Re-addressing the biosafety issues of plant growth promoting rhizobacteria.

To promote agronomic sustainability, extensive research is being carried out globally, investigating biofertilizer development. Recently, it has been realized that some microorganisms used as biofertilizers behave as opportunistic pathogens and belong to the biosafety level 2 (BSL-2) classification. This poses serious risk to the environmental and human health. Evidence presented in various scientific forums is increasingly favoring the merits of using BSL-2 microorganisms as biofertilizers. In this review, we emphasize that partial characterization based on traditional microbiological approaches and small subunit rRNA gene sequences/conserved regions are insufficient for the characterization of biofertilizer strains. It is advised herein, that research and industrial laboratories developing biofertilizers for commercialization or environmental release must characterize microorganisms of interest using a multilateral polyphasic approach of microbial systematics. This will determine their risk group and biosafety characteristics before proceeding with formulation development and environmental application. It has also been suggested that microorganisms belonging to risk-group-1 and BSL-1 category should be used for formulation development and for field scale applications. While, BSL-2 microorganisms should be restricted for research using containment practices compliant with strict regulations.

Application of essential oils as a natural and alternate method for inhibiting and inducing the sprouting of potato tubers.

Use of harmful chemicals and expensive maintenance of cold-storage conditions for controlling sprouting are among the major problems in potato storage. Here, 20 essential oils (EOs) were tested for their sprouting-inhibiting and sprouting-inducing activities. Overall, treatments of lemon grass (LG) and clove (CL) oils could induce sprouting whereas palmarosa (PR) and ajwain (AZ) oils could inhibit sprouting of potato tubers at normal-room-temperature (25 ±â€¯2 °C) storage. Selected-EOs treatments affected sprouting by modulation of accumulation of reducing sugars, ethylene, and expression of genes involved in tuber-sprouting such as ARF, ARP, AIP and ERF. Surprisingly, 7-days AZ-treatments could inhibit sprouting for 30-days which was mediated via damaging apical meristem. However, LG- and CL-treated tubers could produce enhanced potato yield as well. Present work clearly demonstrates that selected-EOs can be used as a promising eco-friendly approach for inducing/inhibiting sprouting of potato tubers during potato storage and those enhancing sprouting can be used for enhancing productivity.

An ex vivo evaluation of effect of dentin pretreatment with various agents for varying time intervals on the shear bond strength of resin.

BACKGROUND: This article highlights the contribution of collagen structure/stability to the bond strength. We hypothesize that induction of cross-linking in dentin collagen fibrils improves dentin collagen stability and thus bond strength with composite also improves. AIMS: The aim of the study was to evaluate the effect of collagen cross-linking agents on the shear bond strength of composite resins. SUBJECTS AND METHODS: One hundred human permanent teeth were randomly divided into three groups: Group I (n = 20) - no dentin pretreatment done, Group II - dentin pretreatment with 10% sodium ascorbate for 5 min (IIa) and 10 min (IIb), and Group III - dentin pretreatment with 6.5% proanthocyanidin (PA) for 5 min (IIIa) and 10 min (IIIb). A composite resin was bonded on prepared surfaces and cured. Thermocycling was done, and shear bond strength of the prepared samples was tested using a universal testing machine. RESULTS: Values of Group I (control) were lowest whereas that of Group II (sodium ascorbate) were highest. The following order of shear bond strength was observed: IIb > IIa > IIIb ~ IIIa > I. Results for sodium ascorbate were found to be time dependent, whereas for PA, differences were nonsignificant. CONCLUSIONS: Treatment of dentinal surfaces with collagen cross-linking agent increases the shear bond strengths.

Draft genome sequence of a biosurfactant producing, Bacillus aquimaris strain SAMM MCC 3014 isolated from Indian Arabian coastline sea water.

Bacillus aquimaris strain SAMM, a biosurfactant producing moderately halophilic marine bacterium was isolated from Indian Arabian coastline sea water. The strain was found to tolerate up to 2.7 M NaCl indicating osmotic stress sustainable physiological systems. We report here the draft genome sequence of B. aquimaris SAMM, as a candidate bacterium for bioactive surfactant producer. The whole genome sequence with 161 scaffolds, 4,414,932 bp and 44.8% of G+C content for SAMM was obtained using Illumina MiSeq sequencing technology. Annotation was added by the PGAP and RAST prokaryotic genome annotation service and shown 4,247 coding sequences, 123 RNAs genes, classified in 453 subsystems. Several genes encoding enzymatic activities against the high molecular weight polysaccharides, osmotic stress response and siderophore synthesis of potential biotechnological importance were identified in the genome.

Plant growth-promoting rhizobacteria enhance wheat salt and drought stress tolerance by altering endogenous phytohormone levels and TaCTR1/TaDREB2 expression.

Abiotic stresses such as salt and drought represent adverse environmental conditions that significantly damage plant growth and agricultural productivity. In this study, the mechanism of the plant growth-promoting rhizo-bacteria (PGPR)-stimulated tolerance against abiotic stresses has been explored. Results suggest that PGPR strains, Arthrobacter protophormiae (SA3) and Dietzia natronolimnaea (STR1), can facilitate salt stress tolerance in wheat crop, while Bacillus subtilis (LDR2) can provide tolerance against drought stress in wheat. These PGPR strains enhance photosynthetic efficiency under salt and drought stress conditions. Moreover, all three PGPR strains increase indole-3-acetic acid (IAA) content of wheat under salt and drought stress conditions. The SA3 and LDR2 inoculations counteracted the increase of abscisic acid (ABA) and 1-aminocyclopropane-1-carboxylate (ACC) under both salt and drought stress conditions, whereas STR1 had no significant impact on the ABA and ACC content. The impact of PGPR inoculations on these physiological parameters were further confirmed by gene expression analysis as we observed enhanced levels of the TaCTR1 gene in SA3-, STR1- and LDR2-treated wheat seedlings as compared to uninoculated drought and salt stressed plants. PGPR inoculations enhanced expression of TaDREB2 gene encoding for a transcription factor, which has been shown to be important for improving the tolerance of plants to abiotic stress conditions. Our study suggest that PGPR confer abiotic stress tolerance in wheat by enhancing IAA content, reducing ABA/ACC content, modulating expression of a regulatory component (CTR1) of ethylene signaling pathway and DREB2 transcription factor.

Optimization of Nutrients and Culture Conditions for Alkaline Protease Production Using Two Endophytic Micrococci: Micrococcus aloeverae and Micrococcus yunnanensis.

An endophytic species of Micrococcus was isolated from Aloe vera leaf (syn. Aloe barbadensis) and screened for protease production with five other species of Micrococcus. Data indicated that endophytic Micrococcus aloeverae AE-6 MCC 2184T and Micrococcus yunnanensis DSM 21948T showed efficient protease production potential and secreted active protease at high salt (10%), temperature (40 °C) and in wide range of pH 8-10. Unlike M. yunnanensis DSM 21948T, protease production by M. aloeverae AE-6 MCC 2184T was stringently controlled by pH. Protease induction study using different group of peptides, peptide carbohydrates and peptide macronutrient combinations showed variable response with both the organisms. Result indicated that the amount of protease was not directly related to cell biomass but it depends on nature of inducible peptides. In this study we also developed a modified agar-well assay for semi-quantitative data from large number of replicates.

Plant growth promoting rhizobacteria Dietzia natronolimnaea modulates the expression of stress responsive genes providing protection of wheat from salinity stress.

Plant growth promoting rhizobacteria (PGPR) hold promising future for sustainable agriculture. Here, we demonstrate a carotenoid producing halotolerant PGPR Dietzia natronolimnaea STR1 protecting wheat plants from salt stress by modulating the transcriptional machinery responsible for salinity tolerance in plants. The expression studies confirmed the involvement of ABA-signalling cascade, as TaABARE and TaOPR1 were upregulated in PGPR inoculated plants leading to induction of TaMYB and TaWRKY expression followed by stimulation of expression of a plethora of stress related genes. Enhanced expression of TaST, a salt stress-induced gene, associated with promoting salinity tolerance was observed in PGPR inoculated plants in comparison to uninoculated control plants. Expression of SOS pathway related genes (SOS1 and SOS4) was modulated in PGPR-applied wheat shoots and root systems. Tissue-specific responses of ion transporters TaNHX1, TaHAK, and TaHKT1, were observed in PGPR-inoculated plants. The enhanced gene expression of various antioxidant enzymes such as APX, MnSOD, CAT, POD, GPX and GR and higher proline content in PGPR-inoculated wheat plants contributed to increased tolerance to salinity stress. Overall, these results indicate that halotolerant PGPR-mediated salinity tolerance is a complex phenomenon that involves modulation of ABA-signalling, SOS pathway, ion transporters and antioxidant machinery.

Halotolerant PGPRs Prevent Major Shifts in Indigenous Microbial Community Structure Under Salinity Stress.

The resilience of soil microbial populations and processes to environmental perturbation is of increasing interest as alteration in rhizosphere microbial community dynamics impacts the combined functions of plant-microbe interactions. The present study was conducted to investigate the effect of inoculation with halotolerant rhizobacteria Bacillus pumilus (STR2), Halomonas desiderata (STR8), and Exiguobacterium oxidotolerans (STR36) on the indigenous root-associated microbial (bacterial and fungal) communities in maize under non-saline and salinity stress. Plants inoculated with halotolerant rhizobacteria recorded improved growth as illustrated by significantly higher shoot and root dry weight and elongation in comparison to un-inoculated control plants under both non-saline and saline conditions. Additive main effect and multiplicative interaction ordination analysis revealed that plant growth promoting rhizobacteria (PGPR) inoculations as well as salinity are major drivers of microbial community shift in maize rhizosphere. Salinity negatively impacts microbial community as analysed through diversity indices; among the PGPR-inoculated plants, STR2-inoculated plants recorded higher values of diversity indices. As observed in the terminal-restriction fragment length polymorphism analysis, the inoculation of halotolerant rhizobacteria prevents major shift of the microbial community structure, thus enhancing the resilience capacity of the microbial communities.

ACC deaminase-containing Arthrobacter protophormiae induces NaCl stress tolerance through reduced ACC oxidase activity and ethylene production resulting in improved nodulation and mycorrhization in Pisum sativum.

Induction of stress ethylene production in the plant system is one of the consequences of salt stress which apart from being toxic to the plant also inhibits mycorrhizal colonization and rhizobial nodulation by oxidative damage. Tolerance to salinity in pea plants was assessed by reducing stress ethylene levels through ACC deaminase-containing rhizobacteria Arthrobacter protophormiae (SA3) and promoting plant growth through improved colonization of beneficial microbes like Rhizobium leguminosarum (R) and Glomus mosseae (G). The experiment comprised of treatments with combinations of SA3, G, and R under varying levels of salinity. The drop in plant biomass associated with salinity stress was significantly lesser in SA3 treated plants compared to non-treated plants. The triple interaction of SA3+G+R performed synergistically to induce protective mechanism against salt stress and showed a new perspective of plant-microorganism interaction. This tripartite collaboration increased plant weight by 53%, reduced proline content, lipid peroxidation and increased pigment content under 200 mM salt condition. We detected that decreased ACC oxidase (ACO) activity induced by SA3 and reduced ACC synthase (ACS) activity in AMF (an observation not reported earlier as per our knowledge) inoculated plants simultaneously reduced the ACC content by 60% (responsible for generation of stress ethylene) in SA3+G+R treated plants as compared to uninoculated control plants under 200 mM salt treatment. The results indicated that ACC deaminase-containing SA3 brought a putative protection mechanism (decrease in ACC content) under salt stress, apart from alleviating ethylene-induced damage, by enhancing nodulation and AMF colonization in the plants resulting in improved nutrient uptake and plant growth.

The greater effectiveness of Glomus mosseae and Glomus intraradices in improving productivity, oil content and tolerance of salt-stressed menthol mint (Mentha arvensis).

BACKGROUND: Mentha arvensis is cultivated in large parts of the world for its menthol-rich essential oil. The study investigates the potential of four mycorrhizal fungi, viz. Glomus mosseae (Gm), Glomus aggregatum (Ga), Glomus fasciculatum (Gf) and Glomus intraradices (Gi) in alleviating NaCl-induced salt stress in Mentha arvensis cv. Kosi and establishes the specificity of interaction between different mycorrhizal species and their effectiveness in mitigating salt stress in Mentha arvensis. Mycorrhizal and non-mycorrhizal Mentha plants were subjected to NaCl-induced salinity. RESULTS: Among the four Glomus species, Gm and Gi reduced salt-induced herb yield losses: a loss of 27.53% and 25.58% respectively under salt stress in comparison to 51.00% in non-mycorrhizal M. arvensis salt-stressed plants. Gm- and Gi-inoculated plants also recorded higher leaf:stem ratio, oil content, and oil yield and menthol concentration in essential oil under both saline and non-saline conditions. CONCLUSION: Better performance in terms of herb yield, and oil content and yield was observed in Gi- and Gm-inoculated M. arvensis plants, suggesting the capability of Gi and Gm in protecting plants from the detrimental effects of salt stress; beneficial effects of arbuscular mycorrhizal fungi, however, may vary with host and environment.

Exiguobacterium oxidotolerans, a halotolerant plant growth promoting rhizobacteria, improves yield and content of secondary metabolites in Bacopa monnieri (L.) Pennell under primary and secondary salt stress.

Brahmi (Bacopa monnieri), an integral component of Indian Ayurvedic medicine system, is facing a threat of extinction owing to the depletion of its natural populations. The present study investigates the prospective of exploitation of halotolerant plant growth promoting rhizobacteria (PGPR) in utilising the salt stressed soils for cultivation of B. monnieri. The effects of two salt tolerant PGPR, Bacillus pumilus (STR2) and Exiguobacterium oxidotolerans (STR36) on the growth and content of bacoside-A, an important pharmaceutical compound in B. monnieri, were investigated under primary and secondary salinity conditions. The herb yields of un-inoculated plants decreased by 48 % under secondary salinization and 60 % under primary salinization than the non salinised plants. Among the rhizobacteria treated plants, E. oxidotolerans recorded 109 and 138 %, higher herb yield than non-inoculated plants subjected to primary and secondary salinity respectively. E. oxidotolerans inoculated plants recorded 36 and 76 % higher bacoside-A content under primary and secondary salinity respectively. Higher levels of proline content and considerably lower levels of lipid peroxidation were noticed when the plants were inoculated with PGPR under all salinity regimes. From the results of this investigation, it can be concluded that, the treatments with salt tolerant PGPR can be a useful strategy in the enhancement of biomass yield and saponin contents in B. monnieri, as besides being an eco-friendly approach; it can also be instrumental in cultivation of B. monnieri in salt stressed environments.

1-Aminocyclopropane-1-carboxylic acid (ACC) deaminase-containing rhizobacteria protect Ocimum sanctum plants during waterlogging stress via reduced ethylene generation.

Ocimum sanctum grown as rain-fed crop, is known to be poorly adapted to waterlogged conditions. Many a times the crop suffers extreme damages because of anoxia and excessive ethylene generation due to waterlogging conditions present under heavy rain. The usefulness of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-containing plant growth promoting rhizobacteria was investigated under waterlogging stress. The comparison of herb yield and stress induced biochemical changes of waterlogged and non-waterlogged plants with and without ACC deaminase-containing microbiological treatments were monitored in this study. Ten plant growth promoting rhizobacteria strains containing ACC-deaminase were isolated and characterized. Four selected isolates Fd2 (Achromobacter xylosoxidans), Bac5 (Serratia ureilytica), Oci9 (Herbaspirillum seropedicae) and Oci13 (Ochrobactrum rhizosphaerae) had the potential to protect Ocimum plants from flood induced damage under waterlogged glass house conditions. Pot experiments were conducted to evaluate the potential of these ACC deaminase-containing selected strains for reducing the yield losses caused by waterlogging conditions. Bacterial treatments protected plants from waterlogging induced detrimental changes like stress ethylene production, reduced chlorophyll concentration, higher lipid peroxidation, proline concentration and reduced foliar nutrient uptake. Fd2 (A. xylosoxidans) induced maximum waterlogging tolerance as treated waterlogged plants recorded maximum growth and herb yield (46.5% higher than uninoculated waterlogged plants) with minimum stress ethylene levels (53% lower ACC concentration as compared to waterlogged plants without bacterial inoculation) whereas under normal non-waterlogged conditions O. rhizosphaerae was most effective in plant growth promotion.