K+ Nutrition Exchange in the Serendipita-Arabidopsis Symbiosis: Study of the Fungal K+ Transporters Involved

There is mounting evidence that the root-colonizing endosymbiotic fungus Serendipita indica improves plant growth. The beneficial effects have been observed when plants are growing in optimal conditions or under nutritionally deficient soils (e.g., phosphate poor soil) or exposed to stressful environmental conditions such as drought or salinity. However, until now its role in the nutrition of other plant essential macronutrient, such as K+, has not been fully clarified. Here, we study the role of the fungus in the K+ nutrition of Arabidopsis thaliana plants, during growth under K+ limiting conditions. As a first step, we studied the high-affinity K+ uptake of the plant and fungus when growing separately and in symbiosis. In the search for putative fungal actors involved in K+ nutrition, we also have cloned and functionally characterized the K+ transporters of S. indica SiHAK1, SiTRK1, SiTRK2, and SiTOK1, among which it has been shown that SiHAK1 is the main transporter involved in the K+ uptake in the high affinity range of concentrations. In addition, a gene expression study of these transporters and other candidates that could participate in the K+ homeostasis of the fungus has been carried out. The results indicated that, contrary to what happens with P nutrition, S. indica seems not to improve neither the growth nor the plant K+ reserves during K+ starvation. Instead, this nutritionally restrictive condition favored fungal colonization, suggesting that the fungus obtains the greatest benefit in K+ supply during symbiosis.

Efflux pump gene expression study using RNA-seq in multidrug-resistant TB

BACKGROUND: The mechanism underlying kanamycin (KM) resistance in Mycobacterium tuberculosis is not well understood, although efflux pump proteins are thought to play a role. This study used RNA-seq data to investigate changes in the expression levels of efflux pump genes following exposure to KM.METHODS: RNA expression of efflux pump and regulatory genes following exposure to different concentrations of KM (minimum inhibitory concentration MIC 25 and MIC50) in rrs wild-type strain and rrs A1401G mutated strain were compared with the control group.RESULTS: The selected strains had differential RNA expression patterns. Among the 71 putative efflux pump and regulatory genes, 46 had significant fold changes, and 12 genes (Rv0842, Rv1146, Rv1258c, Rv1473, Rv1686c, Rv1687c, Rv1877, Rv2038c, Rv3065, Rv3197a, Rv3728 and Rv3789) that were overexpressed following exposure to KM were thought to contribute to drug resistance. Rv3197A (whiB7) showed a distinct fold change based on the concentration of KM.CONCLUSION: The significant changes in the expression of the efflux pump and regulatory genes following exposure to KM may provide insights into the identification of a new resistance mechanism.

Chemosensory Gene Expression for Two Closely Relative Species Rhodnius robustus and R. prolixus (Hemiptera, Reduviidade, Triatominae) Vectors of Chagas Disease

Two closely related species, Rhodnius prolixus and Rhodnius robustus, are the vectors of Trypanosoma cruzi, which is the causative agent of Chagas disease, but clearly exhibit clear-cut differences in their ecological behavior. R. prolixus is considered as a domiciliated species, whereas R. robustus only sporadically visits human houses in Amazonia. We performed a chemosensory gene expression study via RNA-sequencing (RNA-seq) for the two species and also included a laboratory introgressed R. robustus strain. We built an assembled transcriptome for each sample and for both sexes and compiled all in a reference transcriptome for a differential gene expression study. Because the genes specifically expressed in one condition and not expressed in another may also reflect differences in the adaptation of organisms, a comparative study of the presence/absence of transcripts was also performed for the chemosensory transcripts, namely chemosensory proteins (CSPs), odorant-binding proteins (OBPs), odorant receptors (ORs), gustatory receptors (GRs), and ionotropic receptors (IRs), as well as takeout (TO) transcripts because TO proteins have been proposed to be associated with chemosensory perception in both olfactory and taste systems. In this study, 12 novel TO transcripts from the R. prolixus genome were annotated. Among the 199 transcripts, out of interest, annotated in this study, 93% were conserved between R. prolixus and the sylvatic R. robustus. Moreover, 10 transcripts out of interest were specifically expressed in one sex and absent in another. Three chemosensory transcripts were found to be expressed only in the reared R. prolixus (CSP19, OBP9, and OR89) and only one in sylvatic R. robustus (OR22). A large set of transcripts were found to be differentially expressed (DE) between males and females (1,630), with a majority of them (83%) overexpressed in males. Between environmental conditions, 8,596 transcripts were DE, with most (67%) overexpressed in the sylvatic R. robustus samples, including 17 chemosensory transcripts (4 CSPs, 1 OBP, 5 ORs, 1 GR, 4 IR, and 2 TO), but 4 genes (OBP19, OR13, OR40, and OR79) were overexpressed in the reared samples.

Statistical Approach of Gene Set Analysis with Quantitative Trait Loci for Crop Gene Expression Studies

Genome-wide expression study is a powerful genomic technology to quantify expression dynamics of genes in a genome. In gene expression study, gene set analysis has become the first choice to gain insights into the underlying biology of diseases or stresses in plants. It also reduces the complexity of statistical analysis and enhances the explanatory power of the obtained results from the primary downstream differential expression analysis. The gene set analysis approaches are well developed in microarrays and RNA-seq gene expression data analysis. These approaches mainly focus on analyzing the gene sets with gene ontology or pathway annotation data. However, in plant biology, such methods may not establish any formal relationship between the genotypes and the phenotypes, as most of the traits are quantitative and controlled by polygenes. The existing Quantitative Trait Loci (QTL)-based gene set analysis approaches only focus on the over-representation analysis of the selected genes while ignoring their associated gene scores. Therefore, we developed an innovative statistical approach, GSQSeq, to analyze the gene sets with trait enriched QTL data. This approach considers the associated differential expression scores of genes while analyzing the gene sets. The performance of the developed method was tested on five different crop gene expression datasets obtained from real crop gene expression studies. Our analytical results indicated that the trait-specific analysis of gene sets was more robust and successful through the proposed approach than existing techniques. Further, the developed method provides a valuable platform for integrating the gene expression data with QTL data.

Eight genes are necessary and sufficient for biogenesis of Quinohemoprotein amine dehydrogenase

Quinohemoprotein amine dehydrogenase (QHNDH) containing a peptidyl quinone cofactor, cysteine tryptophylquinone, is produced in the periplasm of Gram-negative bacteria through an intricate process of post-translational modification that requires at least eight genes including those encoding three nonidentical subunits and three modifying enzymes. Our heterologous expression study has revealed that the eight genes are necessary and sufficient for the QHNDH biogenesis.