Ms Allele Research Articles

Distribution of Hybrid Necrosis Genes in Genotypes of Winter Soft wheat (<i>Triticum aestivum</i> L.) Cultivars of English Selection

The genetic diversity of 15 cultivars of winter soft wheat of English selection has been studied. It was found that 11 (73.3%) of the cultivars studied carried the Ne2 gene (genotype ne1ne1Ne2Ne2), and four cultivars (26.7%) were free of hybrid necrosis genes (genotype ne1ne1ne2ne2). The Ne1 gene was not detected. Cultivars with the Ne2 gene differed in allele strength. Moderate (m) allele was identified in four cultivars, and moderately strong (ms) allele was identified in the genotypes of seven cultivars.

A Single Nucleotide Polymorphism Unique to the Galanthum-CMS of Onion

Different sources of cytoplasmic male sterility (CMS) are used to produce hybrid onion seed. The most commonly used source of CMS in onion is S cytoplasm (S-CMS), and male fertility is restored by a dominant allele at the nuclear male-fertility restoration locus (Ms). Male-sterile plants possess S cytoplasm and have the homozygous recessive genotype at Ms; seed propagation of male-sterile plants is possible by crossing with a male-fertile maintainer plant or inbred possessing normal (N) male-fertile cytoplasm and the homozygous recessive at the Ms locus (N msms). Some commercially important onion populations possess S-CMS and high frequencies of the dominant Ms allele, eliminating the possibility to develop maintainer lines. An alloplasmic source of CMS (Gal-CMS) was developed by backcrossing the cytoplasm of Allium galanthum into the nuclear background of onion. The advantage of Gal-CMS is that the dominant allele at Ms does not restore male fertility, making this source of CMS useful for the development of male-sterile lines from populations possessing S cytoplasm and dominant allele(s) at Ms. In this research, a single nucleotide polymorphism unique to the cytoplasms of A. galanthum and Gal-CMS was identified, useful to distinguish Gal-CMS from other onion cytoplasms.

Distribution and Allele Strength of Hybrid Necrosis Genes in the Genotypes of Canadian Cultivars of Spring Bread Wheat (<i>Triticum aestivum</i> L.)

The occurrence of two types of cultivars of spring bread wheat of Canada that differ in their necrotic genotypes was shown. The cultivars of one type had the ne1ne1Ne2Ne2 genotype, and the cultivars of the second type had the ne1ne1ne2ne2 genotype. The Ne1Ne1ne2ne2 genotype was not found. The Ne2 genes were represented by medium strength, ms, and strong alleles, s. By pedigree analysis, the donors of the Ne2 gene were identified.

The investigation of the frequency of the alpha-1-antitrypsin phenotype in patients with liver cirrhosis

Alpha-1-antitrypsin (AAT) has different phenotypes. Evidence suggests that the abundance of each of these phenotypes may be associated with a disease. The purpose of this study was to evaluate the frequency of AAT phenotypes in patients with liver cirrhosis as well as in healthy individuals. In this study, 42 patients with liver cirrhosis were selected. The results of the previous research done by the researcher on healthy individuals were used to construct the control group. After obtaining informed consent, 5mL of fasting venous blood sample was taken, and phenotypes were analyzed by isoelectric focusing. Data were analyzed using Chi-square and Fisher's exact tests at a significant level of 0.05. The results of this study indicated that all 42 healthy subjects had an MM allele (100%). However, among 42 patients, 35 (83.3%) had an MM allele, 5 (11.9%) had an MS allele, and 2 (4.8%) had MZ allele. The difference between the two groups was significant (p=0.02). There was no difference between men and women in the allele type (p=0.557). This study revealed that MS and MZ alleles were observed only in patients with liver cirrhosis, and none of these alleles were found in healthy subjects. Therefore, MS and MZ alleles can be further investigated as risk factors for liver cirrhosis.

Variable Penetrance among Different Sources of the Male Fertility Restoration Allele of Onion

Hybrid onion (Allium cepa) seed is produced using cytoplasmic male sterility (CMS). For the most commonly used source of onion CMS, male fertile plants possess male sterile (S) cytoplasm and dominant allele(s) at one nuclear male fertility locus (Ms). Because male fertility restoration is not necessary for bulb production, it is desirable to purge dominant alleles at Ms from populations and breeding lines to facilitate the development of male sterile inbreds for hybrid production. In this research, we used molecular markers to establish the cytoplasms and genotypes at Ms in progenies from testcrosses of male sterile lines with plants from three populations [B2354, Ailsa Craig (AC), and Sapporo-Ki (Ski)] possessing the dominant Ms allele. We scored male fertility of testcross progenies by visual examination of flowers and acetocarmine staining of pollen. Different sources of the dominant Ms allele showed significantly different amounts of male fertility restoration and proportions of stainable pollen, complicating visual selection against the dominant Ms allele. For AC and Ski, molecular markers correctly predicted male sterility vs. male fertility of progenies in the greenhouse and field. However, for B2354, male fertility restoration was less clear and especially difficult to score under field conditions, consistent with reduced penetrance of male fertility restoration for this source of the dominant Ms allele. These results will be of interest to onion breeders selecting S-cytoplasmic male sterile lines for hybrid onion development.

Searching for sterility genes in bulb onion breeding accessions with the use of DNA markers

Relevance.Sterility is a very important trait that is indispensable for hybrid production. Genetic factor underlying in plant sterility can be now identified in large plant populations by DNA markers with high effectiveness and reliability. The evaluation of such markers enables to define their current applicability in breeding program.Methods.The markers from different publications that had been successfully used were taken to test their effectiveness on 19 accessions of bulb onion (Allium cepaL.).Results.Mitochondrial genes5’cob, orf725andorfA501and alleles of fertility restoring locus Ms were also identified. Four breeding accessions had S-cytoplasm, nine accessions were with T-cytoplasm and six shared normal cytoplasm not showing any sterility gene in the analysis. As a result of marker testing, the all compositions of the genes in cytoplasm and Ms alleles in nucleus affecting the sterility had been revealed, such as one sterility maintainer, one male sterile accession, and two fertility restorers. However, it should be noted that not all markers tested were in accordance with each other, where the markers originated from chloroplast DNA of did not confirmed the results obtained with those cytoplasm-origins. As it was shown the practical use of the set of markers makes it possible to reveal necessary accessions with required gene composition for hybrid production in bulb onion.

Fertility restoration locus and cytoplasm types in onion.

The objective of this study was the identification of the cytoplasmic types and the genotyping for the fertility restoration nuclear locus (Ms) in 59 onion accessions, aiming at the selection of 'A' and 'B' lines essential for the obtainment of hybrids. Three markers were used to identify the cytoplasm 5' cob, orfA501, and orf725, and two were used for the Ms locus (AcSKP1 and AcPMS1). The two types of male-sterile cytoplasm ('S' and 'T'), as well as fertile cytoplasm ('N'), and the Ms and ms alleles in both homozygosity and heterozygosity were detected in the 59 genotypes evaluated in the experiment. The frequencies of the 5' cob/orfA501 and orf725 markers, as well as of the markers AcSKP1 and AcPMS1, were close in the onion accessions evaluated in this study. In the Brazilian germplasm, the frequencies of the 'N', 'S', and 'T' cytoplasm were approximately 0.47, 0.28, and 0.25, respectively, whereas the allele frequencies of Ms and ms were 0.52 and 0.48, respectively. The accessions Régia, EHCEB 20146, EHCEB 201427, Alvorada, Serrana, Crioula Mercosul, EHCEB 20142, BRS 367, Rainha, Juporanga, and Alfa SF C-XI have potential for the identification of 'A' and 'B' lines, since they presented mixtures of cytoplasm and different allele frequencies for Ms. All the plants of the accessions EHCEB 20142040/EHCEB 20141040, EHCEB 20142028/EHCEB 20141028, and EHCEB 20112006/EHCEB 20111006 were in the Nmsms and Smsms conditions, and have the potential for 'B' and 'A' lines, respectively, for the CMS-S system. All the plants of the accessions EHCEB 20142027/EHCEB 20141027, EHCEB 20102019/EHCEB 20101019, and Alfa SF 'B'/Alfa SF 'A' were in Nmsms and Tmsms conditions, and have the potential for 'B' and 'A' lines, respectively, for the CMS-T system.

Isolation of Male Sterile and Maintainer Lines from North-Indian Onion (Allium cepa L.) Populations with the Aid of PCR-Based Molecular Marker

Isolation of Male Sterile and Maintainer Lines from NorthIndian Onion (Allium cepa L.) Populations with the Aid of PCRBased Molecular Marker Marker assisted selection (MAS) using mitochondrial DNA based marker cytochrome b (cob) was integrated with phenotypic evaluation to isolate male sterile and maintainer lines from openpollinated onion varieties adapted to North Indian agro-climatic region. Cytotype (N/S) determination by cob marker followed by morphological and microscopic study of pollen discovered male sterile plants (Smsms) at frequencies of 0.015 in Punjab Naroya, 0.020 in Punjab Selection, and 0.006 in Punjab White. The progeny scoring of test-crosses between male sterile and N-cytoplasmic plants isolated the maintainers (Nmsms) at frequencies of 0.133 in Punjab Naroya, 0.231 in Punjab Selection and 0.182 in Punjab White. As a novel approach, Trait Recovery Programme was demonstrated to reduce the population size required to recover a male sterile plant by 91.08% in Punjab Naroya, 92.99% in Punjab Selection and 97.66% in Punjab White. For recovering a maintainer, 10% reduction in Punjab Naroya and 9.10% in Punjab Selection was calculated. However, no reduction was observed in Punjab White. This analysis also validated that in a randomly mating onion population, frequency of recessive ms allele squared is equal to the frequency of male sterile plants among S-cytotype and frequency of maintainers among N-cytotype (fms2 = fSmsms/fS = fNmsms/fN)....

Limitations of PCR‐based molecular markers to identify male‐sterile and maintainer plants from Indian onion (Allium cepa L.) populations

AbstractThe main aim of this study was to validate PCR markers for determining cytoplasm and genotypes at the Ms locus in short‐day onion. Three cytoplasmic (OSN, MKFR and accD) and four nuclear (OPT, jnurf13, AcSKP1 and AcPMS1) markers were employed. Sel. 121‐1 had 100% S cytoplasm, whereas Sel. 121‐2, ‘Pusa Red’ and ‘Pusa Madhavi’ had 88%, 33% and 17% S cytoplasm, respectively. ‘Early Grano’ and ‘Pusa Riddhi’ did not possess S cytoplasm. Observations in 33 commercial varieties revealed two with sterile (S) cytoplasm. Nuclear markers were not found in linkage disequilibrium with the Ms locus, and the constitution of Ms alleles by OPT was different from other three markers, which were in conformity with each other. The other three markers predicted that most of the plants should be homozygous recessive. Anther colour also did not confirm the sterility status. It can be concluded that accD may be used for cytoplasm determination based on the ease of its use. For the Ms locus tagging, more markers are needed to be evaluated to isolate maintainer lines from open‐pollinated populations.

Abstract LB-134: A novel model to investigate Kras mutation specificity in the skin

Abstract The selection and specificity of mutations occurring in genes that drive cancer is a highly complex process that remains poorly understood for even the most well studied oncogenes. The specific pattern of RAS mutations across different cancers is thought to be modulated by differences in carcinogen exposure and metabolism, local DNA features and repair mechanisms, tissue-specific expression patterns and functional differences of the RAS isoforms1. Less is known about functional differences between the various activating mutations in RAS that may lead to selection of particular mutants in mouse or human cancers. Studies of mouse cancer models have provided evidence for strong genetic background effects on allele-specificity of Ras mutations2 as well as on selection of particular mutations at Kras codon 61 in lung adenomas3. We have also recently demonstrated a major role of germline Kras status in mutation selection during initiation, where carcinogen-induced lung tumors from Kras WT mice carry mostly (94%) Q61R Kras mutations, while those from Kras heterozygous mice carry mostly (92%) Q61L mutations4. In a M. Musculus (Mm) x M. Spretus (Ms) backcrossed population of HrasKO mice treated with DMBA/TPA, a model of Kras-driven tumorigenesis in the skin, we observed a wide spectrum of Kras mutations. Interestingly, genotype at the Kras locus significantly influenced Kras mutation specificity, with a switch from predominantly G13R mutations in Mm/Mm carcinomas to G12 and Q61L mutations in Mm/Ms carcinomas. Importantly, total Kras expression driven by the Mm Kras allele is higher than that driven by the Ms allele in the skin, and Kras mutations occur with high specificity for the Mm allele, suggesting that Q61L and G12 mutations may be suppressed by higher expression of the WT Kras allele in Kras Mm/Mm animals. We hypothesize that the ability of WT Kras to suppress certain mutants may be dependent on upstream activation by EGFR, and that this phenomenon may underlie the divergent responses to EGFR inhibition of colorectal cancer patients harboring Kras G12 versus G13 mutations5. Experiments to test this in mice by extended EGFR inhibition following carcinogen treatment are ongoing.

Single Nucleotide Polymorphisms in Linkage Disequilibrium with the Male-fertility Restoration (Ms) Locus in Open-pollinated and Inbred Populations of Onion

Maintainer lines are used to seed propagate male-sterile lines for the development of hybrid onion (Allium cepa L.) cultivars. The identification of maintainer lines would be more efficient with molecular markers distinguishing genotypes at the nuclear male-fertility restoration (Ms) locus. Ms has been mapped to chromosome 2 of onion and linked genetic markers identified. However, linkages between these markers and Ms were detected using F2 or BC1 families at maximum linkage disequilibrium and, for many markers, their efficacy to predict genotypes at Ms in onion populations at or near linkage equilibrium remains unknown. In this research, near isogenic lines homozygous-dominant and -recessive at Ms were developed and screened for 930 single nucleotide polymorphisms (SNPs). Three SNPs tightly linked on chromosome 2 remained in linkage disequilibrium with genotypes at Ms among randomly selected plants from three open-pollinated populations of onion as well as among a collection of inbred lines. These SNPs should be useful for selection of the recessive ms allele to aid in the development of maintainer lines for hybrid onion development.

Construction of high-resolution linkage map of the Ms locus, a restorer-of-fertility gene in onion (Allium cepa L.)

For the purpose of developing closely-linked molecular markers to the Ms locus, a restorer-of-fertility gene in onions (Allium cepa L.), bulked segregant analysis and randomly amplified polymorphic DNA (RAPD) analyses were utilized. Five RAPD markers polymorphic between male-fertile and male-sterile bulks were identified. These RAPD markers were converted into a simple PCR marker or cleaved amplified polymorphic sequence (CAPS) markers after sequencing the RAPD products and obtaining flanking sequences of the RAPD markers by genome walking. A linkage map was constructed with the Ms locus and flanking markers using a F2 population. There was no recombinant between the Ms locus and two CAPS markers, jnurf05 and jnurf17. To increase resolution among these closely linked molecular markers and the Ms locus, a total of 1,346 F2:3 and 2,927 F2:4 plants were analyzed with two flanking markers for detection of recombinants. Segregation of male-fertility phenotypes in large-sized populations confirmed allelic segregation distortion in favor of the recessive Ms allele. Analysis of the recombinants with closely linked markers revealed only two recombinants between the Ms locus and the jnurf05 markers among 4,273 segregating plants, showing very tight linkage between the two loci. However, linkage disequilibrium between the two loci was not too strong among the breeding lines. Despite weak linkage disequilibrium, these tightly linked markers are useful in accurate marker-assisted selection of the Ms alleles and ultimate isolation of the Ms gene by map-based cloning approach.

Identification of two SCAR markers co-segregated with the dominant Ms and recessive ms alleles in onion (Allium cepa L.)

Cytoplasmic genetic male-sterility is used to produce hybrid onion (Allium cepa L.) seeds worldwide. In this paper, we present the results of research aimed toward identifying PCR-based markers linked to the Ms locus through amplified fragment length polymorphism (AFLP). After screening 512 AFLP primer combinations, only one AFLP fragment was identified as being flanking linked to the dominant Ms allele. Subsequently, the AFLP marker was converted into a sequence-characterized amplified region (SCAR) marker, designated as DNF-566, co-segregated with the dominant Ms allele in first backcross (BC1) segregated populations. Furthermore, we designed another molecular marker (RNS-357) co-segregated with the ms allele to identify different genotypes (i.e., MsMs, Msms, or msms). Both markers could be used for evaluating onion lines with different genetic backgrounds (including male-sterile lines, maintainer lines, male-fertile lines, and commercial based F1 hybrid cultivars). The results of this study indicate that maintainer plants could be directly selected by using these 2 SCAR markers in the onion breeding process, and this may contribute significantly toward breeding onion F1 hybrid cultivars.

A locally congenic backcross design in pig: a new regional fine QTL mapping approach miming congenic strains used in mouse.

BackgroundIn previous studies, a major QTL affecting fatness and growth has been mapped to pig chromosome 1q (SSC1q) using Large White - Meishan intercrosses. A higher fat depth and a larger growth rate have been reported for the allele of MS origin. Additionally the LW allele showed partial dominance effects over the MS allele for both traits. In order to refine the QTL mapping interval, advanced backcross generations were produced. Recombinant heterozygous sires were mated to LW sows in order to progeny test the sire segregation of the QTL and refine the QTL localisation. However due to the partial dominance of the LW allele, BC scheme using LW as the receiving population was not optimal.ResultsTo overcome the difficulties related to the dominance of the LW QTL allele, a population of dams locally homozygous for the MS haplotype in the QTL region, but with an overall 29/32 LW genetic background, has been set up. Progeny testing results, using these receiver dams, were much more significant than those previously obtained with LW dams, and the SSC1 QTL interval was refined to 8 cM. Considering the results obtained, a powerful experimental design for farm animals is proposed, mimicking locally genetically identical strains used in mouse for QTL fine mapping.ConclusionsWe have further characterized the fatness QTL on pig chromosome 1 and refined its map position from a 30 cM interval to a 8 cM interval, using a locally congenic BC design. We have obtained highly significant results and overcome difficulties due to the dominance of the LW allele. This design will be used to produce additional, advanced BC families to further refine this QTL localization.

The Dominant Ms Allele in Onion Shows Reduced Penetrance

The most commonly used source of cytoplasmic male sterility in onion (Allium cepa) is controlled by the interaction of the cytoplasm [male-sterile (S) or normal male-fertile (N)] and one nuclear male-fertility-restoration locus (Ms). Scoring of genotypes at Ms is generally done by testcrossing male-fertile to male-sterile (S msms) plants, followed by scoring of testcross progenies for male-fertility restoration. We identified two N-cytoplasmic families, one that was homozygous dominant and the other segregating at Ms. Plants from each of these two families were individually testcrossed to male-sterile onion. Nuclear restoration of male fertility in testcross progenies was evaluated in the field over 4 years. For male plants homozygous dominant at Ms, we expected testcross families to show 100% male-fertility restoration, but observed mean values between 46% and 100%. For plants segregating at Ms, we again observed lower than expected frequencies of male-fertility restoration. These results demonstrate that the dominant Ms allele shows reduced penetrance, requiring that male-fertility restoration be scored over years to more confidently assign genotypes at Ms.

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.

Petaloid-type cms in carrot is not associated with expression of atp8 (orfB)

Two different atp8 reading frames with divergent C-terminal extensions were identified in a sterility-inducing mitochondrial type (mitotype) of carrot; examination of three fertility-inducing mitotypes revealed atp8 reading frames that were either terminated at a position conserved among other plant atp8 genes or continued with unique C-terminal extensions. Similarities between the C-terminal extensions suggest ancient common ancestry, but the level of sequence divergence implies lack of functional conservation. Northern analysis indicated that the C-terminal extensions are transcribed and are present within the primary atp8 transcripts. Western analysis revealed that the products of the two extended atp8 reading frames encoded by the sterility-inducing mitotype are significantly smaller than those predicted by complete translation of their C-terminal extensions, but are slightly larger than the products from unextended reading frames. No alterations were observed in atp8 transcripts or proteins isolated from flowers of the sterility-inducing mitotype that were phenotypically fertile as a result of segregation for nuclear fertility-restoring Ms or Rf alleles, and thus, there is no evidence that expression of atp8 is directly involved in the petaloid cms phenotype in carrot.

Selection at the Ms locus in open pollinated onion (Allium cepa L.) populations possessing S-cytoplasm or mixtures of N- and S-cytoplasms

The bulb onion (Allium cepa L.) was historically seed propagated by open pollination (OP). Cytoplasmic-genic male sterility (CMS) and protandry encourage outcrossing among individual onion plants. The most common source of CMS in onion is conditioned by the interaction of sterile (S) cytoplasm with a single nuclear male-fertility restoration (Ms) locus. We previously reported that the majority of OP onion populations possess normal (N) male-fertile cytoplasm and varying frequencies of the dominant Ms allele. It was unclear why N-cytoplasmic onion populations often possess relatively high frequencies of the Ms allele, which has no obvious function. We used computer simulations to estimate changes in allelic frequencies at Ms for onion populations possessing S-cytoplasm or a mixture of N- and S-cytoplasms, and to determine if frequencies of the Ms allele stay constant or change due to failure of male gamete production from male-sterile (S msms) plants. The models revealed selection against the recessive ms allele over generations in onion populations possessing S-cytoplasm and varying amounts of self pollination and inbreeding depression. These models were consistent with field and molecular analyses documenting that N-cytoplasm and the dominant Ms allele predominate in OP onion populations.

Isoelectric Focusing and PCR-RFLP Joined Techniques for Alpha1-antitrypsin Deficiency Detection

Summary 53 persons suspected to alpha1-antitrypsin deficiency detection (AATD) were investigated for ZZ, MZ, ZS, SS, and MS alleles analysis by serum protein electrophoresis (SPE), measurement of trypsin inhibiting capacity (TIC), isoelectric focusing (IEF), polymerase chain reaction (PCR), and IEF/PCRRFLP techniques. The result clearly shows by using SPE and TIC techniques only 35.85 % and 50.08% of AATD cases can detect respectively while, IEF/PCR-RFLP joined technique can detect 100% of AATD cases. However, since IEF/PCR-RFLP joined technique can determine the protein structure and alleles gene mutation together, they can detect up to 100% of AAT. Hence for detection of AATD the IEF/PCR-RFLP technique is recommended.

Methionine Synthase: High-Resolution Mapping of the Human Gene and Evaluation as a Candidate Locus for Neural Tube Defects

Periconceptual folate supplementation has been found to prevent the occurrence of many neural tube defects (NTDs). Consequently, genetic variation in folate metabolism genes is expected to contribute to the risk for neural tube defects. Methionine synthase catalyzes the vitamin B12-dependent conversion of homocysteine and 5-methyltetrahydrofolate to methionine and tetrahydrofolate. The observation that homocysteine and vitamin B12 levels are independent predictors of NTD risk suggested that methionine synthase could be a candidate gene for NTDs. To assess the role of the MS gene in NTDs, we performed high-resolution physical mapping of the MS locus, isolated highly polymorphic markers linked to the MS gene, and tested for an association between specific MS alleles and NTDs. We mapped the MS gene to a position between 909 and 913 cR10000 on chromosome 1 by radiation hybrid mapping. Polymorphic markers D1S1567 and D1S1568 map to locations no more than 900 and 194 kb from the MS gene, respectively. The segregation of these polymorphic markers was measured in 85 Irish NTD families. No allele of either marker showed a significant association with NTDs using the transmission disequilibrium test. A lack of association was also observed for the D1919G missense mutation within the gene. Our results suggest that inherited variation in the MS gene does not contribute to NTD risk in this population.