Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome

The shortage of polymorphic markers for the regions of wheat chromosomes that encode commercially valuable traits determined the need for studying wheat microsatellite loci. In this work, SSR markers for individual regions in the short arm of bread wheat chromosome 5B (5BS) were designed based on se...

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Main Authors: M. A. Nesterov, D. A. Afonnikov, E. M. Sergeeva, L. A. Miroshnichenko, M. K. Bragina, A. O. Bragin, G. V. Vasiliev, E. A. Salina
Format: Article
Language:English
Published: Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders 2016-01-01
Series:Вавиловский журнал генетики и селекции
Subjects:
Online Access:https://vavilov.elpub.ru/jour/article/view/488
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author M. A. Nesterov
D. A. Afonnikov
E. M. Sergeeva
L. A. Miroshnichenko
M. K. Bragina
A. O. Bragin
G. V. Vasiliev
E. A. Salina
author_facet M. A. Nesterov
D. A. Afonnikov
E. M. Sergeeva
L. A. Miroshnichenko
M. K. Bragina
A. O. Bragin
G. V. Vasiliev
E. A. Salina
author_sort M. A. Nesterov
collection DOAJ
description The shortage of polymorphic markers for the regions of wheat chromosomes that encode commercially valuable traits determined the need for studying wheat microsatellite loci. In this work, SSR markers for individual regions in the short arm of bread wheat chromosome 5B (5BS) were designed based on sequencing data for BAC clones, and the regions of the corresponding chromosome were saturated with these markers. Totally, 130 randomly selected BAC clones from the 5BS library were sequenced on the Ion Torrent platform and assembled in contigs using MIRA software. The assembly characteristics (N50 = 4 136 bp) are comparable to the recently obtained data for wheat and relative species and acceptable for identification of microsatellite loci. An algorithm utilizing the properties of complexity decompositions in  he sliding-window mode was used to detect DNA sequences with a repeat unit of 2–4 bp. Analysis of 17 770 contigs with the total length of 25 879 921 bp allowed for designing 113, 79, and 67 microsatellite (SSR) loci with a repeat unit of 2, 3, and 4 bp, respectively. The SSR markers with a motif of 3 bp were tested using nullitetrasomic lines of Chinese Spring wheat homoeologous group 5. Thus, 21 markers specific for chromosome 5B were detected. Eight of these markers were mapped to the distal region of this chromosome (bin 5BS6) using a set of Chinese Spring deletion lines for 5BS. Eight and four markers were mapped to the interstitial region (bins 5BS5 and 5BS4, respectively). One marker was mapped to a pericentromeric bin. A comparative analysis of the distribution of trinucleotide microsatellites over wheat chromosome 5B and in different cereal species suggests that the (AAG)n repeat has proliferated and has been maintained during the evolution of cereals.
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spelling doaj.art-fab56512258e48b7851d164c6e2089672024-04-11T15:30:56ZengSiberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and BreedersВавиловский журнал генетики и селекции2500-32592016-01-0119670771410.18699/VJ15.086431Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosomeM. A. Nesterov0D. A. Afonnikov1E. M. Sergeeva2L. A. Miroshnichenko3M. K. Bragina4A. O. Bragin5G. V. Vasiliev6E. A. Salina7Institute of Cytology and Genetics SB RA S, Novosibirsk, RussiaInstitute of Cytology and Genetics SB RA S, Novosibirsk, RussiaInstitute of Cytology and Genetics SB RA S, Novosibirsk, RussiaSobolev Institute of Mathematics SB RA S, Novosibirsk, RussiaInstitute of Cytology and Genetics SB RA S, Novosibirsk, RussiaInstitute of Cytology and Genetics SB RA S, Novosibirsk, RussiaInstitute of Cytology and Genetics SB RA S, Novosibirsk, RussiaInstitute of Cytology and Genetics SB RA S, Novosibirsk, RussiaThe shortage of polymorphic markers for the regions of wheat chromosomes that encode commercially valuable traits determined the need for studying wheat microsatellite loci. In this work, SSR markers for individual regions in the short arm of bread wheat chromosome 5B (5BS) were designed based on sequencing data for BAC clones, and the regions of the corresponding chromosome were saturated with these markers. Totally, 130 randomly selected BAC clones from the 5BS library were sequenced on the Ion Torrent platform and assembled in contigs using MIRA software. The assembly characteristics (N50 = 4 136 bp) are comparable to the recently obtained data for wheat and relative species and acceptable for identification of microsatellite loci. An algorithm utilizing the properties of complexity decompositions in  he sliding-window mode was used to detect DNA sequences with a repeat unit of 2–4 bp. Analysis of 17 770 contigs with the total length of 25 879 921 bp allowed for designing 113, 79, and 67 microsatellite (SSR) loci with a repeat unit of 2, 3, and 4 bp, respectively. The SSR markers with a motif of 3 bp were tested using nullitetrasomic lines of Chinese Spring wheat homoeologous group 5. Thus, 21 markers specific for chromosome 5B were detected. Eight of these markers were mapped to the distal region of this chromosome (bin 5BS6) using a set of Chinese Spring deletion lines for 5BS. Eight and four markers were mapped to the interstitial region (bins 5BS5 and 5BS4, respectively). One marker was mapped to a pericentromeric bin. A comparative analysis of the distribution of trinucleotide microsatellites over wheat chromosome 5B and in different cereal species suggests that the (AAG)n repeat has proliferated and has been maintained during the evolution of cereals.https://vavilov.elpub.ru/jour/article/view/488bread wheattriticum aestivumbac clonesion torrentmiramicrosatelliteschromosome 5bssr markers(aag)n.
spellingShingle M. A. Nesterov
D. A. Afonnikov
E. M. Sergeeva
L. A. Miroshnichenko
M. K. Bragina
A. O. Bragin
G. V. Vasiliev
E. A. Salina
Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome
Вавиловский журнал генетики и селекции
bread wheat
triticum aestivum
bac clones
ion torrent
mira
microsatellites
chromosome 5b
ssr markers
(aag)n.
title Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome
title_full Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome
title_fullStr Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome
title_full_unstemmed Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome
title_short Identification of microsatellite loci according to BAC sequencing data and their physical mapping to the bread wheat 5B chromosome
title_sort identification of microsatellite loci according to bac sequencing data and their physical mapping to the bread wheat 5b chromosome
topic bread wheat
triticum aestivum
bac clones
ion torrent
mira
microsatellites
chromosome 5b
ssr markers
(aag)n.
url https://vavilov.elpub.ru/jour/article/view/488
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