Dissecting the nutrient partitioning mechanism in rice grain using spatially resolved gene expression profiling.

Rice, a staple food worldwide, contains varying amounts of nutrients in different grain tissues. The underlying molecular mechanism of such distinct nutrient partitioning remains poorly investigated. Here, an optimized rapid laser capture microdissection (LCM) approach was used to individually collect pericarp, aleurone, embryo and endosperm from grains 10 days after fertilization. Subsequent RNA-Seq analysis in these tissues identified 7760 differentially expressed genes. Analysis of promoter sequences of tissue-specific genes identified many known and novel cis-elements important for grain filling and seed development. Using the identified differentially expressed genes, comprehensive spatial gene expression pathways were built for accumulation of starch, proteins, lipids, and iron. The extensive transcriptomic analysis provided novel insights about nutrient partitioning mechanisms; for example, it revealed a gradient in seed storage protein accumulation across the four tissue types analysed. The analysis also revealed that the partitioning of various minerals, such as iron, is most likely regulated through transcriptional control of their transporters. We present the extensive analysis from this study as an interactive online tool that provides a much-needed resource for future functional genomics studies aimed to improve grain quality and seed development.

Identification and molecular characterization of rice bran-specific lipases.

KEY MESSAGE: Among the 113 lipases present in rice genome, bran and endosperm-specific lipases were identified and lipase activity for one of the selected lipase gene is demonstrated in yeast. Rice bran is nutritionally superior than endosperm as it has major reservoirs of various minerals, vitamins, essential mineral oils and other bioactive compounds, however it is often under-utilized as a food product due to bran instability after milling. Various hydrolytic enzymes, such as lipases, present in bran causes degradation of the lipids present and are responsible for the bran instability. Here, in this study, we have systematically analyzed the 113 lipase genes present in rice genome, and identified 21 seed-specific lipases. By analyzing the expression of these genes in different seed tissues during seed development, we have identified three bran-specific and three endosperm-specific lipases, and one lipase which expresses in both bran and endosperm tissues. Further analysis of these genes during seed maturation and seed germination revealed that their expression increases during seed maturation and decreases during seed germination. Finally, we have shown the lipase activity for one of the selected genes, LOC_Os05g30900, in heterologous system yeast. The bran-specific lipases identified in this study would be very valuable for engineering designer rice varieties having increased bran stability in post-milling.

Mapping of Genomic Regions for Biochemical and Physiological Parameters Contributing Towards Drought Tolerance in Horsegram (Macrotyloma uniflorum (Lam.) Verdc.).

Horsegram (Macrotyloma uniflorum (Lam.) Verdc.) is a drought hardy legume which can be grown in varied soil and temperature regimes. Though it has numerous, nutritive and medicinal benefits, it still lags behind other legumes in terms of genomic resources and genetic improvement. This crop is mostly cultivated on marginal and drought-prone area; thus, genetics of drought stress tolerance can be understood by studying the various drought parameters. To get insight, quantitative trait loci for drought-tolerant traits were identified using an intraspecific mapping population of 162 F8 recombinant inbred lines derived from a cross between HPKM249 and HPK4. The linkage map already developed was used along with the phenotypic data for biochemical and physiological parameters to identify genomic regions which are linked to drought tolerance. In the study, a total of seven QTLs were identified for ten different drought-related traits. One QTL for malondialdehyde content on linkage group 2, two QTLs for root length on linkage groups 3 and 9, one QTL each for proline and chlorophyll content under drought stress on linkage group 4, and one QTL each for root dry weight and root fresh weight on linkage group 5 were identified using composite interval mapping. The identified QTLs will be utilized in marker-assisted breeding and increase our understanding on the physiology of drought stress tolerance.

Identification of QTLs Linked to Phenological and Morphological Traits in RILs Population of Horsegram (Macrotyloma uniflorum).

Horsegram [Macrotyloma uniflorum (Lam.) Verdc.] is an important legume but understudied in terms of its genetic improvement. Genetic information on various phenological and morphological traits may help in the utilization of new genes for breeding in horsegram and thus affect agronomic practices and crop yield. A total of 162 recombinant inbred lines derived from intraspecific crosses between HPKM249 × HPK4 was used to construct a genetic linkage map and to identify quantitative trait loci (QTLs) associated with phenological and morphological traits. Of the total 2011 molecular markers, which were screened on parental lines for polymorphism survey, 493 markers were found to be polymorphic and used for genotyping of recombinant inbred line population. Out of 493 polymorphic markers, 295 were mapped on ten linkage groups at LOD 3.5 spanning a total distance of 1,541.7 cM with an average distance between markers of 5.20 cM. Phenotypic data of two years at two different locations were used to identify QTLs by composite interval mapping A total of four QTLs (LOD ≥2.5) for phenological traits (days to 50% flowering, reproductive period and days to maturity) and seven QTLs (LOD ≥2.5) for morphological traits (plant height, primary branches and secondary branches) were detected across different environments. The phenotypic variation explained by QTLs ranged from 6.36 to 47.53%. The present study will help to augment scanty genomic information in this orphan crop that would provide genomics tools to breeders for its genetic enhancement through molecular-assisted selection.

Identification of genomic regions associated with early plant vigour in lentil (Lens culinaris).

Lentil is one of the most important food legume species, however its genetic and genomic resources remained largely uncharacterized and unexploited. In the past few years, a number of genetic maps have been constructed and marker resources have been developed in lentil. These resources could be exploited for understanding the extent and distribution of genetic variation in genus Lens and also for developing saturated and consensus genetic maps suitable for quantitative trait loci (QTL) mapping and marker-assisted selection. The present study aims to enrich polymerase chain reaction-based linkage map of F10 recombinant inbred lines (RILs) population of 94 individuals derived from cross WA8649090 9 Precoz and identification of QTLs linked to early plant vigour traits. Of the 268 polymorphic markers (93 simple sequence repeats (SSR), three inter-simple sequence repeats (ISSRs) and 172 random amplified polymorphic DNA (RAPDs)), 265 (90 SSRs, three ISSRs and 172 RAPDs) were mapped on seven linkage groups, varying in length between 25.6 and 210.3 cM, coverage of 809.4 cM with an average marker spacing of 3.05 cM. The study also reported assigning of 24 new cross-genera SSRs of Trifolium pratense on the present linkage map. The RILs along with the parents were screened for shoot length, root length, seedling length, dry weight, number of leaves and number of branches based on two replications under polyhouse conditions. A QTLhotspot consisting of six QTLs for shoot length (cm), root length (cm) and seedling length (cm) was observed between a map distances of56.61 and 86.81 cM on LG1.