Phytobiotic-Prebiotic Feed Additive Containing a Combination of Carob Pulp, Chicory, and Fenugreek Improve Growth Performance, Carcass Traits, and Fecal Microbiota of Fattening Pigs.

The purpose of this study was to determine the effectiveness of a phytobiotic-prebiotic feed additive (PPFA, which contains a combination of chicory and extracts of carob pulp and fenugreek) in the diets of fattening pigs on growth indicators, carcass characteristics, and fecal microbiota. A total of 329 crossbred pigs were randomly divided into two dietary treatments, including a basal diet without additives as the control group and a basal diet supplemented with 1 kg/T PPFA as the trial group. The PPFA supplementation led to a significant increase in the body weight gain and average daily gain of the trial group compared to those of the control group after 70 days of feeding. Through the S-EUROP evaluation system, we also found that the fattening pigs fed PPFA significantly improved their carcass indicators. Furthermore, it was shown that PPFA regulated porcine intestinal microbiota, including promoting the growth of the beneficial commensal bacteria (i.e., Bifidobacterium and Lactobacillus) while inhibiting some potential pathogen bacteria (i.e., Bacteroidaceae and Campylobacteraceae). Our work revealed that the phytobiotic-prebiotic feed additive containing carob pulp, chicory, and fenugreek positively influences the intestinal microbiota, growth performance, and carcass traits in fattening swine.

[Possible applications of poultry immunoglobulins focusing on mycotoxin environmental loads and human influence]. / Baromfi-immunglobulinok lehetséges alkalmazásai a mikotoxin-környezetterhelések és a humánérintettség fókuszában.

In addition to their role, immunoglobulins can be used in animal and human diagnostic (immunoassay-based) measurements, prophylaxis and (immuno)therapy. For these purposes, today's "alternative" that is advantageous from an animal ethical point of view is the bird immunoglobulin Y isolated from egg yolk. Its development and production are cost-effective, the complexity is low, and due to its advantageous properties, it can be used in assays or even more so in medical therapies (primarily passive immunization). It is widely used (against pathogens or their toxins) in treatments of intestinal or metabolic diseases and inflammations. Its application in human diagnostics is limited, some markers are measured using immunoglobulin Y as assay component. In this study, a possible application, which is less common today, is presented. The problem of environmental impacts is becoming significant. Due to human activities, industrialization, environmental changes increase the appearance of natural environmental pollutants, including the effects of mycotoxins produced by molds locally and/or globally, which (mainly through nutrition) affect humans. Such agents often appear together, several mycotoxins affect the individual. As a result of their persistence, mycotoxins absorbed in the intestinal tract and accumulated in organs, can already reach levels that can cause physiological and/or behavioral effects. Although the examination of sources (contaminated foods) is regulated by law, the extent of accumulation has not been or cannot be examined and is often insufficiently taken into account. Due to the nature of the technique, the anti-mycotoxin avian immunoglobulin Y could be used both for detection of (deposited) mycotoxin(s) and/or even for immunotherapy (e.g., mycotoxin neutralization). Demonstrating the endocrine-disrupting mycotoxins using the example of zearalenone (with an explanation of its reproductive and immunological effects), we present generation of zearalenone (and mycotoxin-specific) avian immunoglobulin developments, advocate its use in human detection, urging the development of measurements that are suitable for detecting (multiple) accumulation. Orv Hetil. 2023; 164(39): 1527-1536.

Distinct impact of arbuscular mycorrhizal isolates on tomato plant tolerance to drought combined with chronic and acute heat stress.

Arbuscular mycorrhizal (AM) fungi could mitigate individual drought and heat stress in host plants. However, there are still major gaps in our understanding of AM symbiosis response to the combined stresses. Here, we compared seven AM fungi, Rhizophagus irregularis, Funneliformis mosseae, Funneliformis geosporum, Funneliformis verruculosum, Funneliformis coronatum, Septoglomus deserticola, Septoglomus constrictum, distributed to many world regions in terms of their impacts on tomato endurance to combined drought and chronic heat as well as combined drought and heat shock. A multidisciplinary approach including morphometric, ecophysiological, biochemical, targeted metabolic (by ultrahigh-performance LC-MS), and molecular analyses was applied. The variation among AM fungi isolates in the enhancement in leaf water potential, stomatal conductance, photosynthetic activity, and maximal PSII photochemical efficiency, proline accumulation, antioxidant enzymes (POD, SOD, CAT), and lowered ROS markers (H2O2, MDA) in host plants under combined stresses were observed. S. constrictum inoculation could better enhanced the host plant physiology and biochemical parameters, while F. geosporum colonization less positively influenced the host plants than other treatments under both combined stresses. F. mosseae- and S. constrictum-associated plants showed the common AM-induced modifications and AM species-specific alterations in phytohormones (ABA, SA, JA, IAA), aquaporin (SlSIP1-2; SlTIP2-3; SlNIP2-1; SlPIP2-1) and abiotic stress-responsive genes (SlAREB1, SlLEA, SlHSP70, SlHSP90) in host plants under combined stresses. Altogether, mycorrhizal mitigation of the negative impacts of drought + prolonged heat and drought + acute heat, with the variation among different AM fungi isolates, depending on the specific combined stress and stress duration.

Isolation and Characterization of Lactic Acid Bacteria With Probiotic Attributes From Different Parts of the Gastrointestinal Tract of Free-living Wild Boars in Hungary.

Lactic acid bacteria (LAB) in the microbiota play an important role in human and animal health and, when used as probiotics, can contribute to an increased growth performance in livestock management. Animals living in their native habitat can serve as natural sources of microorganisms, so isolation of LAB strains from wild boars could provide the opportunity to develop effective probiotics to improve production in swine industry. In this study, the probiotic potential of 56 LAB isolates, originated from the ileum, colon, caecum and faeces of 5 wild boars, were assessed in vitro in details. Their taxonomic identity at species level and their antibacterial activity against four representative strains of potentially pathogenic bacteria were determined. The ability to tolerate low pH and bile salt, antibiotic susceptibility, bile salt hydrolase activity and lack of hemolysis were tested. Draft genome sequences of ten Limosilactobacillus mucosae and three Leuconostoc suionicum strains were determined. Bioinformatic analysis excluded the presence of any known acquired antibiotic resistance genes. Three genes, encoding mesentericin B105 and two different bacteriocin-IIc class proteins, as well as two genes with possible involvement in mesentericin secretion (mesE) and transport (mesD) were identified in two L. suionicum strains. Lam29 protein, a component of an ABC transporter with proved function as mucin- and epithelial cell-adhesion factor, and a bile salt hydrolase gene were found in all ten L. mucosae genomes. Comprehensive reconsideration of all data helps to select candidate strains to assess their probiotic potential further in animal experiments.

Chemical Profile and Biological Activities of Fungal Strains Isolated from Piper nigrum Roots: Experimental and Computational Approaches.

The current report describes the chemical investigation and biological activity of extracts produced by three fungal strains Fusarium oxysporum, Penicillium simplicissimum, and Fusarium proliferatum isolated from the roots of Piper nigrum L. growing in Vietnam. These fungi were namely determined by morphological and DNA analyses. GC/MS identification revealed that the EtOAc extracts of these fungi were associated with the presence of saturated and unsaturated fatty acids. These EtOAc extracts showed cytotoxicity towards cancer cell lines HepG2, inhibited various microbacterial organisms, especially fungus Aspergillus niger and yeast Candida albicans (the MIC values of 50-100 µg/mL). In α-glucosidase inhibitory assay, they induced the IC50 values of 1.00-2.53 µg/mL were better than positive control acarbose (169.80 µg/mL). The EtOAc extract of F. oxysporum also showed strong anti-inflammatory activity against NO production and PGE-2 level. Four major compounds linoleic acid (37.346 %), oleic acid (27.520 %), palmitic acid (25.547 %), and stearic acid (7.030 %) from the EtOAc extract of F. oxysporum were selective in molecular docking study, by which linoleic and oleic acids showed higher binding affinity towards α-glucosidase than palmitic and stearic acids. In subsequent docking assay with inducible nitric oxide synthase (iNOS), palmitic acid, oleic acid and linoleic acid could be moderate inhibitors.

Volatile organic compounds shape belowground plant-fungi interactions.

Volatile organic compounds (VOCs), a bouquet of chemical compounds released by all life forms, play essential roles in trophic interactions. VOCs can facilitate a large number of interactions with different organisms belowground. VOCs-regulated plant-plant or plant-insect interaction both below and aboveground has been reported extensively. Nevertheless, there is little information about the role of VOCs derived from soilborne pathogenic fungi and beneficial fungi, particularly mycorrhizae, in influencing plant performance. In this review, we show how plant VOCs regulate plant-soilborne pathogenic fungi and beneficial fungi (mycorrhizae) interactions. How fungal VOCs mediate plant-soilborne pathogenic and beneficial fungi interactions are presented and the most common methods to collect and analyze belowground volatiles are evaluated. Furthermore, we suggest a promising method for future research on belowground VOCs.

Detection of Acquired Antibiotic Resistance Genes in Domestic Pig (Sus scrofa) and Common Carp (Cyprinus carpio) Intestinal Samples by Metagenomics Analyses in Hungary.

The aim of this study was metagenomics analyses of acquired antibiotic-resistance genes (ARGs) in the intestinal microbiome of two important food-animal species in Hungary from a One Health perspective. Intestinal content samples were collected from 12 domestic pigs (Sus scrofa) and from a common carp (Cyprinus carpio). Shotgun metagenomic sequencing of DNA purified from the intestinal samples was performed on the Illumina platform. The ResFinder database was applied for detecting acquired ARGs in the assembled metagenomic contigs. Altogether, 59 acquired ARG types were identified, 51 genes from domestic pig and 12 genes from the carp intestinal microbiome. ARG types belonged to the antibiotic classes aminoglycosides (27.1%), tetracyclines (25.4%), ß-lactams (16.9%), and others. Of the identified ARGs, tet(E), a blaOXA-48-like ß-lactamase gene, as well as cphA4, ampS, aadA2, qnrS2, and sul1, were identified only in carp but not in swine samples. Several of the detected acquired ARGs have not yet been described from food animals in Hungary. The tet(Q), tet(W), tet(O), and mef(A) genes detected in the intestinal microbiome of domestic pigs had also been identified from free-living wild boars in Hungary, suggesting a possible relationship between the occurrence of acquired ARGs in domestic and wild animal populations.

Biomonitoring and assessment of toxic element contamination in floodplain sediments and soils using fluorescein diacetate (FDA) enzymatic activity measurements: evaluation of possibilities and limitations through the case study of the Drava River floodplain.

The EU Water Framework Directive requires the monitoring and evaluation of surface water sediment quality based on the assessment of risk posed by contamination on the biotic receptors. Floodplain sediments are important receptors of potentially toxic element (PTE) contamination from the upstream catchment areas, and floodplains host climate-sensitive riverine ecosystems and fertile agricultural areas at the same time. This study investigates the effect of PTE contamination on microbial communities in floodplain sediments and soils using the fast, inexpensive and reliable fluorescein diacetate (FDA) method in order to estimate its applicability for sediment quality monitoring and preliminary toxicity-based risk assessment. Sediment and soil samples were collected from the actively flooded alluvial plain and the river terrace areas along a 130-km stretch of the large Drava River floodplain known to be widely contaminated by historical mining, smelting and the associated industry in the upstream Alpine region. Results of detailed data analysis show that the total microbial activity represented by the measured FDA values is related to PTE (As, Cu, Zn, Cd, Pb) concentrations, but this relationship shows significant heterogeneity and depends on the spatial location and on the soil properties such as organic matter content, dissolved salt and nutrient content, and it is specific to the toxic elements. Results show that some microbe species appear to be able to adapt to the elevated PTE concentrations in toxic soil micro-environments, over time. Despite the observed heterogeneity of microbial activity, the results revealed a breakpoint in the FDA dataset around the FDA = 3 FC (fluorescein concentration) value suggesting that microbial activity is controlled by thresholds.

Alternative to ZnO to establish balanced intestinal microbiota for weaning piglets.

A wide range of phytobiotic feed additives are available on the market claiming to have beneficial effects on the growth of the host animal and to promote the development of a balanced microflora. The present study investigated the effects of the phytobiotic-prebiotic mixture of curcumin, wheat germ, and chicory on the growth performance and on the intestinal microflora composition of weaning piglets. Post weaning diarrhea causes significant losses for the producers, most commonly it is prevented by feeding high doses of zinc oxide (ZnO). The effect of a phytobiotic-prebiotic feed additive (1 kg T-1) was compared to a positive control (3.1 kg T-1 ZnO) and to a negative control (no feed supplement) in an in vivo animal trial. There was no significant difference in the final body weight and average daily gain of the trial and positive control groups, and both groups showed significantly (P<0.05) better results than the negative control. The feed conversion ratio of the phytobiotic-prebiotic supplemented group was significantly improved (P<0.05) compared to both controls. Both phytobiotic-prebiotic mixture and ZnO were able to significantly reduce (P<0.05) the amount of coliforms after weaning, even though ZnO reduced the amount of coliforms more efficiently than the trial feed additive, it also reduced the amount of potentially beneficial bacteria. Metagenomic data also corroborated the above conclusion. In the trial and positive control groups, the relative abundance of Enterobacteriaceae decreased by 85 and 88% between 3 weeks and 6 weeks of age, while in the negative control group a slight increase occurred. Lactobacillaceae were more abundant in the trial group (29.98%) than in the positive (8.67%) or in the negative (22.45%) control groups at 6 weeks of age. In summary, this study demonstrated that a phytobiotic-prebiotic feed additive may be a real alternative to ZnO for the prevention of post weaning diarrhea and promote the development of a balanced gut system.

Metagenomic Analysis of Bacterial Communities in Agricultural Soils from Vietnam with Special Attention to Phosphate Solubilizing Bacteria.

Bacterial communities can promote increased phosphorus (P) availability for plants and microbes in soil via various mechanisms of phosphate solubilization. The production of extracellular phosphatases releases available P through the hydrolysis of organic P. Examining the abundance and diversity of the bacterial community, including phosphate solubilizing bacteria in soil, may provide valuable information to overcome P scarcity in soil ecosystems. Here, the diversity and relative abundance of bacterial phyla and genera of six agricultural soil samples from Vietnam were analysed by next generation sequencing of the 16S rRNA gene. Phosphatase activities of each soil were compared with physico-chemical parameters and the abundance of the alkaline phosphatase gene phoD. We showed the dominance of Chloroflexi, Proteobacteria, Actinobacteria, Acidobacteria and Firmicutes. Total nitrogen positively correlated with phyla Proteobacteria, Acidobacteria, Firmicutes and Planctomycetes. The abundance of several genera of Proteobacteria showed positive relationship with the copy number of the phoD gene. The abundance of several taxa positively correlated with silt content, while a negative relationship of Proteobacteria was found with sand content. Our results demonstrated the clear influence of soil physico-chemical properties on the abundance of various bacterial taxa including those potentially involved in phosphate solubilization.

Differences in the effects of single and mixed species of AMF on the growth and oxidative stress defense in Lolium perenne exposed to hydrocarbons.

Arbuscular mycorrhizal fungi (AMF) are ubiquitous mutualistic plant symbionts that promote plant growth and protect them from abiotic stresses. Studies on AMF-assisted phytoremediation have shown that AMF can increase plant tolerance to the presence of hydrocarbon contaminants by improving plant nutrition status and mitigating oxidative stress. This work aimed to evaluate the impact of single and mixed-species AMF inocula (Funneliformis caledonium, Diversispora varaderana, Claroideoglomus walkeri), obtained from a contaminated environment, on the growth, oxidative stress (DNA oxidation and lipid peroxidation), and activity of antioxidative enzymes (superoxide dismutase, catalase, peroxidase) in Lolium perenne growing on a substrate contaminated with 0/0-30/120 mg phenol/polynuclear aromatic hydrocarbons (PAHs) kg-1. The assessment of AMF tolerance to the presence of contaminants was based on mycorrhizal root colonization, spore production, the level of oxidative stress, and antioxidative activity in AMF spores. In contrast to the mixed-species AMF inoculum, single AMF species significantly enhanced the growth of host plants cultured on the contaminated substrate. The effect of inoculation on the level of oxidative stress and the activity of antioxidative enzymes in plant tissues differed between the AMF species. Changes in the level of oxidative stress and the activity of antioxidative enzymes in AMF spores in response to contamination also depended on AMF species. Although the concentration of phenol and PAHs had a negative effect on the production of AMF spores, low (5/20 mg phenol/PAHs kg-1) and medium (15/60 mg phenol/PAHs kg-1) substrate contamination stimulated the mycorrhizal colonization of roots. Among the studied AMF species, F. caledonium was the most tolerant to phenol and PAHs and showed the highest potential in plant growth promotion. The results presented in this study might contribute to the development of functionally customized AMF-assisted phytoremediation strategies with indigenous AMF, more effective than commercial AMF inocula, as a result of their selection by the presence of contaminants.

Adsorption of Remazol Brilliant Violet-5R Textile Dye from Aqueous Solutions by Using Eggshell Waste Biosorbent.

Based on the well-known excellent adsorbent ability of chicken eggshells, the adsorptive capacity and mechanism of Remazol Brilliant Violet-5R (RBV-5R) dye by eggshell was investigated. Exploiting the high surface-area-to-volume ratio and porous structure of this natural adsorbent, the developed procedure showed to be useful for the efficient adsorption of RBV-5R dye from contaminated water. The protocol was thoroughly optimized by investigating the effect of the dye concentration, biomass-contaminated water ratio, particle size of the adsorbent, pH and temperature, as they are key factors in the efficiency of the dye removal process. The eggshell material was characterized by different types of microscopy techniques (stereo, polarization, SEM) as well as elemental analysis (element distribution mapping, EDX), Raman spectroscopy and BET-surface density measurements. EDX, FTIR and Raman spectroscopy proved the presence of the adsorbed dye on the surface of the biomaterial. It was shown that under optimal conditions, the environmentally friendly and inexpensive eggshell could be a reliable adsorbent for Remazol dye removal from wastewater.

Arbuscular Mycorrhizal Fungi Improve Tolerance of the Medicinal Plant Eclipta prostrata (L.) and Induce Major Changes in Polyphenol Profiles Under Salt Stresses.

Eclipta prostrata (L.) is an important and well-known medicinal plant due to its valuable bioactive compounds. Microorganisms, including arbuscular mycorrhizal fungi (AMF), and salinity could directly impact plant metabolome, thus influencing their secondary metabolites and the efficacy of herbal medicine. In this study, the role of different single AMF species (Funneliformis mosseae, Septoglomus deserticola, Acaulospora lacunosa) and a mixture of six AMF species in plant growth and physio-biochemical characteristics of E. prostrata under non-saline conditions was investigated. Next, the most suitable AM treatment was chosen to examine the impact of AMF on physio-biochemical features and polyphenol profiles of E. prostrata under saline conditions (100 and 200 mM NaCl). The findings indicated that AMF mixture application resulted in more effective promotion on the aboveground part of non-saline plants than single AMF species. AM mixture application improved growth and salt tolerance of E. prostrata through increasing the activity of catalase, peroxidase (at 4 weeks), proline, and total phenolic content (at 8 weeks). Such benefits were not observed under high salinity, except for a higher total phenolic concentration in mycorrhizal plants at 8 weeks. Through high-performance liquid chromatography, 14 individual phenolic compounds were analyzed, with wedelolactone and/or 4,5-dicaffeoylquinic acid abundant in all treatments. Salinity and mycorrhizal inoculation sharply altered the polyphenol profiles of E. prostrata. Moderate salinity boosted phenolic compound production in non-AM plants at 4 weeks, while at 8 weeks, the decline in the content of phenolic compounds occurred in uncolonized plants subjected to both saline conditions. Mycorrhization augmented polyphenol concentration and yield under non-saline and saline conditions, depending on the growth stages and salt stress severity. Plant age influenced polyphenol profiles with usually a higher content of phenolic compounds in older plants and changed the production of individual polyphenols of both non-AM and AM plants under non-stress and salt stress conditions. A better understanding of factors (involving mycorrhiza and salinity) affecting the phenolic compounds of E. prostrata facilitates the optimization of individual polyphenol production in this medicinal plant.

Genotyping of Acantamoeba spp. from rhisophere in Hungary.

The protista Acanthamoeba is a free-living amoeba existing in various environments. A number of species among protista are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK), granulomatous amoebic encephalitis (GAE), and chronic granulomatous lesions. In this study, 10 rhizosphere samples were collected from maize and alfalfa plants in experimental station at Institute of Genetics, Microbiology and Biotechnology, Szent István University. We detected Acanthamoeba based on the quantitative real-time PCR assay and sequence analysis of the 18S rRNA gene. All studied molecular biological methods are suitable for the detection of Acanthamoeba infection in humans. The quantitative real-time PCR-based methods are more sensitive, simple, and easy to perform; moreover, these are opening avenue to detect the effect of number of parasites on human disease. Acanthamoeba species were detected in five (5/10; 50%) samples. All Acanthamoeba strains belonged to T4 genotype, the main AK-related genotype worldwide. Our result confirmed Acanthamoeba strains in rhizosphere that should be considered as a potential health risk associated with human activities in the environment.

Changes in DNA methylation pattern of apple long-term in vitro shoot culture and acclimatized plants.

DNA methylation is a process of epigenetic modification that can alter the functionality of a genome. Using whole-genome bisulfite sequencing, this study quantify the level of DNA methylation in the epigenomes of two diploid apple (Malus x domestica) scion cultivars ('McIntosh' and 'Húsvéti rozmaring') derived from three environmental conditions: in vivo mother plants in an orchard, in vitro culture, and acclimatized in vitro plants. The global DNA methylation levels were not dependent on the source of plant material, and the average level of DNA methylation was 49.77%, 34.65% and 8.77% in CpG, CHG and CHH contexts, respectively. Significant differences in DNA methylation were identified in 586 (specifically 334, 201 and 131 in CpG, CHG and CHH contexts, respectively) out of 45,116 genes, including promoter and coding sequences. These were classified as differentially methylated genes (DMGs). This is a 1.3% difference in the level of DNA methylation of genes in response to a change in the environment. Differential methylation was visualised by MA plots and functional genomic maps were established for biological processes, molecular functions and cellular components. When the DMGs were considered, in vitro tissue culture resulted in the highest level of methylation, but it was lower in acclimatized in vitro plants which was similar to that in the mother tree. Methylation patterns of the two scions differed, indicating cultivar-specific epigenetic regulation of gene expression during adaptation to various environments. After selecting genes that displayed differences larger than ±10% in CpG and CHG contexts, or larger than ±1.35% in the CHH context from among the DMGs, they were annotated in Blast2 GO v5.1.12 for Gene Ontology. DMGs identified as MD07G1113000 (protein transport), MD08G1041600 (extracellular space), MD09G1054800 (phosphatidic acid binding), and MD10G1265800 (not annotated) were methylated in all three contexts in in vitro shoots. These DNA methylation results suggest that epigenetic changes may contribute to the adaptation of apple to environmental changes by modifying the epigenome and thereby gene expression.

Adsorptive Removal of Remazol Brilliant Violet-5R Dye from Aqueous Solutions using Calcined Eggshell as Biosorbent.

As a result of industrial development water pollution has become a major issue. Providing clean water of sufficient quality and quantity is essential. According to literature, 100 000 different dyes are used worldwide in the printing, food and textile industries. Remazol Brilliant Violet-5R belongs to the group of azo dyes used in the textile industry. The purpose of this paper is to study the removal of anionic dye RBV-5R using calcined eggshell. We investigated the effect of initial dye concentration (20-100 mg/L RBV-5R), biomass amount, contact time, temperature, and pH on the adsorption process. We calculated the amounts of adsorbed material in equilibrium and the efficiency of the adsorption process. Various studies were performed on the adsorption process and the adsorbent morphology (EDX, SEM, FTIR, Raman), as well as isotherm (Langmuir, Freundlich, Temkin and Dubinin-Radushkevich), kinetic and diffusion models were used to characterize the process. Calcined eggshell was also characterized by thermogravimetric, BET surface methods. Based on the results obtained, it can be said that the calcined eggshell can be used effectively to remove the dye from the aqueous solution.

Arbuscular mycorrhizal fungi mitigate negative effects of combined drought and heat stress on tomato plants.

Arbuscular mycorrhizal (AM) symbiosis can alleviate drought and temperature stresses in plants, but it is unknown whether the benefits can be maintained when the plants are exposed to combined drought and heat stress. In this study, the impacts of AM fungi, Septoglomus deserticola and Septoglomus constrictum on tomato plant tolerance to combined drought and heat stress were investigated. No substantial differences in physiological parameters were found in all plants under non-stress conditions, except a higher expression of SlLOXD and SlPIP2.7 in plants + S. constrictum. Under drought, heat and drought + heat stress, both fungal symbionts could moderate oxidative stress by decreasing the lipid peroxidation, hydrogen peroxide level and improving leaf and root antioxidant enzyme activities, however better performance in plants + S. constrictum. Under drought and the combined stress, inoculation with S. constrictum enhanced stomatal conductance, leaf water potential and relative water content, elevated Fv/Fm and biomass production of the hosts as compared to non-inoculated plants whilst these improvements in plants + S. deserticola were not obvious. Under the combined stress inoculation of S. constrictum did not change the expression of SlNCED and SlPIP2.7 in roots as under heat stress. Expression of SlLOXD in root were upregulated in plants + S. contrictum under drought + heat stress as in mycorrhizal roots under drought stress. Altogether, our results indicated that AM inoculation, particularly with S. constrictum had a positive influence on the tomato plant tolerance to drought + heat stress. Further studies are essential to add some light on molecular mechanisms of mycorrhizal plant tolerance to this combined stress.

Mycorrhiza-induced alleviation of plant disease caused by Clavibacter michiganensis subsp. michiganensis and role of ethylene in mycorrhiza-induced resistance in tomato.

The protective role of arbuscular mycorrhizal fungi (AMF) against the phytopathogen Clavibacter michiganensis subsp. michiganensis (Cmm) was examined in tomato plants. Seven different AMF isolates were used to determine which ones were able to induce effectively resistance against Cmm. Stems of seven-week tomato plants were infected with Cmm, then a disease severity index (DSI) was determined during the next three weeks. In addition to different responses to mycorrhizal inoculation, three levels of responses to the bacterial disease were recognized in treatments. Plants inoculated with Rhizophagus irregularis (Ri) showed both the highest colonization and the highest induced resistance to Cmm while the effect of Funneliformis mosseae, Gigaspora margarita and Claroideoglomus claroideum on mycorrhizal colonization and on the induced resistance were intermediate and high, respectively. Subsequently, Ri was chosen to inoculate ethylene-insensitive tomato mutant line Never ripe (Nr) and its background (Pearson) to investigate the possible role of ethylene (ET) in the mycorrhiza-induced resistance (MIR). The results showed that Ri could induce systemic resistance against Cmm in the Pearson background, whereas ET-insensitivity in Nr plants impaired MIR. These results suggest that ET is required for Ri-induced resistance against Cmm. To our knowledge, this is the first study to examine the effect of different AMF isolates on the response of tomato plants to Cmm and involvement of ET in MIR against Cmm.

How arbuscular mycorrhizal fungi influence the defense system of sunflower during different abiotic stresses.

The association between terrestrial plants and arbuscular mycorrhizal (AM) fungi is one of the most common and widespread mutualistic plant-fungi interaction. AM fungi are of beneficial effects on the water and nutrient uptake of plants and increase plant defense mechanisms to alleviate different stresses. The aim of this study was to determine the level of polyphenol oxidase (PPO), guaiacol peroxidase (POX) and glutathione S-transferase (GST) enzyme activities and to track the expression of glutathione S-transferase (GST) gene in plant-arbuscular mycorrhizal system under temperature- and mechanical stress conditions. Our results suggest that induced tolerance of mycorrhizal sunflower to high temperature may be attributed to the induction of GST, POX and PPO enzyme activities as well as to the elevated expression of GST. However, the degree of tolerance of the plant is significantly influenced by the age which is probably justified by the energy considerations.

The chemical inducer, BTH (benzothiadiazole) and root colonization by mycorrhizal fungi (Glomus spp.) trigger resistance against white rot (Sclerotinia sclerotiorum) in sunflower.

White rot caused by Sclerotinia sclerotiorum (SS) is one of the most devastating plant diseases of sunflower. Controlling this pathogen by available tools hardly result in acceptable control. The aim of this study was to elucidate the effects of plant resistance inducers, BTH (benzothiadiazole in Bion 50 WG) and arbuscular mycorrhizal fungi (AMF) on disease development of white rot in three sunflower genotypes. Defence responses were characterized by measuring the disease severity and identifying cellular/histological reactions (e.g. autofluorescence) of host plants upon infection. Depending on the host genotype, a single application of inducers reduced disease symptoms. Histological examination of host responses revealed that BTH and/or AMF pre-treatments significantly impeded the development of pathogenic hyphae in Iregi szürke csíkos and P63LE13 sunflower plants and it was associated with intensive autofluorescence of cells. Both localized and systemic induction of resistance was observed. Importantly, the frequency of mycorrhization of hybrid P63LE13 and PR64H41 was significantly increased upon BTH treatment, so it had a positive effect on the formation of plant-mycorrhiza interactions in sunflower. To our knowledge, this is the first report on the additive effect of BTH on mycorrhization and the positive effect of these inducers against SS in sunflower.