Dominant Genic Male Sterility Research Articles

An epi-allele of SMS causes Sanming dominant genic male sterility in rice.

Male sterility is an important trait in rice for hybrid rice (Oryza sativa) breeding. However, the factors involved in dominant male sterility are largely unknown. Here, we identified a gene from Sanming dominant genic male sterile rice, named Sanming dominant male sterility (SMS), and reported that an epi-allele of this locus contributes to male sterility. Segregation analysis attributed dominant male sterility to a single locus, SMS, which we characterized using a male-sterile near isogenic line (NIL) of rice cultivar 93-11. The SMS locus was heterozygous in the male-sterile 93-11 NIL, containing an epi-allele identical to that in 93-11, and an epi-allele identical to that in rice cultivar Nipponbare, which we refer to as SMS9 and SMSN, respectively. SMS9 is silent and hyper-methylated, whereas SMSN is expressed and hypo-methylated in the 93-11 NIL. Overexpressing SMSN led to male sterility. Mutations in SMS rescued the male sterility of the 93-11 NIL. Interestingly, we observed the duplication of SMSN in Nipponbare, but did not observe the duplication of SMS9 in 93-11. Together, these findings suggest that the reduced methylation and enhanced expression of the SMSN epi-allele in the 93-11 NIL is responsible for its role in conferring dominant male sterility.

A natural mutation in the promoter of Ms-cd1 causes dominant male sterility in Brassica oleracea

Male sterility has been used for crop hybrid breeding for a long time. It has contributed greatly to crop yield increase. However, the genetic basis of male sterility has not been fully elucidated. Here, we report map-based cloning of the cabbage (Brassica oleracea) dominant male-sterile gene Ms-cd1 and reveal that it encodes a PHD-finger motif transcription factor. A natural allele Ms-cd1PΔ−597, resulting from a 1-bp deletion in the promoter, confers dominant genic male sterility (DGMS), whereas loss-of-function ms-cd1 mutant shows recessive male sterility. We also show that the ethylene response factor BoERF1L represses the expression of Ms-cd1 by directly binding to its promoter; however, the 1-bp deletion in Ms-cd1PΔ−597 affects the binding. Furthermore, ectopic expression of Ms-cd1PΔ−597 confers DGMS in both dicotyledonous and monocotyledonous plant species. We thus propose that the DGMS system could be useful for breeding hybrids of multiple crop species.

Spontaneous movement of a retrotransposon generated genic dominant male sterility providing a useful tool for rice breeding.

Male sterility in plants provides valuable breeding tools in germplasm innovation and hybrid crop production. However, genetic resources for dominant genic male sterility, which hold great promise to facilitate breeding processes, are extremely rare in natural germplasm. Here we characterized the Sanming Dominant Genic Male Sterility in rice and identified the gene SDGMS using a map-based cloning approach. We found that spontaneous movement of a 1978-bp long terminal repeat (LTR) retrotransposon into the promoter region of the SDGMS gene activates its expression in anther tapetum, which causes abnormal programmed cell death of tapetal cells resulting in dominant male sterility. SDGMS encodes a ribosome inactivating protein showing N-glycosidase activity. The activation of SDGMS triggers transcription reprogramming of genes responsive to biotic stress leading to a hypersensitive response which causes sterility. The results demonstrate that an ectopic gene activation by transposon movement can give birth to a novel trait which enriches phenotypic diversity with practical utility.

Improvement of Salinity Tolerance in Water-Saving and Drought-Resistance Rice (WDR)

Rice is one of the most economically important staple food crops in the world. Soil salinization and drought seriously restrict sustainable rice production. Drought aggravates the degree of soil salinization, and, at the same time, increased soil salinity also inhibits water absorption, resulting in physiological drought stress. Salt tolerance in rice is a complex quantitative trait controlled by multiple genes. This review presents and discusses the recent research developments on salt stress impact on rice growth, rice salt tolerance mechanisms, the identification and selection of salt-tolerant rice resources, and strategies to improve rice salt tolerance. In recent years, the increased cultivation of water-saving and drought-resistance rice (WDR) has shown great application potential in alleviating the water resource crisis and ensuring food and ecological security. Here, we present an innovative germplasm selection strategy of salt-tolerant WDR, using a population that is developed by recurrent selection based on dominant genic male sterility. We aim to provide a reference for efficient genetic improvement and germplasm innovation of complex traits (drought and salt tolerance) that can be translated into breeding all economically important cereal crops.

Kenyan farmers appreciate the higher yield of 50% non-pollen producing Maize (Zea mays) hybrids

Summary Incorporating the dominant male sterile gene, Ms44, in new maize varieties results in 50% non-pollen producing (FNP) varieties. This makes the varieties more nitrogen efficient and increases yield directly by an average of 200 kg ha−1 across yield levels. However, as half of the plants do not shed pollen, the presence of Ms44 in an FNP variety is clearly visible. This technology can improve food production and security in the African maize-based agri-food systems, but only if accepted by farmers. Farmers were therefore invited to 11 on-farm, researcher managed trial sites of FNP varieties in Kenya over 2 years. They were asked to identify the traits they find important in evaluating maize varieties and to score the FNP varieties, as well as their conventional counterparts, on these criteria (including yield, resistance to pests, and cob size) and overall, using a five-point hedonic scale. In total, 2,697 farmers participated, of which 62% were women. Farmers mentioned many traits they find important, especially yield and related traits, early maturity, and drought resistance, but also tassel and pollen formation. In 2017, mid-season, participants scored FNP varieties lower than conventional varieties on tassel and pollen formation, indicating that farmers could distinguish the trait. FNP varieties still received higher scores for yield and overall evaluation. In mid-season 2018, participants no longer scored FNP varieties lower for pollen formation as they now understood the technology. In both years, at the end-season evaluation, scores for tassel formation were not different, but participants scored FNP varieties higher for yield and overall. We conclude that farmers recognized the FNP trait but did not mind it as they clearly favored its yield advantage. The FNP technology, therefore, has high potential not only to increase maize yields, food production, and food security in the agricultural systems of Africa but also to increase varietal turnover and the adoption of new, high-yielding, climate-smart maize hybrids.

Global transcriptome analysis reveals potential genes associated with genic male sterility of rapeseed (Brassica napus L.).

Rapeseed is the third leading source of edible oil in the world. Genic male sterility (GMS) lines provide crucial material for harnessing heterosis for rapeseed. GMS lines have been widely used successfully for rapeseed hybrid production. The physiological and molecular mechanism of pollen development in GMS lines of rapeseed (Brassica napus L.) need to be determined for the creation of hybrids and cultivation of new varieties. However, limited studies have focused on systematically mining genes that regulate the pollen development of GMS lines in B. napus. In the present study, to determine the stage at which pollen development begins to show abnormality in the GMS lines, we performed semi-thin section analysis of the anthers with five pollen development stages. The results indicated that the abnormal pollen development in DGMS lines might start at the meiotic stage, and abnormal pollen development in RGMS lines probably occurred before the tetrad stage. To investigate the critical genes and pathways involved in pollen development in GMS lines, we constructed and sequenced 24 transcriptome libraries for the flower buds from the fertile and sterile lines of two recessive GMS (RGMS) lines (6251AB and 6284AB) and two dominant GMS (DGMS) lines (4001AB and 4006AB). A total of 23,554 redundant DEGs with over two-fold change between sterile and fertile lines were obtained. A total of 346 DEGs were specifically related to DGMS, while 1,553 DEGs were specifically related to RGMS. A total of 1,545 DEGs were shared between DGMS and RGMS. And 253 transcription factors were found to be differentially expressed between the sterile and fertile lines of GMS. In addition, 6,099 DEGs possibly related to anther, pollen, and microspore development processes were identified. Many of these genes have been reported to be involved in anther and microspore developmental processes. Several DEGs were speculated to be key genes involved in the regulation of fertility. Three differentially expressed genes were randomly selected and their expression levels were verified by quantitative PCR (qRT-PCR). The results of qRT-PCR largely agreed with the transcriptome sequencing results. Our findings provide a global view of genes that are potentially involved in GMS occurrence. The expression profiles and function analysis of these DEGs were provided to expand our understanding of the complex molecular mechanism in pollen and sterility development in B. napus.

Wheat male-sterile 2 reduces ROS levels to inhibit anther development by deactivating ROS modulator 1

Ms2 is an important dominant male-sterile gene in wheat, but the biochemical function of Ms2 and the mechanism by which it causes male sterility remain elusive. Here, we report the molecular basis underlying Ms2-induced male sterility in wheat. We found that activated Ms2 specifically reduces the reactive oxygen species (ROS) signals in anthers and thereby induces termination of wheat anther development at an early stage. Furthermore, our results indicate that Ms2 is localized in mitochondria, where it physically interacts with a wheat homolog of ROS modulator 1 (TaRomo1). Romo1 positively regulates the ROS levels in humans but has never been studied in plants. We found that single amino acid substitutions in the Ms2 protein that rescue the ms2 male-sterile phenotype abolish the interaction between Ms2 and TaRomo1. Significantly, Ms2 promotes the transition of TaRomo1 proteins from active monomers to inactive oligomers. Taken together, our findings unravel the molecular basis of Ms2-induced male sterility and reveal a regulatory mechanism in which ROS act as essential signals guiding the anther development program in wheat.

Development of the PARMS Marker of the Dominant Genic Male Sterility (DGMS) Line and Its Utilization in Rapeseed (Brassica napus L.) Breeding.

The 8029AB line is a dominant genic male sterility (DGMS) two-type line in Brassica napus L., which can be used in a three-line approach for the seed production of rapeseed hybrids. Genetic analyses have demonstrated that the sterility of 8029A is controlled by a single dominant nuclear gene (BnMS5e) interacting with one recessive gene (BnMS5c). Six pairs of penta-primer amplification refractory mutation system (PARMS) markers were designed according to the sequence of BnMS5a, BnMS5c and BnMS5e. Two pairs of these PARMS markers were successfully identified and validated. The PARMS markers MS5-1Fc/MS5-1Ft/MS5-1R12 could distinguish BnMS5c from BnMS5a/BnMS5e, and the PARMS markers MS5-2Ft/MS5-2Fa/MS5-1R12 could genotype BnMS5a and BnMS5c/BnMS5e. The combination of these two pairs of PARMS markers could be used to identify the presence or absence of BnMS5a/BnMS5c/BnMS5e effectively. Consequently, marker-assisted selection can be carried out in the early generation to shorten the breeding period and improve the breeding efficiency.

Identification of miRNAs and their target genes in genic male sterility lines in Brassica napus by small RNA sequencing

BackgroundBrassica napus is the third leading source of edible oil in the world. Genic male sterility (GMS) lines provide crucial material for harnessing heterosis for rapeseed. GMS lines have been used successfully for rapeseed hybrid production in China. MicroRNAs (miRNAs) play crucial regulatory roles in various plant growth, development, and stress response processes. However, reports on miRNAs that regulate the pollen development of GMS lines in B. napus are few.ResultsIn this study, 12 small RNA and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two recessive GMS (RGMS) lines, namely, “6251AB” and “6284AB”. At the same time, 12 small RNA and transcriptome libraries were also constructed and sequenced for the flower buds from the fertile and sterile lines of two dominant GMS (DGMS) lines, namely, “4001AB” and “4006AB”. Based on the results, 46 known miRNAs, 27 novel miRNAs on the other arm of known pre-miRNAs, and 44 new conserved miRNAs were identified. Thirty-five pairs of novel miRNA-3p/miRNA-5p were found. Among all the identified miRNAs, fifteen differentially expressed miRNAs with over 1.5-fold change between flower buds of sterile and fertile lines were identified, including six differentially expressed miRNAs between “4001A” and “4001B”, two differentially expressed miRNAs between “4006A” and “4006B”, four differentially expressed miRNAs between “6251A” and “6251B”, and ten differentially expressed miRNAs between “6284A” and “6284B”. The correlation analysis of small RNA and transcriptome sequencing was conducted. And 257 candidate target genes were predicted for the 15 differentially expressed miRNAs. The results of 5′ modified RACE indicated that BnaA09g48720D, BnaA09g11120D, and BnaCnng51960D were cleaved by bna-miR398a-3p, bna-miR158-3p and bna-miR159a, respectively. Among the differentially expressed miRNAs, miR159 was chosen to analyze its function. Overexpression of bna-miR159 in Arabidopsis resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate the fertility and silique development in rapeseed.ConclusionsOur findings provide an overview of miRNAs that are potentially involved in GMS and pollen development. New information on miRNAs and their related target genes are provided to exploit the GMS mechanism and reveal the miRNA networks in B. napus.

Breeding with dominant genic male-sterility genes to boost crop grain yield in the post-heterosis utilization era

Global food security is facing severe challenges from an ever-growing population, limited resources, and various stresses. Dominant genic male sterility (DGMS) technology combined with modern breeding strategies may create novel cultivation models with ~50% DGMS F1 hybrids for field production of cross-pollinated crops, boosting crop grain yield to ensure global food security and sustainable agriculture in the post-heterosis utilization era.

Fertility recovery of wheat male sterility controlled by Ms2 using CRISPR/Cas9.

Fertility recovery of wheat male sterility controlled by Ms2 using CRISPR/Cas9.

Cytological and genetic characterisation of dominant GMS line Shaan-GMS in Brassica napus L.

Genic male sterility (GMS) is an effective pollination control system applied in the hybrid breeding of Brassica napus L. Shaan-GMS is a spontaneous mutant of dominant GMS in B. napus. In this research, anther abortion in the homozygous two-type line 9A15AB derived from Shaan-GMS was characterised with the combined use of light microscopy and transmission electron microscopy. Results indicated that the most striking differences between the fertile and sterile plants occurred in the tapetum in the early microsporocyte stage. In sterile plants, the tapetal cells were irregularly arranged, multi-layered and occupied the growing space of microsporocytes. When entering into meiosis, the tapetum cells degraded and the cytoplasm fused. Some oval monolayer or bilayer membrane organelles existed in the tapetal cells in sterile anthers. Mitochondria in the tapetal cells were abnormal, and middle layer cells degraded early. Pollen mother cells of Shaan-GMS degenerated at the start of meiosis and ceased at the anaphase I stage, with no dyads or tetrads formed. The combined effects of the abnormal development of the tapetum, the middle layer cells and meiosis lead to male sterility in Shaan-GMS. Inheritance of male sterility of Shaan-GMS is controlled by a monogenically multiallelic locus with three different alleles (Ms, ms and Mf), with a relationship expressed as Mf > Ms and Ms > ms. The findings help lay the foundation for illustrating the mechanism of male sterility and the utilisation of Shaan-GMS in rapeseed.

Characterization and transcriptome analysis of a dominant genic male sterile cotton mutant

BackgroundMale sterility is an efficient trait for hybrid seed production and germplasm innovation. Until now, most studies on male sterility were on cytoplasmic and recessive genic sterility, with few on dominant genic male sterility, especially in cotton, due to lack of such mutant.ResultsWe discovered a natural male sterile (MS) Sea Island cotton (G. barbadense) mutant. Genetic analysis showed the mutation was caused by a dominant mutation in a single nuclear gene. Comparative cytological observation of anther sections from MS and wild-type (WT) uncovered cellular differences in anther at and after the tetrad stage of pollen mother cells (PMC). In the MS anthers, the outer wall of pollen grains was free of spinules, the tapetum was vacuolated and showed delayed degradation, consequently, no functional pollen grains. Comparison of transcriptomes from meiosis, tetrad, mononuclear and binuclear pollen, and pollen maturation stages identified 13,783 non-redundant differentially expressed genes (DEGs) between MS and WT. Based on the number of DEGs, analyses of enriched GO terms and KEGG pathways, it was evident that significant transcriptomic changes occurred at and after the tetrad stage, consistent with cytological observation, and that the major differences were on metabolism of starch, sucrose, ascorbate, aldarate, alanine, aspartate and glutamate, and biosynthesis of cutin, suberine and wax. WGCNA analysis identified five modules containing 920 genes highly related to anther development, especially the greenyellow module with 54 genes that was highly associated with PMC meiosis and tetrad formation. A NAC transcription factor (Gh_D11G2469) was identified as a hub gene for this module, which warrants further functional characterization.ConclusionsWe demonstrated that the MS trait was controlled by a single dominant nuclear gene and caused by delayed tapetum degradation at the tetrad stage. Comparative transcriptome analysis and gene network construction identified DEGs, enriched GO terms and metabolic pathways, and hub genes potentially associated with anther development and the MS trait. These results contribute to our understanding of dominant genic male sterility (DGMS) and provided source for innovation of cotton germplasm.

Identification of brown planthopper resistance gene Bph32 in the progeny of a rice dominant genic male sterile recurrent population using genome-wide association study and RNA-seq analysis

Brown planthopper (BPH) is one of the most damaging pests of rice. Improving the resistance of rice varieties is considered a safe and effective way to control this pest. Here, a field experiment was conducted to test BPH resistance in a rice population developed by recurrent selection of a variety with dominant genic male sterility. Three high-resistance lines were identified and subjected to a genome-wide association study (GWAS), which mapped the BPH resistance gene to the short arm of chromosome 6. One highly resistant line (14CF2426) was further crossed with Taichuang Native 1 (TN1) to construct an F2 segregating population. Artificial insect infestation identified three quantitative trait loci (QTLs), which were mapped to chromosomes 1, 6, and 10; the QTL on chromosome 6 of 14CF2426 contained a known resistance gene, Bph32. Near-isogenic lines NIL-Bph32 and NIL-bph32 of Bph32 were constructed using molecular marker-assisted selection. RNA sequencing (RNA-seq) detected 1268 differentially expressed genes (DEGs) between the two near-isogenic lines. Compared with NIL-Bph32, 1064 genes were upregulated and 204 were downregulated in NIL-bph32. Gene ontology (GO) enrichment analysis detected 46 significantly enriched terms, and KEGG analysis revealed four significantly enriched metabolic pathways. This robust multi-method strategy showed that combining GWAS analysis with a dominant genic male sterile recurrent rice population can significantly improve the efficiency of gene identification. RNA-seq revealed some candidate genes with a potential regulatory function in BPH resistance, thus providing insights into the transcriptional regulatory network and the molecular mechanisms of resistance.

Wheat Ms2 encodes for an orphan protein that confers male sterility in grass species

Male sterility is a valuable trait for plant breeding and hybrid seed production. The dominant male-sterile gene Ms2 in common wheat has facilitated the release of hundreds of breeding lines and cultivars in China. Here, we describe the map-based cloning of the Ms2 gene and show that Ms2 confers male sterility in wheat, barley and Brachypodium. MS2 appears as an orphan gene within the Triticinae and expression of Ms2 in anthers is associated with insertion of a retroelement into the promoter. The cloning of Ms2 has substantial potential to assemble practical pipelines for recurrent selection and hybrid seed production in wheat.

Map-based cloning of the dominant genic male sterile Ms-cd1 gene in cabbage (Brassica oleracea).

Using map-based cloning, we delimited the Ms - cd1 gene responsible for the male sterile phenotype in B. oleracea to an approximately 39-kb fragment. Expression analysis suggests that a new predicted gene, a homolog of the Arabidopsis SIED1 gene, is a potential candidate gene. A dominant genic male sterile (DGMS) mutant 79-399-3 in Brassica oleracea (B. oleracea) is controlled by a single gene named Ms-cd1, which was genetically mapped on chromosome C09. The derived DGMS lines of 79-399-3 have been successfully applied in hybrid cabbage breeding and commercial hybrid seed production of several B. oleracea cultivars in China. However, the Ms-cd1 gene responsible for the DGMS has not been identified, and the molecular basis of the DGMS is unclear, which then limits its widespread application in hybrid cabbage seed production. In the present study, a large BC9 population with 12,269 individuals was developed for map-based cloning of the Ms-cd1 gene, and Ms-cd1 was mapped to a 39.4-kb DNA fragment between two InDel markers, InDel14 and InDel24. Four genes were identified in this region, including two annotated genes based on the available B. oleracea annotation database and two new predicted open reading frames (ORFs). Finally, a newly predicted ORF designated Bol357N3 was identified as the candidate of the Ms-cd1 gene. These results will be useful to reveal the molecular mechanism of the DGMS and develop more practical DGMS lines with stable male sterility for hybrid seed production in cabbage.

A generic SSR marker closely linked to a dominant genic male sterility gene (DGMs79-399-3) in broccoli (Brassica oleracea var. italica)

As an important pollination system, male sterility has been used widely for broccoli (Brassica oleracea var. italica) hybrid production. New male sterile lines are important for broccoli breeding. We transferred the dominant genic male sterility (DGMs) 79-399-3, which was developed from a spontaneous mutation in cabbage, to broccoli. In this study, molecular markers linked to the DGMs79-399-3 gene in broccoli were identified by screening a segregating population DGMs8554 with 747 individuals, obtained by ten backcrosses (BC10) using a cabbage-dominant genic male sterility line containing DGMs79-399-3 as the donor and broccoli inbred line 8554 as the recipient. Bulked segregation analysis with respect to male sterility and fertility for the BC10 population of DGMs8554 was conducted by scanning 2585 different types of primers. Three SSR and one SCAR marker were identified, and a map surrounding DGMs79-399-3 was constructed. Two flanking markers, scaffold10312a and scaffold129_2012, were linked to the dominant male sterile DGMs79-399-3 gene at distances of 0.563 and 0.328 cM, respectively, which mapped the DGMs79-399-3 gene to a 0.891 cM range. The markers were verified in another three broccoli groups, and a generic SSR marker, scaffold10312a, was obtained with an accuracy ≥96.43 %. The markers developed here will accelerate the breeding of new elite DGMs lines in broccoli and provide a solid basis for cloning of the DGMs79-399-3 gene.

Inheritance and molecular mapping of a novel dominant genic male-sterile gene in Sesamum indicum L.

Exploiting heterosis using pollen control systems such as genic male sterility (GMS) can significantly improve the seed yield of sesame. In this study, we developed a novel GMS line (W1098A) by backcrossing and sib-mating the wild accession Yezhi2 (Sesamum mulayanum Nair, 2n = 26) plants with the cultivar Ezhi1 (S. indicum L., 2n = 26). The sterility of this line is mainly controlled by one dominant GMS gene (Ms). Cytological investigation showed that pollen abortion in W1098A plants began in pollen mother cells, continued throughout pollen development, and peaked at the late microspore stage. The abortion process was closely associated with the abnormal behaviour of the tapetum. We employed bulked segregation analysis to identify SSR markers linked to the Ms gene in a segregating population derived from sib-mating between the near-isogenic lines W1098A and W1098B. Among 1,500 SSR primer pairs screened against the sterile and fertile bulks of DNA, 13 markers were found to be linked to the Ms locus. The closest markers on both sides of Ms were SBM298 and GB50, at a respective genetic distance of 0.15 and 0.70 cM. These markers will be valuable for marker-aided breeding of GMS hybrids and map-based cloning of Ms.

Fine mapping of a dominant thermo-sensitive genic male sterility gene (BntsMs) in rapeseed (Brassica napus) with AFLP- and Brassica rapa-derived PCR markers.

A new thermo-sensitive dominant genic male sterility (TSDGMS) line of Brassica napus was found and mapped in this paper. Our result will greatly accelerate the map-based cloning of the BntsMs gene. TE5A is a thermo-sensitive dominant genic male sterility line originating from spontaneous mutation of the inbred line TE5 in Brassica napus and provides a promising system for the development of hybrid cultivars. Genetic analysis has revealed that the BntsMs mutant is controlled by a single, dominant gene. Here, we describe the fine mapping of BntsMs using amplified fragment length polymorphism (AFLP) and intron polymorphism (IP) methodologies. We screened 1,024 primer combinations and then identified five AFLP markers linked to the BntsMs gene, two of which were successfully converted into sequence-characterised amplified region (SCAR) markers. The linkage of the markers was identified by analysing a large BC2 population of 700 recessive-fertility individuals. Two SCAR markers were found in the flanking region of the BntsMs gene at distance of 3.5 and 4.8 cm. Based on sequence information from the previously screened AFLP markers and on genome organisation comparisons of the A genome of Brassica rapa and Arabidopsis, seven IP markers linked to the BntsMs gene were developed. By analysing the 700 recessive-fertility individuals, two IP markers, IP004 and IP470, were localised to the flanking region of the BntsMs gene at a distance of 0.3 and 0.2 cm, respectively. A comparison of the B. rapa and Arabidopsis genomes revealed 27 genes of B. rapa in the flanking region of these two IP markers. It is likely that the molecular markers developed from these investigations will greatly accelerate the positional cloning of the BntsMs gene.

A Co-Dominant Marker BoE332 Applied to Marker-Assisted Selection of Homozygous Male-Sterile Plants in Cabbage (Brassica oleracea var. capitata L.)

The dominant genic male sterility (DGMS) gene CDMs399-3 derived from a spontaneous mutation in the line 79-399-3 of spring cabbage (Brassica oleracea var. capitata L.), has been successfully applied in hybrid seed production of several cabbage cultivars in China. During the development of dominant male sterility lines in cabbage, the conventional identification of homozygous male-sterile plants (CDMs399-3/CDMs399-3) is a laborious and time-consuming process. For marker-assisted selection (MAS) of the gene CDMs399-3 transferred into key spring cabbage line 397, expressed sequence tag-simple sequence repeats (EST-SSR) and SSR technology were used to identify markers that were linked to CDMs399-3 based on method of bulked segregant analysis (BSA). By screening a set of 978 EST-SSRs and 395 SSRs, a marker BoE332 linked to the CDMs399-3 at a distance of 3.6 cM in the genetic background of cabbage line 397 were identified. 7 homozygous male-sterile plants in population P1170 with 20 plants were obtained finally via MAS of BoE332. Thus, BoE332 will greatly facilitate the transferring of the gene CDMs399-3 into the key spring cabbage line 397 and improve the application of DGMS in cabbage hybrid breeding.