Dominant Genic Male Sterility Research Articles

Cytological investigation of anther development in DGMS line Shaan-GMS in Brassica napus L.

The cytological mechanism of male sterility of Shaan-GMS, a natural mutant dominant genic male sterile (DGMS) line in Brassica napus L., is not well studied. Cytological observation was made on different-size buds of DGMS line 0A30A derived from Shaan-GMS line. The pollen mother cells (PMCs) of DGMS line 0A30A were degenerating at the beginning of meiosis and could not pass the anaphase I stage, with no dyads or tetrads formed, suggesting that the DNA damage checkpoint and spindle assembly checkpoint were activated in sterile anthers. During the meiosis process of sterile anthers in the sterile plants, several kinds of abnormal meiotic cells could be observed: nuclei condensed PMCs, cells with micronuclei, collapsed cells, plasmolysis cells, cells connected with nucleoplasmic bridge, and microspore analogue developed from PMCs without meiosis but enclosed by the exine wall. The results suggested Shaan-GMS to be a new type of DGMS line in B. napus.

A Dominant Gene for Male Sterility in Salvia miltiorrhiza Bunge

A natural male sterile mutant of Salvia miltiorrhiza (Labiatae, Sh-B) was found during field survey in 2002. Our objective was to analyze its genetic mechanism for producing F1 hybrid seeds and to develop a molecular marker linked to male sterile gene for selection of a hybrid parent line. The segregation ratios of male sterile plants to fertile plants in the progenies of both testcross and backcross were 1:1 in continuous experiments conducted in 2006–2009. The male sterile Sh-B was heterozygous (Msms). The male sterile plants could capture most pollen (2 granule/cm2·24 h) with row ratio (female : male 2 : 1) within 45-cm distance and harvest the largest amount of 6495 g hybrid seeds per hectare. We also developed DNA markers linked to the male sterile gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 128 primer combinations. One out of fourteen AFLP markers (E11/M4208) was identified as tightly linked to the dominant male sterile gene with a recombination frequency of 6.85% and at a distance of 6.89 cM. This marker could be converted to PCR-based assay for large-scale selection of fertile plants in MAS (marker-assisted selection) at the seedling stage. Blastn analysis indicated that the male sterile gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1–5, and was more likely to encode S-adenosylmethionine-dependent methyltransferase, in which DNA methylation regulated the development of plant gametogenesis.

Mapping of Sanming dominant genic male sterility gene in rice

The mutant of "Sanming Dominant Genic Male Sterile Rice" was found from an F2 population of cross "SE2lS/Basmati370" by Sanming Institute of Agricultural Science in 2001. It has proven that the male sterility of this mutant is controlled by a dominant gene (named as SMS). By multiple backcrosses, this dominant male sterile allele was introduced into the genetic background of an indica rice cultivar Jiafuzhan (which was known as Jiabuyu). In order to map SMS, a mapping population was constructed by crossing Jiabuyu with a japonica cultivar Nipponbare and further crossing the F1 with Jiafuzhan. By bulked segregant analysis and linkage analysis using SSR and INDEL markers, SMS was mapped to a 99 kb interval between INDEL markers ZM30 and ZM9 on chromosome 8. This result will facilitate cloning of SMS.

EST-Aided Conversion of AFLP Marker Linked to Dominant Male Sterility Gene in <I>Brassica napus</I> into SCAR

EST-Aided Conversion of AFLP Marker Linked to Dominant Male Sterility Gene in <I>Brassica napus</I> into SCAR

Meiotic abnormality in dominant genic male sterile Brassica napus

Rs1046AB is a dominant genic male sterile (DGMS) Brassica napus line derived from Yi-3A. Until now the molecular mechanism of its male sterility is still unknown. In this paper, cytological observations demonstrated that all cells in sterile plants contained condensed nuclei at the beginning stage of meiosis; this implied that meiotic cells were degenerating. Although 31% (93/300) cells escaped from the state of nuclei condensation in buds about 3 mm in length (in such length, normal plants are at tetrade stage), no cells could pass the pachytene stage. Then pachytene or zygotene like chromatin/chromosomes sometimes congregated into two or more groups with different size, which resulted in the formation of micronuclei. Nucleoplasmic bridge could also be found in some meiotic cells. Even when the "microspore's analogue" appeared in sterile buds about 4 mm in length (in such length, mature pollens could be detected in normal buds), the nuclei condensation and escaped cells with pachytene like chromosome still could be found in the sterile anthers. So it could be concluded that male sterility was caused by meiotic abnormality. According to our previous research, four genes related to cell cycle/DNA processing were identified in fertile plants. RT-PCR further confirmed that three DNA repair genes were partially or completely repressed in the sterile plants, and were only expressed in the early stage fertile flower buds, i.e. the buds <3 mm in length. Therefore, DGMS of rapeseed was probably caused by the abnormality in DNA damage repair system during meiosis. According to these results, some possible mechanisms of fertility control were discussed.

Dominant male-sterile populations for association mapping and introgression of exotic wheat germplasm

The introgression of exotic wheat germplasm such as synthetic hexaploid (tetraploid × diploid amphiploid) wheat (Triticum aestivum L.) into an adapted gene pool has the potential to amplify the genetic variation for complex traits. The dominant male-sterile gene can be used to facilitate recurrent selection in wheat, thus increasing the opportunity for genetic recombination between exotic and cultivated genomes and the identification and retention of desirable alleles. Our dominant male-sterile recurrent selection project began in 1983 with the intermating of the source germplasm (Chris hard red spring wheat in Triticum tauschii cytoplasm) with 34 soft winter lines and varieties, intermated without selection for 3 generations, and then selecting male-sterile plants for agronomic type each season. After 20 generations of recurrent selection, we extracted 94 inbred lines without selection during inbreeding except for fertility. Those lines were evaluated for linkage disequilibrium (LD) on the 5A chromosome for comparison with an earlier study involving 95 elite soft winter wheat varieties of similar genetic background. For the male-sterile population-derived lines, LD decayed within 1 cM (r2 &lt; 0.1) compared with 5 cM in the previous report for soft winter wheat varieties. This approach to introgression facilitates the potential use of synthetic wheat and other unadapted germplasm for expanding the gene pool and provides a potential resource for high-resolution association mapping of genes controlling traits of interest.

Global analysis of gene expression in flower buds of Ms-cd1 Brassica oleracea conferring male sterility by using an Arabidopsis microarray

The dominant male sterility gene Ms-cd1 is identified in Brassica oleracea. Electron microscopical observations revealed that abortion of pollen development starts after tetrad formation. This important male sterility phenotype is characterized by lack of degradation of the primary pollen mother cell (PMC) wall and delayed degradation of callose surrounding the tetrads and thus arrest of microspore release. Gene expression of the male sterile and fertile buds was analyzed by heterologous hybridization of Brassica oleracea cRNA onto an Arabidopsis whole genome oligonucleotide microarray. A total of 277 suppressed genes including 40 kinase-, 32 cell wall modification and 29 transport related genes were found to be significantly down regulated >3-fold in the male sterile mutant. The vast majority of the differentially expressed transcripts are found to present late pollen stage specific genes. Kinase genes, cell wall modification genes and ion transport genes were greatly over-represented when compared to their percentage of all flower bud expressed genes and represent 36.5% of the genes suppressed by Ms-cd1. Our results also suggest that Ms-cd1 may blocks an anther developmental pathway with a small number of genes suppressed in tapetum cells which prevent the degradation of callose and PMC wall, which further leads to the suppression of a large number of genes involved in signaling pathways, cell wall modification and ion transport in pollen grains.

Development and characterization of SCAR markers associated with a dominant genic male sterility in rapeseed

AbstractRs1046AB is a dominant genic male sterility (DGMS) line in rapeseed, in which the sterility has always been thought to be conditioned by the interaction of a male sterility gene (Ms) and its non‐allelic restorer gene (Rf). This system provides not only a tool for assisting in recurrent selection but also a promising system for hybrid production. Based on previous studies, two amplified fragment length polymorphism markers linked with the Ms gene were converted into a dominant and a co‐dominant sequence characterized amplified region (SCAR) marker, respectively. The putative linear order relationship of three dominant SCAR markers with the same genetic distance from the Rf gene, was also determined by an examination of whether the homologues of these markers are present or not in different lines carrying Rf. A bigger fragment generated by the closest marker linked to the Rf gene was observed in all lines carrying the recessive allele rf, suggesting that this marker is a co‐dominant marker, which was further confirmed by nucleotide sequence comparison of these fragments. SCAR markers specific for Ms and Rf will be especially valuable in marker‐assisted DGMS three‐line breeding.

Genetic Analysis and Mapping of Genes Involved in Fertility of Pingxiang Dominant Genic Male Sterile Rice

Pingxiang dominant genic male sterile rice (PDGMSR) was the first dominant genic male sterile mutant identified in rice ( Oryza sativa L.), and the corresponding dominant genic male sterile gene was designated as Ms-p. The fertility of PDGMSR can be restored by introduction of a dominant epistatic fertility restoring gene in some rice varieties. In the present study, E823, an indica inbred rice variety, restored the fertility of PDGMSR, and the genetic pattern was found to be consistent with a dominant epistatic model, therefore, the dominant epistatic fertility restorer gene was designated as Rfe. The F2 population from the cross of PDGMSR/E823 was developed to map gene Rfe. The F2 plants with the genotypes Ms-pMs-pRferfe or Ms-pms-pRferfe were used to construct a fertile pool, and the corresponding sterile plants with genotypes Ms-pMs-prferfe or Ms-pms-prferfe were used to construct a sterile pool. The fertility restoring gene Rfe was mapped to one side of the microsatellite markers RM311 and RM3152 on rice chromosome 10, with genetic distances of 7.9 cM and 3.6 cM, respectively. The microsatellite markers around the location of the Ms-p gene were used to finely map the Ms-p gene. The findings of this study indicated that the microsatellite markers RM171 and RM6745 flanked the Ms-p gene, and the distances were 0.3 cM and 3.0 cM, respectively. On the basis of the sequence of rice chromosome 10, the physical distance between the two markers is approximately 730 kb. These findings facilitates molecular marker-assisted selection (MAS) of genes Ms-p and Rfe in rice breeding programs, and cloning them in the future.

Cytological and molecular characterization of a novel monogenic dominant GMS in Brassica napus L.

A novel genic male sterile (GMS) line in Brassica napus L., which was identified in 1999, was found to be controlled by a monogenic dominant gene, which we have designated as MDGMS. The microspores of the MDGMS abort before the degradation of the tapetal cell layer. The F1 fertility from any fertile lines crossed with MDGMS segregated and the ratio was close to 1:1. Bulked segregation analysis (BSA) was employed to identify random amplified polymorphic DNA (RAPD) markers linked to the Ms gene in MDGMS. Among 880 random 10-mer oligonucleotide primers screened against the bulk DNA of sterile and fertile, one primer S243 (5'-CTATGCCGAC-3') gave a repeatable 1500-bp DNA polymorphic segment S243(1500) between the two bulks. Analysis of individual plants of each bulks and other types of GMS and cytoplasmic male sterility (CMS) lines suggest that the RAPD marker S243(1500) is closely linked to the MDGMS locus in rapeseed. This RAPD marker has been converted into sequence characterized amplified region (SCAR) marker to aid identification of male-fertility genotypes in segregating progenies of MDGMS in marker-assisted selection (MAS) breeding programs.

Isolation and analysis of differentially expressed genes in dominant genic male sterility (DGMS) Brassica napus L. using subtractive PCR and cDNA microarray

Dominant genic male sterility (DGMS) is an important approach to utilize the heterosis of Brassica napus, but the molecular mechanism of DGMS is not well understood. As an initial step towards understanding this event, some pilot studies were performed. Using suppression subtractive hybridization (SSH) and cDNA microarray, about 1200 significantly differentially expressed clones were isolated between the fertile and sterile plants of the homozygous DGMS two-type line, Rs1046AB. Northern blot further demonstrated the credibility of the microarray data. Subsequently, about 400 clones were selected for sequencing and they represented 216 unigenes (212 were up-regulated in fertile buds, and the other 4 were up-regulated in sterile ones). Of the 212 up-regulated genes in forward-subtracted library, 178 homologous sequences could be divided into 17 groups excluding those that encode the unclassified proteins; and the other 34 genes had no homology in the databases at the National Center for Biotechnical Information (NCBI). Furthermore, some important pathways related to male gametogenesis were identified using the program of KEGG Orthology (KO)-Based Annotation System (KOBAS), such as nitrogen metabolism, nitrobenzene degradation and starch and sucrose metabolism.

AFLP and SCAR markers linked to the suppressor gene (Rf) of a dominant genetic male sterility in rapeseed (Brassica napus L.)

Rs1046AB is a line which is true breeding for a dominant genetic male sterility gene (Ms) but which is a mixture of male fertile and sterile individuals (a two-type line) because it is segregating for a dominant suppressor gene (Rf). This system provides a promising alternative to the CMS system for hybrid breeding in Brassica napus. In order to identify molecular markers linked to the rf gene, a near-isogenic line (NIL) population from the cross between a sterile individual (MsMsrfrf) and a fertile individual (MsMsRfrf) in Rs1046AB was subjected to amplified fragment length polymorphism (AFLP) analysis, with a combination of comparing near isogenic lines (NILs) and bulked segregant analysis (BSA). From 2,816 pairs of AFLP primers, six fragments showing polymorphism between the fertile and sterile bulks as well as the individuals of the bulks were identified. Linkage analysis indicated that the six AFLP markers are tightly linked to the Rf gene and all are distributed on the same side. The minimum genetic distance between the Rf gene and a marker was 0.7 cM. Since the AFLP markers are not suitable for large-scale application in MAS (marker-assisted selection), our objective was to develop a fast, cheap and reliable PCR-based assay. Consequently, three of the four closest AFLP markers were converted directly to sequence characterized amplified region (SCAR) markers. For the other marker a corresponding SCAR marker was successfully obtained after isolating the adjacent sequences by PCR Walking. The available SCAR markers of the Rf gene will greatly facilitate future breeding programs using dominant GMS to produce hybrid varieties.

Molecular validation of multiple allele inheritance for dominant genic male sterility gene in Brassica napus L

Dominant genic male sterility (DGMS) has been playing an increasingly important role, not only as a tool for assisting in recurrent selection but also as an alternative approach for efficient production of hybrids. Previous studies indicate that fertility restoration of DGMS is the action of another unlinked dominant gene. Recently, through classical genetic analysis with various test populations we have verified that in a DGMS line 609AB the trait is inherited in a multiple allelic pattern. In this study, we applied molecular marker technology to provide further validation of the results. Eight amplified fragment length polymorphism (AFLP) markers tightly linked to the male sterility allele (Ms) were identified in a BC1 population from a cross between 609A (a sterile plant in 609AB) and a temporary maintainer GS2467 as recurrent parent. Four out of the eight markers reproduced the same polymorphism in a larger BC(1) population generated with microspore-derived doubled haploid (DH) parents (S148 and S467). The two nearest AFLP markers SA12MG14 and P05MG15, flanking the Ms locus at respective distances of 0.3 centiMorgan (cM) and 1.6 cM, were converted into sequence characterized amplified region (SCAR) markers designated SC6 and SC9. Based on the sequence difference of the marker P05MG15 between S148 and a DH restorer line S103, we further developed a SCAR marker SC9f that is specific to the restorer allele (Mf). The map distance between SC9f and Mf was consistent with that between SC9 and Ms allele. Therefore, successful conversion of the marker tightly linked to Ms into a marker tightly linked to Mf suggested that the restoration for DGMS in 609AB is controlled by an allele at the Ms locus or a tightly linked gene (regarded as an allele in practical application). The Ms and Mf-specific markers developed here will facilitate the breeding for new elite homozygous sterile lines and allow further research on map-based cloning of the Ms gene.

Fertility behaviour of rice ( Oryza sativa) lines with dominant male sterile gene and inheritance of sterility and fertility restoration

The genetic basis of male sterility and fertility restoration of the Pingxiang male-sterile rice (PMSR) was studied using progeny populations created between PMSR and 11 fertile lines. It was found that the male sterility was determined by two interacting (epistatic) dominant nuclear genes, one for sterility and one for fertility restoration. The dominant sterile gene expresses as male-sterility when existing solely, but as normally fertile when coexisting with the restoration gene. The individuals with only the restoration gene are normal and fertile. A homozygous sterile line developed using PMSR by repeated selfing was characterised for its fertility behaviour under controlled and field conditions. Male fertility was affected by both temperature and photoperiod with temperature being more important. The critical temperature for inducing fertility was 27–28 °C. Blow this critical temperature, plants remained sterile, but become partially fertile at higher temperature. The panicle development stages that are sensitive to temperature were from differentiation of the secondary branch primordium (S3) to the meiotic division of the pollen mother cells (S6). Continuous high temperature (>30 °C) during these sensitive stages is necessary to maintain male fertility. Long photoperiod (15 h) induced partial fertility even under temperature, which could induce sterility. In practice, this line can be regarded as thermo-sensitive. In the sub-tropical zone, this line has complete sterile phase longer than 4 weeks and thus is suitable for hybrid production using the two-line system (a pair of pure sterile and fertile lines). Male sterile line required low temperature and short photoperiod to express male sterility, like the one derived from PMSR, has never been reported in any crop species. This line is also the first dominant nuclear male-sterile line that could be exploited for hybrid seed production using the two-line system.

Linkage mapping of a dominant male sterility geneMs-cd1inBrassica oleracea

The dominant male sterility gene Ms-cd1 (c, cabbage; d, dominant) was identified as a spontaneous mutation in the spring cabbage line 79-399-3. The Ms-cd1 gene is successfully applied in hybrid seed production of several Brassica oleracea cultivars in China. Amplified fragment length polymorphism (AFLP) technology was used to identify markers linked to the Ms-cd1 gene in bulks of male-sterile and male-fertile individuals of a segregating BC3 population and in a near-isogenic population of 25 male-sterile plants. Twelve markers within a 20-cM interval proximal to the Ms-cd1 gene were identified, 5 of which can be used to select homozygous male-sterile Ms-cd1/ Ms-cd1 plants. Three AFLP markers and 3 sequence characterized amplified region markers that were linked to MS-cd1 mapped onto linkage group O9, corresponding to chromosome 3 of B. oleracea. This region corresponds to the top of chromosome 5 in Arabidopsis thaliana.

Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus)

AbstractRs1046AB is a genic male sterile two‐type line in rapeseed that has great potential for hybrid seed production. The sterility of this line is conditioned by the interaction of two genes, i.e. the dominant genic male sterility gene (Ms) and the suppressor gene (Rf). The present study was undertaken to identify DNA markers for the Ms locus in a BC1 population developed from a cross between a male‐sterile plant in Rs1046AB and the fertile canola‐type cultivar ‘Samourai’. Bulked segregant analysis was performed using the amplified fragment length polymorphism (AFLP) methodology. From the survey of 480 AFLP primer combinations, five AFLP markers (P10M13350, P13M8400, P6M6410, E7M1230 and E3M15100) tightly linked to the target gene were identified. Two of them, E3M15100 and P6M6410, located the closest, at either side of Ms at a distance of 3.7 and 5.9 cM, respectively. The Ms locus was subsequently mapped on linkage group LG10 in the map developed in this laboratory, adding two additional markers weakly linked to it. This suite of markers will be valuable in designing a marker‐assisted genic male sterility three‐line breeding programme.

High-density physical maps reveal that the dominant male-sterile gene Ms3 is located in a genomic region of low recombination in wheat and is not amenable to map-based cloning

In wheat it is essential to know whether a gene is located in a high or low recombination region of the genome before initiating a map-based cloning approach. The objective of this study was to explore the potential feasibility of map-based cloning of the dominant male-sterile gene Ms3 of wheat. High-density physical maps of the short arms of the group-5 chromosomes (5AS, 5BS, and 5DS) of Triticum aestivum L. were constructed by mapping 40 DNA markers on a set of 17 homozygous deletion lines. One hundred RFLP loci were mapped: 35 on 5AS, 37 on 5BS, and 28 on 5DS. A consensus physical map was colinearly aligned with a consensus genetic map of the group-5 short arms. Sixteen of the 17 markers in the consensus genetic map encompass a genetic distance of 25 cM and correspond to the distal region (FL 0.56–0.97) of the consensus physical map. Two rice probes, RG463 and RG901, previously identified to be linked to markers CDO344 and CDO749 (group-5 short arm of wheat), respectively, in the genetic map of rice chromosome 12, map between FL 0.56 and 0.63 in the consensus map. Thus at least a part of the group-5 short arm is homoeologous to a region of chromosome 12 of rice. The genetic map of chromosome arm 5AS was constructed using a population of 139 BC1 plants derived from a cross between the euploid wheat ”Chris” carrying a dominant male-sterile gene Ms3 and a disomic substitution line in which chromosome 5A of T. aestivum cv Chinese Spring was substituted by chromosome 5A from Triticum turgidum ssp. dicoccoides. The map has a genetic length of 53.4 cM with 11 DNA markers. The initial map showed that the gene Ms3 cosegregated with three markers, WG341, BCD1130 and CDO677. High-resolution mapping using an additional 509 BC1 plants indicated that the marker WG341 was closely linked to Ms3 at a genetic distance of 0.8 cM. The Ms3 was mapped physically in the region spanning 40% of the arm length from the centromere of 5AS. Therefore, map-based cloning of the Ms3 is not feasible, although WG341 can be used as a useful tag for the Ms3 gene for breeding purposes.

Molecular tagging of a genic male-sterile gene in rice

The sterility of Pingxiang male-sterile rice (Pms), possibly derided from a spontaneous mutation in Pingxiang fertile rice (Pmf), was previously reported to be controlled by a single dominant nuclear gene. It can be restored to fertility either by a dominant epistatic gene or by higher temperature treatment at the early stage of inflorescence development. In order to tag the genic male-sterile gene, Pms, Pmf and Ce 64, a cytoplasmic male-sterile restoring line without the epistatic gene for Pms, were used to construct mapping populations. Two segregation populations, “(Pms/Ce 64) F1s (sterile plant)//Pmf” F1 and “Pms//(Pmf/Ce 64) Fl” F1, were simultaneously developed. Subsequently, the genic malesterile gene was mapped between a simple sequence length polymorphism marker, RM228, and a restriction fragment length polymorphism marker, G2155, with distances of 14.9 and 2.6 cM, respectively. The tagged dominant genic male-sterile gene is temporarily designated Ms-p.

Male sterility derived from Cajanus sericeus × Cajanus cajan: Confusion of cytoplasmic male sterility with dominant genic male sterility

The male sterile plants that segregated in a BC5F2 of `C. sericeus × C. cajan var. TT-5' population were maintained by sib mating. The male sterile plants were crossed with ICPL-85012.Approximately 50% of the F1 plants were sterile. F2 plants derived from the fertile F1 plants did not segregate for male sterility. The reciprocal hybrid i.e. ICPL-85012 × Fertile derivatives from C. sericeus × TT-5, did not express male sterility. However, among the 12 F2 plant to row progenies, two segregated 25% male sterile plants and remaining 10 did not segregate. The segregation pattern in subsequent progenies revealed that the sterility was under control of a single recessive allele. Studies on the backcross and their BC1F2 and BC1F3progenies revealed another sterility gene which was found to be dominant in inheritance. This paper shows that what was thought to be cytoplasmic male sterility from C. sericeus cytoplasm is actually a single dominant gene possibly acting in concert with a single recessive gene to mimic cytoplasmic male sterility.

An extended random primer amplified region (ERPAR) marker linked to a dominant male sterility gene in cabbage (Brassica oleracea var. capitata).

Similar to SCAR, an extended random primer amplified region (ERPAR) marker is a PCR amplified genomic DNA fragment at a single genetically defined locus. However, ERPAR uses specific primer pairs derived from RAPD primers by adding bases sequentially to their 3′-ends. As an example, an ERPAR marker was derived from a RAPD marker (OT11900) linked to a dominant male sterility gene in cabbage (Brassica oleracea var. capitata). After two cycles of base adding and primer pair screening, a primer pair (5′-TTCCCCGCGACT-3′and 5′-TTCCCCGCGAGA-3′) amplified a single intense band with the same size as OT11900. The identity of the new marker and OT11900 was verified by segregation analysis. The new marker amplified by this extended primer pair was named as EPT11900. The development of ERPAR exploits the importance of 3′-end bases of primers in PCR ERPAR shares advantages of SCAR, but eliminates the need for cloning and sequencing. It is a fast and universal way of converting RAPD markers into stable markers.