scholarly journals Identification of ‘Haryejosaeng’ mandarin by multiplex SNP genotyping

2020 ◽  
Author(s):  
Seong-Beom Jin ◽  
Ho Bang Kim ◽  
Suk Man Park ◽  
Min Ju Kim ◽  
Seok-Beom Kang ◽  
...  

Abstract BackgroundMost of the satsuma mandarin (Citrus unshiu Marc.) cultivars grown on Jeju Island farms, Korea, are difficult to improve through hybridization because of polyembryony and male sterility. Therefore, their improvement has mostly been based on the selection of nucellar embryo and bud mutation. These cultivars are supplied to breeders and farms at the seedling stage, which renders their identification based only on morphological traits. In addition, because these seedlings originate from nucellar embryo and bud mutation selection, they are genetically very similar. Therefore, the present study was carried out to develop markers that can specifically and rapidly distinguish ‘Haryejosaeng,’ which is generally supplied to breeders, from other satsuma mandarin cultivars that are planted on farms.ResultsPolymerase chain reaction (PCR) was performed to distinguish ‘Haryejosaeng’ from other 8 cultivars (‘Haryejosaeng’- breeder’s stock, ‘Miyagawa wase,’ ‘Okitsu wase,’ ‘Yura wase,’ ‘Miyamoto wase,’ ‘Ueno wase,’ ‘Yonezawa wase,’ and ‘Nichinan 1 gou’) using 6 single nucleotide polymorphism (SNP) markers specific for ‘Haryejosaeng’ and one SNP primer pair, which was used as the negative control. Using a multiplex PCR, SNP markers P1 (HL-SNP-SCAF_2-23997586-F and HL-SNP-SCAF_2-23997586-R), P2 (HL-SNP-SCAF_2-36059523-F and HL-SNP-SCAF_2-36059523-R), and P5 (HL-SNP-SCAF_9-30793978-F and HL-SNP-SCAF_9-30793978-R) simultaneously yielded 165, 150, and 526 bp amplicons, respectively, for Haryejosaeng only. The SNP markers were further validated by high-resolution melting analysis. The multiplex PCR based on P1/P5 and P2/P5 was also used to identify ‘Haryejosaeng’ on a farm growing 17 different cultivars of satsuma mandarin.ConclusionsWe developed specific molecular markers for accurate identification of ‘Haryejosaeng,’ which can be performed by multiplex PCR to save time and cost associated with the supply of ‘Haryejosaeng’ to farms.

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0242940
Author(s):  
Mark Winfield ◽  
Amanda Burridge ◽  
Matthew Ordidge ◽  
Helen Harper ◽  
Paul Wilkinson ◽  
...  

Accurate identification of named accessions in germplasm collections is extremely important, especially for vegetatively propagated crops which are expensive to maintain. Thus, an inexpensive, reliable, and rapid genotyping method is essential because it avoids the need for laborious and time-consuming morphological comparisons. Single Nucleotide Polymorphism (SNP) marker panels containing large numbers of SNPs have been developed for many crop species, but such panels are much too large for basic cultivar identification. Here, we have identified a minimum set of SNP markers sufficient to distinguish apple cultivars held in the English and Welsh national collections providing a cheaper and automatable alternative to the markers currently used by the community. We show that SNP genotyping with a small set of well selected markers is equally efficient as microsatellites for the identification of apple cultivars and has the added advantage of automation and reduced cost when screening large numbers of samples.


Author(s):  
Kotaro Dokan ◽  
Sayu Kawamura ◽  
Kosuke M Teshima

Abstract Single nucleotide polymorphism (SNP) data are widely used in research on natural populations. Although they are useful, SNP genotyping data are known to contain bias, normally referred to as ascertainment bias, because they are conditioned by already confirmed variants. This bias is introduced during the genotyping process, including the selection of populations for novel SNP discovery and the number of individuals involved in the discovery panel and selection of SNP markers. It is widely recognized that ascertainment bias can cause inaccurate inferences in population genetics and several methods to address these bias issues have been proposed. However, especially in natural populations, it is not always possible to apply an ideal ascertainment scheme because natural populations tend to have complex structures and histories. In addition, it was not fully assessed if ascertainment bias has the same effect on different types of population structure. Here we examine the effects of bias produced during the selection of population for SNP discovery and consequent SNP marker selection processes under three demographic models: the island, stepping-stone, and population split models. Results show that site frequency spectra and summary statistics contain biases that depend on the joint effect of population structure and ascertainment schemes. Additionally, population structure inferences are also affected by ascertainment bias. Based on these results, it is recommended to evaluate the validity of the ascertainment strategy prior to the actual typing process because the direction and extent of ascertainment bias vary depending on several factors.


2020 ◽  
Vol 98 (12) ◽  
Author(s):  
Garrett M See ◽  
Benny E Mote ◽  
Matthew L Spangler

Abstract Numerous methods have been suggested to incorporate crossbred (CB) phenotypes and genotypes into swine selection programs, yet little research has focused on the implicit trade-off decisions between generating data at the nucleus or commercial level. The aim of this study was to investigate the impact of altering the proportion of purebred (PB) and CB phenotypes and genotypes in genetic evaluations on the response to selection of CB performance. Assuming CB and PB performance with moderate heritabilities (h2=0.4), a three-breed swine crossbreeding scheme was simulated and selection was practiced for six generations, where the goal was to increase CB performance. Phenotypes, genotypes, and pedigrees for three PB breeds (25 and 175 mating males and females for each breed, respectively), F1 crosses (400 mating females), and terminal cross progeny (2,500) were simulated. The genome consisted of 18 chromosomes with 1,800 quantitative trait loci and 72k single nucleotide polymorphism (SNP) markers. Selection was performed in PB breeds using estimated breeding value for each phenotyping/genotyping strategy. Strategies investigated were: 1) increasing the proportion of CB with genotypes, phenotypes, and sire pedigree relationships, 2) decreasing the proportion of PB phenotypes and genotypes, and 3) altering the genetic correlation between PB and CB performance (rpc). Each unique rpc scenario and data collection strategy was replicated 10 times. Results showed that including CB data improved the CB performance regardless of  rpc or data collection strategy compared with when no CB data were included. Compared with using only PB information, including 10% of CB progeny per generation with sire pedigrees and phenotypes increased the response in CB phenotype by 134%, 55%, 33%, 23%, and 21% when rpc was 0.1, 0.3, 0.5, 0.7, and 0.9, respectively. When the same 10% of CB progeny were also genotyped, CB performance increased by 243%, 54%, 38%, 23%, and 20% when the rpc was 0.1, 0.3, 0.5, 0.7, and 0.9, respectively, compared with when no CB data were utilized. Minimal change was observed in the average CB phenotype when PB phenotypes were included or proportionally removed when CB were genotyped. Removal of both PB phenotypes and genotypes when CB were genotyped greatly reduced the response in CB performance. In practice, the optimal inclusion rate of CB and PB data depends upon the genetic correlation between CB and PB animals and the expense of additional CB data collection compared with the economic benefit associated with increased CB performance.


2020 ◽  
Vol 23 (8) ◽  
pp. 993-998 ◽  
Author(s):  
N. V. Dementeva ◽  
A. B. Vakhrameev ◽  
T. A. Larkina ◽  
O. V. Mitrofanova

In the poultry industry, indicators reflecting the growth rate of young stock and the exterior characteristics of chickens are important benchmarks for breeding. Traditional selection based on phenotypic evaluation is characterized by low efficiency with a low character inheritance ratio and is difficult to apply in small groups of animals and birds bred in bioresource collections. The use of molecular genetic markers associated with economically important traits makes it possible to carry out early selection of birds. This entails an increase in the profitability of the poultry industry. Recently, single nucleotide polymorphisms (SNPs) have served as convenient markers for selection purposes. For five generations (P1–P5), an experimental selection of hens of the Pushkin breed was carried out for live weight. It was based on selection for single nucleotide polymorphism rs313744840 in the MSTN gene. As a result, a significant increase in the frequency of allele A in this gene, from 0.11 to 0.50, took place. The association of SNP markers with meat qualities in the experimental group led to changes in the exterior profile of an adult bird at 330 days of age. The individuals with the AA and AG genotypes had the greatest live weight and longest body. As a result of selection, the bird on average became larger due to an increase in the number of heterozygous individuals with long bodies and large chest girths. The depth of the chest and the width of the pelvis increased due to an increase in the frequency of allele A in the experimental population. A tendency towards an increase in these indicators with the substitution of G with A in the genotype was found. Saturation of the population with desirable alleles led to an increase in the average live weight of the chickens. Analysis of the exterior parameters of adult birds showed that this growth is achieved by increasing the depth and volume of the bird body, and not by increasing the length of the limbs. Thus, marker selection carried out for five generations in the experimental population of Pushkin breed chickens to increase body weight has reliably (p < 0.001) changed the exterior profile of adult birds.


2018 ◽  
Vol 48 (8) ◽  
Author(s):  
Leiri Daiane Barili ◽  
Naine Martins do Vale ◽  
Fabyano Fonseca e Silva ◽  
José Eustáquio de Souza Carneiro ◽  
Hinayah Rojas de Oliveira ◽  
...  

ABSTRACT: We aimed to apply genomic information based on SNP (single nucleotide polymorphism) markers for the genetic evaluation of the traits “stay-green” (SG), plant architecture (PA), grain aspect (GA) and grain yield (GY) in common bean through Bayesian models. These models were compared in terms of prediction accuracy and ability for heritability estimation for each one of the mentioned traits. A total of 80 cultivars were genotyped for 377 SNP markers, whose effects were estimated by five different Bayesian models: Bayes A (BA), B (BB), C (BC), LASSO (BL) e Ridge regression (BRR). Although, prediction accuracies calculated by means of cross-validation have been similar within each trait, the BB model stood out for the trait SG, whereas the BRR was indicated for the remaining traits. The heritability estimates for the traits SG, PA, GA and GY were 0.61, 0.28, 0.32 and 0.29, respectively. In summary, the Bayesian methods applied here were effective and ease to be implemented. The used SNP markers can help in the early selection of promising genotypes, since incorporating genomic information increase the prediction accuracy of the estimated genetic merit.


2021 ◽  
Author(s):  
Guiyun Huang ◽  
Fengying Gao ◽  
Zhigang Liu ◽  
Jianmeng Cao ◽  
Gang Chen ◽  
...  

Abstract The dojo loach Misgurnus anguillicaudatus is an endemic freshwater species to Asia. The effective conservation and molecular-aided selection of M. anguillicaudatus have been limited without sufficient molecular markers. In this study, 112 novel single nucleotide polymorphisms (SNPs) were screened based on 2b-RAD sequencing database, and 57 SNP markers were developed and characterized by genotyping 40 individuals using SNaPshot method. The observed heterozygosity (Ho) ranged from 0.025 to 0.675, while the expected heterozygosity (He) varied from 0.025 to 0.500. The minor allele frequency (MAF) ranged from 0.013 to 0.500. Among these SNPs, 18 loci were found to deviate significantly from the Hardy–Weinberg equilibrium after Bonferroni correction (P < 0.05). The first set of SNP markers developed from M. anguillicaudatus will provide valuable information in further population genetic analysis and natural resource conservation.


2021 ◽  
Vol 8 ◽  
Author(s):  
Yue Wang ◽  
Xiaoting Xu ◽  
Yuanfeng Hao ◽  
Yelun Zhang ◽  
Yuping Liu ◽  
...  

Deficiency of micronutrient elements, such as zinc (Zn) and iron (Fe), is called “hidden hunger,” and bio-fortification is the most effective way to overcome the problem. In this study, a high-density Affymetrix 50K single-nucleotide polymorphism (SNP) array was used to map quantitative trait loci (QTL) for grain Zn (GZn) and grain Fe (GFe) concentrations in 254 recombinant inbred lines (RILs) from a cross Jingdong 8/Bainong AK58 in nine environments. There was a wide range of variation in GZn and GFe concentrations among the RILs, with the largest effect contributed by the line × environment interaction, followed by line and environmental effects. The broad sense heritabilities of GZn and GFe were 0.36 ± 0.03 and 0.39 ± 0.03, respectively. Seven QTL for GZn on chromosomes 1DS, 2AS, 3BS, 4DS, 6AS, 6DL, and 7BL accounted for 2.2–25.1% of the phenotypic variances, and four QTL for GFe on chromosomes 3BL, 4DS, 6AS, and 7BL explained 2.3–30.4% of the phenotypic variances. QTL on chromosomes 4DS, 6AS, and 7BL might have pleiotropic effects on both GZn and GFe that were validated on a germplasm panel. Closely linked SNP markers were converted to high-throughput KASP markers, providing valuable tools for selection of improved Zn and Fe bio-fortification in breeding.


2021 ◽  
Vol 12 ◽  
Author(s):  
Jinkwan Jo ◽  
Youngin Kim ◽  
Geon Woo Kim ◽  
Jin-Kyung Kwon ◽  
Byoung-Cheorl Kang

Genotyping by sequencing (GBS) enables genotyping of multiple loci at low cost. However, the single nucleotide polymorphisms (SNPs) revealed by GBS tend to be randomly distributed between individuals, limiting their direct comparisons without applying the various filter options to obtain a comparable dataset of SNPs. Here, we developed a panel of a multiplex targeted sequencing method, genotyping-in-thousands by sequencing (GT-seq), to genotype SNPs in Capsicum spp. Previously developed Fluidigm® SNP markers were converted to GT-seq markers and combined with new GT-seq markers developed using SNP information obtained through GBS. We then optimized multiplex PCR conditions: we obtained the highest genotyping rate when the first PCR consisted of 25 cycles. In addition, we determined that 101 primer pairs performed best when amplifying target sequences of 79 bp. We minimized interference of multiplex PCR by primer dimer formation using the PrimerPooler program. Using our GT-seq pipeline on Illumina Miseq and Nextseq platforms, we genotyped up to 1,500 (Miseq) and 1,300 (Nextseq) samples for the optimum panel size of 100 loci. To allow the genotyping of Capsicum species, we designed 332 informative GT-seq markers from Fluidigm SNP markers and GBS-derived SNPs. This study illustrates the first application of GT-seq in crop plants. The GT-seq marker set developed here will be a useful tool for molecular breeding of peppers in the future.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Grimar Abdiel Perez ◽  
Pumipat Tongyoo ◽  
Julapark Chunwongse ◽  
Hans de Jong ◽  
Anucha Wongpraneekul ◽  
...  

AbstractThis study explored a germplasm collection consisting of 112 Luffa acutangula (ridge gourd) accessions, mainly from Thailand. A total of 2834 SNPs were used to establish population structure and underlying genetic diversity while exploring the fruit characteristics together with genetic information which would help in the selection of parental lines for a breeding program. The study found that the average polymorphism information content value of 0.288 which indicates a moderate genetic diversity for this L. acutangula germplasm. STRUCTURE analysis (ΔK at K = 6) allowed us to group the accessions into six subpopulations that corresponded well with the unrooted phylogenetic tree and principal coordinate analyses. When plotted, the STRUCTURE bars to the area of collection, we observed an admixed genotype from surrounding accessions and a geneflow confirmed by the value of FST = 0.137. AMOVA based on STRUCTURE clustering showed a low 12.83% variation between subpopulations that correspond well with the negative inbreeding coefficient value (FIS =  − 0.092) and low total fixation index (FIT = 0.057). There were distinguishing fruit shapes and length characteristics in specific accessions for each subpopulation. The genetic diversity and different fruit shapes in the L. acutangula germplasm could benefit the ridge gourd breeding programs to meet the demands and needs of consumers, farmers, and vegetable exporters such as increasing the yield of fruit by the fruit width but not by the fruit length to solve the problem of fruit breakage during exportation.


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