Identification of candidate genes associated with fertility restoration of cytoplasmic male-sterility in onion (Allium cepa L.) using a combination of bulked segregant analysis and RNA-seq

2015 ◽  
Vol 128 (11) ◽  
pp. 2289-2299 ◽  
Author(s):  
Sunggil Kim ◽  
Cheol-Woo Kim ◽  
Minkyu Park ◽  
Doil Choi
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Candidate Genes ◽  
Allium Cepa ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Rna Seq ◽  
Allium Cepa L

scholarly journals Identification of Ms2, a Novel Locus Controlling Male-fertility Restoration of Cytoplasmic Male-sterility in Onion (Allium Cepa L.), and Development of Tightly Linked Molecular Markers

2021 ◽  
Author(s):  
Nari Yu ◽  
Sunggil Kim
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Allium Cepa ◽  
Male Fertility ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Chromosome 2 ◽  
Rna Seq ◽  
Male Sterile ◽  
Male Fertility Restoration

Abstract Cytoplasmic male-sterility (CMS) has been exclusively used to produce F1 hybrid seeds of onion (Allium cepa L.). A single nuclear locus, Ms, is known to restore male-fertility of CMS in onions. Unstable male-sterile onions producing a small amount of pollen grains have been identified in a previous study. When such unstable male-sterile onions were crossed with stable male-sterile onions containing CMS-T cytoplasm, male-fertility was completely restored, although genotypes of the Ms locus were homozygous recessive. Inheritance patterns indicated that male-fertility restoration was controlled by a single locus designated as Ms2. A combined approach of bulked segregant analysis and RNA-seq was used to identify candidate genes for the Ms2 locus. High resolution melting (HRM) markers were developed based on single nucleotide polymorphisms (SNPs) detected by RNA-Seq. Comparative mapping of the Ms2 locus showed that Ms2 was positioned at the end of chromosome 2 with a distance of approximately 70 cM away from the Ms locus. Although 38 contigs containing reliable SNPs were analyzed using recombinants selected from 1,344 individuals, no contig showed perfect linkage to Ms2. Interestingly, transcription levels of orf725, a CMS-associated gene in onions, were significantly reduced in male-fertile individuals of segregating populations. However, no significant change in its transcription level was observed in individuals of a segregating population with male-fertility phenotypes determined by the Ms locus, suggesting that male-fertility restoration mechanism of Ms2 might be different from that of the Ms locus.

Alloplasmic male sterile Brassica napus with Enarthrocarpus lyratus cytoplasm: introgression and molecular mapping of an E. lyratus chromosome segment carrying a fertility restoring gene

Genome ◽  
2003 ◽  
Vol 46 (5) ◽  
pp. 792-797 ◽  
Author(s):  
H S Janeja ◽  
S K Banga ◽  
P B Bhaskar ◽  
S S Banga
Keyword(s):  
Brassica Napus ◽  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Molecular Mapping ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Hybrid Seed Production ◽  
Male Sterile ◽  
Fertility Restorer ◽  
Donor Species

A cytoplasmic male sterility (CMS) system for Brassica napus (2n = 38; AACC) was developed by backcross substitution of its nucleus into the cytoplasm of a wild crucifer, Enarthrocarpus lyratus. Male sterility was complete, stable, and expressed in small flowers with rudimentary anthers. Since the B. napus germplasm lines were complete or partial maintainers of male sterility, the required fertility restorer gene (Rfl) was introgressed from the cytoplasm donor species. Inheritance studies carried out on F1 and F2 populations derived from hybridizing cytoplasmic male sterile and male fertile near-isogenic (PNILs) lines of B. napus 'Westar', revealed a monogenic dominant control for fertility restoration. Bulked segregant analysis with 215 RAPD primers helped in the identification of putative primers associated with fertility restoration. Co-segregation analysis of eight such primers with Rfl gene revealed two markers, OPK 15700 and OPZ 061300, which flank the Rfl locus on either side at a distance of 8.2 and 2.5 cM, respectively. These DNA markers will be useful in marker-assisted selection for improving the commercial potential of this newly developed CMS-fertility-restorer system for hybrid seed production programs in rapeseed.Key words: oilseed rape, hybrids, cytoplasmic male sterility, fertility restoration, RAPD mapping.

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