Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing
Background: Persimmon (Diospyros kaki Thunb.) is the most widely cultivated species of the genus Diospyros. In this study, genetic diversity and variations in persimmon genotypes were investigated using single nucleotide polymorphism (SNP) markers identified by genotyping-by-sequencing (GBS) analysi...
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doaj-c9d69077a2cf44179b294b52a4d8879d2020-12-31T04:40:57ZengElsevierElectronic Journal of Biotechnology0717-34582021-01-01497281Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencingKyeong Bok Ma0Sang-Jin Yang1Ye-Seul Jo2Sam Seok Kang3Moon Nam4Pear Research Institute, National Institute of Horticultural & Herbal Science, 121, Byeongnyugil, Geumcheon-Myeon, Naju-si, Jeollanam-do 58216, Republic of Korea; Corresponding author.Pear Research Institute, National Institute of Horticultural & Herbal Science, 121, Byeongnyugil, Geumcheon-Myeon, Naju-si, Jeollanam-do 58216, Republic of KoreaPear Research Institute, National Institute of Horticultural & Herbal Science, 121, Byeongnyugil, Geumcheon-Myeon, Naju-si, Jeollanam-do 58216, Republic of KoreaPear Research Institute, National Institute of Horticultural & Herbal Science, 121, Byeongnyugil, Geumcheon-Myeon, Naju-si, Jeollanam-do 58216, Republic of KoreaSEEDERS Inc., N219, 11-3, Techno 1-ro, Yuseong-gu, Daejeon 34015, Republic of KoreaBackground: Persimmon (Diospyros kaki Thunb.) is the most widely cultivated species of the genus Diospyros. In this study, genetic diversity and variations in persimmon genotypes were investigated using single nucleotide polymorphism (SNP) markers identified by genotyping-by-sequencing (GBS) analysis. Results: Ninety-five persimmon accessions grown in the Pear Research Institute, National Institute Horticultural and Herbal Science, were sequenced using the Illumina Hiseq2500 platform and polymorphic SNPs were detected to develop molecular markers. These reliable SNPs were analyzed using the Kompetitive Allele Specific PCR (KASP) assay to discriminate among persimmon genotypes. GBS generated a total of 447,495,724 trimmed reads, of which 89.7% were raw reads. After demultiplexing and sequence quality trimming, 108,876,644 clean reads were mapped to the reference transcriptome. An average of 1,146,070 genotype reads were mapped. Filtering of raw SNPs in each sample led to selection of a total of 1,725,401 high-quality SNPs. The number of homozygous and heterozygous SNPs ranged from 1,933 to 6,834 and from 846 to 5,927, respectively. Conclusions: Of the 49 SNPs selected for development of an identification system for persimmons, 15 SNPs were used in the KASP assay to analyze 32 persimmon accessions. These KASP markers discriminated among all accessions.How to cite: Ma KB, Yang SJ, Jo YS, et al. Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing. Electron J Biotechnol 2021;49.https://doi.org/10.1016/j.ejbt.2020.11.003http://www.sciencedirect.com/science/article/pii/S0717345820300592AccessionGenotyping-by-sequencingHomozygousKASP markersKompetitive AlleleSpecific PCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kyeong Bok Ma Sang-Jin Yang Ye-Seul Jo Sam Seok Kang Moon Nam |
spellingShingle |
Kyeong Bok Ma Sang-Jin Yang Ye-Seul Jo Sam Seok Kang Moon Nam Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing Electronic Journal of Biotechnology Accession Genotyping-by-sequencing Homozygous KASP markers Kompetitive Allele Specific PCR |
author_facet |
Kyeong Bok Ma Sang-Jin Yang Ye-Seul Jo Sam Seok Kang Moon Nam |
author_sort |
Kyeong Bok Ma |
title |
Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing |
title_short |
Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing |
title_full |
Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing |
title_fullStr |
Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing |
title_full_unstemmed |
Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing |
title_sort |
development of kompetitive allele specific pcr markers for identification of persimmon varieties using genotyping-by-sequencing |
publisher |
Elsevier |
series |
Electronic Journal of Biotechnology |
issn |
0717-3458 |
publishDate |
2021-01-01 |
description |
Background: Persimmon (Diospyros kaki Thunb.) is the most widely cultivated species of the genus Diospyros. In this study, genetic diversity and variations in persimmon genotypes were investigated using single nucleotide polymorphism (SNP) markers identified by genotyping-by-sequencing (GBS) analysis. Results: Ninety-five persimmon accessions grown in the Pear Research Institute, National Institute Horticultural and Herbal Science, were sequenced using the Illumina Hiseq2500 platform and polymorphic SNPs were detected to develop molecular markers. These reliable SNPs were analyzed using the Kompetitive Allele Specific PCR (KASP) assay to discriminate among persimmon genotypes. GBS generated a total of 447,495,724 trimmed reads, of which 89.7% were raw reads. After demultiplexing and sequence quality trimming, 108,876,644 clean reads were mapped to the reference transcriptome. An average of 1,146,070 genotype reads were mapped. Filtering of raw SNPs in each sample led to selection of a total of 1,725,401 high-quality SNPs. The number of homozygous and heterozygous SNPs ranged from 1,933 to 6,834 and from 846 to 5,927, respectively. Conclusions: Of the 49 SNPs selected for development of an identification system for persimmons, 15 SNPs were used in the KASP assay to analyze 32 persimmon accessions. These KASP markers discriminated among all accessions.How to cite: Ma KB, Yang SJ, Jo YS, et al. Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing. Electron J Biotechnol 2021;49.https://doi.org/10.1016/j.ejbt.2020.11.003 |
topic |
Accession Genotyping-by-sequencing Homozygous KASP markers Kompetitive Allele Specific PCR |
url |
http://www.sciencedirect.com/science/article/pii/S0717345820300592 |
work_keys_str_mv |
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