analyzing genetic diversity and determining cultivar identities, a molecular marker is a useful tool. Using 30 SSR (simple sequence repeat) and 30 RAPD (randomly amplified polymorphic DNA) markers, this study evaluated the genetic divergence of 17 mango cultivars. The effectiveness of the two marker systems was evaluated using their genetic diversity characteristics. Additionally, the effects of SM (simple matching) and Dice similarity coefficients and their effects on mango clustering were evaluated. The findings showed that SSR markers generated 192 alleles, all of which were polymorphic (100%). With RAPD markers, 434 bands were obtained, 361 of which were polymorphic (83%). The average polymorphic information content (PIC) for RAPD and SSR was 0.378 and 0.735, respectively. Using SSR markers resulted in much higher values for other genetic diversity parameters compared to RAPD markers. Furthermore, grouping the genotypes according to the two similarity coefficients without detailed consideration of these coefficients could not influence the study results. The RAPD markers OPA_01, OPM_12 followed by OPO_12 and SSR markers MIAC_4, MIAC_5 followed by mMiCIR_21 were the most informative in terms of describing genetic variability among the cultivars under study; they can be used in further investigations such as genetic mapping or marker-assisted selection. Overall, ‘Zebda’ cultivar was the most diverse of the studied cultivars.

Species identification is the first step in the examination of newly isolated microorganism strains, including the cases when they are intended for application in the development of probiotic preparations or starters for different food products. The thorough identification process of newly isolated strains combines the application of different physiological, biochemical, and molecular genetic methods. The aim of the present study was to identify the species-level lactic acid bacteria (LAB) strains isolated from spontaneously fermented sourdoughs prepared from different flour types: Khorosan wheat, wheat, barley, buckwheat, spelled, spelt, and corn. Both classical phenotypic (cellular morphology characterization, and API 50 CHL) and molecular genetic methods (RAPD, ARDRA-analysis, 16S rDNA sequencing, and species-specific PCR) were applied. It was found that cultures with a short-rod morphology predominated among the 30 sourdough isolates. According to the RAPD profiles obtained, the isolates were divided into nine genotypes corresponding to nine genetically distinct strains. It was determined that individual sourdoughs made with different flour types shared cultures with a common genotype. The analysis of the physiological and biochemical profiles of the LAB isolates performed with the API 50 CHL system divided them into two groups according to their identification: Lactiplantibacillus plantarum (Lp. plantarum) 1 and Levilactobacillus brevis (Lv. brevis) 3. According to the 16S rDNA restriction profile, the LAB isolates showed two profiles corresponding to the Lp. plantarum and Lv. brevis groups. 16S rDNA sequencing and a comparison of the partially read 16S rDNA sequences of the studied isolates confirmed that some of them belonged to the Lv. Brevis species, but did not provide sufficient evidence that the rest of the cultures belonged to the Lp. Plantarum species. The species-specific PCR clearly separated the isolates from the Lp. plantarum group into two groups: isolates of the Lp. plantarum species and isolates of the Lp. paraplantarum species. The summary of the results of the conducted polyphasic taxonomic study determined the investigated LAB strains isolated from spontaneously fermented sourdoughs as representatives of the Lv. brevis, Lp. plantarum ssp. paraplantarum, and Lp. paraplantarum species.

The objective of the study is to determine the genetic relationships and diversity among 20 genotypes of greengram. Ten randomly chosen decamers in total were screened, and all of them were amplified. There were 1071 amplified bands in total, 110 of which were polymorphic, 10.27 percent polymorphism was the average. The band size of the final PCR-amplified products varied from 100 bp to 1000 bp. According to the presence or absence of unique alleles were found in sixteen genotypes of greengram. The values of the Jaccard’s similarity coefficient for RAPD primers ranged from 0.0 to 0.5. The majority of the genotypes were divided into six major clusters based on the dendrogram produced by using the UPGMA method. The genetic distance between genotypes VRM-1 and MH 421was the smallest, while the genetic distance between genotypes CO 1 and CO 8, VBN 3 and CO 2 (99%) was the greatest.Hence, these findings provides valuable information for molecular classification and marker-assisted breeding for crop improvement in the Vigna germplasm pool.

Wine quality is determined by the particular yeast strains prevailing at various stages of fermentation. Therefore, the ability to make an easy, fast, and unambiguous discrimination of yeasts at the strain level is of great importance. Here, the tandem repeat-tRNA (TRtRNA) method with the 5GAC or ISSR-MB primer sets and random amplified polymorphic DNA (RAPD) analysis with (GTG)3, R5, and RF2 oligonucleotides were tested on various non-Saccharomyces wine yeast species. The TRtRNA-PCR employing ISSR-MB showed the highest capacity in discriminating Lachancea thermotolerans and Metschnikowia pulcherrima isolates. RAPD with RF2 was the most efficient method in resolving Starmerella bacillaris isolates, although it produced few polymorphic bands. RAPD with R5 showed the highest capacity to discriminate among the Issatchenkia orientalis, Hanseniaspora guilliermondii, and Pichia anomala isolates. RAPD with either R5 or RF2 exhibited the highest ability to discriminate among the Torulaspora delbrueckii isolates. RAPD with (GTG)3 was the most discriminating method for the H. uvarum isolates. Here we concluded that both TRtRNA-PCR and RAPD-PCR offer rapid means for typing non-Saccharomyces species. However, each method performs better for a given species when paired with a particular primer set. The present results can be useful in wine research for the fast fingerprinting of non-Saccharomyces yeasts.

Centella asiatica (L.) Urb., belonging to the Apiaceae family, grows widely in shady, moist or marshy places. C. asiatica has been used in traditional medical systems in Asia for over 100 years. A better understanding of genetic diversity is essential for its conservation and use. In this study, we performed the genetic diversity analysis for 24 cultivars of Centella asiatica using 24 selected RAPD markers. There are genetic differences among the studied samples. However, the molecular screening with RAPD markers shows that genetic difference (0.0025–0.2194) and genetic diversity indices (%P = 42.02%, h = 0.1272, I = 0.1938) are relatively low. Besides, HUIB_CA06 and HUIB_CA26 (from Thua Thien Hue province) are genetically different from the remaining varieties, which warrants further analysis of these varieties.

The aim of this study was to investigate the prevalence of antibiotic resistance genes in bacteria isolated from Saudi Arabian patients in the Al-Qassim region. We also examine the application of Random Amplified Polymorphic DNA- Polymerase chain reaction (RAPD-PCR) in genetic diversity of the strains in use. In this study, we have used 29 different clinical samples from Al-Qassim hospitals which consists of wound swabs, blood, sputum and urine. Clinical samples were streaked onto sterile Petri dishes containing nutrient agar media using sterile dry swabs. Additionally antibiotic sensitivity tests, micro scan walkway susceptibility tests and extraction of genomic DNA, amplification of RAPD and Inter simple sequence repeat assay were performed in this study. This study results confirmed from the clinical samples were streaked onto sterile Petri dishes containing nutrient agar media using sterile dry swabs. The PCR analysis in ISSR was confirmed that 30–70% of E. coli in Asia and Africa carried ESBLs in the GLASS 2020 study, which is on par with the 59% in VINARES 2016–2017. Comparing VINARES 2016–2017 to other nations, the prevalence of ESBL carriage among K. pneumoniae was 35%. However, RAPD results showed that all employed strains and primers could successfully fingerprint DNA. Between isolated bacterial strains, similarity dendrograms were produced. The technique generated bands with the same intensity as the typical PCR carried out using pure DNA, and it worked for all 29 bacterial strains examined. In conclusion, the results show that all of the isolates have at least one antibiotic-resistance gene, and that the PCR approach is a quick, easy, and reliable way to identify genes.

Bacillus species are among the most researched and frequently applied biocontrol agents. To estimate their potential as environmentally friendly microbial-based products, reliable and rapid plant colonization monitoring methods are essential. We evaluated repetitive element-based (rep) and Random Amplified Polymorphic DNA (RAPD) PCR (Polymerase Chain Reaction) genotyping in a diversity assessment of 251 strains from bulk soil, straw, and manure samples across Serbia, highlighting their discriminative force and the presence of unique bands. RAPD 272, OPG 5, and (GTG)5 primers were most potent in revealing the high diversity of a sizable environmental Bacillus spp. collection. RAPD 272 also amplified a unique band for a proven biocontrol strain, opening the possibility of Sequence Characterized Amplified Region (SCAR) marker design. That will enable colonization studies using the SCAR marker for its specific detection. This study provides a guide for primer selection for diversity and monitoring studies of environmental Bacillus spp. isolates. [ABSTRACT FROM AUTHOR]

An investigation of the phenotypic characteristics of 19 rhizobial strains, isolated from root nodules of different plant legumes grown in the soil of agriculture farms in Ismailia governorate, transpired. Most isolates were creamy or white opaque, mucoid, with a convex elevation, translucent, and smooth margined. Microscopic investigation revealed that all bacterial isolates were rod-shaped and had no positive affinity for Gram-stain. Identifying rhizobial cultures from any bacterial contaminants employed confirmatory tests based on prepared special media, including YMA supplement with Congo red, glucose peptone agar, Kit-lactose agar, and Hoffer's alkaline test. Based on an infectivity test, all isolates proved their ability to reinfect their host. These rhizobial isolates, classified into two categories, included fast and slow-growing rhizobia according to their growth in the YEM medium containing bromothymol blue (BTB). Meanwhile, the assessment of the genetic diversity among these isolates proceeded using ISSR and RAPD markers, which ISSR marker proved a more powerful tool in discriminating among the tested isolates than the RAPD marker. The cluster analysis, with the RAPD marker, classified the isolates into two main groups. The first group included the isolates (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 13), while the second group contained the isolates (12, 14, 15, 16, 17, 18, and 19). Moreover, using ISSR markers also showed a cluster of two main groups with diverse categories; the first cluster included isolates 1 to 11, and the second group contained isolates 12 to 19. [ABSTRACT FROM AUTHOR]

Ceiba pentandra (L.) Gaertn. is a plant whose fruit is used to produce fiber and seeds for biofuel, which has the potential to be developed in critical land because it is adaptive to grow in critical land that is less fertile and lacks water. In West Sumatra, several critical land locations are overgrown by C. pentandra plants. This study aims to determine the genetic variation of C. pentandra in five critical land populations in West Sumatra as the basis for selecting superior seeds for development in critical land. The research was conducted by the descriptive method using molecular data with the molecular marker RAPD (random amplified polymorphic DNA). The results showed that the primers OPA-01, OPA-02, and OPB-10 could detect polymorphisms. Pangkalan Koto Baru in the regency Lima Puluh Kota (H = 0.1212) was the population with the highest intrapopulation genetic variation value. Interpopulation genetic variation (DST = 0.0321) was lower than intrapopulation genetic variation (HS = 0.1021), with a low genetic differentiation value (GST = 0.2392) and a high gene flow value (Nm = 1.5894). The genetic variation of C. pentandra can be used to select the parent in plant breeding programs. [ABSTRACT FROM AUTHOR]

Gymnema inodorum (Lour.) Decne or Chiang Da, the local northern Thai vegetable, is renowned for its anti-diabetic and hypoglycemic properties and is highly sought after by the functional food industry. This research investigated the relationship between the physiological characteristics, genetic variation, and chemical compositions of different Chiang Da accessions. Two commercial lines (COM1 and COM2) and eight local accessions (BAC1-8) of Chiang Da were gathered and maintained in the same study plot for the investigation of their morphological characteristics, genetic variation, and nutritional and phytochemical constituents. From the morphological data, the two commercial lines were clearly distinctive with their closely related leaf structures. Random amplification of the polymorphic illustrated the genetic diversity between the local accessions and commercial strains in a similar pattern as described by their morphology characteristics and proximate contents. The phytochemical characteristics, nonetheless, projected the two commercial lines in separate groups. The outcome of this study could be beneficial toward the selection of the Chiang Da strains with specific traits for plant breeding programs of functional food interest. [ABSTRACT FROM AUTHOR]

The aim of the study was to evaluate the genetic stability of lavender cultivars during long-term clonal micropropagation using RAPD and ISSR markers. The material for the investigation was three lavender (Lavandula angustifolia Mill.) cultivars of Crimean breeding: Vdala, Sineva, and Stepnaya. The biological objects were the original donor plants (grown under controlled conditions) as well as microshoots after six and 16 subcultivations during in vitro propagation. Two RAPD (OPA 10, OPO 13) and five ISSR primers (HB 13, HB 15, ISSR 1, ISSR 2, ISSR 3) were used. Significant differences in the number and length of shoots, the number of nodes on the shoot, and the multiplication index after six and 16 subcultivations for all cultivars were not found. According to the morphology, the microshoots of the three studied cultivars after different periods of cultivation also did not differ from each other. Using seven markers, we identified 62 loci. All primers used in the work were polymorphic (41.7–88.9%), and the amplification products reliably identified lavender cultivars. The length of the amplified fragments varied from 378 to 2177 base pairs. The microshoots obtained using clonal micropropagation after six and 16 subcultivations, were identical in genetic profile to the original lavender cultivars. As a result, the possibility of long-term (at least 16 subcultivations) micropropagation of lavender was shown while maintaining their genetic stability. [ABSTRACT FROM AUTHOR]

Genetic diversity and relationship among 10 indigenous cattle breeds (Sahiwal, Red Sindhi, Cholistani, Dajal, Dhanni, Rojhan, Lohani, Hissar, Hariana and Tharparkar) were analysed using RAPD markers. A total of 80 RAPD primers were used for PCR amplification of DNA from the cattle breeds. The PCR products were analyzed by agarose gel electrophoresis. Scoring of bands were performed to generate dendrogram using unweighted pair group method of arithmetic means (UPGMA). Six hundred and four amplified bands were observed from successful amplifications of 73 primers (average 8.2 bands per prime). Only seven primers amplified no product or unidentifiable (smeared) bands. A total of 68 polymorphic bands were observed, showing 11.25 % polymorphism on average across cattle breeds. Results revealed a low genetic variation among the cattle breeds. A RAPD primer OPF-07 was able to discriminate more than one cattle breed. Phylogenetic analysis showed the relatively high similarity indices among 10 cattle breeds (ranging from 83.85% to 89.62%). The highest similarity was observed between Dhanni and Lohani (89.62%), followed by Sahiwal and Red Sindhi with 89.44% similarity. However, the lowest similarity was observed between Cholistani and Tharparkar (83.85%). The cluster analysis of UPGMA showed two major clusters: Cluster A having Hissar and Hariana (similarity coefficient 0.87) grouping with Tharparkar; and cluster B having Dajal and Rojhan (0.88) grouped together, Dhanni clustered with Lohani while Cholistani grouped with Dajal and Rojhan. The Sahiwal breed clustered with Red Sindhi having a similarity coefficient of 0.89. The findings revealed close genetic relationships among the cattle breeds which need to be refined by an increased sample size of each breed from their respective home tracts. Moreover, further studies are warranted by using latest genomics tools to get insights into the evolution and domestication of indigenous cattle breeds. [ABSTRACT FROM AUTHOR]

As the diversity and characteristics of Trichoderma species are difficult to determine using morphological methods, henceforth molecular tools are crucial. This study utilized random amplified polymorphic DNA (RAPD) technique to investigate the genetic diversity of Trichoderma with sexual phase Hypocrea and to identify similarities and differences in the phylogenetic tree. Nine Iraqi Trichoderma strains (four strains of T. atroviride, one strain of Hypocrea lixii, two strains of T. gamsii and two strains of T. longibriantium) were examined in this research. The genomic DNA of each species was extracted and amplified with each of the five primers. 197 bands were obtained by using five oligodeoxynucleotide primers of which 98.47% were polymorphic and about 1.52% were monomorphic. When primers OPA4, OPA8, and OPA10 were used, the genetic variability was about 100%. Whereas, after using primers OPA7 and OPA5, the obtained genetic variability was 95.7% and 92.6% respectively. Gel images of the RAPD were processed with photo-cap program which detected the bands of isolates based on the ladder. The detected bands were then clustered based on the Jaccard method in Past software that showed T. atroviride, T. gamsii, Hypocrea lixii, and T. longibrachiatum isolates were grouped as clades and lineages. Although all strains belonged to the same species and group in one clade, they differed in size and number of bands. The Jaccard cluster analysis showed that the three isolates of T. atroviride were closely related to each other, while the four isolates of T. atroviride in one cluster were same as Hypocrea lixii, the isolates of T. gamsii and two strains of T. longibrachiatum formed one cluster. Thus, the high reliability of RAPD markers could be applied to identify Trichoderma species and create genetic maps instead of other DNA-based methods which are not only costly but time-consuming too. [ABSTRACT FROM AUTHOR]

Information on the genetic structure of fish is a useful means for optimizing identification of stocks, stock enhancement, breeding programs, management of sustainable yield and preservation genetic diversity. The population genetic structure of African Catfish, Clarias gariepinus (Burchell, 1822) in cultured population and feral populations from Rivers Guzuguzu, Fada and Magaga (Kano State) were investigated using RAPD (Random Amplified Polymorphic DNA). Using a CTAB protocol, genomic DNA was extracted from the caudal peduncle of 157 samples of live specimen collected from each population. Five RAPD primers were used to amplify different loci on the extracted genomic DNA by Polymerase Chain Reaction (PCR) and the resultant DNA fragments were analyzed on agarose gel. A total of 406 reproducible bands were obtained in four populations for five primers. The dendrogram separated the four C. gariepinus populations into two distinct clades, Guzuguzu and Fada populations being in one clade, while Magaga and Cultured populations belonged to the other clade. The results based on RAPD-PCR profile ranged from 0.012 in Magaga/Cultured to 0.089 in Guzuguzu/Cultured. The genetic identity of C. gariepinus from four populations also ranged between 0.885 in Magaga/Guzguzu to 0.998 in Magaga/Cultured. The Nei's genetic distance and identity also confirmed the above information with the following ranges: 0.002 in Fada/Cultured to 0.102 in Magaga/Guzuguzu, and 0.903 in Magaga/Guzuguzu to 1.000 in Magaga/Cultured respectively. In conclusion, the genetic diversity and allele richness of the feral and domestic fish populations were comparable. [ABSTRACT FROM AUTHOR]

Zingiberaceae is widely distributed in Malaysia, with 750 species and 31 genera. This family comprises a different number of subfamilies and genera according to different taxonomic classification methods — classical taxonomy: one subfamily and four tribes vs molecular taxonomy: four subfamilies and six tribes. However, the taxonomic classification of Zingiberaceae is still debated, especially the classical taxonomy. It is due to some Zingiberaceae species showing cryptic morphologies that make it difficult to classify them through classical taxonomy, which refers to the unique morphological characteristics of a tribe/species. Therefore, accurate taxonomic classification is required by using a molecular approach. In this study, 20 selected species of Zingiberaceae collected from the Agricultural Conservatory Park, Institute of Bioscience (IBS), Universiti Putra Malaysia (UPM) were taxonomically classified using a molecular method with the help of three random amplified polymorphic DNA (RAPD) and three inter simple sequence repeat (ISSR) markers until the tribe level. The combined RAPD and ISSR unweighted pair group method with arithmetic mean (UPGMA) phylogenetic tree was comparable to Zingiberaceae’s current molecular and classical taxonomy. The 20 selected species were grouped into three tribes (Alpinieae, Zingiberaceae, and Globbeae). This finding has contributed additional biological information to better manage the 20 Zingiberaceae species in the Agricultural Conservatory Park, IBS UPM. Further studies are needed to explore the genetic diversities and properties of Zingiberaceae species. [ABSTRACT FROM AUTHOR]

Background: Multidrug-resistant bacteria represent a potential threat to patients in healthcare facilities, with higher abundance and more profound consequences in the developing world. Controlling the spread of these bacteria requires timely identification of clonally related isolates that are cross transmitted from one patient to another within healthcare facilities. The relatively high cost of sequencing-based typing methods hinders their routine use in low-income countries. This highlights the need for an alternative easily applicable molecular typing method that can be routinely used in laboratories with basic equipment and low financial resources, for epidemiologic investigation of outbreaks. Aim and methods: This study aimed to evaluate molecular typing of 30 multidrug-resistant (MDR) Klebsiella pneumoniae (K. pneumoniae) isolates, that were isolated in a previous hospital outbreak and were of previously identified clonal relatedness, by 2 non-sequencing-based typing methods: Plasmid Profile Analysis and Random Amplified Polymorphic DNA (RAPD). The obtained banding patterns were converted into binary data and phylogenetically analyzed by DARwin 6.0.21 software. Results: There was a statistically significant fair agreement between the 2 methods in grouping of the isolates into distinct clusters (Kappa=.23, p=.025). Both methods proved to be of high typeability of near 100% and discriminatory power of more than 0.9. Both methods fulfilled the performance and convenience criteria evaluated. Conclusion: Any of the two methods may be applicable for routine investigation of outbreaks by MDR bacteria in the developing world, when more precise molecular typing methods are not applicable for shortage of resources. [ABSTRACT FROM AUTHOR]

Abstract Marine seaweeds are known to have a potential role against microbial and pesticidal activities. Ulva lactuca, a green macroalgae extract analysed through gas chromatography mass spectrometry reveals 31 compounds. Resistance of mosquito vectors to synthetic insecticides remains a major problem. Discovering and applying natural agents to act against disease vectors is challenging. The activities of the extract and nano‐fabricated green synthesised silver nanoparticles were checked for use against Aedes aegypti and Culex pipiens. The crude extract and synthesised silver nanoparticles exhibited a notable larvicidal effect, and very effective inhibition of pupal and adult emergence. Inhibition of adult emergence of Ae.aegypti was 97.7% and in Cu.pipiens, it was 93.3%. Our genotypic study of Deoxyribonucleic acid from treated larvae utilising random primers MA‐09, MA‐12 and MA‐26 revealed damaged nucleotide sequences when compared with the controls. The antimicrobial activity of both the extract and green synthesised nanomaterials showed prominent activity against pathogenic drug resistant bacteria. Our results contribute to further development of eco‐friendly insecticides with lower cost of preparation. This could further contribute to further research helping future generations to be free from these deadly disease‐causing vectors and pathogenic microbes.

Abstract The present work was to study the genetic variability between the major carps Labeo rohita and Cirrhinus mrigala and their hybrids of L. rohita (male♂) and C. mrigala (female♀). Genetic variability was studied by employing RAPD molecular markers. 25 samples of each target species having different sizes with the same age group for the determination of interspecific variation were collected. The morphometric parameters such as body weight, total length, tail length, and lengths of dorsal and anal fins of each individual were recorded and results showed that wet body weight, total length, dorsal fin, anal fin, and tail fin length are positively correlated and then the DNA was extracted using the inorganic salt-based method and conformed by Gel electrophoresis. Twenty-four arbitrary decamer primers were used to get species-specific RAPD analysis Distinct and highly reproducible RAPD profiles with significant genetic variability was detected among species. Only five primers showed amplification. The RAPAD primer OPB-05 produced a total of seven bands out of these 5 monomorphic and 2 polymorphic, so in this case, the percentage polymorphism was 28.57%. The Hybrid show more than a 50% difference from the Labeo rohita. This shows that the Hybrid more resembles C.mrigala. Phylogenetic analysis demonstrated that hybrid (L. rohita ♂ X Cirrhinus mrigala ♀) is the closest to C. mrigala and the farthest from L. rohita. Overall data are presented concerning the applications of RAPD markers for hybrid identification, genetic diversity assessment, and studying taxonomic relationships at a molecular level.

The relationship among six Jordanian almond landraces was studied using morphological traits and RAPD analyses. Across six almond landraces studied, nut shape, kernel size, nut length, nut size, and shell length That showed a high level of variation (CV>30%), while kernel length, shell width, internodes length, and nut width showed comparatively low values (CV<20%). Principal component analysis showed that the first five components explained all morphological variation among the landraces investigated. Kernel and nut traits were predominant in the first three components contributing to most of the total variation that existed among landraces. Euclidean distance was used to construct clusters from morphological data which allocated individuals into two main groups with a distance ranging from 5.5 to 10.14. Hajari, Hami Hallo, and Mukhmaly with small fruit sizes composed one main cluster, while the other three landraces (Oga, Fark, and Abu Dabos) with large fruit sizes composed the other main cluster. Out of 62 pre-screened RAPD primers, 12 with reproducible bands and maximum polymorphism were selected for diversity analysis. Seventy-one bands were scored with 28 of them being the polymorphic. Average value of polymorphism/primer ranged from 20% to 74.2%. Nei's genetic distance coefficient ranged from 0.5 to 0.85 with an average of 0.70. Molecular analysis revealed inconsistent separation among the landraces compared with that based on morphological traits. Although landraces found during the screening in the Ajloun area are limited in number, but considerable variation was observed both at morphological and DNA levels indicating that Jordanian almond landraces are rich and valuable genetic materials for almond improvement. [ABSTRACT FROM AUTHOR]

Objective: The Anopheles maculupennis complex is known to be a vector of malaria in Türkiye. Anopheles sacharovi is a member of the Anopheles maculipennis complex and is a major vector of malaria in Europe and the Middle East capable of transmitting both Plasmodium vivax and P. falciparum. The genetic diversity and population structure of the six populations of Anopheles sacharovi from Türkiye are studied here using the random amplified polymorphic DNA (RAPD) marker. Materials and Methods: Anopheles sacharovi samples were collected from Muğla, Aydın, İzmir, Çanakkale, Balıkesir, and Denizli. Total DNA was extracted using the Lifton method, and a total of 120 individuals were screened using six RAPD primers. Results: A total of 300 loci were observed in the six primers. Very close genetic diversity was observed in the studied populations. The number of alleles (na) observed for all populations was 1.55 ± 0.49, and the mean number of effective alleles (ne) was 1.23 ± 0.31. The ratio of polymorphic loci for all populations ranged between 50.33%-60.33%, and Nei's genetic diversity (h) ranged between 0.1253-0.1576. The gene flow level (Nm) was 2.08, and the genetic differentiation value (Gst) was 0.19. The conclusions from the unweighted pair group arithmetic mean analysis (UPGMA) was inclined to be homogeneous on the whole, with the İzmir population being clearly separated from the rest. Conclusion: Regional environmental conditions such as human-mediated transport, agricultural implementations, and discrepancies in vector control strategies might be considered agents in forming the genetic structure of this species in İzmir. Understanding the gene flow rates and phylogenetic relationships between vector species are very important for applying sustainable and effective pest management. This study ensures helpful knowledge for better understanding the population genetic structure of An. sacharovi populations in Türkiye. [ABSTRACT FROM AUTHOR]

Melon (Cucumis melo) is one of the most important cultivated cucurbits. In this project, the genetic variability among 14 accessions of melon collected by the authors has been studied using 23 anatomical characteristics across 2 consecutive years as well as 146 random amplified polymorphic DNA (RAPD) markers. Combined analysis of variance revealed significant effects on genotype and year × genotype effects. Among examined anatomical characters, the highest CV% values across two years belonged to fruit store at room temperature (88.96%, 103.6%) and total fruit weight (42.2%, 40.00%), while the lowest CV% values were observed in flower petal width (0.31%, 0.21%), flower petal length (0.28%, 0.49%), and peduncle length (0.40%, 0.19%). Classification of melon accessions based on anatomical characteristics using the Ward method produced three groups. The highest Jaccard's similarity coefficient (0.76) was observed between accessions "Atashi koluche" and "Atashi miyaneh" and the lowest value (0.49) was found between accessions "Bakermellon" and "Mashhadi" with the mean value of 0.59. In this study, OPA06 and OPB13 primers possessed greater efficiency in the genetic evaluation of the studied germplasm. Analysis of population structure, which imply on the existence of admixture in the studied melon germplasm, revealing three subpopulations. Accession "Sabzevari" identified as mixed subgroups. From the breeder's view, the introduced heterotic groups can be utilized in parental selection for the construction of mapping population and the identified population structure can prevent any false positive output in marker-trait association studies of melon. [ABSTRACT FROM AUTHOR]

Micropropagation of five grapevine cultivars (Fetească albă, Cabernet Sauvignon, Merlot, Riesling Italian and Traminer roz) was performed using axillary shoots as a source of explant from field-grown plants. Shoot initiation and proliferation were performed on Murashige and Skoog (MS) medium with 0.5 mg/L NAA and BAP (0.5, 1.0 and 2.5 BAP mg/L). Subsequently, proliferated shoots from V3 were used for grapevine callus induction. The callus was then subcultured on fresh MS medium supplemented with 1.0 mg/L BAP +5.0 (V4) and 10 mg/l NAA (V5). Embryogenic calli grown on V5 were transferred and subcultured on MS medium supplemented with 0.5 mg/L NAA and three different concentrations of TDZ (0.5; 1.0; 2.0 mg/L) for shoot regeneration. RAPD assay was performed after the 12th subculture using in vitro-raised plants from V6 culture medium (0.5 mg/L NAA+0.5 mg/L TDZ) and the mother plants grown in the field and used as control. The results of this study reveal that the highest number of somaclones (5) was regenerated from the Merlot variety and the lowest number (1) from the Feteasca albă and Traminer roz varieties. For practice, the induction of somaclonal variation and the selection of valuable somaclones are important goals for future grapevine breeding programs. [ABSTRACT FROM AUTHOR]

The aim of this work is to isolate the microbes possessing arsenic degrading property from contaminated soil, collected from Cauvery River at Pallipalayam, Erode District. Six microbial strains were grown well in 40Mm sodium arsenate as a sole carbon source amended M9 minimal media. Based on the zone of clearance, three microbial strains were found to be potent arsenic degrading microbes and they are identified as Bacillus spp., Staphylococcus spp., and Pseudomonas spp. They may potentially be used in the bioremediation of arsenic and other contaminants. It infers that the presence of arsenate reductase (ArcC) gene in three of the microbial strain and they were taken for further studies. Genomic DNA isolation protocol was standardized and DNA isolation was performed. ArcC gene-specific primers were designed using Primer3 bioinformatics tool. Genetic diversity among the strains was studied by RAPD analysis using four different primers. Dendrogram was constructed using Unweighted Pair Group using Arithmetic Averages and NJ tools. The presence of genetic diversity was observed from the analysis. Polymerase chain reaction amplification and sequencing of amplified gene products are to be done in future. Background: The aim of this work is to isolate the microbes possessing arsenic degrading property from contaminated soil, collected from Cauvery River at Pallipalayam, Erode District. Six microbial strains were grown well in 40Mm sodium arsenate as a sole carbon source amended M9 minimal media. Based on the zone of clearance, three microbial strains were found to be potent arsenic degrading microbes and they are identified as Bacillus spp., Staphylococcus spp., and Pseudomonas spp. They may potentially be used in the bioremediation of arsenic and other contaminants. It infers that the presence of arsenate reductase (ArcC) gene in three of the microbial strain and they were taken for further studies. Genomic DNA isolation protocol was standardized and DNA isolation was performed. ArcC gene-specific primers were designed using Primer3 bioinformatics tool. Genetic diversity among the strains was studied by RAPD analysis using four different primers. Dendrogram was constructed using Unweighted Pair Group using Arithmetic Averages and NJ tools.The presence of genetic diversity was observed from the analysis. Polymerase chain reaction amplification and sequencing of amplified gene products are to be done in future. Methods: The soil sample was collected from Cauvery River, Pallipalayam. Arsenate, arsenic bioremediation, arsenic reducing gene, RAPD, and genetic diversity were used. Results: With the dilution concentrations, 10−5 and 10−6 microbial population was obtained in M9 minimal media. From the pure colonies of isolates, TA1, TA2, TA4, and TA5 genomic DNA was extracted using the protocol mentioned above. The culture was inoculated in LB broth and kept in incubation at 37°C for overnight. From overnight culture, genomic DNA was extracted. RAPD analysis for the isolates was performed using four different random primers namely RBA-1, RBA-4, RBA-5, and RBA-6. Conclusion: Three of the isolates designated as TA2, TA4, and TA5 were found to be potent arsenic degarding microbes. They are able to degrade sodium arsenate of about 40mM. It infers that they can be potentially used in bioremediation of arsenic. Isolation of ArcC gene from the isolates is in progress. Sequencing will reveal the nature of amplified products. If the amplified genes are cloned and mass production of ArcC gene could be obtained. [ABSTRACT FROM AUTHOR]

Atriplex canescens is a polymorphic species that can be deciduous or evergreen depending on the climate. Having an extensive root system, it controls soil erosion very well and is resistant to drought. The present study examined the morphological and genetic diversity of 13 Iranian populations. Multivariate statistical analyses were performed on 25 morphological characteristics (quantitative and qualitative). To assess genetic diversity, eight primers were applied, resulting in 42 bands. With the ANOVA test, all quantitative morphological characteristics among the studied populations were significantly different. Similarly, the AMOVA test revealed that the genetic content of the studied populations was different. Genetic distance and geographic distance were significantly correlated in Mantel's test. Based on morphological studies and the effects of environmental factors (latitude, longitude, and altitude) on genetic structure, different genetic branches have been proposed for A. canescens. [ABSTRACT FROM AUTHOR]

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Background: Soybean is one of the most important commodities around the globe. Increasing soybean production absolutely continue to be pursued because soybean consumption increased steadily. One way that can be done is assembly technology of new superior varieties that have better and adaptive properties. The aim of this study was to determine molecular diversity of soybean mutants lines resulting from gamma irradiation in M6 generation. Methods: Molecular analysis with RAPD markers was carried out in the Tropical Balitbu Laboratory Sumatera Barat, Indonesia in November 2021. The tested genotypes consisted of 8 genotypes, namely Kipas Putih variety (elderly), Anjasmoro variety (comparison), M.1.1.3, M.5.2.1, M.5.2.3 l, M.1.1.8, M.1.1.9 and M.1.1.17 mutant lines. The primers used were RAPD 2 (5'-GTTTCGCTCC-3'), RAPD 3 (5'-GTAGACCCGT-3'), RAPD 4 (5'-AAGAGCCCGT-3') and RAPD 5 (5'-AACGCGCAAC-3'). Result: The results of the RAPD analysis showed that there were differences in size of amplicon between the mutants and the parents, namely the RAPD 2 primer (1400 bp band) and the RAPD 5 primer (550 bp and 1000 bp band). The parents had a pattern of DNA bands but in the M.5.2.3 and M.1.1.9 mutants the DNA band was absent. The parent RAPD 5 primers (1000 bp, 550 bp) had a DNA banding pattern but in the M.1.1.3 and M.1.1.8 mutants there was no DNA banding pattern. This indicated that the M.1.1.3, M.5.2.3, M.1.1.8 and M.1.1.9 mutants had different genetic diversity from their parent (Kipas Putih variety). [ABSTRACT FROM AUTHOR]

Background: Arecanut (Areca catechu L.) is an important plantation crop of India belongs to the family Arecaceae. As it is a highly cross-pollinated crop there is a significant difference among the genotypes. Morphological traits used for determining diversity and relationships among genotypes or variety are prone to environmental influences. Thus, the usefulness of molecular markers has been investigated as a means of characterizing and discriminating difference more precisely. Therefore, the present study was carried out to assess the genetic relationship and diversity among the arecanut genotypes by employing RAPD, ISSR and SSR markers. Methods: A total of 11 genotypes are being grown in University of Agricultural and Horticultural Sciences (UAHS), Shivamogga, Karnataka, India, which includes both local and exotic collections. These 11 genotypes were evaluated for genetic variations using molecular markers viz., random amplified polymorphic DNA (RAPD), inter-simple sequence repeats (ISSR) and simple sequence repeats (SSR). Result: All the markers studied showed significant polymorphism among the genotypes. Jaccard's genetic similarity coefficient values of RAPD, ISSR and SSR markers were found in the range of 0.56-0.76, 0.55-0.74 and 0.52-0.91. The higher similarity was observed between the genotypes Mohitnagar and Mangala, as well as Theerthahalli Local and Hirehalli Dwarf. The genotypes Maidan Local and Cameroon were placed in very distinct clusters, showing that they are having very distinct traits. The dendrogram prepared using SSR data separately and combining data of RAPD, ISSR, SSR markers showed similar results. Therefore, employing SSR markers for further studies will be more useful for identifying germplasm for varietal improvement programmes. [ABSTRACT FROM AUTHOR]

The objective this study was to determine the markers related to plant height using PCR-RAPD (polymerase chain reaction-random amplified polymorphic DNA) method. As a plant material, dwarf and normal pomegranate parents and their hybrids were used in hybridization breeding and self-breeding methods located in ADU Faculty of Agriculture, Department of Horticulture. In the plant height measurements made in 2016; the height of the average, shortest, and longest plants, respectively, was in the hybridization of P. nana × P. granatum, out of 54 plants, 55 cm, 38 cm, and 79 cm; in P. nana selfing, out of 50 plants, 41 cm, 26 cm, and 57 cm; in P. granatum selfing, out of six plants 37 cm, 30 cm, and 42 cm. Female parent (P. nana) was 85 cm and male parent (P. granatum) was 339 cm. In the plant height measurements made in 2020; the height of the average, shortest, and longest plants, respectively, was in the hybridization of P. nana × P. granatum, out of 52 plants, 184 cm, 129 cm, and 250 cm; in P. nana selfing, out of 44 plants, 113 cm, 59 cm, and 154 cm; in P. granatum selfing, out of two plants 173 cm, 172 cm, and 174 cm; and female parent was 110 cm and male parent was 396 cm. DNA was extracted from the leaves of a total of 16 samples together with the parents using 2016 data. In order to test the RAPD primers, BSA was created using equal amounts of DNA of two different groups corresponding to the height of the hybrid plants. Among the 120 RAPD primers tested, the persistent bands gave polymorphisms in the bulks assessed in the further study which each individual and both parents tested for polymorphisms. The 650 bp band obtained from the OPM07 primer was not observed in the female parent, dwarf pomegranate bulk and self-pollinated dwarf plants, but that was observed in the male parent, normal height pomegranate bulk and the four normal height pomegranate hybrids. As a result of this study, OPM07-650 RAPD marker was able to determine 57% of the polymorphism in plant height characteristic in pomegranate. [ABSTRACT FROM AUTHOR]

• Calligonum comosum of Polygonaceae is one of the most economically important resources of the Tunisian desert. • To better appreciate and further refine the diversity analysis, we have opted for two molecular analysis techniques, RAPD and ISSR, and one technique of biochemical analysis. • HPLC/MS screened methanol extracts of identified phenolic compounds. • Biochemical and molecular polymorphism could provide information regarding genetic background and distribution of diversity of C. comosum. • Genetic diversity help to plan an effective conservation strategy for the species. The species Calligonum comosum of Polygonaceae is one of the most economically important resources of the Tunisian desert, playing an important role in the lives of the local populace. This plant has been reported to possess various chemical constituents and many medicinal effects. A great range of genetic diversity could be seen in diverse populations of this species spread of south Tunisian. To better appreciate and further refine the diversity analysis, we have opted for two molecular analysis techniques, RAPD and ISSR, and one technique of biochemical analysis. The extent of genetic variability was estimated in a set of seven Calligonum genotypes collected in seven Tunisian arid regions using 7 RAPD (random amplified polymorphic DNA) and 7 ISSR (inter simple sequence repeats) markers. ISSR and RAPD markers revealed 67% and 72, 85% polymorphic bands, respectively. The relative efficiencies of these marker systems are compared using various parameters such as effective multiplexing ratio (EMR), marker index (MI), and the polymorphic information content (PIC). RAPD seems to be the most informative and efficient system. A significant correlation is observed between the Jaccard dissimilarity matrices based on the ISSR and RAPD markers (r = 0.66). HPLC/MS screened methanol extracts of the seven genotypes and the identified phenolic compounds were further quantified. Significant differences were observed among the various studied genotypes regarding their phenolic compound content. The overall results showed that C.comosum is an excellent source of diverse bioactive molecules, which could be an interesting material for industrial purposes. The biochemical markers do not correlate with the ISSR and RAPD markers. These findings are important for a better understanding of the adaptive strategy of C. comosum in south Tunisia. They will be useful for conservation managers to work out an effective strategy to protect this important species. [ABSTRACT FROM AUTHOR]

Soybean (Glycine max L.) is an important crop plant which contains a high amount of oil and protein. The main goal of the work was to use 13 RAPD (Random Amplified Polymorphic DNA) markers to study genetic polymorphism in a set of 28 soybean genotypes and to construct a dendrogram from the obtained results, based on which we will recommend genotypes for further breeding. In total, using 13 RAPD markers, we determined 108 fragments in a set of 28 soybean genotypes with an average number of 8.31 fragments per genotype. The number of fragments varied from 6 (OPB-08, OPE-07) to 12 (SIGMA-D-01). Of the total number of 108 fragments, 70 were polymorphic with an average number of 5.38 polymorphic fragments per genotype. The number of polymorphic fragments varied from 3 (OPE-07, OPF-14) to 9 (SIGMA-D-01). The average percentage of fragment polymorphism was 63.68% and ranged from 42.86% (OPF-14) to 83.33% (OPB-08). Diversity index (DI) values ranged from 0.710 (OPA-03) to 0.846 (OPD-08) with an average value of 0.763. A dendrogram prepared based on hierarchical cluster analysis using UPGMA algorithm separated 28 soybean genotypes into four clusters. The applied RAPD markers proved to be a suitable tool for detection of the genetic diversity of soybean genotypes. Based on the constructed dendrogram, it is possible to select suitable varieties for marker-assisted breeding. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

White clover (Trifolium repens L.) is a perennial legume with various traits; it grows in temperate regions and has high potential as an ornamental plant. In the present study, four parental varieties (Ladino, Green Ice, William, and Dragon's Blood) of white clover were classified based on 11 morphological traits [plant form, leaf shape, leaf edge shape, upper lamina color, lower lamina color, white V mark, midrib color, leaf patterns (front and back), flower color, and peduncle color]. Based on these traits, two hybrids were selected as follows: Ladino × Green Ice [hereafter referred to as Jeju Hapiness-1 (JH-1)] and William × Dragon's Blood (JH-5). In addition, random amplified polymorphic DNA (RAPD) analysis was performed to confirm the molecular biological relationship between the parental varieties and hybrids. We confirmed to contain the polymorphic bands of the parental varieties at OPC13, OPD6, and OPG4 operon primers from JH-1, and at OPE10, OPG12, and OPH11 from JH-5. Further, the RAPD patterns from the hybrids were compared with the four parental varieties. A distinguishable RAPD pattern was identified in OPC13 and OPG4 from JH-1 and in OPG12 and OPH11 from JH-5. In addition, polymorphic DNA fragments in which the genetic relationship between the parental varieties and hybrids was confirmed were converted into sequence characterized amplified region (SCAR) markers. The morphological classification criteria, RAPD pattern, and SCAR marker are all considered useful for the classification and breeding of white clover and protecting varieties developed. [ABSTRACT FROM AUTHOR]

Lipaphis erysimi (Lipaphis erysimi erysimi and Lipaphis erysimi pseudobrassicae) group, often known as the mustard or turnip aphid, is one of the most devastating specialist aphid of Rapeseed-mustard, causing yield losses of 9 to 96%. This study was carried out with a goal of precisely identifying the dominant specialist aphid of Rapeseed-mustard and to address the issues such as genetic differentiation from different Rapeseed-mustard growing regions covering all the six zones of India. The dendrogram, Principal component analysis (PCA) and phylogenetic relationships revealed that populations of Jorhat (North Eastern India) were highly diverged. Phylogenetic relationship produced by Tamura 3-parameter with Gamma distribution (T92 + G) model using mtCOI demonstrated the presence of L. e. pseudobrassicae in all of the surveyed locations in India including Chintapalli (Andhra Pradesh) a Southern India, where it was reported for the first time. Neutrality tests viz. Tajima and Fisher test supported neutral theory of evolution. Furthermore, our findings in mapping the distribution of L. e. pseudobrassicae in various locations will pave the way for future research and the development of area-wide management tactics for L. e. pseudobrassicae in India. [ABSTRACT FROM AUTHOR]

A field experiment was carried out on the left side of the Euphrates River, Ramadi district, the center of Anbar Governorate. Five genotypes of barley were used (Buhooth-244, Amal, Samir, BKL88-38 and BKL88-216), which were introduced into reciprocal hybridization in one direction in the winter season of 2019-2020 to produce 10 single hybrids. The seeds of parents and hybrids resulting from half-reciprocal cross were planted in the winter season of 2020-2021 according to the randomized complete block design (RCBD) at three replications, in order to estimate the Heterosis. The BKL88-38 x Buhooth-244 hybrid was significantly superior by giving the highest grain yield (50.06 g), grain weight (50.7 g), number of spikes (384.4 spike m-2), while the Amal x BKL88-38 hybrid was distinguished by giving the highest harvest index (43.82%), whereas the BKL88-38 x Buhooth-244 hybrid was distinguished by giving a highest mean of Heterosis of the 1000-grain weight, grain yield and biological yield (25.50, 28.88 and 14.84%), respectively. Randomly Amplified Polymorphic DNA (RAPD) technique was used by using 10 primers. The highest genetic divergence was 0.70 between Amal and BKL88-38 genotypes and Buhooth-244 and BKL88-38 genotypes. The ten primers gave 233 bands with an average of 23.3 band per primer. The number of identical bands was 24, the dissimilarity was 209 and the percentage of dissimilar bands ranged between 58.62-100%. [ABSTRACT FROM AUTHOR]

Date palm is propagated by the offshoots, the number of which is limited. Therefore, date palms may be produced through the use of tissue culture of shoot tips and axillary shoot meristems. Date palm (Phoenix dactylifera L.) micropropagation still faces many problems, such as reduced growth and development of callus, low multiplication efficiency, and low rooting rate. Here, we established an efficient plant-regeneration system for (Phoenix dactylifera L.) cv. Ashgar by tissue culture. Murashige and Skoog medium containing 6-benzyladenine (BA) (2.0 and 5.0 mg L−1) and a-naphthaleneacetic acid (NAA) or phenylacetic acid (PAA) (0.05–2.00 mg L−1) was used to initiate shoot formation from callus tissues. The maximum number of shoots per jar was produced on a medium containing 5.0 mgL−1 BA and 0.5 mg L−1 PAA, while the medium supplemented with 2.0 mgL−1 BA in combination with 0.05 m L−1 PAA gave the highest callus induction (+++). A decrease in browning percentage was observed in the tissue-cultured media supplied with BA in combination with NAA or PAA compared to those provided with BA alone. In comparison with other treatments, the total amount of phenolic compounds was significantly reduced to 0.45 mg g−1 in buds cultured in the media supplemented with 5.0 mg L−1 BA and 0.5 mg L−1 PAA. The genetic stability of this protocol was confirmed by DNA-based fingerprinting technique RAPD. The RAPD banding patterns showed no variation among the tissue culture-derived plants tested. The in vitro micropropagation protocol reported herein could be served production of genetically stable date palm plants. [ABSTRACT FROM AUTHOR]