Sub-QTLs and multiple intra-QTL genes are hypothesized to underpin large-effect QTLs. Known QTLs over gene families, biosynthetic pathways or certain traits represent functional gene-clusters of genes of the same gene ontology (GO). Gene-clusters containing genes of different GO have not been elaborated, except in silico as coexpressed genes within QTLs. Here we demonstrate the requirement of multiple intra-QTL genes for the full impact of QTL qDTY12.1 on rice yield under drought. Multiple evidences are presented for the need of the transcription factor ‘no apical meristem’ (OsNAM12.1) and its co-localized target genes of separate GO categories for qDTY12.1 function, raising a regulon-like model of genetic architecture. The molecular underpinnings of qDTY12.1 support its effectiveness in further improving a drought tolerant genotype and for its validity in multiple genotypes/ecosystems/environments. Resolving the combinatorial value of OsNAM12.1 with individual intra-QTL genes notwithstanding, identification and analyses of qDTY12.1has fast-tracked rice improvement towards food security.

It is widely known that numerous adaptive responses of drought-stressed plants are stimulated by chemical messengers known as phytohormones. Jasmonic acid (JA) is one such phytohormone. But there are very few reports revealing its direct implication in drought related responses or its cross-talk with other phytohormones. In this study, we compared the morpho-physiological traits and the root proteome of a wild type (WT) rice plant with its JA biosynthesis mutant coleoptile photomorphogenesis 2 (cpm2), disrupted in the allene oxide cyclase (AOC) gene, for insights into the role of JA under drought. The mutant had higher stomatal conductance, higher water use efficiency and higher shoot ABA levels under severe drought as compared to the WT. Notably, roots of cpm2 were better developed compared to the WT under both, control and drought stress conditions. Root proteome was analyzed using the Tandem Mass Tag strategy to better understand this difference at the molecular level. Expectedly, AOC was unique but notably highly abundant under drought in the WT. Identification of other differentially abundant proteins (DAPs) suggested increased energy metabolism (i.e., increased mobilization of resources) and reactive oxygen species scavenging in cpm2 under drought. Additionally, various proteins involved in secondary metabolism, cell growth and cell wall synthesis were also more abundant in cpm2 roots. Proteome-guided transcript, metabolite, and histological analyses provided further insights into the favorable adaptations and responses, most likely orchestrated by the lack of JA, in the cpm2 roots. Our results in cpm2 are discussed in the light of JA crosstalk to other phytohormones. These results together pave the path for understanding the precise role of JA during drought stress in rice.

BackgroundQuantitative reverse transcription PCR (qRT-PCR) has been routinely used to quantify gene expression level. This technique determines the expression of a target gene by comparison to an internal control gene uniformly expressed among the samples analyzed. The reproducibility and reliability of the results depend heavily on the reference genes used. To achieve successful gene expression analyses for drought tolerance studies in rice, reference gene selection should be based on consistency in expression across variables. We aimed to provide reference genes that would be consistent across different tissues, developmental stages and genotypes of rice and hence improve the quality of data in qRT-PCR analysis.FindingsTen candidate reference genes were screened from four ubiquitously expressed gene families by analyzing public microarray data sets that included profiles of multiple organs, developmental stages, and water availability status in rice. These genes were evaluated through qRT-PCR experiments with a rigorous statistical analysis to determine the best reference genes. A ubiquitin isogene showed the best gene expression stability as a single reference gene, while a 3-gene combination of another ubiquitin and two cyclophilin isogenes was the best reference gene combination. Comparison between the qRT-PCR and in-house microarray data on roots demonstrated reliability of the identified reference genes to monitor the differential expression of drought-related candidate genes.ConclusionsSpecific isogenes from among the regularly used gene families were identified for use in qRT-PCR-based analyses for gene expression in studies on drought tolerance in rice. These were stable across variables of treatment, genotype, tissue and growth stage. A single gene and/or a three gene set analysis is recommended, based on the resources available.Electronic supplementary materialThe online version of this article (doi:10.1186/s12284-016-0104-7) contains supplementary material, which is available to authorized users.

Integral subcellular and cellular functions ranging from gene expression, protein targeting and nutrient supply to cell differentiation and cell death require proteases. Plants have unique organelles such as chloroplasts composed of unique proteins that carry out the unique process of photosynthesis. Hence, along with proteases common across kingdoms, plants contain unique proteases. Improved knowledge on proteases can lead to a better understanding of plant development, differentiation and death. Because of their importance in multiple processes, plant proteases are actively studied. However, root proteases specifically are not as well studied. The associated rhizosphere, organic matter and/or inorganic matter make roots a difficult system. Yet recent research conclusively demonstrated the occurrence of endocytosis of proteins, peptides and even microbes by root cells, which, hitherto known for specialized pathogenesis or symbiosis, was unsuspected for nutrient uptake. These results reinforced the importance of root proteases in endocytosis or root exudate-mediated nutrient uptake. Rhizoplane, rhizosphere or in planta protease action on proteins, peptides and microbes generates sources of nitrogen, especially during abiotic stresses such as drought. This article highlights the recent research on root proteases for nitrogen uptake and the connection of the two to drought-tolerance mechanisms. Drought-induced proteases in rice roots, as known from rice expression databases, are discussed for future research on certain M50, Deg, FtsH, AMSH and deubiquitination proteases. The recent emphasis on linking drought and plant hydraulics to nutrient metabolism is illustrated and connected to the value of a systematic study of root proteases in crop improvement.

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There is a widespread consensus that drought will mostly affect present and future agriculture negatively. Generating drought-tolerant crops is thus a high priority. However complicated the underlying genetic and regulatory networks for differences in plant performance under stress are, they would be reflected in straightforward differences in primary metabolites. This is because primary metabolites such as amino acids and sugars form the building blocks of all pathways and processes for growth, development, reproduction, and environmental responses. Comparison of such differences was undertaken between the parental line and a near-isogenic line of qDTY12.1, a QTL for rice yield under drought. The comparison was informative regarding the effect of the QTL in three genetic backgrounds: donor, recipient, and improved recipient, thus illustrating the gene × gene (G × G) interactions. Such a comparison when extended to well-watered and drought conditions illustrated the gene × environment (G × E) interactions. Assessment of such G × G and G × E responses in roots, flag leaves, and spikelets added a yet more informative dimension of tissue-specific responses to drought, mediated by qDTY12.1. Data on variation in primary metabolites subjected to ANOVA, Tukey’s test, Welch’s t test, and PCA underscored the importance of the roots and demonstrated concordance between variation in metabolites and morpho-physiological responses to drought. Results suggested that for gainful insights into rice yield under drought, rather than vegetative stage drought tolerance, multiple tissues and genotypes must be assessed at the reproductive stage to avoid misleading conclusions about using particular metabolites or related genes and proteins as candidates or markers for drought tolerance.Electronic supplementary materialThe online version of this article (doi:10.1007/s11032-015-0322-5) contains supplementary material, which is available to authorized users.

Due to the increasing popularity of Jatropha curcas as a feedstock for biodiesel, generating non-toxic and high yielding varieties of the plant requires genotypic characterization towards identifying breeding lines. There is little information on the phylogenetic relationships between its global accessions and species. Assessing genetic variation by RAPD, AFLP and combinatorial tubulin based polymorphism (cTBP) in 38 J. curcas accessions from 13 countries on 3 continents revealed narrow genetic diversity. However, 6 different species of Jatropha from India exhibited pronounced genetic diversity indicating possibilities of improving J. curcas by interspecific breeding. The relatively unexplored cTBP approach we used was a highly efficient and cost effective genotyping tool. Using such tools towards breeding J. curcas for low phorbol ester (PE) content is highly desirable because of the co-carcinogenic nature of the PEs present in all the commercially relevant parts such as seeds, seed-cake and biodiesel. We report initial success in obtaining interspecific F1 and back cross (BC1) plants with low PE and improved agronomic traits.Further efforts will lead to generating varieties with targeted traits. Despite the limited genetic diversity within J. curcas accessions, appreciable variability was noticed in important phenotypic, physiological and biochemical traits such as seed size, water use efficiency and seed oil content respectively. This implicates fundamental epigenetic regulatory mechanisms and posits J. curcas as a unique system to study them.

Like other crop species, barley, the fourth most important crop worldwide, suffers from the genetic bottleneck effect, where further improvements in performance through classical breeding methods become difficult. Therefore, indirect selection methods are of great interest. Here, genomic prediction (GP) based on 33,005 SNP markers and, alternatively, metabolic prediction (MP) based on 128 metabolites with sampling at two different time points in one year, were applied to predict multi-year agronomic traits in the nested association mapping (NAM) population HEB-25. We found prediction abilities of up to 0.93 for plant height with SNP markers and of up to 0.61 for flowering time with metabolites. Interestingly, prediction abilities in GP increased after reducing the number of incorporated SNP markers. The estimated effects of GP and MP were highly concordant, indicating MP as an interesting alternative to GP, being able to reflect a stable genotype-specific metabolite profile. In MP, sampling at an early developmental stage outperformed sampling at a later stage. The results confirm the value of GP for future breeding. With MP, an interesting alternative was also applied successfully. However, based on our results, usage of MP alone cannot be recommended in barley. Nevertheless, MP can assist in unravelling physiological pathways for the expression of agronomically important traits.