Search Results (828)

Kaempferia parviflora, a medicinal plant widely used in Southeast Asia, has been validated clinically for its diverse pharmaceutical applications. Despite extensive research in pharmacology, there is a notable lack of cytogenetic and genomic research, primarily due to limited genetic information. Simple Sequence Repeat (SSR) is considered a robust class of molecular markers frequently used in biodiversity studies. In this study, we adopted Microsatellite Capture Sequencing (MiCAPs) to obtain SSR sequences for marker development. We identified 13,644 SSRs and developed and validated ten sets of SSR markers through capillary electrophoresis. The ten primer sets generated 27 alleles, with an average Polymorphism Information Content (PIC) of 0.36. Principle Coordinate Analysis (PCoA) distinguished two types of K. parviflora, consistent with classification by leaf margin color (red and green). A neighbor-joining dendrogram of seven Zingiberaceae species was constructed with the SSR-containing sequences. The 2-c value of K. parviflora is first reported here as 3.16 ± 0.03; the genome size is estimated at 3090.48 Mbp. The newly developed molecular markers are crucial for variety identification and the conservation of wild resources. Additionally, the cytogenetic and phylogenetic information provides valuable insights into the genetic diversity and evolutionary relationships. Full article

Ascochyta blight (AB) is a major threat to Kabuli-type chickpea production worldwide. This study aimed to identify AB-resistant Kabuli-type chickpea lines through combined phenotypic and genotypic screening. Twenty-six Kabuli-type chickpea lines were phenotyped at the seedling stage using spray inoculation with conidial suspension. Genotyping employed marker-aided selection (MAS) with markers linked to quantitative trait loci (QTL) for AB resistance. The allele-specific marker, CaETR, closely linked to QTLAR1, and the sequence-tagged microsatellite (STMS) markers GAA47, TAA146, and TA194 linked to QTLAR1, QTLAR2, and QTLAR3 were used to assess their utility in distinguishing between resistant and susceptible chickpea lines. The study revealed that none of the lines tested were completely resistant (R) phenotypically. However, some lines, such as AVTCPK#6 and AVTCPK#14, were found to be moderately resistant (MR). Of the two MR lines identified phenotypically, only AVTCPK#6 was found to have bands linked to QTLs for adult plant resistance. The other MR line for AB showed the presence of bands in only one or two of the four markers used. These MR lines can be further utilized in chickpea breeding programs for the development of AB-resistant chickpea cultivars. It is recommended that these results be verified through repeat experiments, using more diverse isolates, and including additional chickpea lines as reference checks for resistance and susceptibility. The allele-specific marker, CaETR, closely linked to QTL_AR1 and sequence-tagged microsatellite (STMS) markers GAA47, TAA146 and TA194 linked to QTL_AR1, QTL_AR2, and QTL_AR3 were used to explore these markers’ utility in discriminating between resistant and susceptible chickpea lines. The study showed that phenotypically, none of the lines tested are completely resistant (R). However, some lines, namely AVTCPK#6 and AVTCPK#14, were found to be moderately resistant (MR). Of the two MR lines identified phenotypically, only AVTCPK#6 was identified to have bands linked to QTLs for adult plant resistance. The other MR line for AB showed the presence of bands in only one or two markers among the four markers used. These MR lines can be exploited further in chickpea breeding programs for the development of AB-resistant chickpea cultivars. It is recommended that these results are verified by repeat experiments, using more as well as diverse isolates alongside additional chickpea lines for resistant and susceptible reference checks. Full article

Anthropogenic pressures such as over-urbanization, intensive agriculture/forestry practices, and the development of energy farms alter natural landscapes. Intensive urban development poses the greatest threat to natural ecosystems. Habitat degradation, fragmentation, and loss are among the key factors behind the current rise of biodiversity loss. In this study, we hypothesized that urbanization advances the adaptation of forest bird populations to relatively new urban ecosystems. The study was conducted in Kaunas, Lithuania, located in Eastern Europe. Genetic samples were collected in the city, representing urban landscapes, and its surrounding forests. In total, 160 nest boxes were erected, of which 80 were placed in the urban areas and 80 in the forests. Using a set of microsatellite markers, we investigated the genetic differentiation, genetic diversity, gene flow, and population structure of two common forest bird species of the Paridae family, the great tit (Parus major) and blue tit (Cyanistes caeruleus), in forests and urbanized areas. We observed low but significant differences between urban and forest great tit and blue tit populations, proving relatively high population genetic diversity. We determined that cities’ spatial structure and fragmented natural habitats can influence the formation of small and isolated bird populations (subpopulations). Urban blue tits had higher genetic differentiation and a higher tendency to form subpopulations. In conclusion, forest birds can inhabit urbanized landscapes but both great tits and blue tits respond differently to urbanization-related changes. Full article

The conservation biology field underscores the importance of understanding genetic diversity and gene flow within plant populations and the factors that influence them. This study employs Simple Sequence Repeat (SSR) molecular markers to investigate the genetic diversity of the endangered plant species Saussurea involucrata, offering a theoretical foundation for its conservation efforts. Utilizing sequencing results to screen SSR loci, we designed and scrutinized 18 polymorphic microsatellite primers across 112 samples from 11 populations in the Bayinbuluke region. Our findings reveal high genetic diversity (I = 0.837, He = 0.470) and substantial gene flow (Nm = 1.390) among S. involucrata populations (China, Xinjiang), potentially attributed to efficient pollen and seed dispersal mechanisms. Principal Coordinate Analysis (PCoA) indicates a lack of distinct genetic structuring within the Bayinbuluke populations. The cluster analysis using STRUCTURE reflected the genetic structure of S. involucrata to a certain extent compared with PCoA. The results showed that all samples were divided into four groups. To safeguard this species, we advocate for the in situ conservation of all S. involucrata populations in the area. The SSR markers developed in this study provide a valuable resource for future genetic research on S. involucrata. Full article

Colorectal cancer (CRC) remains the second leading cause of cancer-related mortality worldwide. While immune checkpoint inhibitors have significantly improved patient outcomes, their effectiveness is mostly limited to tumors with microsatellite instability (MSI-H/dMMR) or an increased tumor mutational burden, which comprise 10% of cases. Advancing personalized medicine in CRC hinges on identifying predictive biomarkers to guide treatment decisions. This comprehensive review examines established tissue markers such as KRAS and HER2, highlighting their roles in resistance to anti-EGFR agents and discussing advances in targeted therapies for these markers. Additionally, this review summarizes encouraging data on promising therapeutic targets and highlights the clinical utility of liquid biopsies. By synthesizing current evidence and identifying knowledge gaps, this review provides clinicians and researchers with a contemporary understanding of the biomarker landscape in CRC. Finally, the review examines future directions and challenges in translating promising biomarkers into clinical practice, with the goal of enhancing personalized medicine approaches for colorectal cancer patients. Full article

Genetic diversity analysis provides the scientific basis for the preservation, evaluation, and utilization of the germplasm resources of tree species. We explored the genetic diversity and structure of Populus tomentosa elite trees in North China using 13 nuclear microsatellite markers. We compared nine groups of accessions including 20 originating from Beijing (BJ), 122 from Hebei (HB), 20 from Shandong (SD), 113 from Henan (HN), 270 from Shanxi (SX), 54 from Shaanxi (SAX), 8 from Gansu (GS), 10 from Anhui (AH), and 6 from Jiangsu (JS). All of the studied primer pairs were polymorphic and generated 125 alleles. Analyses of molecular variance revealed that 79%, 14%, and 8% of the total variation was due to variations within the individual, among individuals, and among populations, respectively. Based on principal coordinate and STRUCTURE cluster analyses, individuals distributed in the southern region (HN, SAX, AH, and JS) were roughly classified into one group, while those distributed in the northeastern region (BJ, HB, and SD) and northwestern regions (SX) were separately divided into one group each. Moreover, the northwestern region included two-thirds of the SX trees, and the remainder were in the northeast region. By analyzing genetic diversity and structure within populations, individuals with different genetic backgrounds were screened for constituent training populations (TRS), including broad allelic variation for related traits. This ensures that the genomic prediction model can accurately capture genetic effects and provide reliable predictions across a broad spectrum of genetic backgrounds. Therefore, our results will benefit genome breeding technology. Full article

Barley is indeed a versatile cereal crop, valued for its uses in food, animal feed, and increasingly in biofuel production. As interest grows in developing new barley genotypes that are better adapted to diverse environmental conditions and production systems, integrating agro-morphological evaluations with molecular marker analyses in barley breeding programs is essential for developing new genotypes. It is necessary to explore the genetic diversity of those germplasm to predicate their responses to targeted environments and regions. The current study explored the phenotypic and genotypic relations among Saudi advanced germplasm to facilitate the development of superior barley cultivars suitable for desert environments. Molecular microsatellites (SSR) markers revealed considerable wide genetic variation among Saudi germplasm and checks. Population structure analyses revealed four main groups. Those groups were validated using similarity analyses and coefficients. As well, principal components analysis (PCA) and heat map analyses separated the studied genotypes into four main groups. The improved Saudi germplasm, selected from the barley breeding program, revealed considerably wide genetic and phenotypic diversities, indicating the feasibility of selection to improve for semi-arid conditions. The improved line KSU-BR-C/G-2 had the highest grain yield and harvest index in the first season. Rihana/Lignee was followed by the KSU-BR-C/G-2 genotype, with a grain yield averaging 6734.07 (kg ha⁻¹), in the first season. KSU-BR-88-29-10 yielded 20,000 kg ha⁻¹ for biomass yield. In the second year, KSU-BR-30-7 had the highest biomass yield, with 27,037.04 kg ha⁻¹. Full article

Buckwheat is a highly nutritional pseudocereal with antioxidant potential. The aim of this study was to analyze the genetic variability of 21 varieties of common buckwheat (Fagopyrum esculentum Moench.) and 14 varieties of Tartary buckwheat (Fagopyrum tataricum Gaertn.) using microsatellite markers. By analyzing 21 SSR markers, an average of 11.6 alleles per locus were amplified and an average PIC value of 0.711 was determined. We determined the heterozygous status of the individuals and variability in the set using the SSR analysis on the basis of expected heterozygosity (He, 0.477), observed heterozygosity (Ho, 0.675), Shannon’s index (I, 0.820), and fixation indices (FST, FIS, FIT). Based on the SSR analyses, the lower level of expected heterozygosity in the analyzed set of Tartary buckwheat genotypes was observed compared to common buckwheat. With the help of a hierarchical cluster analysis using the UPGMA algorithm, Structure analysis, and PCoA analysis for the SSR markers, we divided the buckwheat varieties in the dendrogram into two main clusters according to the species. The AMOVA analysis showed that genetic variability between the individuals prevails in the analyzed set. The SSR technique proved to be a suitable tool for the determination of intra- and inter-varietal genetic variability and for analysis of diversity. Full article

The aim of this study was to evaluate phenotypic traits and genetic diversity of the 13 tomato (Solanum lycopersicum L.) varieties and 6 hybrids developed at the Institute of Horticulture Lithuanian Research Centre for Agriculture and Forestry (LRCAF IH). For the molecular characterisation, seven previously published microsatellite markers (SSR) were used. A24 and 26 alleles were detected in tomato varieties and hybrids, respectively. Based on the polymorphism information content (PIC) value, the most informative SSR primers for varieties were TMS52, TGS0007, LEMDDNa and Tom236-237, and the most informative SSR primers for hybrids were SSR248 and TMS52. In UPGMA cluster analysis, tomato varieties are grouped in some cases due to genetic relationships, as the same cluster cultivars ‘Viltis’ (the parent of cv. ‘Laukiai’) and ‘Aušriai’ (the progeny of cv. ‘Jurgiai’) are present. The grouping of all hybrids in the dendrogram is related to the parental forms, and it shows the usefulness of molecular markers for tomato breeding, as they can be used to trace the origin of hybrids and, eventually, varieties accurately. The knowledge about the genetic background of Lithuanian tomato cultivars will help plan targeted crosses in tomato breeding programs. Full article

Chimonanthus praecox, a member of the Calycanthaceae family, is a unique, traditional, and famous flowering economic tree species in China. Despite the existence of several varieties, only a few cultivars have been formally named. Currently, expression sequence tag–simple sequence repeat (EST-SSR) markers are extensively used to identify different species and varieties; a large number of microsatellites can be identified from transcriptome databases. A total of 162,638 unigenes were assembled using RNA-seq; 82,778 unigenes were annotated using the Nr, Nt, Swiss-Prot, Pfam, GO, KOG, and KEGG databases. In total, 13,556 SSR loci were detected from 11,691 unigenes, with trinucleotide repeat motifs being the most abundant among the six repeat motifs. To develop the markers, 64,440 pairs of SSR primers with polymorphism potential were designed, and 75 pairs of primers were randomly selected for amplification. Among these markers, seven pairs produced amplified fragments of the expected size with high polymorphism. Using these markers, 12 C. praecox varieties were clustered into two monophyletic clades. Microsatellites in the transcriptome of C. praecox exhibit rich types, strong specificity, and great polymorphism potential. These EST-SSR markers serve as molecular technical methods for identifying different varieties of C. praecox and facilitate the exploration of a large number of candidate genes associated with important traits. Full article

The pink river dolphin, or bufeo, is one of the dolphins which lives in the rivers of the Orinoco and Amazon basins in South America. The Bolivian bufeo population is considered a differentiated species (Inia boliviensis) from the Amazon and Orinoco species (Inia geoffrensis). Until now, no study has completed an extensive population genetics analysis of the bufeo in Bolivian rivers. We analyzed 82 bufeos from different rivers from the Mamoré and Iténez (Guaporé) river basins for the mt control region (CR), nuclear microsatellites, and DQB-1 gene sequences to determine if the inner rapids of these Bolivian river basins have some influence on the genetic structure of this species. The first relevant result was that the genetic diversity for CR, and the microsatellites were substantially lower in the Bolivian bufeos than in the dolphins studied in other areas of the Amazon and Orinoco. However, the DQB-1 gene sequences yielded similar genetic diversity to those found in other areas. The second relevant result is the existence of some significant genetic heterogeneity among the bufeo populations within Bolivia, although in a small degree, but this differentiation is independent of the inner rapids of the Bolivian rivers we sampled. The third relevant result was the existence of significant isolation by distance for the CR, but not for microsatellites and DQB-1 gene sequences. This was related to differential gene flow capacity of females (philopatric) and males (less philopatric and more migrants) and, possibly, to different selective patterns affecting the molecular markers studied. The fourth relevant result was related to diverse demographic changes of these bufeos. At least two or three bottleneck events and one or two population expansions have occurred in the Bolivian bufeo population. The major part of these events occurred during the Pleistocene. Full article

Tertiary lymphoid structures (TLSs) are complex lymphocyte clusters that arise in non-lymphoid tissues due to inflammation or cancer. A mature TLS with proliferating germinal centers is associated with a favorable prognosis in various cancers. However, the effect of TLS maturity on advanced colorectal cancer (CRC) remains unexplored. We analyzed the significance of TLS maturity and tumor Ki-67 expression in surgically resected tumors from 78 patients with pathological T4 CRC. Mature TLS was defined as the organized infiltration of T and B cells with Ki-67-positive proliferating germinal centers. We analyzed the relationship between TLS maturity and intratumoral immune cell infiltration. Mature TLS with germinal center Ki-67 expression was associated with microsatellite instability and improved survival; however, high tumor Ki-67 expression was associated with poor survival in the same cohort. Multivariate analysis identified the absence of mature TLS as an independent predictor of poor post-recurrence overall survival. Intratumoral infiltration of T lymphocytes and macrophages was significantly elevated in tumors with mature TLS compared to those lacking it. High Ki-67 levels and absent mature TLS were identified as poor prognostic factors in advanced CRC. Mature TLS could serve as a promising marker for patients at high-risk of CRC. Full article

Spinocerebellar ataxia type 3/Machado–Joseph disease (SCA3/MJD) is a neurodegenerative disorder caused by the ATXN3 CAG repeat expansion. Preimplantation genetic testing for monogenic disorders (PGT-M) of SCA3/MJD should include reliable repeat expansion detection coupled with high-risk allele determination using informative linked markers. One couple underwent SCA3/MJD PGT-M combining ATXN3 (CAG)_n triplet-primed PCR (TP-PCR) with customized linkage-based risk allele genotyping on whole-genome-amplified trophectoderm cells. Microsatellites closely linked to ATXN3 were identified and 16 markers were genotyped on 187 anonymous DNAs to verify their polymorphic information content. In the SCA3/MJD PGT-M case, the ATXN3 (CAG)_n TP-PCR and linked marker analysis results concurred completely. Among the three unaffected embryos, a single embryo was transferred and successfully resulted in an unaffected live birth. A total of 139 microsatellites within 1 Mb upstream and downstream of the ATXN3 CAG repeat were identified and 8 polymorphic markers from each side were successfully co-amplified in a single-tube reaction. A PGT-M assay involving ATXN3 (CAG)_n TP-PCR and linkage-based risk allele identification has been developed for SCA3/MJD. A hexadecaplex panel of highly polymorphic microsatellites tightly linked to ATXN3 has been developed for the rapid identification of informative markers in at-risk couples for use in the PGT-M of SCA3/MJD. Full article

Background: Colorectal cancer (CRC) ranks among the most prevalent malignancies affecting the gastrointestinal tract. The infiltration of CD8⁺ T cells significantly influences the prognosis and progression of tumor patients. Methods: This study establishes a CRC immune risk model based on CD8⁺ T cell-related genes. CD8⁺ T cell-related genes were identified through Weighted Gene Co-expression Network Analysis (WGCNA), and the enriched gene sets were annotated via Gene Ontology (GO) and Reactome pathway analysis. Employing machine learning methods, including the Least Absolute Shrinkage and Selection Operator (LASSO) algorithm and Random Forest (RF), we identified nine genes associated with CD8⁺ T-cell infiltration. The infiltration levels of immune cells in CRC tissues were assessed using the ssGSEA algorithm. Results: These genes provide a foundation for constructing a prognostic model. The TCGA-CRC sample model’s prediction scores were categorized, and the prediction models were validated through Cox regression analysis and Kaplan–Meier curve analysis. Notably, although CRC tissues with higher risk scores exhibited elevated levels of CD8⁺ T-cell infiltration, they also demonstrated heightened expression of immune checkpoint genes. Furthermore, comparison of microsatellite instability (MSI) and gene mutations across the immune subgroups revealed notable gene variations, particularly with APC, TP53, and TNNT1 showing higher mutation frequencies. Finally, the predictive model’s efficacy was corroborated through the use of Tumor Immune Dysfunction and Exclusion (TIDE), Immune Profiling Score (IPS), and immune escape-related molecular markers. The predictive model was validated through an external cohort of CRC and the Bladder Cancer Immunotherapy Cohort. CLRN3 expression levels in tumor and adjacent normal tissues were assessed using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Subsequent in vitro and in vivo experiments demonstrated that CLRN3 knockdown significantly attenuated the malignant biological behavior of CRC cells, while overexpression had the opposite effect. Conclusions: This study presents a novel prognostic model for CRC, providing a framework for enhancing the survival rates of CRC patients by targeting CD8⁺ T-cell infiltration. Full article

The production of backyard chickens is an activity of great importance in the economy of rural families in Paraguay. The Rustipollos population was created through directed crosses between a commercial meat line and a local population belonging to non-specific breeds but phenotypically assimilated to Creole breeds. The aim of this study was to evaluate the genetic diversity, relationship, and structure of Rustipollos using 29 microsatellite markers. Analysis was performed on 50 Rustipollos animals and 926 other individuals as reference breeds/populations from Europe, Africa, South, and North America. A total of 318 alleles were detected, with a mean of 10.97 per locus. The polymorphic information content indicated that 80% of all loci were highly to moderately informative. Only two breeds/populations showed loci that did not deviate from the Hardy–Weinberg equilibrium. The results of genetic diversity indexes suggested moderate levels of genetic variability in Rustipollos population and low inbreeding level. The genetic differentiation index indicates a high genetic differentiation between populations. The results of the Neighbor-Net tree and STRUCTURE analyses indicate the existence of distinct gene pools, with some genetic relationships between Rustipollos, the commercial chicken strain, and south Spanish breeds. The Discriminant Analysis of Principal Components confirmed the observed genetic distances between breeds/populations. The results will be useful for sustainable use and official recognition of this population. Full article