Marker Techniques Research Articles

Integrated omic techniques and their genomic features for invasive weeds.

Many emerging invasive weeds display rapid adaptation against different stressful environments compared to their natives. Rapid adaptation and dispersal habits helped invasive populations have strong diversity within the population compared to their natives. Advances in molecular marker techniques may lead to an in-depth understanding of the genetic diversity of invasive weeds. The use of molecular techniques is rapidly growing, and their implications in invasive weed studies are considered powerful tools for genome purposes. Here, we review different approach used multi-omics by invasive weed studies to understand the functional structural and genomic changes in these species under different environmental fluctuations, particularly, to check the accessibility of advance-sequencing techniques used by researchers in genome sequence projects. In this review-based study, we also examine the importance and efficiency of different molecular techniques in identifying and characterizing different genes, associated markers, proteins, metabolites, and key metabolic pathways in invasive and native weeds. Use of these techniques could help weed scientists to further reduce the knowledge gaps in understanding invasive weeds traits. Although these techniques can provide robust insights about the molecular functioning, employing a single omics platform can rarely elucidate the gene-level regulation and the associated real-time expression of weedy traits due to the complex and overlapping nature of biological interactions. We conclude that different multi-omic techniques will provide long-term benefits in launching new genome projects to enhance the understanding of invasive weeds' invasion process.

Genetic Similarity from Collections of Seed for Two Peanut (Arachis hypogaea L.) Cultivars in Ghana

Maintenance of genetic purity of crop cultivars is critical to meet the needs of farmers, processors, and consumers. In Ghana where the informal seed sector is popular (e. g., farmer-saved seed), purity of improved cultivars in the farming community can be compromised. The objective of the study was to assess the genetic purity of farmer-saved seed of two peanut (Arachis hypogaea L.) cultivars (Shitaochi and Yenyawoso) using molecular marker techniques. Twenty samples of Shitaochi and eight samples of Yenyawoso were collected from farmers across five regions of the country and were compared with seed provided by the research institutes of the Council for Scientific and Industrial Research (CSIR). Genetic similarity ranged from 43% to 100% when comparing seed collections from farmers to those from research institutes. Only five of the 20 seed samples of Shitaochi (25%) and two of the 8 samples of Yenyawoso (25%) collected from farmers were found to be completely similar to the reference samples provided by CSIR. These results demonstrate the lack of purity among described cultivars in the informal seed sector in Ghana. While diversity between samples of Shitaochi have been reported and would be expected in the informal seed system, the pace at which Yenyawoso has lost genetic purity in a relatively short period of time indicates that a more effective formal seed system is needed to maintain cultivar purity.

Elucidating genetic diversity and population structure in jamun [Syzygium cumini (L.) Skeels] using morpho-physiological traits and CAAT box-derived polymorphism

• Assessment of morpho-physiological and molecular diversity in Syzygium cumini . • Population structure analysis using CBDP markers. • CBDP markers were highly informative in detecting the polymorphism. • Evidence for high levels of gene flow and less genetic differentiation. The use of molecular markers has received little attention in jamun [ Syzygium cumini (L.) Skeels], an underutilized fruit crop highly valued for its medicinal and nutraceutical properties. In this study, 50 jamun accessions from four different populations were analyzed using 21 morpho-physiological traits and 22 polymorphic gene-targeted CAAT box-derived polymorphism (CBDP) markers. The morpho-physiological variability was fairly high among the jamun accessions. The UPGMA analysis based on morpho-physiological traits produced three clusters with a cophenetic correlation of 0.75, reflecting a good fit between the original matrix and the clustering algorithm. The average polymorphism of 91.17% and the wide-ranging heterozygosity values (0.282-0.499) for the CBDP markers revealed considerable genetic variation among jamun genotypes. The average PIC of 0.352 evinced a high discriminatory power of the CBDP markers in distinguishing the accessions. CDBP data-based UPGMA clustering and population structure analysis identified four distinct populations; albeit with overlaps among them. This, together with the existence of admixed types in all four populations, indicated a fairly high degree of gene flow within jamun populations sampled in this study. It is therefore likely that despite widely differing geographic origins, some accessions in all the populations have a shared ancestral origin. Analysis of Molecular Variance also supports this assumption, as an exceedingly higher within-population variance (84%) than among-population variance (16%) reflected a high degree of gene flow and low genetic differentiation for the Jamun populations examined. Our findings improve the existing understanding vis-à-vis genetic diversity and population structure in jamun, and are expected to increase the use of CBDP and similar marker techniques in cultivar development programs.

Abstract 14127: The Role of Sodium Glucose Co-Transporter 1 in Hyperglycemia Ischemia Reperfusion Injury

Introduction: Hyperglycemia is a common finding in ACS patients in both diabetic and non-diabetic, it is considered a powerful predictor of prognosis and mortality. The role of hyperglycemia in ischemia-reperfusion injury is not fully understood, whether the Sodium Glucose Co-Transporter 1(SGLT1) plays a role in increase injury, before and/or after reperfusion, remains to be elucidated. SGLT2 inhibitors clinical trials have shown significant improvements in cardiovascular outcomes in diabetic and non-diabetic, yet the mechanism is not fully understood and whether SGLT1 plays a role in infarct augmentation remains to be elucidated. Hypothesis: High glucose at reperfusion leads to excess myocardial injury and the increased injury is mediated through the activity of SGLT1. Methods: RT-PCR and in-situ hybridization (RNAScope) combined with Immunofluorescence integrated co detection with different cell marker techniques were used to detect SGLT1 mRNA expression in Sprague-Dawley whole myocardium and Zucker diabetic rats. An Ex-vivo Langendorff ischemia-reperfusion perfusion model was used to study the effect of high glucose on myocardium at reperfusion. Canagliflozin a non-selective SGLT inhibitor (1μmoL/L to block the SGLT1 and SGLT2 transporter and 5nmol/L to block only the SGLT2 transposer) and Mizagliflozin a selective SGLT1 inhibitor (100nmol/L) was introduced following ischemia at two different glucose concentration concentrations at reperfusion and its effect on infarct size measured using triphenyltetrazolium chloride (TTC) staining. Results: Our data reveal that SGLT1 is homogenously expressed throughout the myocardium and is particularly evident within the vasculature. We have also demonstrated that high-glucose mediated injury in the isolated, perfused heart model and it is abrogated through the administration of both mixed SGLT2/SGLT1 inhibitor, canagliflozin, at a dose that inhibits both SGLT2 and SGLT1, and through the administration of novel specific SGLT1 inhibitor, Mizagliflozin. Conclusions: We have shown that SGLT1 is present in the myocardium. Hyperglycemia appears to augment myocardial infarction and inhibition of SGLT1 attenuates this increase.

The cardioprotective effect of inhibiting SGLT1 in hyperglycemia ischemia reperfusion injury

Abstract Background Diabetes clinical trials have shown SGLT inhibition improves cardiovascular outcomes, yet the mechanism is not fully understood. Hyperglycemia is a common finding in diabetic and non-diabetic patients presenting with ACS and is a powerful predictor of prognosis and mortality. The role of hyperglycemia in ischemia-reperfusion injury (IRI) is not fully understood, and whether the Sodium Glucose Co-Transporter 1 (SGLT1) plays a role in infarct augmentation, before and/or after reperfusion, remains to be elucidated. Purpose Investigate if SGLT1 is involved in a glucotoxicity injury during IRI and whether inhibiting SGLT1 with an SGLT1 inhibitor may reduce infarct size. Method RT-PCR and in-situ hybridization (RNAScope) combined with Immunofluorescence integrated co detection with different cell marker techniques were used to detect SGLT1 mRNA expression in Sprague-Dawley whole myocardium and isolated primary cardiomyocytes. An Ex-vivo Langendorff ischemia-reperfusion perfusion model was used to study the effect of high glucose (22mmol) on myocardium at reperfusion. Canagliflozin (CANA) a non-selective SGLT inhibitor (1μmoL/L to block the SGLT1 receptor and SGLT2 and 5nmol/L to block only the SGLT2 receptor) and Mizagliflozin a selective SGLT1 inhibitor (100nmol/L) was introduced following ischemia at two different glucose concentration concentrations at reperfusion and its effect on infarct size measured using triphenyltetrazolium chloride (TTC) staining. Results We showed that SGLT1 is homogenously expressed throughout the myocardium and is particularly evident within the vasculature. we demonstrate that hyperglycemia at reperfusion is injurious to myocardium with an increase of myocardial infarction. Our data reveal that glucose exacerbation of injury appears to be mediated via SGLT1. We have also demonstrated that high-glucose mediated injury in the isolated, perfused heart model is abrogated through the administration of a clinically available mixed SGLT2/SGLT1 inhibitor, canagliflozin, at a dose that inhibits both SGLT2 and SGLT1, but by the SGLT2-selective concentration. Conclusion We have shown that SGLT1 is present in the myocardium. Hyperglycemia appears to augment myocardial infarction and inhibition of SGLT1 attenuates this incre Funding Acknowledgement Type of funding sources: Private grant(s) and/or Sponsorship. Main funding source(s): The government of saudi Arabia

Skimming for barcodes: rapid production of mitochondrial genome and nuclear ribosomal repeat reference markers through shallow shotgun sequencing.

DNA barcoding is critical to conservation and biodiversity research, yet public reference databases are incomplete. Existing barcode databases are biased toward cytochrome oxidase subunit I (COI) and frequently lack associated voucher specimens or geospatial metadata, which can hinder reliable species assignments. The emergence of metabarcoding approaches such as environmental DNA (eDNA) has necessitated multiple marker techniques combined with barcode reference databases backed by voucher specimens. Reference barcodes have traditionally been generated by Sanger sequencing, however sequencing multiple markers is costly for large numbers of specimens, requires multiple separate PCR reactions, and limits resulting sequences to targeted regions. High-throughput sequencing techniques such as genome skimming enable assembly of complete mitogenomes, which contain the most commonly used barcoding loci (e.g., COI, 12S, 16S), as well as nuclear ribosomal repeat regions (e.g., ITS1&2, 18S). We evaluated the feasibility of genome skimming to generate barcode references databases for marine fishes by assembling complete mitogenomes and nuclear ribosomal repeats. We tested genome skimming across a taxonomically diverse selection of 12 marine fish species from the collections of the National Museum of Natural History, Smithsonian Institution. We generated two sequencing libraries per species to test the impact of shearing method (enzymatic or mechanical), extraction method (kit-based or automated), and input DNA concentration. We produced complete mitogenomes for all non-chondrichthyans (11/12 species) and assembled nuclear ribosomal repeats (18S-ITS1-5.8S-ITS2-28S) for all taxa. The quality and completeness of mitogenome assemblies was not impacted by shearing method, extraction method or input DNA concentration. Our results reaffirm that genome skimming is an efficient and (at scale) cost-effective method to generate all mitochondrial and common nuclear DNA barcoding loci for multiple species simultaneously, which has great potential to scale for future projects and facilitate completing barcode reference databases for marine fishes.

Gene Flow and Recruitment Patterns among Disjunct Populations of Allocasuarina verticillata (Lam.) L.A.S. Johnson

Allocasuarina verticillata (Lam.) L.A.S. Johnson is a widespread species in south-eastern Australia providing vegetation cover, protecting fragile soils and providing food for birds. Understanding the effects of gene flow on the recruitment patterns, genetic differentiation and structure of fragmented populations provides fundamental guidelines to underpin plant conservation strategies and activities. In this study, four spatially disjunct populations of A. verticillata were sampled to explore the effects of population size, reproductive patterns and pollen and seed dispersal on among-population genetic diversity, genetic differentiation and structure, using field survey and microsatellite marker techniques. It was found that stands of A. verticillata were predominantly sexually reproductive, but asexual reproduction through root suckering was an additional mode of reproduction. The reproductive success of A. verticillata is positively correlated with the effective population size rather than actual population size. The reduction in effective population size and increment of spatial isolation resulted in lower genetic diversity and higher inbreeding coefficient of progenies. Moderate pairwise genetic differentiation and weak genetic structure were identified. The results suggest that exogenous, wind-mediated pollen flow provides some maintenance of genetic diversity in the isolated stands. Seed dispersal appears mainly to be over short distances (i.e., within populations), but the infrequent transport of seeds between disjunct locations cannot be ruled out as another factor that may help maintain genetic diversity.

Investigation of genetic polymorphism of Russian rape and turnip rape varieties using SSR and SRAP markers.

Rapeseed (Brassica napus L.) and turnip rape (B. rapa L. subsp. campestris (L.)) are important agricultural plants widely used for food, fodder and technical purposes and as green manure. Over the past decades, a large number of perspective varieties that are being currently cultivated in every region of Russia have been developed. To increase the breeding eff iciency and facilitate the seed production, modern molecular-genetic techniques should be introduced as means to estimate species and varietal diversity. The objective of the presented research study was to investigate DNA polymorphism of the rapeseed and turnip rape varieties developed at Federal Williams Research Center of Forage Production and Agroecology and detect informative markers for varietal identif ication and genetic certif ication. To genotype 18 gDNA samples, 42 and 25 combinations of respective SSR and SRAP primers were used. The results obtained demonstrate that SRAP markers were more effective for polymorphism analysis: 36 % of the tested markers revealed genetic polymorphism compared with only 16.7 % of microsatellite loci. Molecular markers to detect differences at interspecif ic and intervarietal levels have also been found. For the investigated set, such microsatellite loci as Na12A02, Ni2C12, Ni02-D08a, Ra02-E01, Ni03H07а and SRAP-marker combinations as F13-R9, Me4- R7, F11-Em2, F10-R7, F9-Em2 and F9-R8 proved to be informative. Application of the two marker techniques made it possible to detect a higher level of DNA polymorphism in plants of different types (spring and winter varieties) if compared against the intervarietal differences within a species or a group. According to Nei’s genetic diversity index, in the cluster of winter rapeseed, VIK 2 and Gorizont varieties had the longest genetic distance, and in the spring cluster, these were Novosel and Veles. A high level of similarity was found between Vikros and Bizon winter rapeseed varieties. The results obtained have a high practical value for varietal specif ication of seed material and genetic certif ication of rapeseed and turnip rape varieties.

Use of molecular marker methods in the classification of bamboo taxa: A review

Bamboo plants are currently attractive to researchers because of their versatile uses. Understanding the bamboos’ genetic level is needed to develop new varieties. Taxonomic identification is the basis for plant development. Bamboos were identified as their taxonomical morphological characters which are dependent on environmental factors. Molecular Marker techniques can be used to perform accurate genotype identification, which can be used for genetic diversity analyses. The RFLP, RAPD, AFLP, SSR, ISSR, iPBS, SCARS, SCoT, SRAP marker systems have been shown to be able to efficiently determine the genetic diversity of bamboo species based on genotyping. This paper summarizes research that aims to analyze the genetic diversity of bamboo species on a molecular basis.

Simple sequence repeat marker development and diversity analysis in buffalograss

AbstractBuffalograss [Buchloë dactyloides (Nutt.) Engelm. syn. Bouteloua dactyloides (Nutt.) Columbus] is a stoloniferous low input turfgrass species native to the central Great Plains of the United States. Several molecular marker techniques have been used to compare relationships among elite buffalograss germplasm, but most rely on random amplification, are transferred from other species, or are developed from limited DNA sequence information. The objectives of this study were to develop buffalograss derived simple sequence repeat markers (SSRs) from buffalograss sequence data using an in‐silico bioinformatics pipeline and use high resolution melt analysis to screen the SSRs based on their melt profiles on buffalograss cultivars and germplasm. Transcriptomes from buffalograss cultivars ‘378’ and ‘Prestige’ were mined for SSRs using the MIcroSAtellite tool. There were 259 conserved SSRs that were polymorphic between 378 and Prestige. Of those, 96 were tested by HRM on a panel of eight genotypes and 24 were selected for producing melt profiles that could distinguish the genotypes. The selected reactions produced 356 distinct melt profiles when tested on a panel of 96 genotypes, including 88 buffalograss germplasm selections, six buffalograss cultivars, and two blue grama [Bouteloua gracilis (H. B. K.) Lag. ex Steud.] selections, with melt profile frequencies ranging from 0.01 to 0.97. A subset of 86 melt profiles segregating among the blue grama and elite buffalograss cultivars distinguished each genotype. The buffalograss‐specific SSR markers presented here will be useful to study buffalograss genetic diversity, line purity maintenance, and for marker supported plant breeding strategies.

Polydendrocytes – undifferentiated cells of the nervous system

Oligodendrocyte progenitor cells (OPCs) were first described more than two decades ago. New marker techniques have revealed that they are independent cells with the high nerve tissue repair potential, and they have been classified as the fourth glial cell type in addition to astrocytes, oligodendrocytes and microglial cells. Another term used for this type of cells is polydendrocytes, this is due to both their morphology and developing knowledge of their diverse functions. OPCs play an important role in the development and myelinogenesis in adults giving rise to oligodendrocytes that cover axons providing signal isolation by the myelin sheath, thus accelerating action potential propagation and ensuring high transmission fidelity without the need for an increased axon diameter. Loss or absence of oligodendrocyte precursors and the resulting absence of differentiated oligodendrocytes are associated with lost myelination and subsequent impairment of neurological functions. Demyelination is a feature of various diseases such as multiple sclerosis, Alzheimer's disease, schizophrenia, infantile cerebral palsy and childhood cognitive impairment. Moreover, OPCs express receptors for various neurotransmitters and are exposed to membrane depolarisation to receive synaptic signals from neurons. Numerous research studies have been investigated polydendrocyte functionality and potential for use as target cells in the treatment and prevention of neural tissue diseases. There are no studies related to morphology, functionality and potential of oligodendrocyte precursors in our country. In this review we highlight issues of the polydendrocyte discovery, their localization and migration potential, possibilities of remyelination through OPCs in hypoxic injury in the embryonic and postnatal period.

An Eruption of LTR Retrotransposons in the Autopolyploid Genomes of Chrysanthemum nankingense (Asteraceae).

Whole genome duplication, associated with the induction of widespread genetic changes, has played an important role in the evolution of many plant taxa. All extant angiosperm species have undergone at least one polyploidization event, forming either an auto- or allopolyploid organism. Compared with allopolyploidization, however, few studies have examined autopolyploidization, and few studies have focused on the response of genetic changes to autopolyploidy. In the present study, newly synthesized C. nankingense autotetraploids (Asteraceae) were employed to characterize the genome shock following autopolyploidization. Available evidence suggested that the genetic changes primarily involved the loss of old fragments and the gain of novel fragments, and some novel sequences were potential long terminal repeat (LTR) retrotransposons. As Ty1-copia and Ty3-gypsy elements represent the two main superfamilies of LTR retrotransposons, the dynamics of Ty1-copia and Ty3-gypsy were evaluated using RT-PCR, transcriptome sequencing, and LTR retrotransposon-based molecular marker techniques. Additionally, fluorescence in situ hybridization(FISH)results suggest that autopolyploidization might also be accompanied by perturbations of LTR retrotransposons, and emergence retrotransposon insertions might show more rapid divergence, resulting in diploid-like behaviour, potentially accelerating the evolutionary process among progenies. Our results strongly suggest a need to expand the current evolutionary framework to include a genetic dimension when seeking to understand genomic shock following autopolyploidization in Asteraceae.

Breeding Potentials of Bambara Groundnut for Food and Nutrition Security in the Face of Climate Change.

Constant production of quality food should be a norm in any community, but climate change, increasing population, and unavailability of land for farming affect food production. As a result, food scarcity is affecting some communities, especially in the developing world. Finding a stable solution to this problem is a major cause of concern for researchers. Synergistic application of molecular marker techniques with next generation sequencing (NGS) technologies can unlock the potentials hidden in most crop genomes for improving yield and food availability. Most crops such as Bambara groundnut (BGN), Winged bean, and African yam bean are underutilized. These underutilized crops can compete with the major crops such as cowpea, soybean, maize, and rice, in areas of nutrition, ability to withstand drought stress, economic importance, and food production. One of these underutilized crops, BGN [Vigna subterranea (L.), Verdc.], is an indigenous African legume and can survive in tropical climates and marginal soils. In this review, we focus on the roles of BGN and the opportunities it possesses in tackling food insecurity and its benefits to local farmers. We will discuss BGN’s potential impact on global food production and how the advances in NGS technologies can enhance its production.

Continental-scale measurements of soil pyrogenic carbon in Europe

Pyrogenic carbon (PyC), the product of incomplete biomass combustion, is a key component of soil organic carbon (SOC) because it can persist in soils for centuries to millennia. Quantifying PyC across large spatial scales remains a significant challenge in constraining the global carbon cycle. We measured PyC in topsoils across Europe using molecular marker (benzene polycarboxylic acids, BPCA) and spectroscopic techniques (Diffuse Reflectance Infrared Fourier Transform Spectroscopy, DRIFTS). We developed a calibration between BPCA and DRIFTS, but the calibration was less reliable (Y-variance explained = 0.62) than previous reports due to low soil PyC content and heterogeneity of soil matrices. Thus, we performed multiple regressions to identify drivers of PyC distribution using only the measured BPCA data. PyC content varied widely among soils, contributing 0–24% of SOC. Organic carbon was the strongest predictor of soil PyC content, but mean annual temperature, clay, and cation exchange capacity also emerged as predictors. PyC contributes a smaller proportion of SOC in European soils compared to other geographic regions. Comparing soil PyC measurements to PyC production rates in high latitude and Mediterranean regions suggests that transport, degradation, and recombustion are important mechanisms regulating soil PyC accumulation.

Genetic Polymorphisms of Four Egyptian Plant Species using some Molecular Markers Techniques RAPD, ISSR, and SCoT

Genetic Polymorphisms of Four Egyptian Plant Species using some Molecular Markers Techniques RAPD, ISSR, and SCoT

Evaluation of Allelic Variation in Cedrus deodara (Roxb.) G. Don to identify Region Specific Nuclear Microsatellite Markers (nSSRs) in Uttarakhand, India

Cedrus deodara is an excellent timber species extensively prone to illicit felling and smuggling of timber. Knowledge of molecular marker techniques can help in tracing the origin of the stolen timber back to the forest area for legal purposes. The present study demonstrates the use of nSSR markers for timber tracking in Cedrus deodara. In this research twentyfive individual trees were randomly selected from two geographically distinct populations Kathiyan and Jageshwar forest of Uttarakhand. Out of 47 nuclear SSR primers, only nine showed positive amplification out of which five (pdms009, pm01, pm05, SSRPtctg4698 and RPTest9) were finally selected to reveal allelic variation. Total of 10 alleles of individuals were detected at all five loci for both populations. For Kathiyan, population effective number of alleles ranged from 1.26 to 2 while for Jageshwar, it ranged from 1.14 to 1.98. Expected heterozygosity (HE) for Chakrata population ranged from 0.21 to 0.51 while for Jageshwar it was between 0.13 to 0.5. Analysis of molecular variance (AMOVA) shows that most of the variations were found within a population (97.81 %) as compared to among the population (2.19 %) with an FST value of 0.02. In this research, we found that the same primer pairs gave a different product length for both geographically distinct populations of C. deodara, for Kathiyan population locus pdms009, pm01, pm05, SSRPtctg4698 and RPTest9 product lengths for both alleles a and b were 197 and 125, 304 and 221,402 and 317,261 and 208, 332 and 246 base pair respectively, while for Jageshwar population, it were 339 and 238, 319 and 203, 456 and 283, 245 and 197, 309 and 235 base pair respectively.

Evaluation of genetic diversity by morphological, biochemical and molecular markers in sour cherry genotypes.

Turkey's ecological and geographical diversity allows the cultivation of most plant species that have economic importance throughout the country. One of these species, sour cherry, is among the stone fruits that are very important for human health. This study aimed to determine genetic diversity of 21 sour cherry genotypes grown from seed in Kelkit Valley and standard Kutahya cultivars. Morphological, biochemical, and molecular marker techniques [ISSR (Inter Simple Sequence Repeat)] was used to determine genetic diversity. G11 in terms of fruit characteristics, particularly weight with 4.81g, and G20 in terms of leaf width produced better results than the other genotypes with 40.10mm. Molecular marker analysis, a total of 70 bands were obtained from 12 ISSR primers and 49 of them were polymorphic. The mean polymorphism rate in the study was 70%, and the DICE's similarity coefficient of genotypes differed between 0.81 and 0.95. Polymorphism percentages of the ISSR primers varied from 20 to 100%. In biochemical analyzes, G17 with 140.18mg QE/100g in total flavonoids content, G19 with 208.63mg GAE/100g in total phenolics content, G19 with 8.33mg cyn-3-gluc/100g in total anthocyanin content compared to other genotypes prominent genotypes. The use of the obtained results in breeding studies to be carried out on this species, as well as the detection and protection of gene sources, may be beneficial for researchers.

Morphological Characterization and Estimation of Genetic Diversity of Sorghum Accessions by SSR Markers

Sorghum ranks 5th in the cereal’s crops having 60 million tonnes annual production. it is important food, feed as well as forage crop and provides raw material for fibbers, biofuel, starch and alcohol. It is very diverse crop; therefore, genetic fingerprints require for the estimation of diversity. The present study was conducted to develop fingerprints and for the estimation of genetic diversity through morphological and molecular characterization. Genetic divergence pattern was assessed among 82 sorghum accessions by evaluating the 9 different parameters for growing season. Analysis of variance (ANOVA) revealed that all parameters had significant differences. The Principal Component analysis showed that 1st three Principal Components (PCs) having eigen value >1 and shared 29.95% variability and 26.57% variation for all parameters. Correlation exhibited that leaf width, plant height, and panicle length had positive correlation with leaf length. Biplot analysis exhibited that plant height, leaf length, grain number/panicle and 100-grain weight had positive and positive coordinates while none of the parameters lies in negative and negative coordinates. The molecular relationship among 96 sorghum accessions was estimated through 16 SSRs markers techniques. Analysis revealed that 99 alleles were analyzed by SSRs. Maximum allelic frequency was observed 1 at locus Sorg6 and Sorg11. Polymorphic information content (PIC) showed that locus Sorg16 had maximum value (0.88) while maximum heterozygosity was 0.85 at locus Sorg5. Dendrogram was constructed on 96 accessions by 16 SSRs and analysis revealed that 30 morphotypes were produced. This study depicted that SSRs had discriminating power for the estimation of genetic diversity. In addition, the sorghum germplasm showed maximum diversity in Pakistan climate season. 
 Keyword: Sorghum, SSRs, Biplot, Genetic diversity, PCA

Morphological Characterization and Estimation of Genetic Diversity of Sorghum Accessions by SSR Markers

Sorghum ranks 5th in the cereal’s crops having 60 million tonnes annual production. it is important food, feed as well as forage crop and provides raw material for fibbers, biofuel, starch and alcohol. It is very diverse crop; therefore, genetic fingerprints require for the estimation of diversity. The present study was conducted to develop fingerprints and for the estimation of genetic diversity through morphological and molecular characterization. Genetic divergence pattern was assessed among 82 sorghum accessions by evaluating the 9 different parameters for growing season. Analysis of variance (ANOVA) revealed that all parameters had significant differences. The Principal Component analysis showed that 1st three Principal Components (PCs) having eigen value >1 and shared 29.95% variability and 26.57% variation for all parameters. Correlation exhibited that leaf width, plant height, and panicle length had positive correlation with leaf length. Biplot analysis exhibited that plant height, leaf length, grain number/panicle and 100-grain weight had positive and positive coordinates while none of the parameters lies in negative and negative coordinates. The molecular relationship among 96 sorghum accessions was estimated through 16 SSRs markers techniques. Analysis revealed that 99 alleles were analyzed by SSRs. Maximum allelic frequency was observed 1 at locus Sorg6 and Sorg11. Polymorphic information content (PIC) showed that locus Sorg16 had maximum value (0.88) while maximum heterozygosity was 0.85 at locus Sorg5. Dendrogram was constructed on 96 accessions by 16 SSRs and analysis revealed that 30 morphotypes were produced. This study depicted that SSRs had discriminating power for the estimation of genetic diversity. In addition, the sorghum germplasm showed maximum diversity in Pakistan climate season. 
 Keyword: Sorghum, SSRs, Biplot, Genetic diversity, PCA

Breeding Aspects of Selected Ornamental Bulbous Crops

This article provides an overview of the origin, genetic diversity and methods and trends in breeding of selected ornamental geophytes (Lilium, Tulipa, Narcissus and Hippeastrum). The role of interspecific hybridisation and polyploidisation in assortment development is reviewed. A great variety of cultivars with traits of interest have been generated over the last century by using classical breeding. Geophyte breeders have been interested in a diversity of traits, including resistance to diseases, flower colour and shape, long lasting flowering and a long vase life. Shortening the long breeding process of many geophytes by reducing the juvenile phase and using in vitro techniques are reviewed. Currently, the breeding process has been enhanced by using modern molecular cytogenetic techniques. Genomic in situ hybridisation is frequently used, among other techniques, for genome differentiation in interspecific hybrids, and for assessment of the extent of intergenomic recombination in backcross progenies. Furthermore, several molecular marker techniques are used for verification of hybrid status, identification of genetic diversity, confirmation of the genetic fidelity of in vitro propagated plants and construction of high-density linkage maps. Recently, a myriad of new plant breeding technologies, such as cisgenetics and genome editing technologies have been used to improve the traits of ornamental geophytes, an endeavour that is discussed here. Breeding trends, cultivar novelties as well a new cultivars registered by international authorities during the last five years are presented in detail.