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Plant Genomics 2019
June 13-14, 2019
Berlin, Germany
Asian Journal of Plant Science & Research
ISSN: 2249-7412 Page 26
Plant Genomics
5th Edition of International Conference on
Marek Szklarczyk, AJPSKY 2019, Volume 09
Conversion of GBS polymorphisms into CAPS markers for a
fertility restoring gene in carrots
Marek Szklarczyk
Krakow Agricultural University, Poland
BreedingofcarrothybridsisbasedonCytoplasmicMale
Sterility (CMS) and CMS lines are reproduced using
the so called maintainer lines which should be devoid
of fertility restoring alleles. Therefore, the development
of DNA markers for such alleles would greatly facilitate
production of new hybrids. For this purpose we mapped
carrot restorers in the context of DNA polymorphisms
detected via Genotyping By Sequencing (GBS). In the
present work we selected a few GBS polymorphisms,
which were linked to fertility restoration, and subjected
them to conversion into Cleaved Amplified Polymorphic
Sequences – CAPS markers.
Methodology and Theoretical Orientation: The GBS
datasets were screened for possible CAPS markers
with the VCF2CAPS software developed by Wojciech
Wesołowski. The respective PCR primers were designed
manually. PCR was followed by restriction with a
tetra-cutter and routine electrophoresis in agarose.
Genotyping was performed for plants from population
170 which segregated into male-sterile and male-fertile
(restored) plants.
Findings: Conversion was performed for four GBS
polymorphisms from chromosome 9 where the
restorer gene from population 170 was mapped.
For two GBS polymorphisms the respective CAPS
polymorphisms were not detected. The remaining two
GBS polymorphisms translated into polymorphic CAPS
markers named pe098/RsaI and pe106/TaqI. Both
polymorphic CAPSmarkers showed co-segregation with
the sterility/fertility phenotype. Co-segregation reached
the value of 84 and 89 % for pe098/RsaI and pe106/TaqI,
respectively. For both markers the male-fertile plants
were homozygous and the male-sterile-plants were
heterozygous.
ConclusionandSignificance: Theresultsshowthatsome
GBS polymorphisms may be negatively verified with the
use of CAPS markers. Likely, it is related to low coverage
of DNA sequencing during GBS which results in low
quality of the identified polymorphisms. Nevertheless,
two GBS polymorphisms were converted into useful
CAPSmarkers which can be used for identification of the
restorer fromchromosome 9 in segregating populations.
Biography
Marek Szklarczyk is a faculty member at the Institute of Plant
Biology and Biotechnology, Krakow Agricultural University, Po-
land. Hismain scientific interests are cytoplasmicmale sterility
(CMS) and fertility restoration. He studies these traits in car-
rots, beets, onions and rye.