Showing papers on "Genetic marker published in 2022"

An Analysis of Genetic Variability and Population Structure in Wheat Germplasm Using Microsatellite and Gene-Based Markers

Abstract: Knowledge of the natural patterns of genetic variation and their evolutionary basis is required for sustainable management and conservation of wheat germplasm. In the current study, the genetic diversity and population structure of 100 individuals from four Triticum and Aegilops species (including T. aestivum, Ae. tauschii, Ae. cylindrica, and Ae. crassa) were investigated using two gene-based markers (start codon targeted (SCoT) polymorphism and CAAT-box derived polymorphism (CBDP)) and simple-sequence repeats (SSRs). The SCoT, CBDP, and SSR markers yielded 76, 116, and 48 polymorphism fragments, respectively. The CBDP marker had greater efficiency than the SCoT and SSR markers due to its higher polymorphism content information (PIC), resolving power (Rp), and marker index (MI). Based on an analysis of molecular variance (AMOVA) performed using all marker systems and combined data, there was a higher distribution of genetic variation within species than among them. Ae. cylindrica and Ae. tauschii had the highest values for all genetic variation parameters. A cluster analysis using each marker system and combined data showed that the SSR marker had greater efficiency in grouping of tested accessions, such that the results of principal coordinate analysis (PCoA) and population structure confirmed the obtained clustering patterns. Hence, combining the SCoT and CBDP markers with polymorphic SSR markers may be useful in genetic fingerprinting and fine mapping and for association analysis in wheat and its germplasm for various agronomic traits or tolerance mechanisms to environmental stresses.

QTL identification, fine mapping, and marker development for breeding peanut (Arachis hypogaea L.) resistant to bacterial wilt

Abstract: Abstract Key message A major QTL, qBWA12 , was fine mapped to a 216.68 kb physical region, and A12.4097252 was identified as a useful KASP marker for breeding peanut varieties resistant to bacterial wilt. Abstract Bacterial wilt, caused by Ralstonia solanacearum , is a major disease detrimental to peanut production in China. Breeding disease-resistant peanut varieties is the most economical and effective way to prevent the disease and yield loss. Fine mapping the QTLs for bacterial wilt resistance is critical for the marker-assisted breeding of disease-resistant varieties. A recombinant inbred population comprising 521 lines was used to construct a high-density genetic linkage map and to identify QTLs for bacterial wilt resistance following restriction-site-associated DNA sequencing. The genetic map, which included 5120 SNP markers, covered a length of 3179 cM with an average marker distance of 0.6 cM. Four QTLs for bacterial wilt resistance were mapped on four chromosomes. One major QTL, qBWA12 , with LOD score of 32.8–66.0 and PVE of 31.2–44.8%, was stably detected in all four development stages investigated over the 3 trial years. Additionally, qBWA12 spanned a 2.7 cM region, corresponding to approximately 0.4 Mb and was fine mapped to a 216.7 kb region by applying KASP markers that were polymorphic between the two parents based on whole-genome resequencing data. In a large collection of breeding and germplasm lines, it was proved that KASP marker A12.4097252 can be applied for the marker-assisted breeding to develop peanut varieties resistant to bacterial wilt. Of the 19 candidate genes in the region covered by qBWA12 , nine NBS-LRR genes should be further investigated regarding their potential contribution to the resistance of peanut against bacterial wilt.

Quantitative Trait Loci Mapping and Development of KASP Marker Smut Screening Assay Using High-Density Genetic Map and Bulked Segregant RNA Sequencing in Sugarcane (Saccharum spp.)

Abstract: Sugarcane is one of the most important industrial crops globally. It is the second largest source of bioethanol, and a major crop for biomass-derived electricity and sugar worldwide. Smut, caused by Sporisorium scitamineum, is a major sugarcane disease in many countries, and is managed by smut-resistant varieties. In China, smut remains the single largest constraint for sugarcane production, and consequently it impacts the value of sugarcane as an energy feedstock. Quantitative trait loci (QTLs) associated with smut resistance and linked diagnostic markers are valuable tools for smut resistance breeding. Here, we developed an F1 population (192 progeny) by crossing two sugarcane varieties with contrasting smut resistance and used for genome-wide single nucleotide polymorphism (SNP) discovery and mapping, using a high-throughput genotyping method called “specific locus amplified fragment sequencing (SLAF-seq) and bulked-segregant RNA sequencing (BSR-seq). SLAF-seq generated 148,500 polymorphic SNP markers. Using SNP and previously identified SSR markers, an integrated genetic map with an average 1.96 cM marker interval was produced. With this genetic map and smut resistance scores of the F1 individuals from four crop years, 21 major QTLs were mapped, with a phenotypic variance explanation (PVE) > 8.0%. Among them, 10 QTLs were stable (repeatable) with PVEs ranging from 8.0 to 81.7%. Further, four QTLs were detected based on BSR-seq analysis. aligning major QTLs with the genome of a sugarcane progenitor Saccharum spontaneum, six markers were found co-localized. Markers located in QTLs and functional annotation of BSR-seq-derived unigenes helped identify four disease resistance candidate genes located in major QTLs. 77 SNPs from major QTLs were then converted to Kompetitive Allele-Specific PCR (KASP) markers, of which five were highly significantly linked to smut resistance. The co-localized QTLs, candidate resistance genes, and KASP markers identified in this study provide practically useful tools for marker-assisted sugarcane smut resistance breeding.

Marker-assisted selection and validation of DNA markers associated with cadmium content in durum wheat germplasm

Abstract: Cadmium (Cd) is a non-essential heavy metal having toxic effects on all living organisms. Durum wheat (Triticum durum Desf.) is widely used in human diets but has the potential to accumulate Cd. It also has a high level of genetic diversity, which may be exploited to develop cultivars with low Cd content. We aimed to perform marker-assisted selection and validate previously identified Cd markers in durum wheat germplasm for use in the investigation of accessions that accumulate low grain Cd content. We assessed 130 durum wheat accessions phenotypically and using three different molecular markers. Grain Cd contents of the studied germplasm varied 4.91-fold (26.2–128.7 μg/kg) with an average of 58.2 μg/kg. Landraces showed lower average values of grain Cd content than cultivars. Three molecular markers (usw47, Cad-5B and KASP marker Cad-5B) were used to differentiate high and low Cd accumulating lines. Results showed high correlation and successfully classified the accessions to the expected high or low Cd level; 87 accessions showed the low Cd alleles, and 43 accessions the high Cd alleles, except for five accessions with the usw47 marker that showed heterozygous status. A significant correlation coefficient (r = 0.944*) was observed among the three molecular markers. Based on molecular markers, 96.2% of the accessions were classified accurately. The KASP assay was highly effective in successfully separating low from high Cd content accessions and could be used as a molecular tool in durum wheat breeding programs, with less cost and time, targeting reduced grain Cd levels. The results of this study will allow durum wheat breeders to accelerate their progress to select suitable genotypes with the desired alleles.

Forensic Applications of Markers Present on the X Chromosome

Abstract: Microsatellite genetic markers are the gold standard for human genetic identification. Forensic analyses around the world are carried out through protocols using the analysis of STR markers in autosomal chromosomes and in the Y chromosome to solve crimes. However, these analyses do not allow for the resolution of all cases, such as rape situations with suspicion of incest, paternity without a maternal sample for comparison, and biological traces with DNA mixture where the profile sought is female, among other situations. In these complex cases, the study of X-chromosome STR markers significantly increases the probability of identification by complementing the data obtained for autosomal and Y-chromosome markers, due to the unique structure of the X chromosome and its exclusive method of inheritance. However, there are currently no validated Brazilian protocols for this purpose, nor are there any population data necessary for statistical analyses that must be included in the issuance of expert reports. Thus, the aim of this article is to provide a literary review of the applications of X-chromosomal markers in population genetics.

KASP Markers Specific for the Fertility Restorer Locus Rf1 and Application for Genetic Purity Testing in Sunflowers (Helianthus annuus L.)

Abstract: Single nucleotide polymorphisms (SNPs) were significantly associated with fertility restoration of cytoplasmic male sterility (CMS) PET1 by the restorer gene Rf1. For these SNPs, four Kompetitive allele-specific PCR (KASP) markers were successfully designed. The KASP markers cover the fertility restorer locus Rf1, spanning about 3 Mb, and clearly differentiate restorer and maintainer lines. For genetic purity testing in sunflower hybrid production, the efficiency for detecting contaminations in samples was simulated using mixtures of hypocotyls or leaves. Contaminations of restorer lines with 1%, 3%, 5%, 10%, and 50% of maintainer lines were screened with all four KASP markers. Contaminations of 10% could be clearly detected in pools of 100 plants. Contaminations below this level require detection on a single plant level. For single plant detections, ethyl methanesulfonate-treated sunflower F1 hybrids, which had been phenotypically evaluated for male sterility (potential mutation in the Rf1 gene) were screened. Nine identified either partially male-sterile or male-sterile plants were analyzed with all four KASP markers and only one proved to be a hybrid with a mutation, seven were male-sterile contaminants in the F1 seeds used (1.6%) and one a recombinant plant. The four KASP markers should be valuable tools for marker-assisted selection (MAS) in sunflower breeding regarding the restorer locus Rf1.

Screening of highly discriminative microhaplotype markers for individual identification and mixture deconvolution in East Asian populations.

Abstract: Microhaplotypes are forensic genetic markers that combine single nucleotide polymorphisms in close proximity to one another. Highly discriminative microhaplotype markers could be superior to short tandem repeats (STRs) in DNA mixture deconvolution investigations because they are not interfered by stutters. In this study, the effective number of alleles (Ae) and discrimination power values of microhaplotypes and STRs were compared. It was found that current microhaplotypes are not as discriminative as commonly used forensic STRs. Effective screening of highly discriminative microhaplotype markers were consequently conducted for East Asian populations. To satisfy different forensic application needs, four sets of microhaplotypes with Ae values ≥ 4 were screened for under different conditions that included marker length and physical distances between markers. While the four sets contained 703, 301, 337, and 190 microhaplotypes, their average Ae values reached 5.38, 6.30, 7.39, and 5.61, respectively. The microhaplotype group containing 301 markers (maximum length of 200 bp and separated by ≥ 5 million bases) was further investigated. The results showed that none of the 301 loci were exactly the same as those previously reported, while seven loci partially overlapped with known markers. While Ae values of 45 loci were ≥ 8, the Ae value of the mh17WL-008 locus reached a maximum of 93.57. Further analysis showed that the newly identified microhaplotype markers were also highly polymorphic in African, American, European, and South Asian populations.

MolMarker: A Simple Tool for DNA Fingerprinting Studies and Polymorphic Information Content Calculation

Abstract: Molecular markers and mapping are used to analyze an organism’s genes. They allow the selection of target genetic areas based on marker genotype (and not trait phenotype), facilitate the study of genetic variability and diversity, create linkage maps, and follow individuals or lines carrying certain genes. They may be used to select parental genotypes, remove linkage drag in back-crossing, and choose difficult-to-measure characteristics. Due to a lack of genetic variety in crops, the gene pools of wild crop relatives for future agricultural production have been examined. The invention of RFLP (Restriction Fragment Length Polymorphism) for linkage mapping allowed for the creation of other traditional approaches such as RAPD (Random Amplified Polymorphic DNA) and AFLP (Amplified Fragment Length Polymorphism). Accordingly, the need to describe the polymorphic information content (PIC) of the ideal marker has been raised. Marker selection reliability depends on the marker’s relationship to the genomic area of interest. Although informativeness must be estimated for genetic study design, there are no readily available tools. Earlier, PICcalc was developed to calculate heterozygosity (H) and PIC to simplify molecular investigations. These two values were corrected for dominant and co-dominant markers (binary and allelic data) to determine polymorphism quality. Due to the popularity of PICcalc web, we developed a downloadable version called MolMarker with extra functionality to reduce server maintenance.

Identification of SSR Markers Associated with Yield-Related Traits and Heterosis Effect in Winter Oilseed Rape (Brassica Napus L.)

Abstract: The identification of markers responsible for regulating important agronomic traits in rapeseed supports breeding and increases the seed yield. Microsatellite (SSR) markers are mainly used as ‘neutral’ genetic markers but are also linked with many biological functions. The objective of this study was identification of microsatellite markers associated with important agronomic traits affecting the seed yield of winter oilseed rape and with the heterosis effect for these traits. The plant material consists of four parental lines, 60 doubled haploid (DH) lines, 60 single cross hybrids, and 60 three-way cross hybrids. The association between molecular markers and observed traits was estimated using regression analysis. Among 89 SSR markers, 43 were polymorphic, and 15 were selected for mapping because they demonstrated stability in both years of observation. These markers were physically mapped in the rapeseed reference genomes and their immediate vicinity was searched to identify candidate genes associated with the studied traits. Six markers (BrGMS3837, BnEMS1119, BrGMS2901, BnGMS0509, BrGMS3688, BrGMS4057), which showed a positive estimation effect in our association analysis, and thus increased the value of a given trait or heterosis effect, turned out to be linked with genes that could be responsible for the development and growth of plants.

Determination of Genetic Diversity in Some Pumpkin Genotypes Using SSR Marker Technique

Abstract: Pumpkin (Cucurbita pepo L.) is one of the important vegetables in the Cucurbita genus of the Cucurbitaceae family. DNA markers can be used in the selection studies carried out on vegetables. Microsatellite DNA sequences, which are a very good source of polymorphisms for eukaryotic genomes, are used in the investigation of genetic diversity, the creation of genetic maps and variety determination. In this study, genetic characterization determined by using 16 SSR markers in 47 pumpkin genotypes. A similarity coefficient between 0.68-1.0 was determined between genotypes. It was determined that three genotypes clustered separately from the others. It was concluded that SSR (Simple Sequence Repeats) markers are a good choice for assessment of genetic diversity and differentiation between genotypes. As a results of this study genetic structures of the pumpkin genotypes, important data were obtained that can shorten the duration of breeding studies.

Novel and Broadly Applicable Microsatellite Markers in Identified Chromosomes of the Philippine Dengue Mosquitoes, Aedes aegypti (Diptera: Culicidae)

Abstract: Abstract Dengue is the leading arboviral infection in the Philippines. Its endemicity in the country is due to the presence of its primary mosquito vector, Aedes aegypti (L.). This species has limited microsatellite markers. This study characterized microsatellite markers screened in silico from intergenic regions of the updated reference genome of Ae. aegypti from Liverpool, U.K. Criteria for good markers are: polymorphic, inherited in a Mendelian codominant manner, no null alleles, selectively neutral, randomly associated, and broadly applicable across different regions. Genotypes were scored using ABI Peak Scanner and were screened for the presence of null alleles. Hardy-Weinberg equilibrium, linkage disequilibrium, and robustness of the markers were determined by GENEPOP using Ae. aegypti samples from selected highland and lowland sites (n = 30 each) in the Philippines and outgroups (Thailand and Vietnam). Mendelian codominant inheritance was examined using F1 offspring of Ae. aegypti family (n = 30 each) derived from samples collected from Cebu city highlands and Maramag, Bukidnon. From the 63 randomly selected markers, nine were polymorphic. Two markers (Aaeg1-3D of chromosome 1 and Aaeg3-4C of chromosome 3) satisfied all criteria, hence, are good broadly useful microsatellite markers. Two other markers (Aaeg2-2E of chromosome 2 and Aaeg3-2A of chromosome 3) met all criteria but deviated from Mendelian codominant inheritance. These new markers of the Philippine Ae. aegypti with their chromosomal locations relative to the other published markers are presented, and will ultimately be useful in a variety of population genetic studies of Ae. aegypti to protect the public health.

Applications of Molecular Markers in Fruit Crops: A Review

Abstract: Markers are any trait of an organism that can be identified with confidence and relative ease, and can be followed in a mapping population or they can be defined as heritable entities associated with the economically important trait under the control of polygenes. Molecular markers have diverse applications in fruit crop improvement, particularly in the areas of genetic diversity and varietal identification studies, disease diagnostics, hybrid detection, sex differentiation and marker assisted selection. Molecular markers provide new directions to the efforts of plant breeders particularly in gene localization, taxonomy, phylogenetic analysis and also play an important role to decrease the time required for development of new and excellent cultivars. The most interesting application of molecular markers is marker-assisted selection (MAS). Suitable DNA markers should be polymorphic in the nature and should be expressed in all tissues, organs, at various developmental stages. Compared with traditional breeding programs, molecular markers can increase the efficiency and effectiveness of fruit breeding programs.

Genome-wide SSR markers in bottle gourd: development, characterization, utilization in assessment of genetic diversity of National Genebank of India and synteny with other related cucurbits

Abstract: Lagenaria siceraria (Molina) Standley is an important cultivated crop with its immense importance in pharmaceutical industry and as vegetable. Its seed, root, stem, leaves, flower, and fruit are used as an ointment for ailment of various diseases throughout Asia. Despite its worldwide importance, informative co-dominant microsatellite markers in the bottle gourd crop are very restricted, impeding genetic improvement, cultivar identification, and phylogenetic studies. Next-generation sequencing has revolutionized the approaches for discovery, assessment, and validation of molecular markers. We conducted a genome-wide analysis, for developing SSR markers by utilizing restriction site-associated DNA sequencing (RAD-Seq) data obtained from NCBI. By performing in silico mining of microsatellite repeat motifs, we developed 45,066 perfect SSR markers. Of which 207 markers were successfully validated and 120 (57.97%) polymorphic primer pairs were utilized for an in-depth genetic diversity and population structure analysis of 96 accessions from the National Genebank of India. Tetranucleotide repeats (∼34.3%) were the most prevalent followed by trinucleotide repeats (∼30.73%), further 21.03%, 9.6%, and 4.3% of di-, penta-, and hexa-nucleotide repeats in the bottle gourd genome, respectively. Synteny of SSR markers on 11 bottle gourd linkage groups was correlated with the 7 chromosomes of cucumber (93.2%), 12 chromosomes of melon (87.4%), and 11 of watermelon (90.8%). The generated SSR markers provide a valuable tool for germplasm characterization, genetic linkage map construction, studying synteny, gene discovery, and for breeding in bottle gourd and other cucurbits species. KEY MESSAGE: Development of 45,066 perfect microsatellite markers as a valuable tool for marker assisted selection (MAS) in plant breeding.

SNP-based identification of QTL for resistance to black point caused by Bipolaris sorokiniana in bread wheat

Abstract: Black point disease caused by Bipolaris sorokiniana is a problem in wheat production worldwide. We aimed to identify major quantitative trait loci (QTL) for resistance to black point and develop molecular markers for marker-assisted selection (MAS). A recombinant inbred line (RIL) population derived from a cross between Wanyuanbai 1 (susceptible) and SN4143 (resistant) was evaluated for black point response at three locations during two years under artificial inoculation with B. sorokiniana, providing data for six environments. Thirty resistant and 30 susceptible RILs were selected to form resistant and susceptible bulks, respectively, that were genotyped by the wheat 660K SNP array; 685 single-nucleotide polymorphisms (SNPs) were identified, among which 385 (56.2%) and 115 (16.8%) were located on chromosomes 4A and 2B, respectively. Bulked segregant RNA-Seq analysis identified candidate regions on chromosomes 4A (4.60–40.28 Mb) and 5A (1.22–48.47 Mb). Genetic linkage maps were constructed for chromosomes 2B, 4A, and 5A using 59 polymorphic dCAPS and SSR markers. Finally, two QTL, designated QBB.hau-4A and QBB.hau-5A, were detected on chromosomes 4A and 5A, respectively. The resistance allele of QBB.hau-4A was derived from SN4143, and that of QBB.hau-5A came from Wanyuanbai 1. QBB.hau-4A with a large and consistent effect (15.1%) is likely to be a new locus for black point resistance. The markers linked to QBB.hau-4A and QBB.hau-5A have potential application in MAS-based breeding.

Retraction note to: ISSR markers and morphometry determine genetic diversity and population structure in Hedera helix L.

Abstract: Retraction to: Czech J. Genet. Plant Breed., 58, 2022 (2): 73–82. https://doi.org/10.17221/93/2021-CJGPBThe article was retracted by the authors based on detected errors in the data processing.

Development of single nucleotide polymorphism‐based functional molecular markers from the Lr22a gene sequence in wheat ( Triticum aestivum )

Abstract: The adult-plant leaf rust resistant gene Lr22a confers broadly effective resistance against the fungal pathogen Puccinia triticina Eriks. (Pt) in wheat that has not been extensively utilized in wheat cultivars. The objective of this study was to develop robust functional molecular markers using the Lr22a gene sequence to facilitate integration of Lr22a in breeding programmes. The Lr22a coding sequence was used to identify isolated SNPs and four kompetitive allele specific polymerase chain reaction (PCR) (KASP) markers were developed. For marker testing, a F2:3 population was developed by crossing a near-isogenic line RL4495 (Lr22a carrier) with Thatcher and phenotyped with Pt race TDBJ and genotyped with a 90K iSelect SNP array, four KASP markers and two SSR markers. A linkage map of chromosome arm 2DS that included Lr22a was constructed. The KASP markers co-segregated with Lr22a and were validated for cross-applicability on a panel of wheat lines. KASP markers Kwh636, Kwh637 and Kwh638 reliably detected the presence or absence of Lr22a. The markers developed in this study will facilitate Lr22a selection in breeding programmes.

Inheritance and identification of ISSR-RGA markers associated with powdery mildew resistance in mungbean for marker-assisted breeding

Abstract: Mungbean ( Vigna radiata (L.) R. Wilczek var. radiata ) yield is dramatically constrained by powdery mildew (PM) caused by Sphaerotheca phaseoli (Z.Y. Zhao) U. Braun (1985), which is considerably prevalent in the cool-dry season of production in South, East, and Southeast Asia countries including Thailand. Exploitation of varieties resistant to the disease is crucial to meet sustainable production. A population of 64 F 2:9 and F 2:10 recombinant inbred lines (RILs) generated by hybridization of the susceptible parent ‘Chai Nat 72’ (CN72) with the resistant parent ‘V4758’ was used to assess genetic resistance and identify inter simple sequence repeat-anchored resistance gene analog (ISSR-RGA) markers linked to the PM resistance gene. The PM response in these RILs was visually scored in the field during the winter seasons, twice in 2015 and 2018, and the segregation pattern was determined by the chi-square test (χ 2 ). The resulting segregation ratios of 1:1 indicated a qualitative nature with a dominantly inherited resistance gene conferring PM resistance. When bulk segregant analysis (BSA) was undertaken using 378 ISSR-RGA primer combinations among both parents and DNA bulks of resistant and susceptible F 2:9 individuals, 11 of these exhibited polymorphisms, and one marker I41tP379 was closest to the PM resistance gene which revealed a highly significant correlation with PM resistance (R 2 (%) = 26; P < 0.001) with a logarithm of odd (LOD) score of 5.85. The closest marker I41tP379 could trace the PM resistance gene in molecular marker assisted breeding for mungbean improvement. parents, 11 ISSR-RGA primer bands the PM resistance gene. These 11 primer combinations a total of scorable DNA bands an average of 26.18 bands/primer pair. we the polymorphic bands between male and female parents of these 11 ISSR-RGA primer combinations, the results revealed that all of these generated 39 polymorphic bands with percentages of the polymorphic bands between male and female parents of 3.70%-26.09% (average 13.60%). The highest percentage of the polymorphic bands between male and female parents was observed in marker I15PL457 (26.09%), while the lowest was observed in marker I84PMR1R600 (3.70%). Each of the primer combination produced one specific DNA band in possible association of these ISSR-RGA markers with the PM resistance gene. simple linear regression analysis to evaluate the relationship between these markers and the PM resistance phenotypes, the that five markers (I41P252, I90PMR1R400, I40R211, I41tP379, and I84PMR1R600) were significantly correlated with PM resistance (R 2 = 21%, 21%, 27%, 27%, and 30%, respectively; P < 0.05) The results revealed that LOD score of the I41tP379 marker was higher than of the other markers. the position of the I41tP379 marker putatively linked to the PM resistance gene. This marker was further observed with individuals of the 64 F 2:9 RIL population and found to be highly significantly correlated with the PM resistance (R 2 = 26%; P < 0.001) with a LOD score of 5.85.

Development of the PARMS Marker of the Dominant Genic Male Sterility (DGMS) Line and Its Utilization in Rapeseed (Brassica napus L.) Breeding

Abstract: The 8029AB line is a dominant genic male sterility (DGMS) two-type line in Brassica napus L., which can be used in a three-line approach for the seed production of rapeseed hybrids. Genetic analyses have demonstrated that the sterility of 8029A is controlled by a single dominant nuclear gene (BnMS5e) interacting with one recessive gene (BnMS5c). Six pairs of penta-primer amplification refractory mutation system (PARMS) markers were designed according to the sequence of BnMS5a, BnMS5c and BnMS5e. Two pairs of these PARMS markers were successfully identified and validated. The PARMS markers MS5-1Fc/MS5-1Ft/MS5-1R12 could distinguish BnMS5c from BnMS5a/BnMS5e, and the PARMS markers MS5-2Ft/MS5-2Fa/MS5-1R12 could genotype BnMS5a and BnMS5c/BnMS5e. The combination of these two pairs of PARMS markers could be used to identify the presence or absence of BnMS5a/BnMS5c/BnMS5e effectively. Consequently, marker-assisted selection can be carried out in the early generation to shorten the breeding period and improve the breeding efficiency.

Genome-wide core sets of SNP markers and Fluidigm assays for rapid and effective genotypic identification of Korean cultivars of lettuce (Lactuca sativa L.)

Abstract: Abstract Lettuce is one of the economically important leaf vegetables and is cultivated mainly in temperate climate areas. Cultivar identification based on the distinctness, uniformity, and stability (DUS) test is a prerequisite for new cultivar registration. However, DUS testing based on morphological features is time-consuming, labor-intensive, and costly, and can also be influenced by environmental factors. Thus, molecular markers have also been used for the identification of genetic diversity as an effective, accurate, and stable method. Currently, genome-wide single nucleotide polymorphisms (SNPs) using next-generation sequencing technology are commonly applied in genetic research on diverse plant species. This study aimed to establish an effective and high-throughput cultivar identification system for lettuce using core sets of SNP markers developed by genotyping by sequencing (GBS). GBS identified 17 877 high-quality SNPs for 90 commercial lettuce cultivars. Genetic differentiation analyses based on the selected SNPs classified the lettuce cultivars into three main groups. Core sets of 192, 96, 48, and 24 markers were further selected and validated using the Fluidigm platform. Phylogenetic analyses based on all core sets of SNPs successfully discriminated individual cultivars that have been currently recognized. These core sets of SNP markers will support the construction of a DNA database of lettuce that can be useful for cultivar identification and purity testing, as well as DUS testing in the plant variety protection system. Additionally, this work will facilitate genetic research to improve breeding in lettuce.

A novel locus conferring resistance to Puccinia hordei maps to the genomic region corresponding to Rph14 on barley chromosome 2HS

Abstract: Barley leaf rust (BLR), caused by Puccinia hordei, is best controlled through genetic resistance. An efficient resistance breeding program prioritizes the need to identify, characterize, and map new sources of resistance as well as understanding the effectiveness, structure, and function of resistance genes. In this study, three mapping populations were developed by crossing Israelian barley lines “AGG-396,” “AGG-397,” and “AGG-403” (carrying unknown leaf rust resistance) with a susceptible variety “Gus” to characterize and map resistance. Genetic analysis of phenotypic data from rust testing F3s with a P. hordei pathotype 5457 P+ revealed monogenic inheritance in all three populations. Targeted genotyping-by-sequencing of the three populations detected marker trait associations in the same genomic region on the short arm of chromosome 2H between 39 and 57 Mb (AGG-396/Gus), 44 and 64 Mb (AGG-397/Gus), and 31 and 58 Mb (AGG-403/Gus), suggesting that the resistance in all three lines is likely conferred by the same locus (tentatively designated RphAGG396). Two Kompetitive allele-specific PCR (KASP) markers, HvGBSv2-902 and HvGBSv2-932, defined a genetic distance of 3.8 cM proximal and 7.1 cM distal to RphAGG396, respectively. To increase the marker density at the RphAGG396 locus, 75 CAPS markers were designed between two flanking markers. Integration of marker data resulted in the identification of two critical recombinants and mapping RphAGG396 between markers- Mloc-28 (40.75 Mb) and Mloc-41 (41.92 Mb) narrowing the physical window to 1.17 Mb based on the Morex v2.0 reference genome assembly. To enhance map resolution, 600 F2s were genotyped with markers- Mloc-28 and Mloc-41 and nine recombinants were identified, placing the gene at a genetic distance of 0.5 and 0.2 cM between the two markers, respectively. Two annotated NLR (nucleotide-binding domain leucine-rich repeat) genes (r2.2HG0093020 and r2.2HG0093030) were identified as the best candidates for RphAGG396. A closely linked marker was developed for RphAGG396 that can be used for marker-assisted selection.

Evidence for hexasomic inheritance in <i>Chrysanthemum morifolium</i> Ramat. based on analysis of EST-SSR markers

Abstract: Chrysanthemums (Chrysanthemum morifolium Ramat.) are ornamental flowers, which are famous worldwide. The mode of inheritance has great implications for the genetic analysis of polyploid species. However, genetic analysis of chrysanthemum has been hampered because of its controversial inheritance mode (disomic or hexasomic). To classify the inheritance mode of chrysanthemums, an analysis of three approaches was carried out in an F1 progeny of 192 offspring using 223 expressed sequence tag-simple sequence repeat (EST-SSR) markers. The analysis included segregation analysis, the ratio of simplex marker alleles linked in coupling to repulsion, as well as the transmission and segregation patterns of EST-SSR marker alleles. After segregation analysis, 204 marker alleles fit hexasomic inheritance and 150 marker alleles fit disomic inheritance, showing that marker alleles were inherited predominantly in a hexasomic manner. Furthermore, the results of the analysis of allele configuration and segregation behavior of five EST-SSR markers also suggested random pairing of chromosomes. Additionally, the ratio of simplex marker alleles linked in coupling to repulsion was 1:0, further supporting hexasomic inheritance. Therefore, it could be inferred that chrysanthemum is a complete or near-complete hexasome.