Root exudate-induced alterations in Bacillus cereus cell wall contribute to root colonization and plant growth promotion

The outcome of an interaction between plant growth promoting rhizobacteria and plants may depend on the chemical composition of root exudates (REs). We report the colonization of tobacco, and not groundnut, roots by a non-rhizospheric Bacillus cereus (MTCC 430). There was a differential alteration in the cell wall components of B. cereus in response to the REs from tobacco and groundnut. Attenuated total reflectance infrared spectroscopy revealed a split in amide I region of B. cereus cells exposed to tobacco-root exudates (TRE), compared to those exposed to groundnut-root exudates (GRE). In addition, changes in exopolysaccharides and lipid-packing were observed in B. cereus grown in TRE-amended minimal media that were not detectable in GRE-amended media. Cell-wall proteome analyses revealed upregulation of oxidative stress-related alkyl hydroperoxide reductase, and DNA-protecting protein chain (Dlp-2), in response to GRE and TRE, respectively. Metabolism-related enzymes like 2-amino-3-ketobutyrate coenzyme A ligase and 2-methylcitrate dehydratase and a 60 kDa chaperonin were up-regulated in response to TRE and GRE. In response to B. cereus, the plant roots altered their exudate-chemodiversity with respect to carbohydrates, organic acids, alkanes, and polyols. TRE-induced changes in surface components of B. cereus may contribute to successful root colonization and subsequent plant growth promotion

Simple sequence repeats showing ‘length preference’ have regulatory functions in humans

Simple sequence repeats (SSRs), simple tandem repeats (STRs) or microsatellites are short tandem repeats of 1–6 nucleotide motifs. They are twice as abundant as the protein coding DNA in the human genome and yet little is known about their functional relevance. Analysis of genomes across various taxa show that despite the instability associated with longer stretches of repeats, few SSRs with specific longer repeat lengths are enriched in the genomes indicating a positive selection. This conserved feature of length dependent enrichment hints at not only sequence but also length dependent functionality for SSRs. In the present study, we selected 23 SSRs of the human genome that show specific repeat length dependent enrichment and analysed their cis-regulatory potential using promoter modulation, boundary and barrier assays. We find that the 23 SSR sequences, which are mostly intergenic and intronic, possess distinct cis-regulatory potential. They modulate minimal promoter activity in transient luciferase assays and are capable of functioning as enhancer-blockers and barrier elements. The results of our functional assays propose cis-gene regulatory roles for these specific length enriched SSRs and opens avenues for further investigations

Accumulation of transcription factors and cell signaling-related proteins in the nucleus during citrus–Xanthomonas interaction

The nucleus is the maestro of the cell and is involved in the modulation of cell signaling during stress. We performed a comprehensive nuclear proteome analysis of Citrus sinensis during interaction with host (Xanthomonas citri pv. citri-Xcc) and non-host (Xanthomonas oryzae pv. oryzae-Xoo) pathogens. The nuclear proteome was obtained using a sequential method of organelle enrichment and determined by nano-LC–MS/MS analysis. A total of 243 proteins accumulated differentially during citrus–Xanthomonas interaction, belonging to 11 functional groups, with signaling and transcription-related proteins dominating. MADS-box transcription factors, DEAD-box RNA helicase and leucine aminopeptidase, mainly involved in jasmonic acid (JA) responses, were in high abundance during non-host interaction (Xoo). Signaling-related proteins like serine/threonine kinase, histones (H3.2, H2A), phosphoglycerate kinase, dynamin, actin and aldolase showed increased accumulation early during Xoo interaction. Our results suggest that there is a possible involvement of JA-triggered defense responses during non-host resistance, with early recognition of the non-host pathogen

Warriors at the gate that never sleep: non-host resistance in plants

The native resistance of most plant species against a wide variety of pathogens is known as non-host resistance (NHR), which confers durable protection to plant species. Only a few pathogens or parasites can successfully cause diseases. NHR is polygenic and appears to be linked with basal plant resistance, a form of elicited protection. Sensing of pathogens by plants is brought about through the recognition of invariant pathogen-associated molecular patterns (PAMPs) that trigger downstream defense signaling pathways. Race-specific resistance, (R)-gene mediated resistance, has been extensively studied and reviewed, while our knowledge of NHR has advanced only recently due to the improved access to excellent model systems. The continuum of the cell wall (CW) and the CW–plasma membrane (PM)-cytoskeleton plays a crucial role in perceiving external cues and activating defense signaling cascades during NHR. Based on the type of hypersensitive reaction (HR) triggered, NHR was classified into two types, namely type-I and type-II. Genetic analysis of Arabidopsis mutants has revealed important roles for a number of specific molecules in NHR, including the role of SNARE-complex mediated exocytosis, lipid rafts and vesicle trafficking. As might be expected, R-gene mediated resistance is found to overlap with NHR, but the extent to which the genes/pathways are common between these two forms of disease resistance is unknown. The present review focuses on the various components involved in the known mechanisms of NHR in plants with special reference to the role of CW–PM components

Proteins associated with oxidative burst and cell wall strengthening accumulate during citrus-xanthomonas non-host interaction

Citrus proteome changes at 8 and 48 h post inoculation (hpi) were analysed by both 2D gel electrophoresis and nano-LC-MS/MS proteomic approaches during interaction with Xanthomonas axonopodis pv. citri (Xac) and Xanthomonas oryzae pv. oryzae (Xoo) as host and non-host pathogens, respectively. A total of 256 proteins, 72 at 8 hpi and 184 at 48 hpi, differentially accumulated during citrus-Xanthomonas interaction. Of these, 67 and 115 proteins were specific to Xac and Xoo interaction, respectively. In addition, 64 proteins, 10 at 8 hpi and 54 at 48 hpi, variedly accumulated during both the interactions. Proteins related to photosynthesis, carbohydrate metabolism and protein synthesis were in low abundance during both the interactions resulting in reduced rate of photosynthesis. Proteins related to defence response, cell wall (CW) strengthening, lignin deposition and generation of reactive oxygen species (ROS) were in high abundance only during Xoo interaction. Whereas, during Xac interaction, proteins involved in antioxidant metabolism and CW loosening and/or elongation were in high abundance. The precise increase in abundance of these proteins during non-host interaction suggested an important role for CW fortification and ROS accumulation in non-host resistance in plants

Effect of non-rhizospheric bacterial strains on growth of crop plants.

<div><p>Seeds bacterized with respective bacterial strains (approximately1x10⁷cfu/seed, unless otherwise mentioned) were grown <i>in </i><i>vitro</i> in MS medium. After 30 days of growth, shoot height and root length were measured in centimeters, while fresh weight and dry weight of entire plant were measured in milligrams after 30 days of growth. Data represent the mean of the three independent experiments. The vertical line indicates standard error.</p> <p>(A) Effect of five different bacterial strains on growth of tobacco. Treatments included 1. <i>Bacillus </i><i>cereus</i>, 2. <i>B. subtilis</i>, 3. <i>Paenibacillus </i><i>elgii</i>, 4. <i>Stenotrophomonas </i><i>maltophilia</i>, 5. <i>Serratia </i><i>marcescens</i>, and 6. Control, (n=20). Different letters on each bar represent values that were significantly different (p_0.05). (B) Effect of <i>B. cereus</i> on growth of tobacco, tomato, pigeon pea and groundnut (n=24). Data represents percent increase over control. </p> <p>(C) Colonization of <i>B. cereus</i> on tobacco and groundnut roots. Number of days (d) for tobacco: 10, 20, 30 and 40 days of growth and for groundnut: 5, 10, 15 and 20 days of growth (n=20). Students’ t-test of each growth parameter against control for each crop was performed. ** indicate statistically significant at p<0.01, NS =indicate not significant.</p></div

Representative 2DE gels of <i>B. cereus</i> cell-wall proteome.

<p><i>B. cereus</i> grown in MM media amended with (A) tobacco root exudates or (B) groundnut root exudates. In the first dimension (IEF), 500 μg of protein was loaded on an 18 cm IPG strip with a linear gradient of pH 4-7 and 12.5% SDS-PAGE gels were used in the second dimension. Proteins were visualized by Coomasie blue staining. Arrows point towards the differentially expressed proteins. </p

Key Residues Affecting Transglycosylation Activity in Family 18 Chitinases: Insights into Donor and Acceptor Subsites

Understanding features that determine transglycosylation (TG) activity in glycoside hydrolases is important because it would allow the construction of enzymes that can catalyze controlled synthesis of oligosaccharides. To increase TG activity in two family 18 chitinases, chitinase D from Serratia proteamaculans (SpChiD) and chitinase A from Serratia marcescens (SmChiA), we have mutated residues important for stabilizing the reaction intermediate and substrate binding in both donor and acceptor sites. To help mutant design, the crystal structure of the inactive SpChiD-E153Q mutant in complex with chitobiose was determined. We identified three mutations with a beneficial effect on TG activity: Y28A (affecting the −1 subsite and the intermediate), Y222A (affecting the intermediate), and Y226W (affecting the +2 subsite). Furthermore, exchange of D151, the middle residue in the catalytically important DXDXE motif, to asparagine reduced hydrolytic activity ≤99% with a concomitant increase in apparent TG activity. The combination of mutations yielded even higher degrees of TG activity. Reactions with the best mutant, SpChiD-D151N/Y226W/Y222A, led to rapid accumulation of high levels of TG products that remained stable over time. Importantly, the introduction of analogous mutations at the same positions in SmChiA (Y163A equal to Y28A and Y390F similar to Y222A) had similar effects on TG efficiency. Thus, the combination of the decreasing hydrolytic power, subsite affinity, and stability of intermediate states provides a powerful, general strategy for creating hypertransglycosylating mutants of retaining glycoside hydrolases