Identification of Rfd1, a novel restorer-of-fertility locus for cytoplasmic male-sterility caused by DCGMS cytoplasm and development of simple PCR mar

[Tae-Ho Han]

2010 8 김성길교수님 양파 공동저자.pdf

Identification of Rfd1, a novel restorer-of-fertility locus for cytoplasmic male-sterility caused by DCGMS cytoplasm and development of simple PCR markers linked to the Rfd1 locus in radish (Raphanus sativus L.)

Abstract Previously, novel cytoplasmic male-sterility (CMS) caused by DCGMS cytoplasm was
discovered in radish (Raphanus sativus L.) introduced from Uzbekistan. We performed extensive progeny
tests and identified two fertility restorer lines (‘R171’ and ‘R121’) for this new CMS. Two F1 hybrid
populations were self-pollinated and backcrossed to produce F2 and BC populations. Inheritance patterns
of male-sterility in segregating populations varied depending on paternal lines. Segregation of malesterility
in F2 populations originating from the cross between MS19 and R121 showed that a single locus
was involved in fertility restoration. However, populations originating from the cross between MS15
and R171 showed the involvement of more than one restorer-of-fertility genes. The single fertility restorer
locus identified in the cross between MS19 and R121 was designated Rfd1 locus. Bulked segregant analysis
was performed using RAPD and AFLP, which identified one marker each. Both RAPD and AFLP
markers were converted into simple PCR-based co-dominant markers after their isolated flanking
sequences were analyzed. Indels 773-bp and 67-bp in length were identified between two Rfd1 allele-linked
flanking sequences of the RAPD and AFLP fragments, respectively, then utilized to develop simple
PCR markers. In addition, we prove that the newly identified Rfd1 locus is independent of the Rfo locus,
another radish fertility restorer for CMS caused by Ogura cytoplasm.

Keywords Radish (Raphanus sativus L.)  Cytoplasmic male-sterility  Restorer-of-fertility genes  Molecular markers  DCGMS cytotype

Euphytica (2010) 175:79–90
DOI 10.1007/s10681-010-0190-3

Kyuhyun Kim • Young-Pyo Lee • Heerae Lim •
Taeho Han • Soon-Kee Sung • Sunggil Kim