Contribution of Serendipita indica on growth improvement, antioxidative capacity of Dracocephalum kotschyi, and its resistance against cadmium stress / Contribución de Serendipita indica a la mejora del crecimiento, la capacidad antioxidante de Dracocephalum kotschyi y su resistencia al estrés por cadmio

Cadmium pollution is a severe issue worldwide which causes an elevated concern in agriculture. The utilization of plant–microbial interactions offers a promising approach for the remediation of cadmium-polluted soils. To elucidate the mechanism of Serendipita indica-mediated cadmium stress tolerance, a potting experiment was conducted to study the impact of S. indica on Dracocephalum kotschyi plants grown under different cadmium concentrations (0, 5, 10, and 20 mg/kg). The effects of cadmium and S. indica on plant growth, antioxidant enzyme activities, and accumulation of cadmium were investigated. The results showed that cadmium stress significantly decreases biomass, photosynthetic pigments, and carbohydrate content concomitantly with increasing antioxidant activities, electrolyte leakage, and hydrogen peroxide, proline, and cadmium content. Inoculation with S. indica alleviated the adverse effect of cadmium stress by enhancing shoot and root dry weight, photosynthetic pigments, and carbohydrate, proline, and catalase activity. Unlike cadmium stress, the presence of fungus led to a reduction in electrolyte leakage and hydrogen peroxide content as well as the content of cadmium in D. kotschyi leaf which mitigates cadmium-induced oxidative stress. Our findings demonstrated S. indica inoculation alleviates the adverse effects of cadmium stress in D. kotschyi plants which could prolong their survival under stressful conditions. Due to the importance of D. kotschyi and the effect of biomass increase on the amount of its medicinal substances, exploiting S. indica not only promotes plant growth, but also may be used as a potential eco-friendly method for relieving the phytotoxicity of Cd and remediating Cd-contaminated soil.(AU)

Combined Effect of Trehalose and Serendipita indica Inoculation Might Participate in Solanum lycopersicum Induced Cold Tolerance.

The exploitation of symbiotic interactions between fungi and plants, coupled with the application of osmoprotectants such as trehalose (Tre), presents a promising strategy for mitigating environmental stress. To determine the mechanism of Serendipita indica and Tre-mediated cold stress tolerance, a comparative experiment was designed to study the impact of S. indica, Tre and their combination on tomato plants grown under cold stress. The results showed that cold stress significantly decreased biomass, relative water content, photosynthetic pigments and elements concomitantly with increasing antioxidant activities, malondialdehyde (MDA), electrolyte leakage, hydrogen peroxide and proline content. Meanwhile, S. indica and Tre treatments promoted biomass and enhanced carbohydrate, protein, proline, potassium, phosphorous, antioxidant enzymes and photosynthetic pigments content under cold stress. Furthermore, single or dual application of endophyte and Tre mitigated physiological disorders induced by cold stress and increased the integrity of cell membranes by decreasing hydrogen peroxide, MDA, and electrolyte leakage (EL). Our findings suggest that S. indica and Tre combination could significantly promote cold stress tolerance compared with single treatment. This study is novel in showing the cold adaptation of tomato plants by combination use of S. indica and Tre, which can be a promising strategy for improving cold tolerance. The underlying molecular mechanisms of sugar-fungus interaction must be further investigated.

Contribution of Serendipita indica on growth improvement, antioxidative capacity of Dracocephalum kotschyi, and its resistance against cadmium stress.

Cadmium pollution is a severe issue worldwide which causes an elevated concern in agriculture. The utilization of plant-microbial interactions offers a promising approach for the remediation of cadmium-polluted soils. To elucidate the mechanism of Serendipita indica-mediated cadmium stress tolerance, a potting experiment was conducted to study the impact of S. indica on Dracocephalum kotschyi plants grown under different cadmium concentrations (0, 5, 10, and 20 mg/kg). The effects of cadmium and S. indica on plant growth, antioxidant enzyme activities, and accumulation of cadmium were investigated. The results showed that cadmium stress significantly decreases biomass, photosynthetic pigments, and carbohydrate content concomitantly with increasing antioxidant activities, electrolyte leakage, and hydrogen peroxide, proline, and cadmium content. Inoculation with S. indica alleviated the adverse effect of cadmium stress by enhancing shoot and root dry weight, photosynthetic pigments, and carbohydrate, proline, and catalase activity. Unlike cadmium stress, the presence of fungus led to a reduction in electrolyte leakage and hydrogen peroxide content as well as the content of cadmium in D. kotschyi leaf which mitigates cadmium-induced oxidative stress. Our findings demonstrated S. indica inoculation alleviates the adverse effects of cadmium stress in D. kotschyi plants which could prolong their survival under stressful conditions. Due to the importance of D. kotschyi and the effect of biomass increase on the amount of its medicinal substances, exploiting S. indica not only promotes plant growth, but also may be used as a potential eco-friendly method for relieving the phytotoxicity of Cd and remediating Cd-contaminated soil.

Impact of Piriformospora indica on various characteristics of tomatoes during nickel nitrate stress under aeroponic and greenhouse conditions.

As an essential nutrient for plant growth, nickel's (Ni) requirement is very low, and its augmented level causes environmental pollution and toxicity. Being a root endophytic fungus, Piriformospora indica (P. indica) can be beneficial to many plants under stress and non-stress conditions, particularly in terms of their improved growth performance. P. indica, as evidenced, enhances tolerance and resistance in most plants once they experience a range of stresses caused by biotic and abiotic factors, e.g., diseases and heavy metals. Against this background, the positive effects of P. indica on the tomato plants under Ni-induced stress (300, 600, and 900 mg L-1) were analyzed in three experiments at labs, at greenhouses, and via aeroponics in this study. The growth traits of the tomato plants, such as root length (RL) and root dry weight (RDW), were accordingly found to be positively boosted in the cases treated with P. indica compared to the non-treated ones. Treating with P. indica also thwarted the negative effects of Ni on some biochemical traits, including anthocyanin (Anth), proline (Pro), catalase (CAT), and glutathione peroxidase (GPx), while significantly minimizing the adverse impacts of this heavy metal at different levels on hydrogen peroxide (H2O2). Despite this, the Ni-stressed plants indicated much better traits in the presence of this fungus, compared with the non-treated ones, in most of the cases measured. Moreover, the photosynthetic pigments, i.e., chlorophyll a and b (Chl a & b) and carotenoid content (Carrot), were significantly higher in the tomato plants treated with P. indica under high Ni-induced stress as compared with the non-treated ones under non-Ni conditions, in which these pigments were low. The pro-production was further observed all through the P. indica inoculation, which could aid the treated plants in becoming Ni-stress-tolerant. Finally, the current study contributed to a better understanding of how to use the P. indica symbiosis to induce heavy metal tolerance in tomato plants, such as Ni, to meet the goals of sustainable agriculture.

Biochemical changes and quality characterization of cold-stored 'Sahebi' grape in response to postharvest application of GABA.

In this study, the effect of γ-aminobutyric acid (GABA) at 0 (control), 20 and 40 mM on maintaining postharvest quality, chilling tolerance and fungal decay of 'Sahebi' grapevine (Vitis vinifera L.) was investigated during 60 days storage at 1 °C. GABA-treated fruits especially at 40 mM showed less weight loss (35%), rachis browning (30%) and decay incident (63%) compared to the control. GABA-induced abscisic acid was linked to lower membrane electrolyte leakage (13%) in treated grapes. Moreover, at the end of 60 days, GABA treatment at 40 mM resulted in higher activities of antioxidant enzymes including superoxide dismutase (50%), catalase (35%), guaiacol peroxidase (65%), and ascorbate peroxidase (47%) and lower malondialdehyde (21%) compared to control samples. The highest soluble sugars and organic acids were related to 40 mM GABA-treated grape clusters. Phenolic compounds (phenolic acids, stilbenes, flavonoids and anthocyanidins) and antioxidant capacity increased in 40 mM GABA-treated grape due to lower polyphenol oxidase activity. Therefore, GABA is recommended for maintaining internal quality and reduction in fungal decay and chilling injury of grapes during postharvest storage.

Salinity-associated microRNAs and their potential roles in mediating salt tolerance in rice colonized by the endophytic root fungus Piriformospora indica.

Piriformospora indica (P. indica), an endophytic root fungus, supports the growth and enhanced tolerance of plants to biotic and abiotic stresses. Several recent studies showed the significant role of small RNA (sRNA) molecules including microRNAs (miRNAs) in plant adaption to environmental stress, but little is known concerning the symbiosis-mediated salt stress tolerance regulated at miRNAs level. The overarching goal of this research is to elucidate the impact of miRNAs in regulating the P. indica-mediated salt tolerance in rice. Applying sRNA-seq analysis led to identify a set of 547 differentially abundant miRNAs in response to P. indica inoculation and salt stress. These included 206 rice-specific and 341 previously known miRNAs from other plant species. In silico analysis of miRNAs predictions of the differentially abundant miRNAs led to identifying of 193 putatively target genes, most of which were encoded either genes or transcription factors involved in nutrient uptake, sodium ion transporters, growth regulators, and auxin- responsive proteins. The rice-specific miRNAs targeted the transcription factors involved in the import of potassium ions into the root cells, the export of sodium ions, and plant growth and development. Interestingly, P. indica affected the differential abundance of miRNAs regulated genes and transcription factors linked to salt stress tolerance. Our data helps to understand the molecular basis of salt stress tolerance mediated by symbionts in plant and the potential impact of miRNAs for genetic improvement of rice varieties for tolerance to salt stress.

Inoculation and co-inoculation of alfalfa seedlings with root growth promoting microorganisms (Piriformospora indica, Glomus intraradices and Sinorhizobium meliloti) affect molecular structures, nutrient profiles and availability of hay for ruminants.

Inoculation of alfalfa seedlings with root growth promoting microorganisms under semi-arid climate condition may improve biomass production and nutritive value. The current study aimed to investigate the effect of inoculation of alfalfa seedlings with Piriformospora indica (Pi) and co-inoculating Pi with Glomus intraradices (Gi + Pi) or Sinorhizobium meliloti (Sm + Pi) on hay yield, chemical composition, molecular structures by Fourier transformed infrared (FTIR) spectroscopy, in situ ruminal degradability and in vitro gas production. Seedlings were grown in experimental pots in a greenhouse until first cut and then transferred outside and cut a further 4 times. Biomass yield was similar across the treatments. Acid detergent fiber (ADF) concentration was higher in Pi than in control hay, and ADF decreased further with co-inoculation (P < 0.05). The ether extract (EE) concentration was lower for Pi and Gi + Pi compared with control hay, and control, Pi and Gi + Pi hays had lower EE concentration compared with Sm + Pi (P < 0.05). The FTIR spectroscopic vibration peak height ratio related to proteins (amide 1 + amide 2): total carbohydrate ratio was lower for the inoculation treatments compared with control hay (P < 0.05). In situ ruminal degradability of dry matter and organic matter were higher for hay of inoculated and co-inoculated seedlings than for control hay (P < 0.05). In conclusion, hay of alfalfa seedlings inoculated and co-inoculated with root growth promoting microorganisms had improved nutritional value compared with hay from non-treated alfalfa seedlings, and co-inoculation was the most effective, however, changes were relatively minor.

Recombinant production of bovine Lactoferrin-derived antimicrobial peptide in tobacco hairy roots expression system.

LFchimera is a chimerical peptide containing Lactoferricin and Lactoferrampin antimicrobial peptides of bovine lactoferrin, and it has stronger bactericidal activity. Antimicrobial peptides (AMPs) like LFchimera have great potential as an alternative candidate for conventional antibiotics. Plant hairy roots provide suitable platform for fast, easy and cost-effective production of various recombinant proteins. The aim of this study was to express recombinant LFchimera in Nicotiana tabacum hairy roots and investigate its antimicrobial activity. The integration and expression of the transgene in hairy roots were confirmed by PCR and RT-PCR, respectively. LFchimera levels were quantified by ELISA and the presence of LFchimera was verified by SDS-PAGE analysis of root extracts. Biological activity of the plant derived LFchimera was confirmed by investigating the antimicrobial activity of total solution protein against Escherichia coli (ATCC 8739). Hairy root biomass reached to 4.6 g and LFchimera accumulate about 4.8 µg/g fresh weight in Erlenmeyer flasks.

Metabolic and transcriptional response of central metabolism affected by root endophytic fungus Piriformospora indica under salinity in barley.

The root endophytic fungus Piriformospora indica enhances plant adaptation to environmental stress based on general and non-specific plant species mechanisms. In the present study, we integrated the ionomics, metabolomics, and transcriptomics data to identify the genes and metabolic regulatory networks conferring salt tolerance in P. indica-colonized barley plants. To this end, leaf samples were harvested at control (0 mM NaCl) and severe salt stress (300 mM NaCl) in P. indica-colonized and non-inoculated barley plants 4 weeks after fungal inoculation. The metabolome analysis resulted in an identification of a signature containing 14 metabolites and ions conferring tolerance to salt stress. Gene expression analysis has led to the identification of 254 differentially expressed genes at 0 mM NaCl and 391 genes at 300 mM NaCl in P. indica-colonized compared to non-inoculated samples. The integration of metabolome and transcriptome analysis indicated that the major and minor carbohydrate metabolism, nitrogen metabolism, and ethylene biosynthesis pathway might play a role in systemic salt-tolerance in leaf tissue induced by the root-colonized fungus.

Proteomics study reveals the molecular mechanisms underlying water stress tolerance induced by Piriformospora indica in barley.

Piriformospora indica is a mutualistic root endophytic fungus, which transfers several benefits to hosts including enhance plant growth and increase yield under both normal and stress conditions. It has been shown that P. indica root-colonization enhances water stress tolerance based on general and non-specific plant-species mechanism. To better understand the molecular mechanism of P. indica-mediated drought stress tolerance, we designed a set of comparative experiments to study the impact of P. indica on barely plants cultivar "Golden Promise" grown under different drought levels [Filed capacity (F.C.) and 25% F.C.]. P. indica enhanced root and shoot biomass of colonized plants under both well-watered and water-deficit conditions. Proteome analysis of P. indica-colonized barley leaves under well-treated and water-deficit conditions resulted in detection of 726 reproducibly protein spots. Mass spectrometry analysis resulted in the identification of 45 differentially accumulated proteins involved in photosynthesis, reactive oxygen scavenging, metabolisms, signal transduction, and plant defense responses. Interestingly, P. indica increased the level of proteins involved in photosynthesis, antioxidative defense system and energy transport. We propose that P. indica-mediated drought stress tolerance in barely is through photosynthesis stimulation, energy releasing and enhanced antioxidative capacity in colonized plants. BIOLOGICAL SIGNIFICANCE: Plant mutualistic symbionts offer long-term abiotic stress tolerance through the host adaptation to environmental stress. There have been a few published proteomic studies of plant symbionts to drought, and this is thought to be the first proteomic analysis, demonstrating the impact of endophyte on barley plant under drought stress. For some of identified proteins like TCTP and PCNA, a connection to physiological function in plants is novel, and can be the best candidates for sources of drought tolerance in future studies.