BackgroundSalinity is a major environmental factor limiting productivity of crop plants including rice in which wide range of natural variability exists. Although recent evidences implicate epigenetic mechanisms for modulating the gene expression in plants under environmental stresses, epigenetic changes and their functional consequences under salinity stress in rice are underexplored. DNA methylation is one of the epigenetic mechanisms regulating gene expression in plant’s responses to environmental stresses. Better understanding of epigenetic regulation of plant growth and response to environmental stresses may create novel heritable variation for crop improvement.Methodology/Principal FindingsMethylation sensitive amplification polymorphism (MSAP) technique was used to assess the effect of salt stress on extent and patterns of DNA methylation in four genotypes of rice differing in the degree of salinity tolerance. Overall, the amount of DNA methylation was more in shoot compared to root and the contribution of fully methylated loci was always more than hemi-methylated loci. Sequencing of ten randomly selected MSAP fragments indicated gene-body specific DNA methylation of retrotransposons, stress responsive genes, and chromatin modification genes, distributed on different rice chromosomes. Bisulphite sequencing and quantitative RT-PCR analysis of selected MSAP loci showed that cytosine methylation changes under salinity as well as gene expression varied with genotypes and tissue types irrespective of the level of salinity tolerance of rice genotypes.Conclusions/SignificanceThe gene body methylation may have an important role in regulating gene expression in organ and genotype specific manner under salinity stress. Association between salt tolerance and methylation changes observed in some cases suggested that many methylation changes are not “directed”. The natural genetic variation for salt tolerance observed in rice germplasm may be independent of the extent and pattern of DNA methylation which may have been induced by abiotic stress followed by accumulation through the natural selection process.

BackgroundFinger millet (Eleusine coracana (L.) Gaertn.) is an important staple food crop widely grown in Africa and South Asia. Among the millets, finger millet has high amount of calcium, methionine, tryptophan, fiber, and sulphur containing amino acids. In addition, it has C4 photosynthetic carbon assimilation mechanism, which helps to utilize water and nitrogen efficiently under hot and arid conditions without severely affecting yield. Therefore, development and utilization of genomic resources for genetic improvement of this crop is immensely useful.ResultsExperimental results from whole genome sequencing and assembling process of ML-365 finger millet cultivar yielded 1196 Mb covering approximately 82% of total estimated genome size. Genome analysis showed the presence of 85,243 genes and one half of the genome is repetitive in nature. The finger millet genome was found to have higher colinearity with foxtail millet and rice as compared to other Poaceae species. Mining of simple sequence repeats (SSRs) yielded abundance of SSRs within the finger millet genome. Functional annotation and mining of transcription factors revealed finger millet genome harbors large number of drought tolerance related genes. Transcriptome analysis of low moisture stress and non-stress samples revealed the identification of several drought-induced candidate genes, which could be used in drought tolerance breeding.ConclusionsThis genome sequencing effort will strengthen plant breeders for allele discovery, genetic mapping, and identification of candidate genes for agronomically important traits. Availability of genomic resources of finger millet will enhance the novel breeding possibilities to address potential challenges of finger millet improvement.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3850-z) contains supplementary material, which is available to authorized users.

Seed shattering is an important trait that distinguishes crop cultivars from the wild and weedy species. The genetics of seed shattering was investigated in this study to provide insights into rice domestication and the evolution of weedy rice. Quantitative trait locus (QTL) analysis, conducted in 2 recombinant inbred populations involving 2 rice cultivars and a weedy rice accession of the southern United States, revealed 3-5 QTLs that controlled seed shattering with 38-45% of the total phenotypic variation. Two QTLs on chromosomes 4 and 10 were consistent in both populations. Both cultivar and weedy rice contributed alleles for increased seed shattering. Genetic backgrounds affected both QTL number and the magnitude of QTL effects. The major QTL qSH4 and a minor QTL qSH3 were validated in near-isogenic lines, with the former conferring a significantly higher degree of seed shattering than the latter. Although the major QTL qSH4 overlapped with the sh4, the presence of the nonshattering single nucleotide polymorphism allele in the weedy rice accession suggested involvement of a linked locus or an alternative molecular genetic mechanism. Overlapping of several QTLs with those from earlier studies indicated that weedy rice may have been derived from the wild species Oryza rufipogon. Natural hybridization of rice cultivars with the highly variable O. rufipogon present in different geographic regions might be responsible for the evolution of a wide range of phenotypic and genotypic variabilities seen in weedy rice populations worldwide.

Seed dormancy (SD) is a key domestication trait closely related to preharvest sprouting tolerance. Wild and weedy rices ißryza spp.) exhibit higher degrees of seed dormancy compared to the cultivated rice. Red rice {Oryza sativa L.), a major weed in the rice growing areas of the southern United States, was used to elucidate the genetic architecture of SD. Quantitative trait loci (QTL) analysis conducted in two recombinant inbred line (RIL) populations developed from the crosses involving two rice cultivars (Bengal and Cypress) and a red rice accession (PSRR-1) revealed six to seven QTL for seed dormancy, which accounted for 49 to 52% of the total phenotypic variance. The magnitude of the QTL contribution to phenotypic variance was influenced by genetic backgrounds. The majority of QTL had minor effects, except the QTL linked to Re and Sdr4. The genetic architecture for seed dormancy in U.S. red rice was distinct compared with the earlier reported weedy accessions. Four QTL were mapped onto similar positions in both populations. Both cultivars and red rice contributed alleles for increased SD. Most of the digenic epistatic interactions involved loci other than the QTL with main effects. The nucleotide polymorphisms at the Sdr4 locus could not explain the phenotypic variation for seed dormancy in our materials. The variation in SD among the rice cuitivars could be attributed to segregation of minor QTL, which may be exploited to improve preharvest sprouting tolerance.

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The transgenic technology using a single gene has been widely used for crop improvement. But the transgenic pyramiding of multiple genes, a promising alternative especially for enhancing complexly inherited abiotic stress tolerance, has received little attention. Here, we developed and evaluated transgenic rice lines with a single Salt Responsive Protein 3-1 (SaSRP3-1) gene as well as pyramids with two-genes SaSRP3-1 and Vacuolar H+-ATPase subunit c1 (SaVHAc1) derived from a halophyte grass Spartina alterniflora L. for salt tolerance at seedling, vegetative, and reproductive stages. The overexpression of this novel gene SaSRP3-1 resulted in significantly better growth of E. coli with the recombinant plasmid under 600 mM NaCl stress condition compared with the control. During early seedling and vegetative stages, the single gene and pyramided transgenic rice plants showed enhanced tolerance to salt stress with minimal wilting and drying symptoms, improved shoot and root growth, and significantly higher chlorophyll content, relative water content, and K+/Na+ ratio than the control plants. The salt stress screening during reproductive stage revealed that the transgenic plants with single gene and pyramids had better grain filling, whereas the pyramided plants showed significantly higher grain yield and higher grain weight compared to control plants. Our study demonstrated transgenic pyramiding as a viable approach to achieve higher level of salt tolerance in crop plants.

Reproductive-stage drought-stress is a major production constraint in rainfed rice ecosystem. Emergence of marker-assisted breeding strategies for developing drought-tolerant rice varieties are being optimized through exploiting adaptive-traits for their increased contribution towards grain-yield under recurring-drought. Grain-yield is a complex-trait; requires knowledge of genetics and association among yield contributing component-traits. Current study was undertaken using 21 marker-assisted multidrought-traits pyramided genotypes responses for genetic variability and association of traits for grain-yield under aerobic and reproductive-stage drought conditions. Field evaluation was carried-out in two seasons and data was collected on various parameters. Path-coe cient analysis was used as a selection criterion to select yield contributing-traits and found nine phenotypic traits were having a positive direct-effect on grain-yield during both and/or at least one season under both moistureregimes. The data from summer and Kharif seasons have been pooled within their respective moisture-regimes due to the nonsigni cance of Levene's test of homogeneity of variances and estimated BLUP values. ANOVA based on BLUP values revealed signi cant differences for moisture-conditions and also among genotypes. Phenotypic variation via. box-plots and histogram depicted mean phenotypic differences of traits under two moisture-regimes. Majority of the traits possessed high PCV and GCV with high heritability and GAM indicating higher trait expression and additive gene action lead to effectiveness of selection under drought/moisture stress. Grain-yield possessed a positive correlation with all the component-traits under consideration during both moisture-regimes. Selection of genotypes based on these component-traits were rewarding and seems to be better selection-criteria. Finally, we can end-up with superior-genotypes suitable for intermittent-drought conditions.

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