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Microsatellites, also known as SSR or STR, are essential molecular markers in genomic research, playing crucial roles in genetic mapping, population genetics, and evolutionary studies. Their applications range from plant breeding to forensics, highlighting their diverse utility across disciplines. Despite their widespread use, traditional methods for SSR analysis are often laborious and time-consuming, requiring significant resources and expertise. To address these challenges, a variety of computational tools for SSR analysis have been developed, offering faster and more efficient alternatives to traditional methods. However, selecting the most appropriate tool can be daunting due to rapid technological advancements and the sheer number of options available. This study presents a comprehensive review and analysis of 74 SSR tools, aiming to provide researchers with a valuable resource for SSR analysis tool selection. The methodology employed includes thorough literature reviews, detailed tool comparisons, and in-depth analyses of tool functionality. By compiling and analyzing these tools, this study not only advances the field of genomic research but also contributes to the broader scientific community by facilitating informed decision-making in the selection of SSR analysis tools. Researchers seeking to understand SSRs and select the most appropriate tools for their projects will benefit from this comprehensive guide. Overall, this study enhances our understanding of SSR analysis tools, paving the way for more efficient and effective SSR research in various fields of study. [ABSTRACT FROM AUTHOR]

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a globally significant fungal disease that seriously threatens wheat yield, particularly in China. This study investigates the genetic structure and reproductive patterns of Pst populations in Ili, Xinjiang, using 12 pairs of Simple Sequence Repeat (SSR) molecular markers. Analyses of 79 Pst isolates from either spring or winter wheat areas in Ili revealed three primary genetic clusters, indicating notable differences between populations associated with spring and winter wheat. The STRUCTURE results, complemented by UPGMA and PCoA analyses, highlight significant genetic diversity within these populations, with evidence of genetic recombination and sexual reproduction in certain areas. Pst populations in Ili exhibit a mixed mode of reproduction, predominantly sexual in Qapqal and Xinyuan D and primarily asexual within the spring wheat populations. The gene flow analysis underscores extensive inter-population communication, which facilitates the spread and adaptation of the pathogen across diverse wheat-growing environments. This study marks the first documentation of sexual reproduction in Pst within Xinjiang, providing new insights into its spread and genetic variation. These findings suggest that sexual reproduction may play a role in the regional adaptation and evolution of Pst, impacting future management strategies for wheat stripe rust in Xinjiang and potentially in broader Central Asian contexts. [ABSTRACT FROM AUTHOR]

The Amazonian shrub Myrciaria dubia (camu-camu) produces vitamin C-rich fruits of growing commercial interest. However, sustainable utilization requires assessment and protection of the genetic diversity of the available germplasm. This study aimed to develop and apply microsatellite markers to assess genetic diversity and construct a core collection of M. dubia germplasm from the Peruvian Amazon. Sixteen polymorphic microsatellite loci were developed using an enrichment approach. The evaluation of 336 genotypes from 43 accessions of the germplasm bank, originating from eight river basins, was conducted using these newly developed markers. Genetic diversity parameters, including observed and expected heterozygosity, were calculated. Analysis of molecular variance (AMOVA) was performed to assess the distribution of genetic variation within and among accessions and river basins. Bayesian clustering analysis was employed to infer population structure. A core collection was constructed to maximize allelic richness. High genetic diversity was observed, with heterozygosity values ranging from 0.468 to 0.644 (observed) and 0.684 to 0.817 (expected) at the river basin level. AMOVA indicated significant genetic variation within (73–86%) compared to among (14–27%) accessions and river basins. Bayesian clustering detected ten genetic clusters, with several degrees of admixture among river basins, except for the genetically homogeneous Putumayo River basin. A core collection comprising 84 plant genotypes (25% of the full collection) was established, representing 90.82% of the overall allelic diversity. These results have important implications for M. dubia conservation strategies and breeding programs, in demonstrating a need for genetic connectivity between populations but preserving unique genetic resources in isolated basins. These results validate the expected levels of diversity and population subdivision in a crop and stress the need to secure genetically diverse germplasms, underscoring the importance of thorough genetic characterization for ex situ germplasm management. [ABSTRACT FROM AUTHOR]

Interspecific hybridization between two different Brassicaceae species, namely Brassica rapa ssp. pekinensis (♀) (AA, 2n = 2x = 20) and genetically modified Brassica napus (♂) (AACC, 2n = 4x = 38), was performed to study the transmission of a herbicide resistance gene from a tetraploid to a diploid Brassica species. Initially, four different GM B. napus lines were used for hybridization with B. rapa via hand pollination. Among the F1 hybrids, the cross involving the B. rapa (♀) × GM B. napus (♂) TG#39 line exhibited the highest recorded crossability index of 14.7 ± 5.7. However, subsequent backcross progenies (BC1, BC2, and BC3) displayed notably lower crossability indices. The F1 plants displayed morphological characteristics more aligned with the male parent B. napus, with significant segregation observed in the BC1 generation upon backcrossing with the recurrent parent B. rapa. By the BC2 and BC3 generations, the progeny stabilized, manifesting traits from both parents to varying degrees. Cytogenetic analysis revealed a substantial reduction in chromosome numbers, particularly in backcrossing progenies. BC1 plants typically exhibited 21–25 chromosomes, while BC2 progenies showed 21–22 chromosomes, and by the BC3 generation, stability was achieved with an average of 20 chromosomes. SSR marker analysis confirmed the progressive reduction of C-genome regions, retaining minimal C-genome-specific bands throughout successive backcrossing. Despite the extensive elimination of C-genome-specific genomic regions, the glyphosate resistance gene from the male parent B. napus was introgressed into BC3 progenies, suggesting that the glyphosate resistance gene located and introgressed in A-chromosome/genome regions of the Brassica plants. [ABSTRACT FROM AUTHOR]

Abstract The study was designed to validate the previously reported 34 SSR markers using 78 chilli genotypes to detect significant trait specific markers as well as superior genotypes resistant to Phytophthora capsici root rot (PcRR). In this context, the identification of germplasm with higher yield per plant (YPP) leads to hype in stress tolerance index (STI) in genotypes, Chakwal3 (11.98), Greenfire (10.14), Advanta5017 (9.94) and Chakwal4 (7.8). The identified genotypes were also found as resistant and moderately resistant due to existence of below 50% of disease incidence. Moreover, biplot showed the interrelation of STI with YPP through the formation of acute angle by their respective vectors. In the current study, the markers Hpms1172 and CAMS177 was found significant for STI. However, the marker CAMS066 was found associated with relative cell injury, CA06g27450 with disease incidence and CAMS173 with relative leaf damage. The bright bands on gel pictures of significant markers showed the association of these markers with resistant genotypes i.e. Chakwal3, Advanta-5017 and Chakwal4 as well as with a single moderately resistant genotype i.e. Greenfire. The markerstudes confirmed the phenotypic data by showing association of markers i.e. Hpms1172 and CAMS177, r with stress tolerance index. The principal coordinate analysis aligned with the results obtained from marker-assisted selection. Thus, currently practiced marker assisted selection detected high yielding genotypes in PcRR disease stress condition that will be helpful in progressing breeding programs in chilli.

Genetic markers are powerful tools for understanding genetic diversity and the molecular basis of traits, ushering in a new era of molecular breeding in crops. Over the past 50 years, DNA markers have rapidly changed, moving from hybridization-based and second-generation-based to sequence-based markers. Simple sequence repeats (SSRs) are the ideal markers in plant breeding, and they have numerous desirable properties, including their repeatability, codominance, multi-allelic nature, and locus specificity. They can be generated from any species, which requires prior sequence knowledge. SSRs may serve as evolutionary tuning knobs, allowing for rapid identification and adaptation to new circumstances. The evaluations published thus far have mostly ignored SSR polymorphism and gene evolution due to a lack of data regarding the precise placements of SSRs on chromosomes. However, NGS technologies have made it possible to produce high-throughput SSRs for any species using massive volumes of genomic sequence data that can be generated fast and at a minimal cost. Though SNP markers are gradually replacing the erstwhile DNA marker systems, SSRs remain the markers of choice in orphan crops due to the lack of genomic resources at the reference level and their adaptability to resource-limited labor. Several bioinformatic approaches and tools have evolved to handle genomic sequences to identify SSRs and generate primers for genotyping applications in plant breeding projects. This paper includes the currently available methodologies for producing SSR markers, genomic resource databases, and computational tools/pipelines for SSR data mining and primer generation. This review aims to provide a 'one-stop shop' of information to help each new user carefully select tools for identifying and utilizing SSRs in genetic research and breeding programs. [ABSTRACT FROM AUTHOR]

Microsatellites, also known as SSR or STR, are essential molecular markers in genomic research, playing crucial roles in genetic mapping, population genetics, and evolutionary studies. Their applications range from plant breeding to forensics, highlighting their diverse utility across disciplines. Despite their widespread use, traditional methods for SSR analysis are often laborious and time-consuming, requiring significant resources and expertise. To address these challenges, a variety of computational tools for SSR analysis have been developed, offering faster and more efficient alternatives to traditional methods. However, selecting the most appropriate tool can be daunting due to rapid technological advancements and the sheer number of options available. This study presents a comprehensive review and analysis of 74 SSR tools, aiming to provide researchers with a valuable resource for SSR analysis tool selection. The methodology employed includes thorough literature reviews, detailed tool comparisons, and in-depth analyses of tool functionality. By compiling and analyzing these tools, this study not only advances the field of genomic research but also contributes to the broader scientific community by facilitating informed decision-making in the selection of SSR analysis tools. Researchers seeking to understand SSRs and select the most appropriate tools for their projects will benefit from this comprehensive guide. Overall, this study enhances our understanding of SSR analysis tools, paving the way for more efficient and effective SSR research in various fields of study.

Farmers in Garut, West Java, grow mixed varieties of Arabica coffee (Coffea arabica L.). Subsequently, they use harvested beans as seeds. Intercrossing among varieties may result in hybrid progenies and harvesting hybrid progenies as seed results in genetic variations. This research aims to evaluate the genetic diversity of Arabica coffee grown in a mixed population. Ninety-one Arabica coffees comprised 37 Arabica cv. “Ahernt GRT KN” (yellow-), 45 “Sigararutang” (red-), and nine “S795” (red berries) were sampled. Twenty SSR primer pairs were validated using 15 samples representing three varieties; six were polymorphic and used to genotype 91 Arabica accessions. Genetic data were analyzed using PowerMaker 3.25 and Darwin version 6 software. The results showed that the six SSR loci generated from 2 – 3 alleles, with an average of 2.17 alleles per locus. Genetic analysis of Arabica coffee from Garut, West Java, generated SSR markers with an average PIC of 0,33 across loci and varieties. The PIC within Arabica coffee cv. “Ahernt GRT KN” and “Sigararutang” were low, and within “S795” was moderate. Those PICs indicate the presence of more genetic diversity within “S795” than the other two cultivars. The Ho across Arabica coffee cv. “Sigararutang” and “S795” were lower than the He values, confirming their self-pollination nature. However, the Ho values of Arabica coffee cv. “Ahernt GRT KN” was larger than the others, indicating the presence of residual heterozygosity and a low percentage of recent outcrossing. The low Ho values of “Sigararutang” suggest that Arabica coffee is homozygous. Arabica coffee cv. “S795” also showed a low Ho value, but its moderate He value indicates the presence of more genetic diversity than the other cultivars.

Abstract Background Tandem repeats are specific sequences in genomic DNA repeated in tandem that are present in all organisms. Among the subcategories of TRs we have Satellite repeats, that is divided into macrosatellites, minisatellites, and microsatellites, being the last two of specific interest because they can identify polymorphisms between organisms due to their instability. Currently, most mining tools focus on Simple Sequence Repeats (SSR) mining, and only a few can identify SSRs in the coding regions. Results We developed a microsatellite mining software called SATIN (Micro and Mini SATellite IdentificatioN tool) based on a new sliding window algorithm written in C and Python. It represents a new approach to SSR mining by addressing the limitations of existing tools, particularly in coding region SSR mining. SATIN is available at https://github.com/labgm/SATIN.git . It was shown to be the second fastest for perfect and compound SSR mining. It can identify SSRs from coding regions plus SSRs with motif sizes bigger than 6. Besides the SSR mining, SATIN can also analyze SSRs polymorphism on coding-regions from pre-determined groups, and identify SSRs differentially abundant among them on a per-gene basis. To validate, we analyzed SSRs from two groups of Escherichia coli (K12 and O157) and compared the results with 5 known SSRs from coding regions. SATIN identified all 5 SSRs from 237 genes with at least one SSR on it. Conclusions The SATIN is a novel microsatellite search software that utilizes an innovative sliding window technique based on a numerical list for repeat region search to identify perfect, and composite SSRs while generating comprehensible and analyzable outputs. It is a tool capable of using files in fasta or GenBank format as input for microsatellite mining, also being able to identify SSRs present in coding regions for GenBank files. In conclusion, we expect SATIN to help identify potential SSRs to be used as genetic markers.

Background: Tandem repeats are specific sequences in genomic DNA repeated in tandem that are present in all organisms. Among the subcategories of TRs we have Satellite repeats, that is divided into macrosatellites, minisatellites, and microsatellites, being the last two of specific interest because they can identify polymorphisms between organisms due to their instability. Currently, most mining tools focus on Simple Sequence Repeats (SSR) mining, and only a few can identify SSRs in the coding regions. Results: We developed a microsatellite mining software called SATIN (Micro and Mini SATellite IdentificatioN tool) based on a new sliding window algorithm written in C and Python. It represents a new approach to SSR mining by addressing the limitations of existing tools, particularly in coding region SSR mining. SATIN is available at https://github.com/labgm/SATIN.git. It was shown to be the second fastest for perfect and compound SSR mining. It can identify SSRs from coding regions plus SSRs with motif sizes bigger than 6. Besides the SSR mining, SATIN can also analyze SSRs polymorphism on coding-regions from pre-determined groups, and identify SSRs differentially abundant among them on a per-gene basis. To validate, we analyzed SSRs from two groups of Escherichia coli (K12 and O157) and compared the results with 5 known SSRs from coding regions. SATIN identified all 5 SSRs from 237 genes with at least one SSR on it. Conclusions: The SATIN is a novel microsatellite search software that utilizes an innovative sliding window technique based on a numerical list for repeat region search to identify perfect, and composite SSRs while generating comprehensible and analyzable outputs. It is a tool capable of using files in fasta or GenBank format as input for microsatellite mining, also being able to identify SSRs present in coding regions for GenBank files. In conclusion, we expect SATIN to help identify potential SSRs to be used as genetic markers. [ABSTRACT FROM AUTHOR]

Introduction: The Cucurbitaceae family comprises many agronomically important members, that bear nutritious fruits and vegetables of great economic importance. Coccinia grandis, commonly known as Ivy gourd, belongs to this family and is widely consumed as a vegetable. Members of this family are known to display an impressive range of variation in fruit morphology. Although there have been studies on flower development in Ivy gourd, fruit development remains unexplored in this crop. Methods: In this study, comparative transcriptomics of two Ivy gourd cultivars namely "Arka Neelachal Kunkhi" (larger fruit size) and "Arka Neelachal Sabuja" (smaller fruit size) differing in their average fruit size was performed. A de novo transcriptome assembly for Ivy gourd was developed by collecting fruits at different stages of development (5, 10, 15, and 20 days after anthesis i.e. DAA) from these two varieties. The transcriptome was analyzed to identify differentially expressed genes, transcription factors, and molecular markers. Results: The transcriptome of Ivy gourd consisted of 155205 unigenes having an average contig size of 1472bp. Unigenes were annotated on publicly available databases to categorize them into different biological functions. Out of these, 7635 unigenes were classified into 38 transcription factor (TF) families, of which Trihelix TFs were most abundant. A total of 11,165 unigenes were found to be differentially expressed in both the varieties and the in silico expression results were validated through real-time PCR. Also, 98768 simple sequence repeats (SSRs) were identified in the transcriptome of Ivy gourd. Discussion: This study has identified a number of genes, including transcription factors, that could play a crucial role in the determination of fruit shape and size in Ivy gourd. The presence of polymorphic SSRs indicated a possibility for markerassisted selection for crop breeding in Ivy gourd. The information obtained can help select candidate genes that may be implicated in regulating fruit development and size in other fruit crops. [ABSTRACT FROM AUTHOR]

The present investigation unveiled the first complete chloroplast (cp) genome of Tribulus macropterus var. arabicus (Hosni) Al-Hemaid & J. Thomas (Zygophyllaceae), a medicinal herb, indigenous to Saudi Arabia. The cp genome, comprising a length of 158,179 bp and a GC content of 35.8%, exhibited the typical circular quadripartite arrangement of flowering plants, including two inverted repeat regions (25842 bp) separated by a large single-copy (88873 bp) and a small single-copy region (17622 bp). Genome annotation unraveled 132 genes, comprising of 87 protein-coding genes, 37 tRNAs and eight rRNAs. A comparative plastomics approach demonstrated a very similar genome structure and gene organization in closely related taxa. Whole-genome alignment indicated that the inverted repeat regions exhibited greater conservation when compared to the single copy zones. Repeat analysis of the cp genome identified 80 simple sequence repeats, with the majority (64) being mononucleotides. Among the longer repeats, forward repeats were frequent (20) followed by palindromic and reverse repeats. The nucleotide diversity endeavor identified some hypervariable sites (rpl22, cemA, trnLUAG) in the small and large single copies which would offer opportunities to design molecular markers for potential application in taxonomic identification. Phylogenetic analysis with rbcL barcode elucidated the distinct position of T. macropterus var. arabicus compared to T. macropterus within Zygophyllaceae and further validated the assembly. The findings of this investigation provide significant insights into biological fields, particularly enhancing the current understanding of the genetic and evolutionary variations within Zygophyllaceae. [ABSTRACT FROM AUTHOR]

Wood-apple (Limonia acidissima L., Rutaceae) is an underutilized fruit tree species in the Indian subcontinent. The edible fruit and other parts of the wood-apple tree have immense medicinal and nutritional properties. In this study, we used 24 polymorphic simple sequence repeats (SSRs), developed de novo through RNA sequencing, to estimate the genetic diversity and population structure in 96 wood-apple samples collected from 16 states of India. We identified 11,401 SSRs from 215,859 unigenes. The dinucleotide repeat motifs were the most abundant SSRs (56.89%), followed by trinucleotide (37.61%) and tetranucleotide repeats (3.63%). SSR profiling detected 129 alleles, with an average of 5.37 per locus and an average PIC value of 0.62. The Jaccard's genetic dissimilarity among 96 samples ranged from 0.39 to 1.0 (average 0.94 ± 0.03). Cluster analyses by Neighbour-joining (NJ) and principal coordinates analysis (PCoA) grouped all samples into three clusters. The Bayesian model-based structure analysis also identified three subpopulations. Population genetic analysis showed moderate gene diversity (0.226 ± 0.007) and Shannon's information index (0.369 ± 0.009) in the species. Analysis of molecular variance (AMOVA) revealed higher genetic variance within the populations than among populations, while the FST statistics (0.134, p < 0.010) indicated low but significant genetic differentiation. The overall results indicated moderate to high genetic diversity among wood-apple samples. This study identified 14 diverse samples and nine highly informative SSRs that could be useful for developing breeding programs to improve the species and conserve its genetic resources. [ABSTRACT FROM AUTHOR]

Simple sequence repeats (SSRs) are present across both coding & non-coding regions of the genomes of all organisms with diversity in incidence, complexity, and repetition. The present study is focused on Adenoviridae which are non-enveloped viruses consisting of linear double-stranded DNA genome. The members of Adenoviridae are known to cause respiratory illnesses like the common cold (cough, runny nose, mild fever), pneumonia (occasionally), keratoconjunctivitis (infection in the eye known as pink eye), croup and cystitis (inflammation of the bladder). Our investigation aims to extract and analyse SSRs from 68 Adenoviridae genomes. Virus genome sequences were retrieved from NCBI and SSRs extracted from MISA. ETE3 and iTOL were used for the phylogenetic tree construction, annotation and visualization. The genome length of Adenoviridae genomes ranged from 26163 bp to 45667 bp while GC content varied from 33.6 % to 66.9 %. Genome wide analysis revealed a total incidence of 9861 SSRs and 793 cSSRs. The minimum and maximum range of SSR incidence is 112 (A67) to 203 (A61) respectively. The most prevalent mono, di and tri-SSR motif is “A”, “GC/CG” and “GAG/CTC” comprised of 1177, 1859 and 201 occurrences respectively. About 78 % SSRs are present in the coding region in the studied genomes. In terms of protein specific distribution, DNA polymerase enzyme had the highest incidence of 485 SSRs. The presence of mono-SSRs in the A/T region is a marker for host determination and divergence. The average mono-SSRs present in A/T region is 67.14 % and it ranged from 16.22 % to 99.11 %. The high prevalence of mono-SSRs in the A/T region was associated with human and related species as hosts. Further, the clustering of viruses as per their hosts was observed in the phylogenetic tree suggesting the role of host in viral evolution. The presence of unique and conserved cSSRs as genome markers has also been highlighted.

Interspecific hybridization between two different Brassicaceae species, namely Brassica rapa ssp. pekinensis (♀) (AA, 2n = 2x = 20) and genetically modified Brassica napus (♂) (AACC, 2n = 4x = 38), was performed to study the transmission of a herbicide resistance gene from a tetraploid to a diploid Brassica species. Initially, four different GM B. napus lines were used for hybridization with B. rapa via hand pollination. Among the F1 hybrids, the cross involving the B. rapa (♀) × GM B. napus (♂) TG#39 line exhibited the highest recorded crossability index of 14.7 ± 5.7. However, subsequent backcross progenies (BC1, BC2, and BC3) displayed notably lower crossability indices. The F1 plants displayed morphological characteristics more aligned with the male parent B. napus, with significant segregation observed in the BC1 generation upon backcrossing with the recurrent parent B. rapa. By the BC2 and BC3 generations, the progeny stabilized, manifesting traits from both parents to varying degrees. Cytogenetic analysis revealed a substantial reduction in chromosome numbers, particularly in backcrossing progenies. BC1 plants typically exhibited 21–25 chromosomes, while BC2 progenies showed 21–22 chromosomes, and by the BC3 generation, stability was achieved with an average of 20 chromosomes. SSR marker analysis confirmed the progressive reduction of C-genome regions, retaining minimal C-genome-specific bands throughout successive backcrossing. Despite the extensive elimination of C-genome-specific genomic regions, the glyphosate resistance gene from the male parent B. napus was introgressed into BC3 progenies, suggesting that the glyphosate resistance gene located and introgressed in A-chromosome/genome regions of the Brassica plants.

Introduction: Simple sequence repeats (SSR), also known as microsatellites, are crucial molecular markers in both animals and plants. Despite extensive previous research on SSRs, the development of microsatellite markers in Brassica crops remains limited and inefficient. Methods: Krait software was used to identify microsatellites by genome-wide and marker development based on three recently sequenced basic species of Brassica crops in the triangle of U (Brassica rapa, B. nigra and B. oleracea), as well as three allotetraploids (B. juncea, B. napus and B. carinata) using public databases. Subsequently, the primers and the characteristics of microsatellites for most of them were accordingly designed on each chromosome of each of the six Brassica species, and their physical locations were identified, and the cross-transferability of primers have been carried out. In addition, a B-genome specific SSR marker was screened out. Results: A total of 79341, 92089, 125443, 173964, 173604, and 222160 SSR loci have been identified from the whole genome sequences of Brassica crops within the triangle of U crops, B. rapa (AA), B. nigra (BB), B. oleracea (CC), B. napus (AACC), B. juncea (AABB) and B. carinata (BBCC), respectively. Comparing the number distribution of the three allotetraploid SSR loci in the three subgenomes AA, BB and CC, results indicate that the allotetraploid species have significant reduction in the number of SSR loci in the genome compared with their basic diploid counterparts. Moreover, we compared the basic species with their corresponding varieties, and found that the microsatellite characters between the allotetraploids and their corresponding basic species were very similar or almost identical. Subsequently, each of the 40 SSR primers was employed to investigate the polymorphism potential of B. rapa (85.27%), B. nigra (81.33%) and B. oleracea (73.45%), and B. rapa was found to have a higher cross-transfer rate among the basic species in the triangle of U. Meanwhile, a B-genome specific SSR marker, BniSSR23228 possessing the (AAGGA)3 sequence characteristics was obtained, and it located in chromosome B3 with a total length of 97 bp. Discussion: In this study, results suggest that the pattern of distribution may be highly conserved during the differentiation of basic Brassica species and their allotetraploid counterparts. Our data indicated that the allotetraploidization process resulted in a significant reduction in SSR loci in the three subgenomes AA, BB and CC. The reasons may be partial gene dominated chromosomal homologous recombination and rearrangement during the evolution of basic diploid species into allotetraploids. This study provides a basis for future genomics and genetic research on the relatedness of Brassica species. [ABSTRACT FROM AUTHOR]

Simple sequence repeat (SSR) markers are valuable genetic and genomic research tools and are extensively used in major crops. However, a paucity of available molecular markers in lentils (Lens culinaris Medik.) has significantly hindered genomic studies in this vital legume crop. In this study, we developed 33 new SSR markers for lentils using an enriched genomic library and tested their polymorphism in 10 lentil cultivars. We found that 16 (48.5%) SSR markers were polymorphic. The expected heterozygosity values of the polymorphic SSR markers ranged from 0.095 to 0.820, while observed heterozygosity values varied between 0.100 and 1.00. In addition, we tested the transferability of 86 SSR markers, including the 33 newly developed ones, to other legume species. Transferability rates of lentil SSR markers to other species varied between 13.76% (Vicia sativa L.) and 2.58% (Phaseolus vulgaris L. and Trifolium pratense L.). These new SSR markers could be used in further molecular breeding, population genetics, and genetic mapping studies in lentils and other legume species. [ABSTRACT FROM AUTHOR]

Abstract Dicliptera tinctoria is a member of Acanthaceae, which has a wide distribution and contains potentially medicinal species, and exhibited pharmaceutical potentials. This study sequenced and characterized the complete chloroplast genome of Dicliptera tinctoria. The newly sequenced cpDNA of D. tinctoria was 150,733 bp in length and had a typical quadripartite structure consisting of a large single copy (LSC, 82,895 bp), a small single copy (SSC, 17,249 bp), and two inverted repeat (IRs, 25,295 bp each) regions. This genome also contained 80 protein-coding genes, 30 transfer RNAs, and four ribosomal RNAs, which is identical to other chloroplast genomes in Acanthaceae family. Nucleotides diversity analysis among chloroplast genomes of Acanthaceae species revealed eight hypervariable regions, including trnK_UUU-matK, trnC_GCA-petN, accD, rps12-clpP, rps3-rps19, ycf1-ndhF, ccsA-ndhD, and ycf1. Phylogenetic analysis revealed the paraphyly of Dicliptera species and monophyly in four Acanthaceae subfamilies. These results provide an overview of genomic variations in Acanthaceae chloroplast genome, which is helpful for further genomic studies.

IntroductionThe Cucurbitaceae family comprises many agronomically important members, that bear nutritious fruits and vegetables of great economic importance. Coccinia grandis, commonly known as Ivy gourd, belongs to this family and is widely consumed as a vegetable. Members of this family are known to display an impressive range of variation in fruit morphology. Although there have been studies on flower development in Ivy gourd, fruit development remains unexplored in this crop.MethodsIn this study, comparative transcriptomics of two Ivy gourd cultivars namely “Arka Neelachal Kunkhi” (larger fruit size) and “Arka Neelachal Sabuja” (smaller fruit size) differing in their average fruit size was performed. A de novo transcriptome assembly for Ivy gourd was developed by collecting fruits at different stages of development (5, 10, 15, and 20 days after anthesis i.e. DAA) from these two varieties. The transcriptome was analyzed to identify differentially expressed genes, transcription factors, and molecular markers.ResultsThe transcriptome of Ivy gourd consisted of 155205 unigenes having an average contig size of 1472bp. Unigenes were annotated on publicly available databases to categorize them into different biological functions. Out of these, 7635 unigenes were classified into 38 transcription factor (TF) families, of which Trihelix TFs were most abundant. A total of 11,165 unigenes were found to be differentially expressed in both the varieties and the in silico expression results were validated through real-time PCR. Also, 98768 simple sequence repeats (SSRs) were identified in the transcriptome of Ivy gourd.DiscussionThis study has identified a number of genes, including transcription factors, that could play a crucial role in the determination of fruit shape and size in Ivy gourd. The presence of polymorphic SSRs indicated a possibility for marker-assisted selection for crop breeding in Ivy gourd. The information obtained can help select candidate genes that may be implicated in regulating fruit development and size in other fruit crops.

AbstractThe family Orchidaceae is renowned for its extensive diversity. Within this family, the genus Goodyera R. Br. is classified under the subtribe Goodyerinae, comprising approximately 99 species. In this study, a species Goodyera yunnanensis Schltr., its plastid genome was characterized. The plastid genome of G. yunnanensis is 146,197 bp in size and exhibits a typical quadripartite structure with a pair of inverted repeat regions (IRs) of 25,611 bp, a large single-copy region (LSC) of 81,300 bp and a small single-copy region (SSC) of 13,675 bp. A total of 126 genes were identified, containing 80 protein-coding genes, 38 tRNA genes and 8 rRNA genes. The overall GC content is 37.2%, with corresponding values of 43.3%, 34.7% and 29.1% in IR, LSC and SSC regions, respectively. Forty-seven simple sequence repeats (SSRs) are found in G. yunnanensis plastome, and the frequency of mononucleotide repeats is significantly higher than other repeat types. Phylogenetic analysis indicates that Goodyera is resolved into four clades. G. yunnanensis belongs to the monophyletic clade A, and its phylogenetic position can be reasonably supported by morphological and molecular data.

Simple sequence repeat (SSR) markers are valuable genetic and genomic research tools and are extensively used in major crops. However, a paucity of available molecular markers in lentils (Lens culinaris Medik.) has significantly hindered genomic studies in this vital legume crop. In this study, we developed 33 new SSR markers for lentils using an enriched genomic library and tested their polymorphism in 10 lentil cultivars. We found that 16 (48.5%) SSR markers were polymorphic. The expected heterozygosity values of the polymorphic SSR markers ranged from 0.095 to 0.820, while observed heterozygosity values varied between 0.100 and 1.00. In addition, we tested the transferability of 86 SSR markers, including the 33 newly developed ones, to other legume species. Transferability rates of lentil SSR markers to other species varied between 13.76% (Vicia sativa L.) and 2.58% (Phaseolus vulgaris L. and Trifolium pratense L.). These new SSR markers could be used in further molecular breeding, population genetics, and genetic mapping studies in lentils and other legume species.

Genetic breeding is an important approach to improve the economic traits of aquaculture animals. Unfortunately, the genetic basis for important economic traits of red swamp crayfish, including growth-related traits, has not yet been reported. In the present study, two full-sib families and two natural populations were employed to identify quantitative trait loci (QTLs) for growth-related traits of red swamp crayfish. In total, 28 QTLs for five growth-related traits, including body weight, body length, carapace length, carapace width, and abdomen length, were repeatedly identified in more than two families/populations, by means of association analysis; nine of these displayed a heterosis effect. The phenotypic variations of the five investigated traits explained by the 28 QTLs ranged from 4.2% to 19.0%. Eight additive and three heterosis QTLs were further successfully validated using comparative analysis of favoured alleles between the small and big size crayfish groups in the natural population. In conclusion, this study investigated the genetic basis of growth-related traits in red swamp crayfish, which is of great significance for their application in the future molecular breeding of the species.

Genetic markers are powerful tools for understanding genetic diversity and the molecular basis of traits, ushering in a new era of molecular breeding in crops. Over the past 50 years, DNA markers have rapidly changed, moving from hybridization-based and second-generation-based to sequence-based markers. Simple sequence repeats (SSRs) are the ideal markers in plant breeding, and they have numerous desirable properties, including their repeatability, codominance, multi-allelic nature, and locus specificity. They can be generated from any species, which requires prior sequence knowledge. SSRs may serve as evolutionary tuning knobs, allowing for rapid identification and adaptation to new circumstances. The evaluations published thus far have mostly ignored SSR polymorphism and gene evolution due to a lack of data regarding the precise placements of SSRs on chromosomes. However, NGS technologies have made it possible to produce high-throughput SSRs for any species using massive volumes of genomic sequence data that can be generated fast and at a minimal cost. Though SNP markers are gradually replacing the erstwhile DNA marker systems, SSRs remain the markers of choice in orphan crops due to the lack of genomic resources at the reference level and their adaptability to resource-limited labor. Several bioinformatic approaches and tools have evolved to handle genomic sequences to identify SSRs and generate primers for genotyping applications in plant breeding projects. This paper includes the currently available methodologies for producing SSR markers, genomic resource databases, and computational tools/pipelines for SSR data mining and primer generation. This review aims to provide a ‘one-stop shop’ of information to help each new user carefully select tools for identifying and utilizing SSRs in genetic research and breeding programs.

Next-generation sequencing technologies have opened new avenues for using genomic data to study and develop molecular markers and improve genetic resources. Simple Sequence Repeats (SSRs) as genetic markers are increasingly used in molecular diversity and molecular breeding programs that require bioinformatics pipelines to analyze the large amounts of data. Therefore, there is an ongoing need for online tools that provide computational resources with minimal effort and maximum efficiency, including automated development of SSR markers. These tools should be flexible, customizable, and able to handle the ever-increasing amount of genomic data. Here we introduce MegaSSR (https:// bioinformatics.um6p.ma/MegaSSR), a web server and a standalone pipeline that enables the design of SSR markers in any target genome. MegaSSR allows users to design targeted PCR-based primers for their selected SSR repeats and includes multiple tools that initiate computational pipelines for SSR mining, classification, comparisons, PCR primer design, in silico PCR validation, and statistical visualization. MegaSSR results can be accessed, searched, downloaded, and visualized with user-friendly web-based tools. These tools provide graphs and tables showing various aspects of SSR markers and corresponding PCR primers. MegaSSR will accelerate ongoing research in plant species and assist breeding programs in their efforts to improve current genomic resources. [ABSTRACT FROM AUTHOR]

The family Orchidaceae is renowned for its extensive diversity. Within this family, the genus Goodyera R. Br. is classified under the subtribe Goodyerinae, comprising approximately 99 species. In this study, a species Goodyera yunnanensis Schltr., its plastid genome was characterized. The plastid genome of G. yunnanensis is 146,197 bp in size and exhibits a typical quadripartite structure with a pair of inverted repeat regions (IRs) of 25,611 bp, a large single-copy region (LSC) of 81,300 bp and a small single-copy region (SSC) of 13,675 bp. A total of 126 genes were identified, containing 80 protein-coding genes, 38 tRNA genes and 8 rRNA genes. The overall GC content is 37.2%, with corresponding values of 43.3%, 34.7% and 29.1% in IR, LSC and SSC regions, respectively. Forty-seven simple sequence repeats (SSRs) are found in G. yunnanensis plastome, and the frequency of mononucleotide repeats is significantly higher than other repeat types. Phylogenetic analysis indicates that Goodyera is resolved into four clades. G. yunnanensis belongs to the monophyletic clade A, and its phylogenetic position can be reasonably supported by morphological and molecular data. [ABSTRACT FROM AUTHOR]

Background Microsatellites or simple sequence repeats (SSR) consist of 1-6 nucleotide motifs of DNA or RNA which are ubiquitously present in tandem repeated sequences across genome in viruses: prokaryotes and eukaryotes. They may be localized to both the coding and non-coding regions. SSRs play an important role in replication, gene regulation, transcription, and protein function. The Caliciviridae (CLV) family of viruses have ss-RNA, non-enveloped, icosahedral symmetry 27-35 nm in diameter in size. The size of the genome lies between 6.4 and 8.6 kb. Results The incidence, composition, diversity, complexity, and host range of different microsatellites in 62 representatives of the family of Caliciviridae were systematically analyzed. The full-length genome sequences were assessed from NCBI (https://www.ncbi.nlm.nih.gov), and microsatellites were extracted through MISA software. The average genome size is about 7538 bp ranging from 6273 (CLV61) to 8798 (CLV47) bp. The average GC content of the genomes was ~ 51%. There are a total of 1317 SSRs and 53 cSSRs in the studied genomes. CLV 41 and CLV 49 contain the highest and lowest value of SSRs with 32 and 10 respectively, while CLV16 had maximum cSSR incidence of 4. There were 29 species which do not contain any cSSR. The incidence of mono-, di-, and tri-nucleotide SSRs was 219, 884, and 206, respectively. The most prevalent mono-, di-, and tri-nucleotide repeat motifs were "C" (126 SSRs), AC/CA (240 SSRs), and TGA/ACT (23 SSRs), respectively. Most of the SSRs and cSSRs are biased toward the coding region with a minimum of ~ 90% incident SSRs in the genomes' coding region. Viruses with similar host are found close to each other on the phylogenetic tree suggesting virus host being one of the driving forces for their evolution. Conclusions The Caliciviridae genomes does not conform to any pattern of SSR signature in terms of incidence, composition, and localization. This unique property of SSR plays an important role in viral evolution. Clustering of similar host in the phylogenetic tree is the evidence of the uniqueness of SSR signature. [ABSTRACT FROM AUTHOR]

The Aegean region makes a prominent contribution to Turkish viticulture. The region, which includes important grape culture centers such as Manisa, Denizli, and İzmir, has extremely favorable conditions for the cultivation of seedless grapes as well as table and wine grape varieties. In this region, which possesses rich grapevine genetic diversity together with its area and production potential, it is seen that there is variety confusion caused by different nomenclature and intravariety variations. In addition, some old grape varieties that are not used in cultivation are at risk of disappearing. In order to address these problems, the viticulture potentials of Manisa, Muğla, İzmir, Kütahya, Aydın, Denizli, Uşak, and Afyon provinces as populations were evaluated using simple sequence repeat (SSR) markers. In and research conducted using 15 SSR markers on 151 grape varieties including 149 native and 2 reference varieties, 3 similar (identical), 7 synonymous, and 13 homonymous cases were determined. The highest numbers of alleles were seen at the VVMD7 (14 alleles) and VVS2 (13 alleles) loci. The highest genetic similarity ratio among the genotypes was found to be 0.96 (96%) and the lowest value was 0.13 (13%). In population structure analysis, it was observed that there was significant gene flow among the Aegean region populations, while in Bayesian analysis, it was determined that the Aegean region populations (except the Aydın and İzmir populations) were highly admixed. The findings obtained in this research, which was carried out for the characterization of grape genetic resources at the SSR level on a regional scale, shed light for other genetic characterization studies. In addition, the results of this research may contribute to other viticulture activities (breeding, reproduction, variety registration, etc.) to be carried out in the region. [ABSTRACT FROM AUTHOR]

Knowing the extent of genetic variability among and within barley accessions provide bases of selection for breeders. However, given the heterogeneous nature of the germplasm, genetic information about barley landraces stored in gene bank of Ethiopia is not well archived. Therefore, genetic diversity study was conducted among and within 20 barley accessions represented by 194 barley plants. The accessions originate from 10 different geographic regions of Ethiopia covering a wide range of altitudes. The accessions were tested using 15 simple sequence repeat markers (SSRs). The SSR markers detected 57 alleles ranging from two (GBM1042) to seven (Bmac0040) alleles per marker indicating the genetic diversity in the barley plants. The allelic diversity across geographic regions and altitude reveals that barley accessions collected from mid and low altitudes of Ethiopia show high allelic diversity compared to accessions originating from high altitudes. Analysis of molecular variance results based on geographic regions (64.39%) and altitude classes (64.04%) also showed the highest variation within accessions compared to among accessions and groups. Both geographic regions and altitude-based genetic structure analysis detected two sub-populations. The individual assignment following the altitude of collection revealed the role of altitude in shaping the genetic diversity observed in barley populations. In general, the result of this study shows that each single barley line within the accessions should be considered as a potential source of genetic material and must be treated distinctly during germplasm collection and utilization. [ABSTRACT FROM AUTHOR]

Columbia root-knot nematode, Meloidogyne chitwoodi, parasitizes potato plants and causes visible small brown dots in the tuber flesh that dramatically reduce the market value of the crop. In the Pacific Northwest, two races of M. chitwoodi, Race 1 and Race 2, and a pathotype, Race 1Roza, exist. The races and pathotype of M. chitwoodi are primarily differentiated based on host tests. Currently, M. chitwoodi can be differentiated from M. hapla and other Meloidogyne spp. based on morphology and molecular markers; however, the different pathotypes of M. chitwoodi cannot be differentiated using the aforementioned techniques. Based on the in silico genome comparison of M. chitwoodi Race 1, Race 2, and Race 1Roza, we developed diagnostic molecular markers that could successfully differentiate between M. chitwoodi races and the pathotype. We also developed three race-specific markers that can specifically amplify Race 1, Race 2, or Race 1Roza. Overall, we have potential markers that can successfully differentiate the two races and one pathotype of M. chitwoodi through simplex PCR, which has a potential application in plant disease diagnostics. [Figure: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Next-generation sequencing technologies have opened new avenues for using genomic data to study and develop molecular markers and improve genetic resources. Simple Sequence Repeats (SSRs) as genetic markers are increasingly used in molecular diversity and molecular breeding programs that require bioinformatics pipelines to analyze the large amounts of data. Therefore, there is an ongoing need for online tools that provide computational resources with minimal effort and maximum efficiency, including automated development of SSR markers. These tools should be flexible, customizable, and able to handle the ever-increasing amount of genomic data. Here we introduce MegaSSR (https://bioinformatics.um6p.ma/MegaSSR), a web server and a standalone pipeline that enables the design of SSR markers in any target genome. MegaSSR allows users to design targeted PCR-based primers for their selected SSR repeats and includes multiple tools that initiate computational pipelines for SSR mining, classification, comparisons, PCR primer design, in silico PCR validation, and statistical visualization. MegaSSR results can be accessed, searched, downloaded, and visualized with user-friendly web-based tools. These tools provide graphs and tables showing various aspects of SSR markers and corresponding PCR primers. MegaSSR will accelerate ongoing research in plant species and assist breeding programs in their efforts to improve current genomic resources.

Abstract The GenoSSRFinder is a new tool enables the research of Simple Sequence Repeats (SSRs) in DNA sequences and genomes much simpler and more precise in short time. The analysis is carried out by targeting a certain SSR in genome and gene sequences. This utility is quick, accurate, and does its function quite well. It quickly goes across the sequence, revealing all of the locations at which the selected SSR may be found. This tool will tell researchers where selected SSR begins and where it stops, how long it is, how often it repeats, and how long each repetition is. GenoSSRFinder gets the findings quickly, and they will be simple to comprehend. Therefore, when studying SSRs, researchers will have more time to use to thorough work as a result of this time savings. In addition, it provides a valuable information since it is highly precise. GenoSSRFinder is simple to use and produces high-quality findings. It is also accelerating SSRs gene research, which is a direct result of the new approach we use to analyse SSRs. Three case studies in this study demonstrated the usefulness of this program by immediately studying a particular SSR that was associated with genetic illness, biodiversity and criminal science in living organisms. This demonstration explains that GenoSSRFinder might be utilized in a wide variety of fields, such as the research of genetic illnesses, the biodiversity and genetic studies, or even in criminal investigations.

Abstract Background Dung beetles recycle organic matter through the decomposition of feces and support ecological balance. However, these insects are threatened by the indiscriminate use of agrochemicals and habitat destruction. Copris tripartitus Waterhouse (Coleoptera: Scarabaeidae), a dung beetle, is listed as a class-II Korean endangered species. Although the genetic diversity of C. tripartitus populations has been investigated through analysis of mitochondrial genes, genomic resources for this species remain limited. In this study, we analyzed the transcriptome of C. tripartitus to elucidate functions related to growth, immunity and reproduction for the purpose of informed conservation planning. Results The transcriptome of C. tripartitus was generated using next-generation Illumina sequencing and assembled de novo using a Trinity-based platform. In total, 98.59% of the raw sequence reads were processed as clean reads. These reads were assembled into 151,177 contigs, 101,352 transcripts, and 25,106 unigenes. A total of 23,450 unigenes (93.40%) were annotated to at least one database. The largest proportion of unigenes (92.76%) were annotated to the locally curated PANM-DB. A maximum of 5,512 unigenes had homologous sequences in Tribolium castaneum. Gene Ontology (GO) analysis revealed a maximum of 5,174 unigenes in the Molecular function category. Further, in Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, a total of 462 enzymes were associated with established biological pathways. Based on sequence homology to known proteins in PANM-DB, representative immunity, growth, and reproduction-related genes were screened. Potential immunity-related genes were categorized into pattern recognition receptors (PRRs), the Toll-like receptor signaling pathway, the MyD88- dependent pathway, endogenous ligands, immune effectors, antimicrobial peptides, apoptosis, and adaptation-related transcripts. Among PRRs, we conducted detailed in silico characterization of TLR-2, CTL, and PGRP_SC2-like. Repetitive elements such as long terminal repeats, short interspersed nuclear elements, long interspersed nuclear elements and DNA elements were enriched in the unigene sequences. A total of 1,493 SSRs were identified among all unigenes of C. tripartitus. Conclusions This study provides a comprehensive resource for analysis of the genomic topography of the beetle C. tripartitus. The data presented here clarify the fitness phenotypes of this species in the wild and provide insight to support informed conservation planning.

Abstract Background Ziziphus species particularly Ziziphus mauritiana and Ziziphus nummularia constitute an important part of genetic resources in India. They contribute economically as a fruit crop with lots of morphological and pomological variability. In current study, 48 accessions belonging to two wild Ziziphus species, i.e., Z. mauritiana and Z. nummularia, were characterized using SSR markers. In addition, external features were also examined using stereomicroscope. Results Present investigation was done to explore the genetic structure of North Indian jujube. In total, 23 SSR markers detected 57 SSR alleles with an average of 2.47 alleles. Highest number of alleles (4) were detected by three primers, namely BFU1178, BFU479, and ZCMS14, while lowest number of alleles (2) were detected by fifteen primers. Highest Polymorphism Information Content (PIC) was 0.500 and shown by two primers, namely BFU528 and BFU1248, while lowest PIC (0.041) was observed in primers BFU286 with mean value of 0.443. Similarly, highest value of marker index (MI) was detected by primer BFU1178 i.e. 1.969, and lowest value of marker index was observed in primer BFU286 i.e. 0.021. Dendrogram generated using SSR markers data and principal component analysis showed two major groups of the analyzed germplasm with intermixing. STRUCTURE analysis also clustered all the accessions into two groups. We did not found correlation between geographic and genetic distances. Conclusions The preliminary results suggest that there is high level of gene pool mixing in these species which can be attributed to their cross-pollination habit. However, more such studies with large numbers of samples are required in future to gain concrete insights of the genetic structure in these species.

The contribution of vine cultivation to human welfare as well as the stimulation of basic social and cultural features of civilization has been great. The wide temporal and regional distribution created a wide array of genetic variants that have been used as propagating material to promote cultivation. Information on the origin and relationships among cultivars is of great interest from a phylogenetics and biotechnology perspective. Fingerprinting and exploration of the complicated genetic background of varieties may contribute to future breeding programs. In this review, we present the most frequently used molecular markers, which have been used on Vitis germplasm. We discuss the scientific progress that led to the new strategies being implemented utilizing state-of-the-art next generation sequencing technologies. Additionally, we attempted to delimit the discussion on the algorithms used in phylogenetic analyses and differentiation of grape varieties. Lastly, the contribution of epigenetics is highlighted to tackle future roadmaps for breeding and exploitation of Vitis germplasm. The latter will remain in the top of the edge for future breeding and cultivation and the molecular tools presented herein, will serve as a reference point in the challenging years to come. [ABSTRACT FROM AUTHOR]

Cissus quadrangularis L., a member of the Vitaceae family, is an important medicinal plant with widespread application in Indian traditional medicines. C. quadrangularis L. whole chloroplast genome of 160,404 bp was assembled using a genome skimming approach from the whole genome library. The assembled chloroplast genome contained a large single-copy region (88,987 bp), a small single-copy region (18,621 bp), and pairs of inverted repeat regions (26,398 bp). It also comprised 133 genes, including 37 tRNAs, eight rRNAs, and 88 protein-coding genes. Aside from that, we annotated three genes atpH, petB, and psbL, as well as one duplicated copy of the ycf1 gene in C. quadrangularis L. that had previously been missing from the annotation of compared Cissus chloroplast genomes. Five divergent hotspot regions such as petA_psbJ (0.1237), rps16_trnQ-UUG (0.0913), psbC_trnS-UGA (0.0847), rps15_ycf1 (0.0788), and rps2_rpoC2 (0.0788) were identified in the investigation that could aid in future species discrimination. Surprisingly, we found the overlapping genes ycf1 and ndhF on the IRb/SSC junction, rarely seen in angiosperms. The results of the phylogenetic study showed that the genomes of the Cissus species under study formed a single distinct clade. The detailed annotations given in this study could be useful in the future for genome annotations of Cissus species. The current findings of the study have the potential to serve as a useful resource for future research in the field of population genetics and the evolutionary relationships in the Cissus genus. [ABSTRACT FROM AUTHOR]

Aim of study: To characterize and compare the genetic resources and gain some insights into the evolutionary history of Aleppo pine (Pinus halepensis Mill.) and Brutia pine (Pinus brutia Ten.) species which are both distributed across more than 8 million hectares of area in the Mediterranean Basin. Study area: Fifty-six populations from eight Mediterranean basin countries where P. halepensis and P. brutia species are located. Materials and methods: We analyzed 1344 seeds belonging to 56 populations using five cpSSR primers (Pt15169, Pt30204, Pt41093, Pt87268, and Pt110048). Main results: The analysis of molecular variance (AMOVA) revealed that the genetic diversity among the Brutia pine populations was slightly higher than that of Aleppo pine (27.06% and 24.27%, respectively). The Aleppo pine populations separately displayed a clear east-west differentiation across the Mediterranean Basin, confirming previous results using other markers. Although the Brutia pine populations showed no spatial genetic pattern, geographically close populations and/or populations from their continual distribution range were genetically closer than the fragmented and/or ecologically marginal populations. Research highlights: The seven Aleppo pine populations from the eastern range (Türkiye, Greece, and Italy) were more than two-fold diverse than the 13 populations from the western range (Spain and Morocco). The eastern range of Aleppo pine and Brutia pine populations had similar levels of genetic diversity parameters. These results suggested that the Eastern Mediterranean Basin is a possible genetic diversity center for the two pine species.

Microsatellite primers are a valuable tool to use for both observational and experimental studies in numerous taxa. Here, we develop 18 and 16 microsatellite markers for the widespread duckweeds Lemna minor and Spirodela polyrhiza, respectively. Only four primers were not polymorphic when tested on samples from Europe and Western Pennsylvania, USA.

The contribution of vine cultivation to human welfare as well as the stimulation of basic social and cultural features of civilization has been great. The wide temporal and regional distribution created a wide array of genetic variants that have been used as propagating material to promote cultivation. Information on the origin and relationships among cultivars is of great interest from a phylogenetics and biotechnology perspective. Fingerprinting and exploration of the complicated genetic background of varieties may contribute to future breeding programs. In this review, we present the most frequently used molecular markers, which have been used on Vitis germplasm. We discuss the scientific progress that led to the new strategies being implemented utilizing state-of-the-art next generation sequencing technologies. Additionally, we attempted to delimit the discussion on the algorithms used in phylogenetic analyses and differentiation of grape varieties. Lastly, the contribution of epigenetics is highlighted to tackle future roadmaps for breeding and exploitation of Vitis germplasm. The latter will remain in the top of the edge for future breeding and cultivation and the molecular tools presented herein, will serve as a reference point in the challenging years to come.

Sporotrichosis is the main subcutaneous mycosis worldwide transmitted by animal or plant vectors and often escalates to outbreaks or epidemics. The current cat-transmitted sporotrichosis driven by Sporothrix brasiliensis has become a significant public health issue in South America. Transmission dynamics remain enigmatic due to the lack of development of polymorphic markers for molecular epidemiological analysis. This study used a high-throughput mining strategy to characterize simple sequence repeat (SSR) markers from Sporothrix genomes. A total of 118,140--143,912 SSR loci were identified (82,841--98,369 unique markers), with a 3651.55--3804.65 SSR/Mb density and a majority of dinucleotides motifs (GC/CG).We developed a panel of 15 highly polymorphic SSR markers suitable for genotyping S. brasiliensis, S. schenckii, and S. globosa. PCR amplification revealed 240 alleles in 180 Sporothrix isolates with excellent polymorphic information content (PIC = 0.9101), expected heterozygosity (H = 0.9159), and discriminating power (D = 0.7127), supporting the effectiveness of SSR markers in uncovering cryptic genetic diversity. A systematic population genetic study estimated three clusters, corresponding to S. brasiliensis (population 1, n = 97), S. schenckii (population 2, n = 49), and S. globosa (population 3, n = 34), with a weak signature of mixed ancestry between populations 1 and 2 or 3 and 2. Partitioning of genetic variation via AMOVA revealed highly structured populations (FPT = 0.539; Nm = 0.213; p < 0.0001), with approximately equivalent genetic variability within (46%) and between (54%) populations. Analysis of SSR diversity supports Rio de Janeiro (RJ) as the center of origin for contemporary S. brasiliensis infections. The recent emergence of cat-transmitted sporotrichosis in northeastern Brazil indicates an RJ-Northeast migration resulting in founder effects during the introduction of diseased animals into sporotrichosis-free areas. Our results demonstrated high cross-species transferability, reproducibility, and informativeness of SSR genetic markers, helping dissect deep and fine-scale genetic structures and guiding decision making to mitigate the harmful effects of the expansion of cat-transmitted sporotrichosis. [ABSTRACT FROM AUTHOR]

Background: Dung beetles recycle organic matter through the decomposition of feces and support ecological balance. However, these insects are threatened by the indiscriminate use of agrochemicals and habitat destruction. Copris tripartitus Waterhouse (Coleoptera: Scarabaeidae), a dung beetle, is listed as a class-II Korean endangered species. Although the genetic diversity of C. tripartitus populations has been investigated through analysis of mitochondrial genes, genomic resources for this species remain limited. In this study, we analyzed the transcriptome of C. tripartitus to elucidate functions related to growth, immunity and reproduction for the purpose of informed conservation planning. Results: The transcriptome of C. tripartitus was generated using next-generation Illumina sequencing and assembled de novo using a Trinity-based platform. In total, 98.59% of the raw sequence reads were processed as clean reads. These reads were assembled into 151,177 contigs, 101,352 transcripts, and 25,106 unigenes. A total of 23,450 unigenes (93.40%) were annotated to at least one database. The largest proportion of unigenes (92.76%) were annotated to the locally curated PANM-DB. A maximum of 5,512 unigenes had homologous sequences in Tribolium castaneum. Gene Ontology (GO) analysis revealed a maximum of 5,174 unigenes in the Molecular function category. Further, in Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, a total of 462 enzymes were associated with established biological pathways. Based on sequence homology to known proteins in PANM-DB, representative immunity, growth, and reproduction-related genes were screened. Potential immunity-related genes were categorized into pattern recognition receptors (PRRs), the Toll-like receptor signaling pathway, the MyD88- dependent pathway, endogenous ligands, immune effectors, antimicrobial peptides, apoptosis, and adaptation-related transcripts. Among PRRs, we conducted detailed in silico characterization of TLR-2, CTL, and PGRP_SC2-like. Repetitive elements such as long terminal repeats, short interspersed nuclear elements, long interspersed nuclear elements and DNA elements were enriched in the unigene sequences. A total of 1,493 SSRs were identified among all unigenes of C. tripartitus. Conclusions: This study provides a comprehensive resource for analysis of the genomic topography of the beetle C. tripartitus. The data presented here clarify the fitness phenotypes of this species in the wild and provide insight to support informed conservation planning. [ABSTRACT FROM AUTHOR]

Exploring elite alleles based on association analysis of economic traits will promote the breeding, development, and utilization of Prunus sibirica resources. This study used 113 P. sibirica clones with diverse provenances to perform association mapping of 20 economic traits based on 149 pairs of highly polymorphic simple sequence repeat (SSR) markers. Population structure analysis split the clones into three subgroups. General linear model (GLM) and mixed linear model (MLM) identified 117 SSR loci associated with 20 P. sibirica economic traits, specifically fruit (62 loci), nuclear (69), and kernel (46) traits. Phenotypic variation in these three sets of traits ranged 8.93–82.12% (GLM) or 0.23–84.82% (MLM), 10.96–56.65% (GLM) or 0.17–52.54% (MLM), and 5.17–59.41% (GLM) or 4.86–59.89% (MLM), respectively. Of the 117 marker SSR loci, 35 were simultaneously associated with two or more economic traits. In addition, phenotypic effects analysis identified 1842 elite alleles and seven typical materials related to key economic traits. We then designed elite parental combinations through crossing the typical materials. In conclusion, our study detected alleles and plant materials that allowed the identification of clones expressing desired traits of interest. These findings can be used for improved, targeted breeding of P. sibirica through marker-assisted selection. [ABSTRACT FROM AUTHOR]

Mutations in simple sequence repeat loci underlie many inherited disorders in humans, and are increasingly recognized as important determinants of natural phenotypic variation. In eukaryotes, mutations in these sequences are primarily repaired by the MutSβ mismatch repair complex. To better understand the role of this complex in mismatch repair and the determinants of simple sequence repeat mutation predisposition, we performed mutation accumulation in yeast strains with abrogated MutSβ function. We demonstrate that mutations in simple sequence repeat loci in the absence of mismatch repair are primarily deletions. We also show that mutations accumulate at drastically different rates in short (<8 bp) and longer repeat loci. These data lend support to a model in which the mismatch repair complex is responsible for repair primarily in longer simple sequence repeats. [ABSTRACT FROM AUTHOR]

Background: Ziziphus species particularly Ziziphus mauritiana and Ziziphus nummularia constitute an important part of genetic resources in India. They contribute economically as a fruit crop with lots of morphological and pomological variability. In current study, 48 accessions belonging to two wild Ziziphus species, i.e., Z. mauritiana and Z. nummularia, were characterized using SSR markers. In addition, external features were also examined using stereomicroscope. Results: Present investigation was done to explore the genetic structure of North Indian jujube. In total, 23 SSR markers detected 57 SSR alleles with an average of 2.47 alleles. Highest number of alleles (4) were detected by three primers, namely BFU1178, BFU479, and ZCMS14, while lowest number of alleles (2) were detected by fifteen primers. Highest Polymorphism Information Content (PIC) was 0.500 and shown by two primers, namely BFU528 and BFU1248, while lowest PIC (0.041) was observed in primers BFU286 with mean value of 0.443. Similarly, highest value of marker index (MI) was detected by primer BFU1178 i.e. 1.969, and lowest value of marker index was observed in primer BFU286 i.e. 0.021. Dendrogram generated using SSR markers data and principal component analysis showed two major groups of the analyzed germplasm with intermixing. STRUCTURE analysis also clustered all the accessions into two groups. We did not found correlation between geographic and genetic distances. Conclusions: The preliminary results suggest that there is high level of gene pool mixing in these species which can be attributed to their cross-pollination habit. However, more such studies with large numbers of samples are required in future to gain concrete insights of the genetic structure in these species. [ABSTRACT FROM AUTHOR]

Abstract Background Hepatitis E virus (HEV) is small (27–34 nm diameter) non-enveloped with positive sense ssRNA genome. Microsatellites or simple sequence repeats (SSR) are short tandem repeat sequences present across coding and non-coding regions of both prokaryotes and eukaryotes. They are involved with genome function and evolution at multiple levels. Results The complete genome sequences of 22 HEV genomes of the family Hepeviridae and genus Orthohepevirus (21 species) and Piscihepevirus (1 species) were extracted from NCBI database ( http://www.ncbi.nlm.nih.gov/ ). The extraction of microsatellites was done using Imperfect Microsatellite Extractor (IMEx) in ‘Advance-Mode’. The average genome size of the studied HEV genomes was 7003nt and it ranged from 6649nt (HEV11) to 7310nt (HEV22). The average GC content of the genomes was ~ 55%. A total of 519 SSRs and 21 cSSRS were extracted from the HEV genomes with an average incidence of 24 per genome ranging from 14 (HEV13) to 34 (HEV19). The cSSR incidence ranged from 0 (eight species) to 4 (HEV19). The genomes with no cSSR incidence had an SSR incidence range from 14 to 28. There were just four hexa-nucleotide repeat motifs and 5 penta-nucleotide repeat motifs observed. The most prevalent mono-, di-, and tri-nucleotide repeat motifs were “C”, “GT/TG”, and “GAC/CTG” respectively. The studied genomes had a minimum of ~ 90% incident SSRs present in the coding regions. Viruses with same or similar hosts are placed together on the phylogenetic tree implicating viral host being one of the driving forces for evolution. Conclusions Host range in viruses is being decided by multiple factors aided by the unique genome SSR signature and genomes of varied compositions need to be analyzed to forge a widely acceptable rule for predicting the same.

Salinity is a challenge in crop production. High salinity affects soil osmotic pressure and the balance of nutrients that inhibit plant growth. In such case, utilization of salt-tolerant rice varieties could be an alternative. This study aims to identify microsatellite markers associated with salt tolerance, compare the Inpari Unsoed 79 Agritan variety with ten other rice genotypes based on microsatellite markers and determine the qualitative composition metabolites in Inpari Unsoed 79 Agritan associated with the plant response to salinity. This research was carried out at the Laboratory of Plant Breeding and Biotechnology Universitas Jenderal Soedirman and Indonesian Institute of Sciences, Bogor. This research used eleven rice varieties and ten microsatellite markers. The identification of microsatellite markers consisted of genomic DNA extraction, quantification and qualification of DNA, amplification of microsatellite DNA and data analysis. Metabolite profiling was conducted on Gas Chromatography-Mass Spectrometry (GC-MS) instrument. The results showed that microsatellite markers RM 241, RM 515, RM 519 and RM 528 differentiate the Inpari Unsoed 79 Agritan from the IR 29 genotype. Microsatellite markers RM 129 and RM 292 distinguished the Nona Bokra from the IR 29 genotypes. The genetic relationship of eleven rice genotypes resulted in two clusters. The GC-MS metabolite compounds in Inpari Unsoed 79 Agritan are β-Alanine and trimethylsilyl ester β-Alanine, a derivative compound of β-Alanine. These findings suggested that microsatellite markers RM 129, RM 292, RM 241, RM 515, RM 519 and RM 528 were associated with salt-tolerant in the seedling stage.

Guava (Psidium guajava L.) is one of the commercially important cultivated fruit crops in India. Guava ranks fifth in production share of fruit crops in the country. During 2014-15, India produced 36.68 lakh tonnes of guava from an area of 2.68 lakh ha with 13.7 t/ha productivity. In the present investigation, a total 33 genotypes including 6 species of Psidium and 28 genotypes of Psidium guajava were characterized using 39 SSR markers. All accessions are being cultivated at IARI research farm, Pusa campus, New Delhi. Samples were collected from research farm to determine the overall diversity and to assess the similarity among genotypes using molecular markers. Out of 39 primers 26 were reproducible. A total of 64 alleles were produced by 26 primers and number of alleles per locus varied from 1 (mPgCIR01, mPgCIR18 and mPgCIR19) to 5 (mPgCIR256) with mean value of 2.34. Gene diversity ranged from 0.0377 to 0.6681 with a mean value of 0.2766. Highest gene diversity was observed for primer mPgCIR256 (0.6681) and lowest for primer mPgCIR24 (0.0377). SSR primer mPgCIR256 was most informative with high rate of polymorphism as well as number of alleles. Phenotypic correlation indicated the correlated traits with dependent yield trait. PCA and cluster analysis showed a significant phenotypic and genetic diversity among existing cultivars. All accessions were grouped into two major groups and under each group four sub groups. Maximum diversity was found in leaf traits, viz. leaf length, leaf breadth, petiole and petiole length. Genetic distance and the dissimilarity matrix showed three major clusters on the basis of genetic similarity. All these accessions were grouped in to three clusters, viz. A, B and C. While cluster A includes only cultivars, cluster B includes all species except P. friedrichsthalianum. This indicates that, the diversity among and between species is at cluster level. Genetic diversity analysis among Psidium species showed that P. guajava had more similarity with Psidium guienensis compared to other species. Present investigation indicated the presence of substantial diversity among guava species which can be utilized for future guava improvement programmes

Wheat (Triticum aestivum L.) encounters 15–20% yield loss due to fungal diseases. A study was carried out to analyse the allelic variations in functional genes associated with multiple fungal disease resistance, viz. rusts, smuts and powdery mildew in 58 contrasting wheat genotypes. The experiments were conducted at Chaudhary Charan Singh Haryana Agricultural University, Hisar, Haryana during 2020–21. A set of 29 simple sequence repeat (SSR) markers was selected for screening, out of which 24 markers showed amplifications (82.7%) and 23 showed polymorphism (95.83%) with a total of 46 alleles. Alleles per locus varied from 1 to 3 with a mean of 1.96 alleles per locus. At a similarity coefficient of 0.66, dendrogram grouped all the genotypes into 2 major clusters. Two and three dimensional plots also confirmed the distribution. Results showed that genotypes PBW 725 and WH 1268 were found to be most diverse at a similarity coefficient of 77%. SSR polymorphism rates were analysed using polymorphism information content, expected heterozygosity, marker index, discriminating power and resolving power values, where first two ranged from 0.03–0.65, and later three ranged from 0.03–1.94, 0.03–0.66 and 0.03–2.00, respectively. Based on these results, 8 proficient markers, viz. Barc232, Swm271, Xbarc124, Xbarc32, Xwmc44, Xgwm296, Gpw5029 and Xwmc557 are suggested for Indian wheat fungal disease resistance profiling. Among these, first two markers (Barc232 and Swm271) were detected in most (57) of the genotypes which are associated with ut6 and Lr75 genes, providing resistance to loose smut and leaf rust, respectively. This study can further help in gene pyramiding for producing multiple disease resistant genotypes.