Reproducible Bands Research Articles

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

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

Unveiling Genetic Diversity Among Bacterial Isolates Using SCoT Markers.

This study aimed to unveil the genetic diversity among 47 bacterial isolates from various species using start codon targeted (SCoT) markers. Six SCoT primers yielded 219 reproducible bands, with 89.04% exhibiting polymorphism. The amplification process generated 28 to 50 fragments per primer, with an average of 36.50. Genetic diversity was quantified using polymorphic information content (PIC) values ranging from 0.11 to 0.14, with SCoT32 showing the highest PIC (0.14) and SCoT23 the lowest (0.11). The resolving power (RP) index, used to assess primer discriminatory power, varied significantly, with SCoT23 demonstrating the highest RP (6.00) and SCoT29 the lowest (4.51). Comparative analysis with conventional markers like M13 and (GTG)5 revealed that certain SCoT primers exhibited superior PIC values, which indicates enhanced utility for interspecies differentiation. The high discrimination level achieved by SCoT primers underscores their effectiveness in genetic differentiation and biodiversity assessment within bacterial populations. This research highlights SCoT markers as powerful tools for microbial genetic studies, which offers valuable insights into bacterial diversity and provides a robust methodological framework for future investigations aimed at elucidating genetic variation and improving species identification. The application of SCoT markers represents a significant advancement in molecular techniques for bacterial characterization and phylogenetic analysis, demonstrating their potential to enhance our understanding of microbial genetics and evolution.

Genetic analysis of banana cultivars and fidelity testing in somatic embryo derived Ney poovan (Elakki bale) using SRAP (sequence related amplification polymorphism).

Abstract Bananas and plantains are a major staple food in many countries. In the current study, the genetic variability of ten banana cultivars (i.e., Grand naine, Monthan, Nendran, Ney poovan, Rasthali, Robusta, Red banana, Kadali, Musa balbisiana , Calcutta 4) were analyzed using SRAP. SRAP amplification resulted in a total of 6028 strong, clear, and reproducible bands from 10 cultivars using 13 forward and 16 reverse SRAP primers (208 SRAP primer combinations), Of 6028 bands 96.77% were polymorphic and 3.221% were monomorphism. In addition, the fidelity testing of plants derived from somatic embryo was carried out. The 24 SRAP primers were employed for DNA analysis which produced a total of 153 clear, distinct and reproducible amplicons of variable size. Out of 24 SRAP primers, 9 primers produced amplification patterns which are identical between the mother plants and plants derived from somatic embryogenesis. Cluster analysis depicted more than 95% genetic similarity between the somatic embryogenesis derived plants and the mother plants indicating a high degree of genetic fidelity.

Carbon-Related Quantum Emitter in Hexagonal Boron Nitride with Homogeneous Energy and 3-Fold Polarization.

Most hexagonal boron nitride (hBN) single-photon emitters (SPEs) studied to date suffer from variable emission energy and unpredictable polarization, two crucial obstacles to their application in quantum technologies. Here, we report an SPE in hBN with an energy of 2.2444 ± 0.0013 eV created via carbon implantation that exhibits a small inhomogeneity of the emission energy. Polarization-resolved measurements reveal aligned absorption and emission dipole orientations with a 3-fold distribution, which follows the crystal symmetry. Photoluminescence excitation (PLE) spectroscopy results show the predictability of polarization is associated with a reproducible PLE band, in contrast with the non-reproducible bands found in previous hBN SPE species. Photon correlation measurements are consistent with a three-level model with weak coupling to a shelving state. Our ab initio excited-state calculations shed light on the atomic origin of this SPE defect, which consists of a pair of substitutional carbon atoms located at boron and nitrogen sites separated by a hexagonal unit cell.

DNA barcodes, ISSR, RAPD and SCAR markers as potential quality control tools for molecular authentication of black and white mulberry

Morus species, especially black mulberry (Morus nigra L.), are well known for their nutritive and economic values. Black mulberry is often a subject for deliberate and undeliberate substitution with the morphologically similar white mulberry (Morus alba L.). In the current study, the genetic diversity of Morus nigra L. and Morus alba L. was assessed using different genetic profiling and fingerprinting techniques namely namely DNA barcoding, Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR). DNA barcoding was implemented using 2 plastid loci (matK, and rbcL-1) and 2 nuclear spacers (ITS and ITS2) aiming at accurate authentication of the studied species. DNA barcoding, targeting the selected loci succeeded to identify and authenticate the studies Morus species but failed to differentiate them, owing to their close genetic resemblance. Out of 52 screened RAPD primers, 27 were able to produce reproducible and clear profiles with 115 amplified fragments of which 37 were polymorphic accounting for 32.17% polymorphism. The number of amplified bands varied from 2 to 7 ranging in size from 100 to 700 bp. Moreover, 55 ISSR primers were evaluated, 108 fragments were produced, using 20 primers, of which 28 were polymorphic accounting for 25.92% polymorphism with size range of 200–800 bp. Sequence Characterized Amplified Region (SCAR) marker was developed based on reproducible banding pattern obtained by RAPD primer (OPG-03), for the purpose of accelerating the initial screening of Morus batches and enabling the preventative rejection of adulterated samples. The polymorphic band obtained was sequenced and specific primers were designed to develop a SCAR marker of approximately 558 bp exclusively for M. nigra, allowing detection of down to 1 ng, of M. nigra. The SCAR marker showed 100% success in differentiating authentic M. nigra and M. alba samples. In addition, the application of the developed SCAR marker was extended to distinguish M. nigra from its four common adulterants namely Ficus carica L., Fragaria vesca L., Ficus racemosa L. and Vitis vinifera L.. The proposed protocol was successfully implemented for the evaluation of 47 commercial M. nigra samples collected from different Egyptian locations.

Molecular phylogeny of plant pathogenic fungi based on start codon targeted (SCoT) polymorphism.

A number of molecular marker systems have been developed to assess genetic diversity, carry out phylogenetic analysis, and diagnose and discriminate plant pathogenic fungi. The start codon targeted (SCoT) markers system is a novel approach used here to investigate intra and interspecific polymorphisms of phytopathogenic fungi. This study assessed genetic variability between and within 96 isolates of ten fungal species associated with a variety of plant species using 36 SCoT primers. The six primers generated 331 distinct and reproducible banding patterns, of which 322 were polymorphic (97.28%), resulting in 53.67 polymorphic bands per primer. All primers produced informative amplification profiles that distinguished all fungal species. With a resolving power of 10.65, SCoT primer 12 showed the highest polymorphism among species, followed by primer 33 and primer 29. Polymorphic loci (PPL), Nei's diversity index (h), and Shannon index (I) percentages were 6.25, 0.018, and 0.028, respectively. UPGMA analysis separated all isolates based on morphological classification and revealed significant genetic variation among fungal isolates at the intraspecific level. PCoA analysis strongly supported fungal species discrimination and genetic variation. The other parameters of evaluation proved that SCoT markers are at least as effective as other DNA markers. SCoT markers were effective in identifying plant pathogenic fungi and were a powerful tool for estimating genetic variation and population structure of different fungi species.

Determination of Genetic Diversity of Tamarind (Tamarindus indica) Accessions in Eastern Region of Kenya Using Inter Simple Sequence Repeat (ISSR) Markers

There is scanty information on tamarind’s genetic diversity in Easter Kenya. The objective of this study was to determine the genetic diversity of 64 tamarind accessions from Eastern Kenya using 12 Inter Simple Sequence Repeat (ISSR) markers. DNA was extracted from the young apical leaves using modified CTAB method and amplified using standard PCR. The data collected were scored as presence (1) or absence (0) of bands then compared to the 100bp ladder and analyzed using GeneAlex and R softwares. Only seven primers produced reproducible bands. A total of 46 alleles were produced for the 7 loci with an average of 6.6 per loci. Polymorphic information content (PIC) varied from 0.72 to 0.89 and genetic diversity varied from 0.74 to 0.9. The ISSR markers revealed effective polymorphism of 40.87 to 99.46% and the band sizes varied from 100 to 1000 bp. Analysis of Molecular Variance (AMOVA) depicted high variation within the tamarind populations at 90% and the least variation of 10% among the population. The first 3 components of Principal coordinate analysis (PCoA) contributed 40.83% of the total variation. Hierarchical cluster analysis grouped the tamarind accessions into seven major distinct groups. Tamarind accessions were different within counties with minimal variations among counties, proving that genetic diversity exists among the tamarind accessions in the Eastern region of Kenya. High genetic diversity was evident among Embu accessions and least among Masinga accessions. Diversity in tamarind can be utilized in marker-assisted breeding and primer ISSR17899A and ISSRHB11 can be explored in studying genes that code for various traits in tamarind accessions.

Genetic Divergence in French Bean Accessions

Background: Crop improvement has paramount importance to meet the global demand of food to feed the growing human population. Methods: A total of 23 French bean genotypes with diverse genetic background were taken to assess their phenotypic attributes and molecular characterization using inter simple sequence repeat (ISSR) markers. The data collected during two growing seasons over four locations on different quantitative characters were pooled and analyzed under randomized complete block design for deciphering the extent of genetic diversity. Similarity value, based on Jaccard’s coefficient and UPGMA (un-weighted pair group method with arithmetic mean) analysis was calculated. The dendrogram was constructed using Jaccard’s similarity matrix for above analysis. Result: The genotypes showed a wide variation in morphological characters like plant height, pods per plant, pod weight, pod length and pod yield per plant. The genotype HAFB-5 showed the maximum potential for pod yield per plant and long pods. Four ISSR primers used in this study amplified 27 reproducible bands with 18 (66.66%) polymorphic. The genotype Kashi Param and HAFB-4 were found diverse among the selected genotypes on the basis of molecular markers.

Deciphering the Genetic Identity and Fidelity of Banana Genotype Musa acuminata through Molecular Fingerprinting

Micropropagation for extensive production of elite planting materials is an effective and superior alternative to suckers and conventional means of propagation of Musa spp. The present investigation was carried out to evaluate the genetic identity of different micropropagated banana genotypes Grand Naine, Shrimantee and Basrai. Further micropropagated banana genotypes Grand Naine comparing with mother plant during different stages of subculture using Simple Sequence Repeat (SSR) and Inter Simple Sequence Repeat (ISSR). Moreover, for micropropagation explants (suckers) were inoculated on initiation medium containing MS (Murashige and Skoog) supplemented with 5 mg/l BAP (Benzyl amino purine) and 39 mg/l AdSO4 (Adenine sulphate). The second, third and fourth subcultures were carried out to separate all multiple shoots. Furthermore, for rooting MS media fortified with growth hormones viz., 2 mg/l BAP, 1mg/l IBA (Indole-3-butyric Acid) and 200mg/l activated charcoal were used under present investigation. Findings revealed that 90 % and 75 % plantlets were successfully established during primary hardening in controlled and natural field condition. Further, 26 primers were screened from that 20 primers (ISSR and SSR) showed 146 clear, reproducible and scorable bands ranging from 100-2000 base pairs produced. The banding pattern of these randomly selected plants was monomorphic with mother plant of all selected genotype that confirmed the clonal uniformity.

Population Genetic Structure of Feral and Cultured African Catfish (Clarias gariepinus) inferred from Random Amplified Polymorphic DNA in Kano, Nigeria

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.

Molecular and Technological Characterization of Saccharomyces cerevisiae from Sourdough

DNA markers help detect the intraspecific genetic diversity of yeast strains. Eight ISSR (Inter Simple Sequence Repeats) primers were used to assess the intraspecific diversity of Saccharomyces cerevisiae (n = 96) from different populations (n = 3), evaluate the technological characteristics, and investigate trait-loci associations. The primers amplified 154 reproducible and scorable bands, of which 79.87% were polymorphic. The UPGMA (unweighted pair group method with arithmetic mean) dendrogram clustered 96 isolates into two main clusters, supported by STRUCTURE HARVESTER results (ΔK = 2). Analysis of molecular variance (AMOVA) indicated significant genetic differences between (15%) and within the populations (85%) (p < 0.001). Twenty-nine genetically distinct strains were selected for the technological characterization. Principal component analysis (PCA) revealed that five strains with high fermentation capacity, leavening activity, high growth index at 37 °C, and harsh growth conditions were technologically relevant. Trait-loci association analyses indicated that the highest correlation (r = 0.60) was recorded for the fermentation capacity on the 8th and 113th loci, amplified by ISSR-1 and ISSR-6 primers, respectively (p < 0.05). The strains yielding high performances and the associated loci amplified by ISSR markers possess a high potential to generate locus-specific primers to target the strains with high fermentation capacity.

Genetic Variability of Different Jordanian Almond Prunus Amygdalus L. Landraces Revealed by Morphological Traits and RAPD Markers

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.

Diversity Analysis of Rhizoctonia solani Kuhn Isolates Causing Sheath Blight of Rice in Eastern Uttar Pradesh, India

The sheath blight of rice is a devastating fungal disease, which is caused by Rhizoctonia solani AG1 IA. Twenty-one sheath blight isolates of rice were collected from different locations in eastern Uttar Pradesh, India, to study the variation in cultural, pathogenic, and molecular characterization. All the R. solani isolates were classified into four groups depending on the arrangement of sclerotia, i.e., peripheral, centre, scattered, and last group includes placing at the centre and peripheral 11, 1, 5, and 4 isolates, respectively. Depending on pathogenicity, isolates are grouped into weakly virulent, virulent, and highly virulent, representing 12, 4, and 5 isolates, respectively. The random amplified polymorphic DNA has been successfully used for molecular characterization. In our study, ITS1/ITS4 and AG-1 IA-specific markers yielded reproducible banding patterns. UPGMA cluster analysis revealed two major clusters, A and B, with a 13.85% dissimilarity value. This study does not correlate with virulence, geographical location, and RAPD profile groupings.

Genetic diversity and population structure of Saccharomyces cerevisiae isolated from Turkish sourdough by iPBS-retrotransposons markers.

Molecular DNA markers are valuable tools for analyzing genetic variation among yeast from different populations to reveal the genetically different autochthonous strains. In this study, we employed inter-primer binding site (iPBS) retrotransposon polymorphism to assess the genetic variation and population structure of 96 Saccharomyces cerevisiae isolates from four different regions in Turkey. The nine selected iPBS primers amplified 102 reproducible and scorable bands, of which 95.10% were polymorphic with an average of 10.78 polymorphic fragments per primer. The average polymorphism information content and the resolving power were 0.26-3.58, respectively. Analysis of molecular variance (AMOVA) revealed significant (P < 0.001) genetic differences within populations (88%) and between populations (12%). The unweighted pair group mean with arithmetic (UPGMA) dendrogram grouped 96 S. cerevisiae strains into two main clusters, where the highest probability of the data elucidating the population structure was obtained at ΔK = 2. There was not an obvious genetic discrimination of the populations according to geographical regions on UPGMA, supported by principal coordinate analysis. However, the individuals of the closer provinces in each population were more likely to group together or closely. The results indicate that iPBS polymorphism is a useful tool to reveal the genetically diverse autochthonous S. cerevisiae strains that may be important for the production of sourdough or baked goods.

The Genetic Diversity Analysis of Tunisian Male Date Palm Cultivars (Phoneix dactylifera L.) Revealed by Phenotypic and Molecular Markers

Tunisian oases reveal an important genetic heritage of date palm cultivars, with various qualities of use. Since the beginning of the century, this heritage has evolved to a selective orientation based on the monoculture of "Deglet Nour," this orientation risks causing the loss of many cultivars. The male of the date palm is part of this heritage and so far remains marginalized. Although they are important for the date palm production cycle, it is in this context that lies our work to study the genetic diversity of a collection of male date palm pollinators from southern Tunisia. The morphological study of the 20 date palm pollinators using 45 IPGRI (International Plant Genetic Resources Institute) descriptors showed significant discrimination, with a similarity index ranging from 0.207 to 0.457, divided them into five similarity groups. The use of 7 ISSR (Inter Simple Sequences Repeat) primers resulted in 64 reproducible bands, of which 57 were 90% polymorphic, and statistical analysis showed a more or less significant genetic diversity with genetic distances 0.491 to 0.873. According to the Mentel test, a non-significant weak correlation (r = 0.015) was noted between the molecular and morphological data. However, the processing of molecular data by various methods generated very significant correlations. Indeed, the correlation between the SM (Simple matching) coefficient and the DICE coefficient showed an important correlation with r = 0.748, which confirms the discriminating power of the ISSR markers in studying the genetic diversity of date palm pollinators.

Genetic Evaluation of In Vitro Micropropagated and Regenerated Plants of Cannabis sativa L. Using SSR Molecular Markers.

Simple sequence repeat (SSR) markers were used to evaluate the genetic stability of the acclimatized micropropagated and regenerated plants of a high cannabidiol (H-CBD) and a high cannabigerol (H-CBG) variety of Cannabis sativa L. Shoot regeneration and proliferation were achieved by culturing calli in Murashige and Skoog basal medium (MS) supplemented with several concentrations of 6-benzyladenine (BA) or thidiazuron (TDZ). Calli derived mostly from stem explants, rather than leaves, cultured on MS supplemented with 2,4-Dichlorophenoxyacetic acid (2,4-D) or combination of kinetin (KIN) with 1-Naphthaleneacetic acid (NAA) or 2,4-D. Rooting of the regenerated plantlets accomplished on half-strength MS medium supplemented with indole-3-butyric acid (IBA). Previous studies performed have developed an efficient in vitro micropropagation protocol for mass production. Both in vitro methodologies can be employed in genetic breeding via molecular techniques. The genetic stability of micropropagated and regenerated plants was accomplished using twelve SSR primer pairs that produced reproducible and clear bands, ranging from 90 to 330 bp in size, and resulted in amplification of one or two alleles, corresponding to homozygous or heterozygous individuals. The SSR amplification products were monomorphic across all the micropropagated and regenerated plants and comparable to mother plants. The monomorphic banding pattern confirmed the genetic homogeneity of the in vitro cultured acclimatized and mother plants as no somaclonal variation was detected in clones for these specific SSRs. Our results evidently suggest that the developed culture protocols for in vitro multiplication is appropriate and applicable for clonal mass propagation of the C. sativa varieties and demonstrate the reliability of this in vitro propagation system.

Genetic Characterization of Salicornia persica Akhani (Chenopodiaceae) Assessed Using Random Amplified Polymorphic DNA

Salicornia is a wild and annual (biennial, in some species) halophytic plant from Chenopodiaceae family that grows near salt marshes and salted wetlands or the vicinity of coastal areas in Asia, North America, and the Middle East. It is also grown in semi-arid areas of Iran such as Isfahan, Fars, and Yazd provinces. Genetic variability and populations, structure were studied in 10 geographical populations of Salicornia persica. Genetic diversity parameters were determined in these populations. 10 of 20 random amplified polymorphic DNA (RAPD) primers produced 146 reproducible bands with average of 14.6 bands per primer and 88.67% of polymorphism. OPA10 primer showed the highest number of effective allele (Ne), Shannon index (I) and genetic diversity (H). The highest values of genetic diversity in RAPD markers were obtained in Esfahan, Nain, it is 70km to Varzaneh populations. WARD trees of RAPD data grouped the populations in two different clusters/groups, indicating their genetic difference which is discussed in details. The results of this study showed that the level of genetic variation in Salicornia persica is relatively low. WARD-based dendrogram showed a close relationship between members of Esfahan, Varzaneh and Esfahan, the river of Zayanderud at Varzaneh while the Yazd, Meybud and Yazd, Nádüshan protected population differ the most from the other populations. Principal component analysis, however, showed some minor differences with WARD-based dendrograms.

Genetic evaluation of Garcinia gummi-gutta L. (Roxb.) accessions based on inter simple sequence repeat markers

Garcinia gummi-gutta (Family: Clusiaceae) is a rich source of a natural anti-obesity agent, hydroxy citric acid (HCA). The present approach aimed to analyse the extent of genetic variability and structure of a population of 50 accessions of G. gummi-gutta from different geographical localities of Kerala state, India using inter simple sequence repeat (ISSR) markers. Among 30 primers tested, 21 showed clear, reproducible and scorable banding patterns. A total of 157 ISSR fragments were observed with an average of 7.48 bands for each primer. An average of 59.52% polymorphic bands were observed among the accessions, with the highest percentage was observed for UBC 809 (87.5%). Polymorphism information content (PIC) value of 0.355 indicating high degree of allele divergence within the individuals. The highest values for Simpson (0.99) and Shannon indices (1.02) substantiated the significant level of polymorphism within the population. Dendrogram with bootstrapping values (100) based on the neighbour joining method distributed the accessions into three main clusters. Relationships exhibited by the accessions discloses continuous geneflow happening between them irrespective of their geographical distances. STRUCTURE analysis revealed the probability of eight subpopulations based on Bayesian clustering. So, ISSR markers can be used as a powerful tool to differentiate closely related individuals.

Micropropagation, encapsulation, physiological, and genetic homogeneity assessment in Casuarina equisetifolia.

Casuarina equisetifolia is an important tree of the forest, cultivated in tropical and subtropical regions, providing fuelwood, land reclamation, dune stabilization, paper production, and nitrogen fixation. We have developed a systematic in vitro propagation protocol in C. equisetifolia using nodal segments (NS). Murashige and Skoog (MS) medium augmented with BA (5.0 μM) and NAA (0.5 μM) gave rise to a maximum of 32.00 ± 0.31 shoots per explant (S/E) with shoot length (SL) of 3.94 ± 0.02 cm, and a maximum of 70% regeneration potential (RP) was recorded after 8 weeks of post inoculation. For root induction, in vitro derived shoots were transferred to the nutrient medium consisting of a half-strength (½) MS medium augmented with 2.5 μM NAA, which produced a maximum of 12.68 ± 0.33 roots/shoot (R/S) with 3.04 ± 0.50 cm root length (RL) in 60% of culture after 6 weeks. Micropropagated plants with healthy shoots and roots were successfully acclimatized in vermicompost + garden soil + sand (1:2:1) and a maximum survival percentage of 95.1% was recorded. NS was taken from a 6-weeks-old in vitro derived plant of C. equisetifolia for synthetic seed production, and it was reported that CaCl2 · 2H2O (100 mM) + Na2-alginate (4%) resulted in clear and uniform beads. Furthermore, the maximum conversion of synthetic seeds into plantlets occurred over a period of 4 weeks of storage at 4°C. Scanning Electron Microscopy (SEM) revealed the formation of direct shoot buds without any intermediate callus formation. In addition, the chlorophyll and carotenoid contents of the direct regenerated and mother plant were compared. Similarly, RAPD and ISSR primers were used for genetic homogeneity assessment of the direct regenerated plants, where a total of 18 and 19, respectively, clear and reproducible bands with 100% monomorphism were recorded. The developed micropropagation protocol can certainly be used for large-scale multiplication and germplasm preservation of C. equisetifolia. It will also help in meeting the growing demands of C. equisetifolia in the forest industry.

Molecular analyses of genetic diversity in thepopulation of Calligonum Comosum L. ‘Her in Hail region in the Kingdom of Saudi Arabia

This paper presents the first molecular study on Calligonum comosum L. ‘Her (Polygonaceae) in the Kingdom of Saudi Arabia, due it’s known to be part of Saudi Arabia’s flora, as a famous medicinal plant, it has several other uses, as it is palatable as animal feed, and dune stabilization, and to increase soil organic matter content. And it has been included on the endangered species list due to its frequent use. Twenty-five C. comosum samples were collected from five different populations from their natural environment and screened using Inter-Simple Sequence Repeat (ISSR) markers. Eleven ISSR primers amplified 975 reproducible bands of which 495 were polymorphic, and polymorphism of genetic variation between species was 50.8%. This ratio is considered high Among C. comosummembers from multiple populations, this ratio is an indication of the quality of genetic diversity within and between C. comosum populations, which is of great importance for the preservation of the plant. As considered ISSR markers are an effective tool to provide good information for the genetic diversity available in C. comosum inputs from different populations, and the results of this study will help to understand genetic variation and evolutionary dynamics in C. comosum and expand the genetic base for breeding wild species. That additional selection of inputs in each species to assess genetic diversity leads to more useful and reliable results.