Bacterial Volatiles Research Articles

Air Ambulance: Antimicrobial Power of Bacterial Volatiles.

We are currently facing an antimicrobial resistance crisis, which means that a lot of bacterial pathogens have developed resistance to common antibiotics. Hence, novel and innovative solutions are urgently needed to combat resistant human pathogens. A new source of antimicrobial compounds could be bacterial volatiles. Volatiles are ubiquitous produced, chemically divers and playing essential roles in intra- and interspecies interactions like communication and antimicrobial defense. In the last years, an increasing number of studies showed bioactivities of bacterial volatiles, including antibacterial, antifungal and anti-oomycete activities, indicating bacterial volatiles as an exciting source for novel antimicrobial compounds. In this review we introduce the chemical diversity of bacterial volatiles, their antimicrobial activities and methods for testing this activity. Concluding, we discuss the possibility of using antimicrobial volatiles to antagonize the antimicrobial resistance crisis.

Volatile composition changes of fruits in a biopolymer-coated polyethylene active packaging: Effects of modified atmosphere and packaging-shaped bacterial community

The potential impacts of active packaging on the volatile composition of fruits during preservations largely associated with consumers’ acceptance, yet was barely investigated. In this work, a biopolymer-coated polyethylene active film was developed and its effects on qualities and volatile compositions of fruits with different respiration rates were comprehensively investigated. Underlying reasons for the effect of active packaging on volatile composition change of fruits were elucidated through revealing ternary relationship among the packaging-shaped bacterial community, modified atmosphere and volatiles. Off-flavor production was reduced and more sesquiterpenes (α-cubebene, copaene, β-caryophyllene, α-caryophyllene, d-amorphene) were reserved for longan packaged with active film, due to its antimicrobial and moisture absorption ability, as leading contributors to the selective inhibition on spoilage bacteria genera including Zymobacter, Gluconobacter and Pantoea. Whereas volatile profile of strawberry with high respiration rate was more actively affected by the modified atmosphere induced by packaging, instead of the bacterial community, where hypoxia (<2% O2) led to the accumulation of ethyl esters (0.154–0.184 μg/g) that conferred off-flavor but decreased production of methyl (0.172–0.367 μg/g) and hexyl esters (1.26–1.89 × 10−2 μg/g) that represented typical fruity aroma. This work adds new knowledges on impacts of active packaging on the volatile profile of fruits and would provide guidance to smart design of active packaging for preservation of aromatic fruits.

IN VITRO ANTAGONISTIC EFFECTS OF SOME BACILLUS SPP. ON THE GROWTH AND MYCOTOXIN PRODUCTION OF TOXIGENIC ASPERGILLI

This study aimed to investigate the antagonistic effect of three strains of the genus Bacillus on toxigenic fungi contaminating food commodities, with emphasis on two ochratoxigenic isolates (A. ochraceus, A. westerdijkiae) and two aflatoxigenic isolates (A. flavus and A. parasiticus). In vitro studies were carried out using two different methods for cultivation tested bacilli with fungal isolates – coincubation and dual culture method. The most sensitive isolate was A. ochraceus by both used methods. B. mycoides (5.72 ± 6.4 mm) and B. subtilis (5.08 ± 2.84 mm) were able to inhibit its growth and sporulation during ten days of cultivation (both inhibited the sporulation of A. ochraceus 100%) in coincubation. The most effective in the dual culture method were B. subtilis and B. thuringiensis against A. ochraceus (growth inhibition rate 84.40%; 90.55%) and A. flavus (growth inhibition rate 91.54%; 92.43%). The most effective sporulation inhibitors were B. subtilis and B. thuringiensis, which completely inhibited the sporulation of A. ochraceus and A. parasiticus after ten days of coincubation. Likewise, all tested bacterial strains showed complete inhibition of ochratoxin A synthesis in A. ochraceus and A. westerdijkiae exposure to bacterial volatiles. So, the current study illustrated that strains of the genus Bacillus could significantly inhibit the growth, sporulation, and mycotoxin production of toxigenic aspergilli and showing the great potential as a biocontrol agent of pre- and post-harvest food diseases caused by microscopic filamentous fungi.

Impact of bacterial volatiles on phytopathogenic fungi: an in vitro study on microbial competition and interaction.

Microorganisms in the rhizosphere are abundant and exist in very high taxonomic diversity. The major players are bacteria and fungi, and bacteria have evolved many strategies to prevail over fungi, among them harmful enzyme activities and noxious secondary metabolites. Interactions between plant growth promoting rhizobacteria and phytopathogenic fungi are potentially valuable since the plant would benefit from fungal growth repression. In this respect, the role of volatile bacterial metabolites in fungistasis has been demonstrated, but the mechanisms of action are less understood. We used three phytopathogenic fungal species (Sclerotinia sclerotiorum, Rhizoctonia solani, and Juxtiphoma eupyrena) as well as one non-phytopathogenic species (Neurospora crassa) and the plant growth promoting rhizobacterium Serratia plymuthica 4Rx13 in co-cultivation assays to investigate the influence of bacterial volatile metabolites on fungi on a cellular level. As a response to the treatment, we found elevated lipid peroxidation, which indirectly reflected the loss of fungal cell membrane integrity. An increase in superoxide dismutase, catalase, and laccase activities indicated oxidative stress. Acclimation to these adverse growth conditions completely restored fungal growth. One of the bioactive bacterial volatile compounds seemed to be ammonia, which was a component of the bacterial volatile mixture. Applied as a single compound in biogenic concentrations ammonia also caused an increase in lipid peroxidation and enzyme activities, but the extent and pattern did not fully match the effect of the entire bacterial volatile mixture.

Prevalence of toxigenic fungi and mycotoxins in Arabic coffee (Coffea arabica): Protective role of traditional coffee roasting, brewing and bacterial volatiles.

Fungal infection and synthesis of mycotoxins in coffee leads to significant economic losses. This study aimed to investigate the prevalence of toxigenic fungi, their metabolites, and the effect of traditional roasting and brewing on ochratoxin A (OTA) and aflatoxins (AFs) contents of naturally contaminated coffee samples. In addition, in vivo biocontrol assays were performed to explore the antagonistic activities of Bacillus simplex 350–3 (BS350-3) on the growth and mycotoxins synthesis of Aspergillus ochraceus and A. flavus. The relative density of A. niger, A. flavus, Penicillium verrucosum and A. carbonarius on green coffee bean was 60.82%, 7.21%, 3.09% and 1.03%, respectively. OTA contents were lowest in green coffee beans (2.15 μg/kg), followed by roasted (2.76 μg/kg) and soluble coffee (8.95 μg/kg). Likewise, AFs levels were highest in soluble coffee (90.58 μg/kg) followed by roasted (33.61 μg/kg) and green coffee (9.07 μg/kg). Roasting naturally contaminated coffee beans at three traditional methods; low, medium and high, followed by brewing resulted in reduction of 58.74% (3.50 μg/kg), 60.88% (3.72 μg/kg) and 64.70% (4.11 μg/kg) in OTA and 40.18% (34.65 μg/kg), 47.86% (41.17 μg/kg) and 62.38% (53.73 μg/kg) AFs contents, respectively. Significant inhibitions of AFs and OTA synthesis by A. flavus and A. carbonarius, respectively, on infected coffee beans were observed in presence of Bacillus simplex BS350-3 volatiles. Gas chromatography mass spectrochemistry (GC-MS/MS) analysis of head-space BS350-3 volatiles showed quinoline, benzenemethanamine and 1-Octadecene as bioactive antifungal molecules. These findings suggest that marketed coffee samples are generally contaminated with OTA and AFs, with a significant level of roasted and soluble coffee contaminated above EU permissible limits for OTA. Further, along with coffee roasting and brewing; microbial volatiles can be optimized to minimize the dietary exposure to mycotoxins.

Identifying volatile in vitro biomarkers for oral bacteria with proton-transfer-reaction mass spectrometry and gas chromatography\u2013mass spectrometry

We have measured the volatile fingerprints of four pathogenic oral bacteria connected to periodontal disease and dental abscess: Porphyromonas gingivalis (three separate strains), Prevotella intermedia, Prevotella nigrescens and Tannerella forsythia. Volatile fingerprints were measured in vitro from the headspace gas of the bacteria cultured on agar. Concrete identification of new and previously reported bacterial volatiles were performed by a combination of solid phase microextraction (SPME) and offline gas chromatography–mass spectrometry (GC–MS). We also studied the effect of the reduced electric field strength (E/N) on the fragmentation patterns of bacterial volatiles in online proton-transfer-reaction time-of-flight mass spectrometry (PTR-ToF-MS). We aimed to discover possible new biomarkers for the studied oral bacteria, as well as to validate the combination of GC–MS and PTR-MS for volatile analysis. Some of the most promising compounds produced include: 1-Methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), indole, and a cascade of sulphur compounds, such as methanethiol, dimethyl disulphide (DMDS) and dimethyl trisulphide (DMTS). We also found that several compounds, especially alcohols, aldehydes and esters, fragment significantly with the PTR-MS method, when high E/N values are used. We conclude that the studied oral bacteria can be separated by their volatile fingerprints in vitro, which could have importance in clinical and laboratory environments. In addition, using softer ionization conditions can improve the performance of the PTR-MS method in the volatile analysis of certain compounds.

N,N-dimethyl-hexadecylamine modulates Arabidopsis root growth through modifying the balance between stem cell niche and jasmonic acid-dependent gene expression

N,N-dimethyl-hexadecylamine (DMHDA) is released as part of volatile blends emitted by plant probiotic bacteria and affects root architecture, defense and nutrition of plants. Here, we investigated the changes in gene expression of transcription factors responsible of maintenance of the root stem cell niche and jasmonic acid signaling in Arabidopsis seedlings in response to this volatile. Concentrations of DMHDA that repress primary root growth were found to alter cell size and division augmenting cell tissue layers in the meristem and causing root widening. DMHDA triggered the division of quiescent center cells, which correlated with repression of SHORT ROOT (SHR), SCARECROW (SCR), and PLETHORA 1 (PLT1) proteins and induction of WUSCHEL-RELATED HOMEOBOX 5 (WOX5) transcription factor. Interestingly, an activation of the expression of the jasmonic acid-related reporter genes JAZ1/TIFY10A-GFP and JAZ10pro::JAZ10-GFP suggests that the halted growth of the primary root inversely correlated with expression patterns underlying the defense reaction, which may be of adaptive importance to protect roots against biotic stress. Our data help to unravel the gene expression signatures upon sensing of a highly active bacterial volatile in Arabidopsis seedlings.

Azospirillum brasilense Sp245 triggers cytokinin signaling in root tips and improves biomass accumulation in Arabidopsis through canonical cytokinin receptors.

The plant growth promoting rhizobacterium Azospirillum brasilense Sp245 enhances biomass production in cereals and horticultural species and is an interesting model to study the physiology of the phytostimulation program. Although auxin production by Azospirillum appears to be critical for root architectural readjustments, the role of cytokinins in the growth promoting effects of Azospirillum remains unclear. Here, Arabidopsis thaliana seedlings were co-cultivated in vitro with A. brasilense Sp245 to assess whether direct contact of roots with bacterial colonies or exposure to the bacterial volatiles using divided Petri plates would affect biomass production and root organogenesis. Both interaction types increased root and shoot fresh weight but had contrasting effects on primary root length, lateral root formation and root hair development. Cell proliferation in root meristems analyzed with the CYCB1;1::GUS reporter decreased over time with direct contact, but was augmented by plant exposure to volatiles. Noteworthy, the expression of the cytokinin-inducible reporters TCS::GFP and ARR5::GUS increased in root tips in response to bacterial contact, without being affected by the volatiles. In A. thaliana having single (cre1-12, ahk2-2, ahk3-3), double (cre1-12/ahk2-2, cre1-12/ahk3-3, ahk2-2/ahk3-3) or triple (cre1-12/ahk2-2/ahk3-3) mutations in canonical cytokinin receptors, only the triple mutant had a marked effect on plant growth in response to A. brasilense. These results show that different mechanisms are elicited by A. brasilense, which influence the cytokinin-signaling pathway.

1‐Undecene from Pseudomonas aeruginosa is an olfactory signal for flight‐or‐fight response in Caenorhabditis elegans

Animals possess conserved mechanisms to detect pathogens and to improve survival in their presence by altering their own behavior and physiology. Here, we utilize Caenorhabditis elegans as a model host to ask whether bacterial volatiles constitute microbe‐associated molecular patterns. Using gas chromatography–mass spectrometry, we identify six prominent volatiles released by the bacterium Pseudomonas aeruginosa. We show that a specific volatile, 1‐undecene, activates nematode odor sensory neurons inducing both flight and fight responses in worms. Using behavioral assays, we show that worms are repelled by 1‐undecene and that this aversion response is driven by the detection of this volatile through AWB odor sensory neurons. Furthermore, we find that 1‐undecene odor can induce immune effectors specific to P. aeruginosa via AWB neurons and that brief pre‐exposure of worms to the odor enhances their survival upon subsequent bacterial infection. These results show that 1‐undecene derived from P. aeruginosa serves as a pathogen‐associated molecular pattern for the induction of protective responses in C. elegans.

C4 Bacterial Volatiles Improve Plant Health.

Plant growth-promoting rhizobacteria (PGPR) associated with plant roots can trigger plant growth promotion and induced systemic resistance. Several bacterial determinants including cell-wall components and secreted compounds have been identified to date. Here, we review a group of low-molecular-weight volatile compounds released by PGPR, which improve plant health, mostly by protecting plants against pathogen attack under greenhouse and field conditions. We particularly focus on C4 bacterial volatile compounds (BVCs), such as 2,3-butanediol and acetoin, which have been shown to activate the plant immune response and to promote plant growth at the molecular level as well as in large-scale field applications. We also disc/ uss the potential applications, metabolic engineering, and large-scale fermentation of C4 BVCs. The C4 bacterial volatiles act as airborne signals and therefore represent a new type of biocontrol agent. Further advances in the encapsulation procedure, together with the development of standards and guidelines, will promote the application of C4 volatiles in the field.

Biocontrol Activity of Bacillus megaterium BM344-1 against Toxigenic Fungi.

Mycotoxins are secondary metabolites of some fungal species and represent important contaminants of food and feed. This study aimed to explore the biological control activity of Bacillus megaterium BM344-1 volatile organic compounds (VOCs) on the growth and mycotoxin production of single representatives of the toxigenic species Aspergillus flavus, Aspergillus carbonarius, Penicillium verrucosum, and Fusarium verticillioides. In vitro co-incubation experiments indicated the P. verrucosum isolate as the most sensitive one, with a growth inhibition ratio of 66.7%, followed by A. flavus (29.4%) and F. verticillioides (18.2%). Exposure of A. flavus, P. verrucosum, and F. verticillioides to BM344-1 VOCs resulted in complete inhibition of aflatoxins (AFB1, AFG1, and AFG2), ochratoxin A, and fumonisin B1 (FB1) synthesis on artificial media, respectively. In vivo experiments on maize kernels showed 51% inhibition of fungal growth on ears simultaneously infected with A. flavus spores and exposed to BM344-1 volatiles. Likewise, AF synthesis by A. flavus was significantly (p < 0.05) inhibited (25.34 ± 6.72 μg/kg) by bacterial volatiles as compared to that in control maize ears (91.81 ± 29.10 μg/kg). Gas chromatography–tandem mass spectrometry-based analysis of headspace volatiles revealed hexadecanoic acid methyl ester (palmitic acid) and tetracosane as bioactive compounds in the BM344-1 volatilome. Bacterial volatiles have promising potential to control the growth and mycotoxin synthesis of toxigenic fungi and may present valuable aid in the efforts to warrant food and feed safety.

Endophytic Bacteria from the Sahara Desert Protect Tomato Plants Against Botrytis cinerea Under Different Experimental Conditions.

Bacteria endophytes are living microorganisms that live inside plant tissues without visible harmful symptoms, providing a mutualistic interaction. In this study, different bacterial endophytic strains were isolated from different plants primed to live in an arid area, namely, the Sahara Desert. Up to 27 of these strains were selected based on their ability to inhibit Botrytis cinerea growth in dual-culture assay and by bacterial volatiles. The results presented in this study show the capacity of most of the bacterial strains to protect Solanum lycopersicum against the pathogenic fungus B. cinerea, under different experimental conditions. Five of these strains induced susceptibility in tomato plants and no callose accumulation upon fungal infection, pointing to callose deposition as a protective mechanism mediated by endophytic bacteria. Moreover, there was a significant correlation between the bacterial strains inducing callose and the level of protection against B. cinerea. On the other hand, hormone production by bacteria does not explain the relationship between protection and the differences between the phenotypic results obtained in vitro and those obtained in plant experiments. Induced resistance is highly specific in the inducer-plant-stress interaction.

Bacterial volatile mediated suppression of postharvest anthracnose and quality enhancement in mango

Anthracnose disease incited by Colletotrichum gloeosporioides reduces the yield and market-quality of mangoes worldwide. Previous studies have indicated the potential of diverse antimicrobial volatile organic compounds emitted by the endophytic Pseudomonas putida BP25 for plant disease suppression. In the present study, we have explored native-volatiles of Pseudomonas putida BP25 and a synthetic-volatile, 2-ethyl-5-methylpyrazine, earlier identified in volatilome of Pseudomonas putida BP25 for reducing anthracnose in mango cultivar, Amrapali. The bacterial volatile compounds displayed fungistatic effects on the mycelial growth of Colletotrichum gloeosporioides in vitro. In planta prophylactic fumigation of mangoes with native-volatiles or the bacterial origin synthetic 2-ethyl-5-methylpyrazine for 24 h at 25 °C showed a reduction of anthracnose severity (>76 % reduction over mock) on fruit. Additionally, physicochemical qualities such as total-soluble solids, total-phenols, total-proline, total-carotenoid, total-flavonoid, and fruit-firmness were increased in fumigated fruit as compared to untreated mangoes. Anthracnose suppression coupled with the fruit quality enhancement by bacterial volatiles and synthetic 2-ethyl-5-methylpyrazine presents a new opportunity for postharvest management of mango during the storage, transit, and trade.

Control of damping-off in tomato seedlings exerted by Serratia spp. strains and identification of inhibitory bacterial volatiles in vitro

Serratia marcescens can be a plant growth promoting bacteria (PGPB) and an opportunistic human and plant pathogen. We have identified and characterized strains of related species of Serratia and evaluated their biological control of damping-off of tomato seeds caused by Pythium cryptoirregulare. Serratia ureilytica, S. bockelmannii and S. nevei were identified by phylogenetic analysis of partial gyrB gene sequence and average nucleotide identity (ANI). Tomato seeds inoculated with S. ureilytica ILBB 145 showed higher germination percentage and reduced damping-off in greenhouse experiment resembling a commercial operation, and volatiles produced by this strain caused the nearly complete inhibition in vitro of P. cryptoirregulare. Analysis of volatile organic compounds (VOCs) showed that ILBB 145 produced dimethyl disulfide (DMDS), which can partially account for this inhibition. Serratia bockelmannii ILBB 162 performance against damping-off was intermediate and the inhibition of P. cryptoirregulare in vitro was lower and explained by volatile and diffusible metabolites. Both strains augmented DMDS production in the presence of P. cryptoirregulare, suggesting this compound may play a role in the context of interspecific competition. Serratia nevei ILBB 219 showed the lowest inhibition of P. cryptoirregulare in vitro, no DMDS production, and no biocontrol in planta. Draft genomes of the three strains were annotated and individual genes and biosynthesis gene clusters were identified in relation with the observed phenotypes. We report S. ureilytica – a low risk species- with activity as a biological control agent and DMDS produced by this bacterial species putatively involved in seed and seedling protection against P. cryptoirregulare.

Oviposition-Site Selection of Phlebotomus papatasi (Diptera: Psychodidae) Sand Flies: Attraction to Bacterial Isolates From an Attractive Rearing Medium

Phlebotomine sand flies are worldwide vectors of Leishmania parasites as well as other bacterial and viral pathogens. Due to the variable impact of traditional vector control practices, a more ecologically based approach is needed. The goal of this study was to isolate bacteria from the most attractive substrate to gravid Phlebotomus papatasi Scopoli sand flies and determine the role of bacterial volatiles in the oviposition attractancy of P. papatasi using behavioral assays. We hypothesized that gravid sand flies are attracted to bacterially derived semiochemical cues associated with breeding sites. Bacteria were isolated from a larvae-conditioned rearing medium, previously shown to be highly attractive to sand flies. The isolated bacteria were identified by amplifying and sequencing 16S rDNA gene fragments, and 12 distinct bacterial species were selected for two-choice olfactometer bioassays. The mix of 12 bacterial isolates elicited strong attraction at the lower concentration of 107 cells per ml and significant repellence at a high concentration of 109 cells per ml. Three individual isolates (SSI-2, SSI-9, and SSI-11) were particularly attractive at low doses. In general, we observed dose-related effects, with some bacterial isolates stimulating negative and some positive dose-response curves in sand fly attraction. Our study confirms the important role of saprophytic bacteria, gut bacteria, or both, in guiding the oviposition-site selection behavior of sand flies. Identifying the specific attractive semiochemical cues that they produce could lead to development of an attractive lure for surveillance and control of sand flies.

Identification and application of bacterial volatiles to attract a generalist aphid parasitoid: from laboratory to greenhouse assays.

Recent studies have shown that microorganisms emit volatile compounds that affect insect behaviour. However, it remains largely unclear whether microbes can be exploited as a source of attractants to improve biological control of insect pests. In this study, we used a combination of coupled gas chromatography-electroantennography (GC-EAG) and Y-tube olfactometer bioassays to identify attractive compounds in the volatile extracts of three bacterial strains that are associated with the habitat of the generalist aphid parasitoid Aphidius colemani, and to create mixtures of synthetic compounds to find attractive blends for A. colemani. Subsequently, the most attractive blend was evaluated in two-choice cage experiments under greenhouse conditions. GC-EAG analysis revealed 20 compounds that were linked to behaviourally attractive bacterial strains. A mixture of two EAG-active compounds, styrene and benzaldehyde applied at a respective dose of 1 μg and 10 ng, was more attractive than the single compounds or the culture medium of the bacteria in Y-tube olfactometer bioassays. Application of this synthetic mixture under greenhouse conditions resulted in significant attraction of the parasitoids, and outperformed application of the bacterial culture medium. Compounds isolated from bacterial blends were capable of attracting parasitoids both in laboratory and greenhouse assays, indicating that microbial cultures are an effective source of insect attractants. This opens new opportunities to attract and retain natural enemies of pest species and to enhance biological pest control.

An invasive beetle\u2013fungus complex is maintained by fungal nutritional-compensation mediated by bacterial volatiles

Mutualisms between symbiotic microbes and animals have been well documented, and nutritional relationships provide the foundation for maintaining beneficial associations. The well-studied mutualism between bark beetles and their fungi has become a classic model system in the study of symbioses. Despite the nutritional competition between bark beetles and beneficial fungi in the same niche due to poor nutritional feeding substrates, bark beetles still maintain mutualistic associations with beneficial fungi over time. The mechanism behind this phenomenon, however, remains largely unknown. Here, we demonstrated the bark beetle Dendroctonus valens LeConte relies on the symbiotic bacterial volatile ammonia, as a nitrogen source, to regulate carbohydrate metabolism of its mutualistic fungus Leptographium procerum to alleviate nutritional competition, thereby maintaining the stability of the bark beetle–fungus mutualism. Ammonia significantly reduces competition of L. procerum for carbon resources for D. valens larval growth and increases fungal growth. Using stable isotope analysis, we show the fungus breakdown of phloem starch into d-glucose by switching on amylase genes only in the presence of ammonia. Deletion of amylase genes interferes with the conversion of starch to glucose. The acceleration of carbohydrate consumption and the conversion of starch into glucose benefit this invasive beetle–fungus complex. The nutrient consumption–compensation strategy mediated by tripartite beetle–fungus–bacterium aids the maintenance of this invasive mutualism under limited nutritional conditions, exacerbating its invasiveness with this competitive nutritional edge.

A Multiple Approach Combined with Portable Electronic Nose for Assessment of Post-harvest Sapota Contamination by Foodborne Pathogens

Sapota (Manilkara Zapota) belongs to the Sapotaceae family and is one of the sweetest, delicious, and popular tropical fruits with high nutritional benefits. Due to its high water content (78%), it is easily damaged, which in turn reduces its shelf life. Also, significant post-harvest losses occur due to insufficient storage, poor handling, and transportation conditions, which also increase the microbial or pathogenic contamination. In this context, we have deployed an electronic nose for the first time to rapidly discriminate fresh sapota from bacterial contaminated sapota samples. The electronic nose response was varied from 0.1 to 2.9 V for sapota samples studied from day 1 to day 6. Fourier transform infrared spectra showed a gradual increase in the intensities of OH and H-O-H peaks at 3320 cm−1 and 1636 cm−1for these samples. Also, Staphylococcus bacteria were detected for the same set of sapota samples. Headspace GC-MS results of day 2, day 4, and day 6 sapota samples revealed the increasing trend in bacterial volatiles, namely ethanol, acetaldehyde, and isopropyl alcohol. The sensor data were provided as input to principal component analysis (PCA), Ward’s method analysis, and response surface methodology (RSM) for distinguishing fresh, half-contaminated, and fully contaminated sapota samples.

Long-Chain Hydrocarbons (C21, C24, and C31) Released by Bacillus sp. MH778713 Break Dormancy of Mesquite Seeds Subjected to Chromium Stress

Volatile organic compounds (VOCs) produced by rhizobacteria have been proven to stimulate plant growth during germination and seedling stages. However, the modulating effect of bacterial volatiles on the germination of seeds subjected to heavy metal stress is scarcely studied. In this work, the ability of volatiles released by Bacillus sp. MH778713 to induce seed dormancy breakage in Prosopis laevigata and Arabidopsis thaliana seeds were examined. The minimal inhibitory concentration of chromium (Cr) VI that prevents seed germination of P. laevigata and A. thaliana on water-Cr-agar plates was 2500 and 100 mg L–1, respectively. Remarkably, partitioned Petri-dish co-cultivation of Bacillus sp. MH778713 and plant seeds under Cr-stress showed the beneficial effect of volatiles emitted by Bacillus sp. MH778713, helping plant seeds to overcome Cr-stress. Among the metabolites emitted by Bacillus sp. MH778713, octadecane, heneicosane, 2,4-di-tert-butylphenol, hexadecane, eicosane, octacosane, and tetratriacontane were the most abundant. To confirm that these long-chain compounds produced by Bacillus sp. MH778713 could be responsible for the seed dormancy breakage, high pure organic compounds (2,4-di-tert-butylphenol, heneicosane, hentriacontane, and tetracosane) were used directly in germination assays of P. laevigata and A. thaliana seeds instead of volatiles emitted by Bacillus sp. MH778713. All organic compounds allowed Prosopis and Arabidopsis seeds to overcome Cr-toxicity and germinate. The results of this study provide new insight into the role of long-chain bacterial compounds produced by Bacillus sp. MH778713 as triggers of seed abiotic stress tolerance, surmounting chromium stress and stimulating seedling development.

Interaction with Soil Bacteria Affects the Growth and Amino Acid Content of Piriformospora indica.

Exploration of the effect of soil bacteria on growth and metabolism of beneficial root endophytic fungi is relevant to promote favorable associations between microorganisms of the plant rhizosphere. Hence, the interaction between the plant-growth-promoting fungus Piriformospora indica and different soil bacteria was investigated. The parameters studied were fungal growth and its amino acid composition during the interaction. Fungus and bacteria were confronted in dual cultures in Petri dishes, either through agar or separated by a Perspex wall that only allowed the bacterial volatiles to be effective. Fungal growth was stimulated by Azotobacter chroococcum, whereas Streptomyces anulatus AcH 1003 inhibited it and Streptomyces sp. Nov AcH 505 had no effect. To analyze amino acid concentration data, targeted metabolomics was implemented under supervised analysis according to fungal-bacteria interaction and time. Orthogonal partial least squares-discriminant analysis (OPLS-DA) model clearly discriminated P. indica–A. chroococcum and P. indica–S. anulatus interactions, according to the respective score plot in comparison to the control. The most observable responses were in the glutamine and alanine size groups: While Streptomyces AcH 1003 increased the amount of glutamine, A. chroococcum decreased it. The fungal growth and the increase of alanine content might be associated with the assimilation of nitrogen in the presence of glucose as a carbon source. The N-fixing bacterium A. chroococcum should stimulate fungal amino acid metabolism via glutamine synthetase-glutamate synthase (GS-GOGAT). The data pointed to a stimulated glycolytic activity in the fungus observed by the accumulation of alanine, possibly via alanine aminotransferase. The responses toward the growth-inhibiting Streptomyces AcH 1003 suggest an (oxidative) stress response of the fungus.