Endophytes Modulate Plant Genes: Present Status and Future Perspectives.

Interactions among endophytes and plants are widespread and can vary from neutral or positive or negative. Plants are continually in a functionally dynamic state due to interactions with diverse endophytic microorganisms, which produce various metabolic substances. Through quorum sensing, these substances not only help endophytes to outcompete other host-associated pathogens or microbes but also allow them to overcome the plant immune system. Manifold interactions between endophytic microbiota cause a reflective impact on the host plant functioning and the development of 'endobiomes,' by synthesizing chemicals that fill the gap between host and endophytes. Despite the advances in the field, specific mechanisms for the endophytes' precise methods to modulate plant genome and their effects on host plants remain poorly understood. Deeper genomic exploration can provide a locked away understanding of the competencies of endophytes and their conceivable function in host growth and health. Endophytes also can modify host metabolites, which could manipulate plants' growth, adaptation, and proliferation, and can be a more exciting and puzzling topic that must be properly investigated. The consequence of the interaction of endophytes on the host genome was analyzed as it can help unravel the gray areas of endophytes about which very little or no knowledge exists. This review discusses the recent advances in understanding the future challenges in the emerging research investigating how endosymbionts affect the host's metabolism and gene expression as an effective strategy for imparting resistance to biotic and abiotic challenges.

Plant Abiotic and Biotic Stress Alleviation: From an Endophytic Microbial Perspective.

Plant abiotic and biotic stresses can change plant-pest synergism by augmenting host plant vulnerability to pests and lessening competitive capability with weed plants. Climate change, such as a shift in precipitation, intensifies the damaging effects of stresses, undesirably impacting plant growth and survival. However, we have yet to reach a clear answer as the outcome usually depends on complex interactions and agro-climatic conditions. To alleviate plant stresses, more in-depth work is required to elucidate the underlying mechanisms and exploit thereof. In this review, we have confined ourselves to the domain of the role played by endophytic microorganisms to alleviate plant stress. In contrast, some biotic stresses may alter plant response to abiotic stress factors. Hence, methodical analyses are indispensable for understanding the effect of abiotic and biotic stress conditions on crop development and agronomic production. Endophytic microbes have drawn interest owing to their plant growth stimulating attributes and valuable performances related to plant responses under abiotic and biotic stress environments. Endophytes produce secondary metabolites to defend the host plant under stressful climatic conditions and against phytopathogens. Understanding plant resilience mechanisms will assist in the commercialized biotechnological development of endophytes in crop improvement. There is still much scope to explore factors and elucidate mechanisms that result in unquestionably recognized beneficial effects of endophytes. This review article bridges the gap mentioned and focuses on the role played by endophytes in plant development and their stimulating diverse mechanisms for tolerating diverse abiotic and biotic stresses in the host.

An insight into the mechanism of antifungal activity of biogenic nanoparticles than their chemical counterparts.

Herein, we describe the enhanced antifungal activity of silver nanoparticles biosynthesized by cell free filtrate of Trichoderma viride (MTCC 5661) in comparison to chemically synthesized silver nanoparticles (CSNP) of similar shape and size. Biosynthesized silver nanoparticles (BSNP) enhanced the reduction in dry weight by 20 and 48.8% of fungal pathogens Fusarium oxysporum and Alternaria brassicicola respectively in comparison to their chemical counterparts (CSNP). Nitroblue tetrazolium and Propidium iodide staining demonstrated the higher generation of superoxide radicals lead to higher death in BSNP treated fungus in comparison to CSNP. Scanning electron microscopy of A. brassicicola revealed the osmotic imbalance and membrane disintegrity to be major cause for fungal cell death after treatment with BSNP. To gain an insight into the mechanistic aspect of enhanced fungal cell death after treatment of BSNP in comparison to CSNP, stress responses and real time PCR analysis was carried out with A. brassicicola. It revealed that generation of ROS, downregulation of antioxidant machinery and oxidative enzymes, disruption of osmotic balance and cellular integrity, and loss of virulence are the mechanisms employed by BSNP which establishes them as superior antifungal agent than their chemical counterparts. With increasing drug resistance and ubiquitous presence of fungal pathogens in plant kingdom, BSNP bears the candidature for new generation of antifungal agent.

Arsenic tolerant Trichoderma sp. reduces arsenic induced stress in chickpea (Cicer arietinum).

Toxic metalloids including arsenic (As) can neither be eliminated nor destroyed from environment; however, they can be converted from toxic to less/non-toxic forms. The form of As species and their concentration determines its toxicity in plants. Therefore, the microbe mediated biotransformation of As is crucial for its plant uptake and toxicity. In the present study the role of As tolerant Trichoderma in modulating As toxicity in chickpea plants was explored. Chickpea plants grown in arsenate spiked soil under green house conditions were inoculated with two plant growth promoting Trichoderma strains, M-35 (As tolerant) and PPLF-28 (As sensitive). Total As concentration in chickpea tissue was comparable in both the Trichoderma treatments, however, differences in levels of organic and inorganic As (iAs) species were observed. The shift in iAs to organic As species ratio in tolerant Trichoderma treatment correlated with enhanced plant growth and nutrient content. Arsenic stress amelioration in tolerant Trichoderma treatment was also evident through rhizospheric microbial community and anatomical studies of the stem morphology. Down regulation of abiotic stress responsive genes (MIPS, PGIP, CGG) in tolerant Trichoderma + As treatment as compared to As alone and sensitive Trichoderma + As treatment also revealed that tolerant strain enhanced the plant's potential to cope with As stress as compared to sensitive one. Considering the bioremediation and plant growth promotion potential, the tolerant Trichoderma may appear promising for its utilization in As affected fields for enhancing agricultural productivity.

Enhanced Cellular Internalization: A Bactericidal Mechanism More Relative to Biogenic Nanoparticles than Chemical Counterparts.

Biogenic synthesis of silver nanoparticles for enhanced antimicrobial activity has gained a lot of momentum making it an urgent need to search for a suitable biocandidate which could be utilized for efficient capping and shaping of silver nanoparticles with enhanced bactericidal activity utilizing its secondary metabolites. Current work illustrates the enhancement of antimicrobial efficacy of silver nanoparticles by reducing and modifying their surface with antimicrobial metabolites of cell free filtrate of Trichoderma viride (MTCC 5661) in comparison to citrate stabilized silver nanoparticles. Nanoparticles were characterized by visual observations, UV-visible spectroscopy, zetasizer, and transmission electron microscopy (TEM). Synthesized particles were monodispersed, spherical in shape and 10-20 nm in size. Presence of metabolites on surface of biosynthesized silver nanoparticles was observed by gas chromatography-mass spectroscopy (GC-MS), energy dispersive X-ray analysis (EDAX), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The antimicrobial activity of both silver nanoparticles was tested against Shigella sonnei, Pseudomonas aeruginosa (Gram-negative) and Staphylococcus aureus (Gram-positive) by growth inhibition curve analysis and colony formation unit assay. Further, it was noted that internalization of biosynthesized nanoparticles inside the bacterial cell was much higher as compared to citrate stabilized particles which in turn lead to higher production of reactive oxygen species. Increase in oxidative stress caused severe damage to bacterial membrane enhancing further uptake of particles and revoking other pathways for bacterial disintegration resulting in complete and rapid death of pathogens as evidenced by fluorescein diacetate/propidium iodide dual staining and TEM. Thus, study reveals that biologically synthesized silver nanoarchitecture coated with antimicrobial metabolites of T. viride was more potent than their chemical counterpart in killing of pathogenic bacteria.

Elucidation of Complex Nature of PEG Induced Drought-Stress Response in Rice Root Using Comparative Proteomics Approach.

Along with many adaptive strategies, dynamic changes in protein abundance seem to be the common strategy to cope up with abiotic stresses which can be best explored through proteomics. Understanding of drought response is the key to decipher regulatory mechanism of better adaptation. Rice (Oryza sativa L.) proteome represents a phenomenal source of proteins that govern traits of agronomic importance, such as drought tolerance. In this study, a comparison of root cytoplasmic proteome was done for a drought tolerant rice (Heena) cultivar in PEG induced drought conditions. A total of 510 protein spots were observed by PDQuest analysis and 125 differentially regulated spots were subjected for MALDI-TOF MS-MS analysis out of which 102 protein spots identified which further led to identification of 78 proteins with a significant score. These 78 differentially expressed proteins appeared to be involved in different biological pathways. The largest percentage of identified proteins was involved in bioenergy and metabolism (29%) and mainly consists of malate dehydrogenase, succinyl-CoA, putative acetyl-CoA synthetase, and pyruvate dehydrogenase etc. This was followed by proteins related to cell defense and rescue (22%) such as monodehydroascorbate reductase and stress-induced protein sti1, then by protein biogenesis and storage class (21%) e.g. putative thiamine biosynthesis protein, putative beta-alanine synthase, and cysteine synthase. Further, cell signaling (9%) proteins like actin and prolyl endopeptidase, and proteins with miscellaneous function (19%) like Sgt1 and some hypothetical proteins were also represented a large contribution toward drought regulatory mechanism in rice. We propose that protein biogenesis, cell defense, and superior homeostasis may render better drought-adaptation. These findings might expedite the functional determination of the drought-responsive proteins and their prioritization as potential molecular targets for perfect adaptation.

A Comprehensive Characterization of Simple Sequence Repeats in the Sequenced Trichoderma Genomes Provides Valuable Resources for Marker Development.

Members of genus Trichoderma are known worldwide for mycoparasitism. To gain a better insight into the organization and evolution of their genomes, we used an in silico approach to compare the occurrence, relative abundance and density of SSRs in Trichoderma atroviride, T. harzianum, T. reesei, and T. virens. Our analysis revealed that in all the four genome sequences studied, the occurrence, relative abundance, and density of microsatellites varied and was not influenced by genome sizes. The relative abundance and density of SSRs positively correlated with the G + C content of their genomes. The maximum frequency of SSRs was observed in the smallest genome of T. reesei whereas it was least in second smallest genome of T. atroviride. Among different classes of repeats, the tri-nucleotide repeats were abundant in all the genomes and accounts for ∼38%, whereas hexa-nuceotide repeats were the least (∼10.2%). Further evaluation of the conservation of motifs in the transcript sequences shows a 49.5% conservation among all the motifs. In order to study polymorphism in Trichoderma isolates, 12 polymorphic SSR markers were developed. Of the 12 markers, 6 markers are from T. atroviride and remaining 6 belong to T. harzianum. SSR markers were found to be more polymorphic from T. atroviride with an average polymorphism information content value of 0.745 in comparison with T. harzianum (0.615). Twelve polymorphic markers obtained in this study clearly demonstrate the utility of newly developed SSR markers in establishing genetic relationships among different isolates of Trichoderma.

Unraveling Aspects of Bacillus amyloliquefaciens Mediated Enhanced Production of Rice under Biotic Stress of Rhizoctonia solani.

Rhizoctonia solani is a necrotrophic fungi causing sheath blight in rice leading to substantial loss in yield. Excessive and persistent use of preventive chemicals raises human health and environment safety concerns. As an alternative, use of biocontrol agents is highly recommended. In the present study, an abiotic stress tolerant, plant growth promoting rhizobacteria Bacillus amyloliquefaciens (SN13) is demonstrated to act as a biocontrol agent and enhance immune response against R. solani in rice by modulating various physiological, metabolic, and molecular functions. A sustained tolerance by SN13 primed plant over a longer period of time, post R. solani infection may be attributed to several unconventional aspects of the plants' physiological status. The prolonged stress tolerance observed in presence of SN13 is characterized by (a) involvement of bacterial mycolytic enzymes, (b) sustained maintenance of elicitors to keep the immune system induced involving non-metabolizable sugars such as turanose besides the known elicitors, (c) a delicate balance of ROS and ROS scavengers through production of proline, mannitol, and arabitol and rare sugars like fructopyranose, ß-D-glucopyranose and myoinositol and expression of ferric reductases and hypoxia induced proteins, (d) production of metabolites like quinazoline and expression of terpene synthase, and (e) hormonal cross talk. As the novel aspect of biological control this study highlights the role of rare sugars, maintenance of hypoxic conditions, and sucrose and starch metabolism in B. amyloliquefaciens (SN13) mediated sustained biotic stress tolerance in rice.

Response of indigenously developed bacterial consortia in progressive degradation of polyvinyl chloride.

Thermoplastic-based materials are recalcitrant in nature, which extensive use affect environmental health. Here, we attempt to compare the response of indigenously produced bacterial consortium-I and consortium-II in degrading polyvinyl chloride (PVC). These consortia were developed by using different combination of bacterial strains of Pseudomonas otitidis, Bacillus cereus, and Acanthopleurobacter pedis from waste disposal sites of Northern India after their identification via 16S rDNA sequencing. The progressive degradation of PVC by consortia was examined via scanning electron microscopy, atomic force microscopy, UV-vis, FT-IR spectra, gel permeation chromatography, and differential scanning calorimetry analysis at different incubations and time intervals. The consortium-II was superior over consortium-I in degrading the PVC. Further, the carbon source utilization analysis revealed that the extensive use of consortia has not any effect on functional diversity of native soil microbes.

Trichoderma inoculation augments grain amino acids and mineral nutrients by modulating arsenic speciation and accumulation in chickpea (Cicer arietinum L.).

Trichoderma reesei is an industrially important fungi which also imparts stress tolerance and plant growth promotion in various crops. Arsenic (As) contamination of field soils is one of the challenging problems in agriculture, posing potential threats for both human health and the environment. Plants in association with microbes are a liable method to improve metal tolerance and enhance crop productivity. Chickpea (Cicer arietinum L.), is an important grain legume providing cheap source of protein in semi-arid regions including As affected areas. In this study we report the role of T. reesei NBRI 0716 (NBRI 0716) in supporting chickpea growth and improving soil quality in As simulated conditions. NBRI 0716 modulated the As speciation and its availability to improve grain yield and quality (amino acids and mineral content) in chickpea (C. arietinum L.) plants grown in As spiked soil (100 mg As kg(-1) soil). Arsenic accumulation and speciation results indicate that arsenate [As(V)] was the dominant species in chickpea seeds and rhizosphere soil. The Trichoderma reduced total grain inorganic As (Asi) by 66% and enhanced dimethylarsonic acid (DMA) and monomethylarsinic acid (MMA) content of seed and rhizosphere soil. The results indicate a probable role of NBRI 0716 in As methylation as the possible mechanism for maneuvering As stress in chickpea. Analysis of functional diversity using carbon source utilization (Biolog) showed significant difference in diversity and evenness indices among the soil microbial rhizosphere communities. Microbial diversity loss caused by As were prevented in the presence of Trichoderma NBRI 0716.

Roles for root iron plaque in sequestration and uptake of heavy metals and metalloids in aquatic and wetland plants.

Toxic metal(loid) contamination of soil and sediment poses long term risk to soil and human health through plant-human or plant-animal-human food chain pathways. Iron plaque (IP) formation is frequent in aquatic and wetland plant species and is responsible for the sequestration of various metal(loids). The presence of IP may act as a buffer or barrier and may thus enhance or reduce the uptake of potentially phytotoxic metals and metalloids by plants. If IP acts as a barrier, then low IP producing macrophytes/aquatic plants may be better accumulators of toxic metals and may find use in constructed wetlands for remediation of pollutants, while high IP forming edible plant species could be safer for human consumption. Conversely, if IP acts as a buffer for mineral nutrients and toxic elements then those cultivars may be rich in nutrients, but may also cause toxicity. However, an ecotoxicological risk is also inevitable if IP rich macrophyte roots containing heavy metals are consumed by herbivores. In this review, we summarize the current understanding about the role of IP in metal and metalloid sequestration, uptake, and transport. Furthermore, we will address the role of root IP in Oryza sativa for arsenic (As) sequestration leading to lower grain As translocation, reducing the risk of human exposure.

Trichoderma inoculation ameliorates arsenic induced phytotoxic changes in gene expression and stem anatomy of chickpea (Cicer arietinum).

Arsenic, a carcinogenic metalloid severely affects plant growth in contaminated areas. Present study shows role of Trichoderma reesei NBRI 0716 (NBRI 0716) in ameliorating arsenic (As) stress on chickpea under greenhouse conditions. Arsenic stress adversely affected seed germination (25%), chlorophyll content (44%) and almost eliminated nodule formation that were significantly restored on NBRI 0716 inoculation. It also restored stem anomalies like reduced trichome turgidity and density, deformation in collenchymatous and sclerenchymatous cells induced by As stress. Semi-quantitative RT-PCR of stress responsive genes showed differential expression of genes involved in synthesis of cell wall degrading enzymes, dormancy termination and abiotic stress. Upregulation of drought responsive genes (DRE, EREBP, T6PS, MIPS, and PGIP), enhanced proline content and shrunken cortex cells in the presence of As suggests that it creates water deficiency in plants and these responses were modulated by NBRI 0716 which provides a protective role. NBRI0716 mediated production of As reductase enzyme in chickpea and thus contributed in As metabolism. The study suggests a multifarious role of NBRI0716 in mediating stress tolerance in chickpea towards As.

Gene expression profiling through microarray analysis in Arabidopsis thaliana colonized by Pseudomonas putida MTCC5279, a plant growth promoting rhizobacterium.

Plant growth promotion is a multigenic process under the influence of many factors; therefore an understanding of these processes and the functions regulated may have profound implications. Present study reports microarray analysis of Arabidopsis thaliana plants inoculated with Pseudomonas putida MTCC5279 (MTCC5279) which resulted in significant increase in growth traits as compared with non-inoculated control. The gene expression changes, represented by oligonucleotide array (24652 genes) have been studied to gain insight into MTCC5279 assisted plant growth promotion in Arabidopsis thaliana. MTCC5279 induced upregulated Arabidopsis thaliana genes were found to be involved in maintenance of genome integrity (At5g20850), growth hormone (At3g23890 and At4g36110), amino acid synthesis (At5g63890), abcissic acid (ABA) signaling and ethylene suppression (At2g29090, At5g17850), Ca⁺² dependent signaling (At3g57530) and induction of induced systemic resistance (At2g46370, At2g44840). The genes At3g32920 and At2g15890 which are suggested to act early in petal, stamen and embryonic development are among the downregulated genes. We report for the first time MTCC5279 assisted repression of At3g32920, a putative DNA repair protein involved in recombination and DNA strand transfer in a process of rapid meiotic and mitotic division.

Solid-state cultivation of Trichoderma harzianum NBRI-1055 for modulating natural antioxidants in soybean seed matrix.

Trichoderma harzianum NBRI-1055 was used, as fungal candidate to enhance the antioxidant activities of soybean matrix by modulating polyphenolic substances during solid-state fermentation. Trichoderma-fermented soybean (TFS) and unfermented soybean (UFS) products were extracted with water (W) and methanol (M). Total phenolic content of TFS-W and TFS-M extracts were significantly higher than that of UFS-W and UFS-M extracts. The effectiveness of extracts for total flavonoid content, antioxidant activity, free radical scavenging activity, reducing power, lipid peroxidation, metal ions-chelation, hydroxyl and superoxide radicals scavenging properties in descending order was TSF-W>TSF-M>USF-M>UFS-W. TFS extracts demonstrated significant protection against oxidative DNA and protein damages caused by hydroxyl radicals. For the specific phenolics profile, HPLC analysis was performed, which showed that the TFS extracts were also higher in isoflavones, flavonoids and phenolic acids, suggesting that this strategy may help to enhance TFS as potential antioxidants for use in preparation of soy food products.