Analysis Of Recombinant Inbred Lines Research Articles

A novel QTL associated with tolerance to cold-induced seed cracking in the soybean cultivar Toyomizuki.

Low temperatures after flowering cause seed cracking (SC) in soybean. Previously, we reported that proanthocyanidin accumulation on the dorsal side of the seed coat, controlled by the I locus, may lead to cracked seeds; and that homozygous IcIc alleles at the I locus confer SC tolerance in the line Toiku 248. To discover new genes related to SC tolerance, we evaluated the physical and genetic mechanisms of SC tolerance in the cultivar Toyomizuki (genotype II). Histological and texture analyses of the seed coat revealed that the ability to maintain hardness and flexibility under low temperature, regardless of proanthocyanidin accumulation in the dorsal seed coat, contributes to SC tolerance in Toyomizuki. This indicated that the SC tolerance mechanism differed between Toyomizuki and Toiku 248. A quantitative trait loci (QTL) analysis of recombinant inbred lines revealed a new, stable QTL related to SC tolerance. The relationship between this new QTL, designated as qCS8-2, and SC tolerance was confirmed in residual heterozygous lines. The distance between qCS8-2 and the previously identified QTL qCS8-1, which is likely the Ic allele, was estimated to be 2-3 Mb, so it will be possible to pyramid these regions to develop new cultivars with increased SC tolerance.

Genome sequencing and genetic analysis of recombinant inbred lines reveals important agronomic traits related loci under different nitrogen fertilization.

The intensive application of inorganic nitrogen has contributed to increasing the crop yield with the risk of environmental damage. High nitrogen fertilizer requirements are a long-standing problem in japonica cultivation. In the present study, 200 recombinant inbred lines derived from two representative japonica varieties of Japanese and Chinese varieties, Akitakomachi and Liaogeng5, respectively, were planted under different nitrogen fertilization conditions. The relationships among nitrogen fertilization, genetic background, and important agronomic traits were surveyed through the phenotypic investigation and next-generation sequencing. The results showed that the yield components of Akitakomachi dramatically decreased along with the nitrogen reduction, whereas those of Liaogeng5 exhibited a slight decrease. The appearance quality and milling quality of both Akitakomachi and Liaogeng5 were improved, especially Liaogeng5. The recombinant inbred lines (RILs) showed similar trends to their parental lines. The QTLs analysis of important agronomic traits detected 17 loci, one QTL clustering located on chromosome 9 that corresponded to the plant height, panicle angle, grain number per panicle, harvest index, and taste value was identified to be synonymous with a previously reported gene, DEP1. The function of DEP1 was verified by a knock-out mutant constructed by CRISPR/Cas9. The dep1 mutant exhibited improved grain quality without severe yield penalty under nitrogen reduction conditions. The results of the present study provide useful information for the improvement of food security by enhancing nitrogen use efficiency.

Genetic diversity of Lathyrus sp collected from different geographical regions.

The demand for grass peas (Lathyrus sativus L.) had increased as high nutritional safe food, but most of the accessions of South Asia and Africa had low grain harvest. Therefore, this study had been undertaken to collect grass pea germplasm for boosting yields and quality improvement. In this study, 400 accessions of grass pea from different geographical regions had characterized by using 56 Simple Sequences Repeat (SSRs) markers. In total 253 alleles were detected, the maximum and minimum polymorphic information content (PIC) indices were 0.70 and 0.34 found in markers G17922 and G18078, correspondingly. The germplasm was split into two main and one sub-group by cluster assay, by SSR assay, and three populations by model-based population structure analysis (Pop1, Pop2 and admixed). Neighbors joining tree assay showed the tested germplasm highly diverse in structure. Three-dimensional principal components analysis (PCA) and two dimensional principles coordinate analysis (PCoA) exhibited two main and one admixed group (P1, P2 and P1P2). In addition, FST population value of pairwise mean and analysis of molecular variance (AMOVA) showed high population structure across all pairs of populations on an average 0.1710 advocating all population structure categories varied significantly. The average predictable heterozygosity distant was 0.4472-0.4542 in same cluster for the individuals. Discovery from this study revealed SSR markers based polymorphic bands showed in the diversified grasspea germplasm which might be utilized as genetic resource of a breeding scheme and prospective uses for mapping analyses of recombinant inbred lines (RIL).

Common bean (Phaseolus vulgaris L.) with increased cysteine and methionine concentration

AbstractIn common bean (Phaseolus vulgaris L.), seed storage protein deficiency is associated with increased total cysteine and methionine concentration. The goal of this study was to generate germplasm lines that combine this characteristic with adaptation to short season conditions in Manitoba, Canada. A recombinant inbred line population was developed by crossing the storage protein deficient genotype, SMARC1N‐PN1 with the cultivar Morden003. Two lines, 2‐37 and 3‐84, with a stable protein profile over 2 years at two locations were identified. Like SMARC1N‐PN1, both lines had a significantly higher cysteine concentration than Morden003, by approximately 35%. Methionine levels were elevated by approximately 15%, while tryptophan levels were also increased by approximately 30%. Line 2‐37 had a significant increase in protein quality, as measured by in vitro protein digestibility corrected amino acid score, by approximately 40%, as compared with Morden003. The increased protein quality for this line is attributable to higher levels of total cysteine and methionine, while having an overall reduction in crude protein concentration. Line 2‐37 had a similar seed yield as SMARC1N‐PN1, with a maturity comparable to Morden003. The results of high‐density single nucleotide polymorphism (SNP) genotyping and quantitative trait locus analysis of recombinant inbred lines indicated that variation in cysteine concentration was determined by the phaseolin locus, while variation in methionine concentration was determined by both the phaseolin and lectin loci. SNP markers that track the introgression of phaseolin and lectin deficiency into the Morden003 background were identified and validated.

Identification of a Potential Gene for Elevating ω-3 Concentration and Its Efficiency for Improving the ω-6/ω-3 Ratio in Soybean.

This present study was to identify a novel candidate gene that contributes to the elevated α-linolenic acid (ALA, ω-3) concentration in PE2166 from mutagenesis of Pungsannamul. Major loci qALA5_1 and qALA5_2 were detected on chromosome 5 of soybean through quantitative trait loci mapping analyses of recombinant inbred lines. With next-generation sequencing of parental lines and Pungsannamul and recombinant analyses, a potential gene, Glyma.05g221500 (HD), controlling elevated ALA concentration was identified. HD is a homeodomain-like transcriptional regulator that may regulate the expression level of microsomal ω-3 fatty acid desaturase (FAD3) genes responsible for the conversion of linoleic acid into ALA in the fatty acid biosynthetic pathway. In addition, we hypothesized that a combination of mutant alleles, HD, and either of microsomal delta-12 fatty acid desaturase 2-1 (FAD2-1) could reduce the ω-6/ω-3 ratio. In populations where HD, FAD2-1A, and FAD2-1B genes were segregated, a combination of a hd allele from PE2166 and either of the variant FAD2-1 alleles was sufficient to reduce the ω-6/ω-3 ratio in seeds.

The effects of a QTL for root development under hypoxia on yield of soybean exposed to 7-day waterlogging at seedling stage

In humid parts of the world, waterlogging is a major environmental stress limiting the yield of soybean and its root development is an important indicator of tolerance. Recently, quantitative trait loci (QTLs) for this trait were detected on the chromosome 12 of soybean through the analyses of recombinant inbred lines (RILs) developed from a cross between a hypoxia-sensitive cultivar, Tachinagaha, and a tolerant landrace, Iyodaizu. In this study, in order to get a detailed understanding of these QTLs, NIL-9-4-5, a near-isogenic line (NIL), was selected from Tachinagaha/Iyodaizu BC6F2 population through marker assisted selection (MAS). NIL-9-4-5 was then tested for yield performance under waterlogging conditions exposed at the seedling stage. The test was conducted in a greenhouse in 2015 and 2018 at the Experimental Farm of Kyushu University, Fukuoka, Japan. Soybean plants were grown in pots containing loam soil. From six days after germination, seedlings were grown for 7 days under two treatments: drained (control) and waterlogging conditions. Pots were kept in containers where the water level was maintained at 2.0 cm depth from bottom in the control and at 2.0 cm above the soil surface in waterlogged condition. Experimental results consistently showed that seed yields of both Iyodaizu and NIL-9-4-5 were not reduced under waterlogging. Compared to Tachinagaha, NIL-9-4-5 showed higher root development and increased seed yield under waterlogging. These indicate that the tested QTLs are able to increase yield by stimulating root development under waterlogging.

Extensive intraspecies cryptic variation in an ancient embryonic gene regulatory network.

Innovations in metazoan development arise from evolutionary modification of gene regulatory networks (GRNs). We report widespread cryptic variation in the requirement for two key regulatory inputs, SKN-1/Nrf2 and MOM-2/Wnt, into the C. elegans endoderm GRN. While some natural isolates show a nearly absolute requirement for these two regulators, in others, most embryos differentiate endoderm in their absence. GWAS and analysis of recombinant inbred lines reveal multiple genetic regions underlying this broad phenotypic variation. We observe a reciprocal trend, in which genomic variants, or knockdown of endoderm regulatory genes, that result in a high SKN-1 requirement often show low MOM-2/Wnt requirement and vice-versa, suggesting that cryptic variation in the endoderm GRN may be tuned by opposing requirements for these two key regulatory inputs. These findings reveal that while the downstream components in the endoderm GRN are common across metazoan phylogeny, initiating regulatory inputs are remarkably plastic even within a single species.

A multi-parent recombinant inbred line population of C. elegans allows identification of novel QTLs for complex life history traits

BackgroundThe nematode Caenorhabditis elegans has been extensively used to explore the relationships between complex traits, genotypes, and environments. Complex traits can vary across different genotypes of a species, and the genetic regulators of trait variation can be mapped on the genome using quantitative trait locus (QTL) analysis of recombinant inbred lines (RILs) derived from genetically and phenotypically divergent parents. Most RILs have been derived from crossing two parents from globally distant locations. However, the genetic diversity between local C. elegans populations can be as diverse as between global populations and could thus provide means of identifying genetic variation associated with complex traits relevant on a broader scale.ResultsTo investigate the effect of local genetic variation on heritable traits, we developed a new RIL population derived from 4 parental wild isolates collected from 2 closely located sites in France: Orsay and Santeuil. We crossed these 4 genetically diverse parental isolates to generate a population of 200 multi-parental RILs and used RNA-seq to obtain sequence polymorphisms identifying almost 9000 SNPs variable between the 4 genotypes with an average spacing of 11 kb, doubling the mapping resolution relative to currently available RIL panels for many loci. The SNPs were used to construct a genetic map to facilitate QTL analysis. We measured life history traits such as lifespan, stress resistance, developmental speed, and population growth in different environments, and found substantial variation for most traits. We detected multiple QTLs for most traits, including novel QTLs not found in previous QTL analysis, including those for lifespan and pathogen responses. This shows that recombining genetic variation across C. elegans populations that are in geographical close proximity provides ample variation for QTL mapping.ConclusionTaken together, we show that using more parents than the classical two parental genotypes to construct a RIL population facilitates the detection of QTLs and that the use of wild isolates facilitates the detection of QTLs. The use of multi-parent RIL populations can further enhance our understanding of local adaptation and life history trade-offs.

Identification of Soybean Genes Whose Expression is Affected by the Ensifer fredii HH103 Effector Protein NopP.

In some legume–rhizobium symbioses, host specificity is influenced by rhizobial nodulation outer proteins (Nops). However, the genes encoding host proteins that interact with Nops remain unknown. We generated an Ensifer fredii HH103 NopP mutant (HH103ΩNopP), and analyzed the nodule number (NN) and nodule dry weight (NDW) of 10 soybean germplasms inoculated with the wild-type E. fredii HH103 or the mutant strain. An analysis of recombinant inbred lines (RILs) revealed the quantitative trait loci (QTLs) associated with NopP interactions. A soybean genomic region containing two overlapping QTLs was analyzed in greater detail. A transcriptome analysis and qRT-PCR assay were used to identify candidate genes encoding proteins that interact with NopP. In some germplasms, NopP positively and negatively affected the NN and NDW, while NopP had different effects on NN and NDW in other germplasms. The QTL region in chromosome 12 was further analyzed. The expression patterns of candidate genes Glyma.12g031200 and Glyma.12g073000 were determined by qRT-PCR, and were confirmed to be influenced by NopP.

Inheritance of aluminum tolerance in the wheat cultivar Toropi and new findings about the introduction of this trait into the Brazilian wheat germplasm

Aluminum (Al) is the most abundant metal in the earth’s crust, and in acidic soils it dissolves into the phytotoxic form Al3+. Analysis of the inheritance of aluminum tolerance in a donor genotype is an important step in the development of new cultivars adapted to acidic soils. The Brazilian wheat cultivars BH1146 and Carazinho have been important donors of aluminum tolerance genes in wheat breeding programs around the world. This trait is under the control of two main genes, TaALMT1 and TaMATE1B. Interestingly, both cultivars have both of these genes, but different alleles. Lineages developed in Brazil from local landraces gave rise to the cultivars BH1146 and Carazinho, as well as other genotypes among which the cultivar Toropi is highlighted herein because of its excellent performance in acidic soils. It is not known if the genes and alleles responsible for aluminum tolerance in Toropi are the same as those carried by BH1146 and Carazinho. We analyzed the recombinant inbred lines (RILs) from Toropi × Anahuac (an Al-susceptible cultivar) and showed that aluminum tolerance in Toropi was controlled by one gene with major effects on the phenotype, although other gene(s) with minor effects also affected this trait. The analysis of RILs from Toropi × BH1146 showed that this major gene was different from the one carried by BH1146. The genealogy and molecular analyses of the TaALMT1 and TaMATE1B genes of the Al-tolerant Brazilian wheat cultivars allowed us to establish that Toropi has the same allele of the TaMATE1B gene as Carazinho, but probably originating from a different source.

Identification of genomic regions regulating ammonium-dependent inhibition of primary root length in Arabidopsis thaliana

ABSTRACTPlant roots grow into the soil for efficient acquisition of various nutrients, such as inorganic nitrogen, ammonium, and nitrate. A previous study has revealed the genetic diversity of foliar functions of Arabidopsis thaliana ecotypes in an environment containing ammonium; however, the function of roots remains unclear. This work focuses on the root system architecture (RSA) of Arabidopsis ecotypes to investigate the genetic factors regulating ammonium-dependent RSA changes. Arabidopsis ecotypes were grown on vertical agar medium containing ammonium as a major nitrogen source, and root growth and RSA were determined. Arabidopsis ecotypes showed differential sensitivity to ammonium. The shoot dry weight of some ecotypes decreased, whereas that of other ecotypes increased in ammonium medium. The RSA changes also varied among the different ecotypes in response to ammonium. The total root length, measured as the sum of primary root and lateral root length, of some ecotypes was reduced, whereas that of other ecotypes showed no significant difference in ammonium medium. Compared with lateral roots, the primary roots showed a sharp response to ammonium supply. Notably, the RSA showed a partial correlation with shoot dry weight in ammonium medium. Because Col-4 and Ler-0 showed opposite RSA responses to ammonium supply, these two ecotypes were selected for further genetic analysis. Quantitative trait locus (QTL) analysis of recombinant inbred lines of Col and Ler showed the involvement of several genetic factors in ammonium-dependent RSA changes. Moreover, QTL analysis revealed that the primary structure of nitrogen-related enzymes do not account for changes in RSA in response to ammonium supply.

Allelic Variation and Transcriptional Isoforms of Wheat TaMYC1 Gene Regulating Anthocyanin Synthesis in Pericarp

Recently the TaMYC1 gene encoding bHLH transcription factor has been isolated from the bread wheat (Triticum aestivum L.) genome and shown to co-locate with the Pp3 gene conferring purple pericarp color. As a functional evidence of TaMYC1 and Pp3 being the same, higher transcriptional activity of the TaMYC1 gene in colored pericarp compared to uncolored one has been demonstrated. In the current study, we present additional strong evidences of TaMYC1 to be a synonym of Pp3. Furthermore, we have found differences between dominant and recessive Pp3(TaMyc1) alleles. Light enhancement of TaMYC1 transcription was paralleled with increased AP accumulation only in purple-grain wheat. Coexpression of TaMYC1 and the maize MYB TF gene ZmC1 induced AP accumulation in the coleoptile of white-grain wheat. Suppression of TaMYC1 significantly reduced AP content in purple grains. Two distinct TaMYC1 alleles (TaMYC1p and TaMYC1w) were isolated from purple- and white-grained wheat, respectively. A unique, compound cis-acting regulatory element had six copies in the promoter of TaMYC1p, but was present only once in TaMYC1w. Analysis of recombinant inbred lines showed that TaMYC1p was necessary but not sufficient for AP accumulation in the pericarp tissues. Examination of larger sets of germplasm lines indicated that the evolution of purple pericarp in tetraploid wheat was accompanied by the presence of TaMYC1p. Our findings may promote more systematic basic and applied studies of anthocyanins in common wheat and related Triticeae crops.

Genetic analysis of recombinant inbred lines for iron deficiency chlorosis and productivity traits in groundnut

Iron deficiency is an important abiotic constraint reducing the growth and yield of groundnut especially under calcareous soils. Foliar application of Fe-chelates can overcome iron deficiency but it is not economical. Evaluation of 318 recombinant inbred lines (RILs) along with parents for iron deficiency chlorosis (IDC) and productivity traits under iron deficient soils over three years indicated significant genotypic and genotypic x environment interaction (GE) component for both IDC and productivity traits. Among the RILs, range of variation was higher than that of the parents for visual chlorotic rating (VCR), SPAD chlorophyll meter reading (SCMR) and productivity parameters across three years indicated the presence of transgressive segregants. VCR had higher phenotypic and genotypic, variances, heritability and genetic advance as per cent of mean (GAM) in all the three years as compared to SCMR. Among productivity traits, pod yield (g plant–1) had higher PCV and GCV compared to shelling per cent and 100 seed weight. Significant negative correlation between VCR and pod yield per plant indicated effect of IDC on productivity in RILs. Nine lines were superior for both IDC tolerance and productivity traits. This extensive phenotyping of RIL population for IDC tolerance under iron deficient conditions can be used for identification of genomic regions associated with IDC tolerance by genotyping of this RIL population.

Heterotaxy in Caenorhabditis: widespread natural variation in left-right arrangement of the major organs.

Although the arrangement of internal organs in most metazoans is profoundly left–right (L/R) asymmetric with a predominant handedness, rare individuals show full (mirror-symmetric) or partial (heterotaxy) reversals. While the nematode Caenorhabditis elegans is known for its highly determinate development, including stereotyped L/R organ handedness, we found that L/R asymmetry of the major organs, the gut and gonad, varies among natural isolates of the species in both males and hermaphrodites. In hermaphrodites, heterotaxy can involve one or both bilaterally asymmetric gonad arms. Male heterotaxy is probably not attributable to relaxed selection in this hermaphroditic species, as it is also seen in gonochoristic Caenorhabditis species. Heterotaxy increases in many isolates at elevated temperature, with one showing a pregastrulation temperature-sensitive period, suggesting a very early embryonic or germline effect on this much later developmental outcome. A genome-wide association study of 100 isolates showed that male heterotaxy is associated with three genomic regions. Analysis of recombinant inbred lines suggests that a small number of loci are responsible for the observed variation. These findings reveal that heterotaxy is a widely varying quantitative trait in an animal with an otherwise highly stereotyped anatomy, demonstrating unexpected plasticity in an L/R arrangement of the major organs even in a simple animal.This article is part of the themed issue ‘Provocative questions in left–right asymmetry’.

Mapping QTLs for Salt Tolerance in Rice (Oryza sativa L.) by Bulked Segregant Analysis of Recombinant Inbred Lines Using 50K SNP Chip

Soil salinity is a major constraint to rice production in large inland and coastal areas around the world. Modern high yielding rice varieties are particularly sensitive to high salt stress. There are salt tolerant landraces and traditional varieties of rice but with limited information on genomic regions (QTLs) and genes responsible for their tolerance. Here we describe a method for rapid identification of QTLs for reproductive stage salt tolerance in rice using bulked segregant analysis (BSA) of bi-parental recombinant inbred lines (RIL). The number of RILs required for the creation of two bulks with extreme phenotypes was optimized to be thirty each. The parents and bulks were genotyped using a 50K SNP chip to identify genomic regions showing homogeneity for contrasting alleles of polymorphic SNPs in the two bulks. The method was applied to ‘CSR11/MI48’ RILs segregating for reproductive stage salt tolerance. Genotyping of the parents and RIL bulks, made on the basis of salt sensitivity index for grain yield, revealed 6,068 polymorphic SNPs and 21 QTL regions showing homogeneity of contrasting alleles in the two bulks. The method was validated further with ‘CSR27/MI48’ RILs used earlier for mapping salt tolerance QTLs using low-density SSR markers. BSA with 50K SNP chip revealed 5,021 polymorphic loci and 34 QTL regions. This not only confirmed the location of previously mapped QTLs but also identified several new QTLs, and provided a rapid way to scan the whole genome for mapping QTLs for complex agronomic traits in rice.

Pullulanase and Starch Synthase III Are Associated with Formation of Vitreous Endosperm in Quality Protein Maize

The opaque-2 (o2) mutation of maize increases lysine content, but the low seed density and soft texture of this type of mutant are undesirable. Lines with modifiers of the soft kernel phenotype (mo2) called “Quality Protein Maize” (QPM) have high lysine and kernel phenotypes similar to normal maize. Prior research indicated that the formation of vitreous endosperm in QPM might involve changes in starch granule structure. In this study, we focused on analysis of two starch biosynthetic enzymes that may influence kernel vitreousness. Analysis of recombinant inbred lines derived from a cross of W64Ao2 and K0326Y revealed that pullulanase activity had significant positive correlation with kernel vitreousness. We also found that decreased Starch Synthase III abundance may decrease the pullulanase activity and average glucan chain length given the same Zpu1 genotype. Therefore, Starch Synthase III could indirectly influence the kernel vitreousness by affecting pullulanase activity and coordinating with pullulanase to alter the glucan chain length distribution of amylopectin, resulting in different starch structural properties. The glucan chain length distribution had strong positive correlation with the polydispersity index of glucan chains, which was positively associated with the kernel vitreousness based on nonlinear regression analysis. Therefore, we propose that pullulanase and Starch Synthase III are two important factors responsible for the formation of the vitreous phenotype of QPM endosperms.

Confirming a major QTL and finding additional loci responsible for field resistance to brown spot (Bipolaris oryzae) in rice.

Brown spot is a devastating rice disease. Quantitative resistance has been observed in local varieties (e.g., ‘Tadukan’), but no economically useful resistant variety has been bred. Using quantitative trait locus (QTL) analysis of recombinant inbred lines (RILs) from ‘Tadukan’ (resistant) × ‘Hinohikari’ (susceptible), we previously found three QTLs (qBS2, qBS9, and qBS11) that conferred resistance in seedlings in a greenhouse. To confirm their effect, the parents and later generations of RILs were transplanted into paddy fields where brown spot severely occurred. Three new resistance QTLs (qBSfR1, qBSfR4, and qBSfR11) were detected on chromosomes 1, 4, and 11, respectively. The ‘Tadukan’ alleles at qBSfR1 and qBSfR11 and the ‘Hinohikari’ allele at qBSfR4 increased resistance. The major QTL qBSfR11 coincided with qBS11 from the previous study, whereas qBSfR1 and qBSfR4 were new but neither qBS2 nor qBS9 were detected. To verify the qBSfR1 and qBSfR11 ‘Tadukan’ resistance alleles, near-isogenic lines (NILs) with one or both QTLs in a susceptible background (‘Koshihikari’) were evaluated under field conditions. NILs with qBSfR11 acquired significant field resistance; those with qBSfR1 did not. This confirms the effectiveness of qBSfR11. Genetic markers flanking qBSfR11 will be powerful tools for marker-assisted selection to improve brown spot resistance.

Identification of QTL for Grain Protein Content and Grain Hardness from Winter Wheat for Genetic Improvement of Spring Wheat

To utilize the favorable gene(s) from winter wheat for genetic improvement of spring wheat, this study was carried out to identify the quantitative trait loci (QTL) associated with grain protein content (GPC) and grain hardness (GH) by analysis of recombinant inbred lines (RILS) derived from a cross between spring wheat and spring version of winter wheat. A genetic map of 334 loci was constructed which covered 1575.30cM on all 21 chromosomes. Two QTLs on 3B and 5B chromosome were detected for GPC. A QTL identified barc77 on chromosome 3B had additive effect of 0.17 and the other QTL identified by gwm499 on chromosome 5B had additive effect of 0.19. There were two major QTLs for GH identified on Chromosome 1B and chromosome 5A. The QTL on 1B was localized within a 18.7cM region flanked by wmc719 and wmc367-1 with 1.75 additive effect. The QTL on chromosome 5A flanked by SNP markers, IWA6573 and IWA2363, had additive effect of 1.44.

Epigenetic and Genetic Influences on DNA Methylation Variation in Maize Populations

DNA methylation is a chromatin modification that is frequently associated with epigenetic regulation in plants and mammals. However, genetic changes such as transposon insertions can also lead to changes in DNA methylation. Genome-wide profiles of DNA methylation for 20 maize (Zea mays) inbred lines were used to discover differentially methylated regions (DMRs). The methylation level for each of these DMRs was also assayed in 31 additional maize or teosinte genotypes, resulting in the discovery of 1966 common DMRs and 1754 rare DMRs. Analysis of recombinant inbred lines provides evidence that the majority of DMRs are heritable. A local association scan found that nearly half of the DMRs with common variation are significantly associated with single nucleotide polymorphisms found within or near the DMR. Many of the DMRs that are significantly associated with local genetic variation are found near transposable elements that may contribute to the variation in DNA methylation. Analysis of gene expression in the same samples used for DNA methylation profiling identified over 300 genes with expression patterns that are significantly associated with DNA methylation variation. Collectively, our results suggest that DNA methylation variation is influenced by genetic and epigenetic changes that are often stably inherited and can influence the expression of nearby genes.

Long-Range Regulatory Polymorphisms Affecting a GABA Receptor Constitute a Quantitative Trait Locus (QTL) for Social Behavior in Caenorhabditis elegans

Aggregation is a social behavior that varies between and within species, providing a model to study the genetic basis of behavioral diversity. In the nematode Caenorhabditis elegans, aggregation is regulated by environmental context and by two neuromodulatory pathways, one dependent on the neuropeptide receptor NPR-1 and one dependent on the TGF-β family protein DAF-7. To gain further insight into the genetic regulation of aggregation, we characterize natural variation underlying behavioral differences between two wild-type C. elegans strains, N2 and CB4856. Using quantitative genetic techniques, including a survey of chromosome substitution strains and QTL analysis of recombinant inbred lines, we identify three new QTLs affecting aggregation in addition to the two known N2 mutations in npr-1 and glb-5. Fine-mapping with near-isogenic lines localized one QTL, accounting for 5%–8% of the behavioral variance between N2 and CB4856, 3′ to the transcript of the GABA neurotransmitter receptor gene exp-1. Quantitative complementation tests demonstrated that this QTL affects exp-1, identifying exp-1 and GABA signaling as new regulators of aggregation. exp-1 interacts genetically with the daf-7 TGF-β pathway, which integrates food availability and population density, and exp-1 mutations affect the level of daf-7 expression. Our results add to growing evidence that genetic variation affecting neurotransmitter receptor genes is a source of natural behavioral variation.