Biocontrol Potential of Trichoderma Ghanense and Trichoderma Citrinoviride toward Pythium aphanidermatum.

Pythium-induced damping-off of cucumber is a major constraint to cucumber production in different parts of the world. Although chemical fungicides are used for managing this disease, they have many drawbacks to the environment. The ability of the antagonistic fungi isolated from the rhizosphere and endosphere of Dactyloctenium robecchii and Moraea sisyrinchium in the control of soilborne pathogen Pythium aphanidermatum was inspected. Native Trichoderma isolates, Trichoderma ghanense and Trichoderma citrinoviride, were isolated from plant stem and soil samples collected from Al-Seeb, Oman. Using a dual culture technique, the antagonistic activity of the fungal isolates against P. aphanidermatum was examined in vitro. Among Trichoderma isolates, T. ghanense was more efficient in restraining the mycelial growth of P. aphanidermatum, causing an inhibition percentage of 44.6%. Further, T. citrinoviride induced significantly lower cessation of P. aphanidermatum mycelial growth (31.3%). Microscopic and electrolyte leakage inspection of the pathogen mycelia depicted extreme morphological malformations in their mycelium, which can be attributed to the antifungal metabolites of antagonists. Greenhouse studies demonstrated the effectivity of T. ghanense in controlling Pythium damping-off of cucumber plants, where the number of surviving plants was over 90% when the biocontrol agents were used compared to 0 in the control plants. Furthermore, treatment of the plants with the antagonists promoted growth characteristics of plants compared to uninoculated plants. This included improvements in shoot and root lengths, leaf length and width, and dry weight. These findings suggest that T. ghanense and T. citrinoviride can be developed as alternatives to synthetic chemical fungicides to manage soilborne pathogens of cucumber. This research is also the first to clarify the biocontrol ability of T. citrinoviride and T. ghanense against cucumber damping-off caused by P. aphanidermatum.

Plant-Disease-Suppressive and Growth-Promoting Activities of Endophytic and Rhizobacterial Isolates Associated with Citrullus colocynthis.

This study was conducted to investigate the antagonistic potential of endophytic and rhizospheric bacterial isolates obtained from Citrullus colocynthis in suppressing Fusarium solani and Pythium aphanidermatum and promoting the growth of cucumber. Molecular identification of bacterial strains associated with C. colocynthis confirmed that these strains belong to the Achromobacter, Pantoea, Pseudomonas, Rhizobium, Sphingobacterium, Bacillus, Sinorhizobium, Staphylococcus, Cupriavidus, and Exiguobacterium genera. A dual culture assay showed that nine of the bacterial strains exhibited antifungal activity, four of which were effective against both pathogens. Strains B27 (Pantoea dispersa) and B28 (Exiguobacterium indicum) caused the highest percentage of inhibition towards F. solani (48.5% and 48.1%, respectively). P. aphanidermatum growth was impeded by the B21 (Bacillus cereus, 44.7%) and B28 (Exiguobacterium indicum, 51.1%) strains. Scanning electron microscopy showed that the strains caused abnormality in phytopathogens' mycelia. All of the selected bacterial strains showed good IAA production (>500 ppm). A paper towel experiment demonstrated that these strains improved the seed germination, root/shoot growth, and vigor index of cucumber seedlings. Our findings suggest that the bacterial strains from C. colocynthis are suppressive to F. solani and P. aphanidermatum and can promote cucumber growth. This appears to be the first study to report the efficacy of these bacterial strains from C. colocynthis against F. solani and P. aphanidermatum.

Effects of the Rhizosphere Fungus Cunninghamella bertholletiae on the Solanum lycopersicum Response to Diverse Abiotic Stresses.

This study examined the efficiency of fungal strain (Cunninghamella bertholletiae) isolated from the rhizosphere of Solanum lycopersicum to reduce symptoms of salinity, drought and heavy metal stresses in tomato plants. In vitro evaluation of C. bertholletiae demonstrated its ability to produce indole-3-Acetic Acid (IAA), ammonia and tolerate varied abiotic stresses on solid media. Tomato plants at 33 days' old, inoculated with or without C. bertholletiae, were treated with 1.5% sodium chloride, 25% polyethylene glycol, 3 mM cadmium and 3 mM lead for 10 days, and the impact of C. bertholletiae on plant performance was investigated. Inoculation with C. bertholletiae enhanced plant biomass and growth attributes in stressed plants. In addition, C. bertholletiae modulated the physiochemical apparatus of stressed plants by raising chlorophyll, carotenoid, glucose, fructose, and sucrose contents, and reducing hydrogen peroxide, protein, lipid metabolism, amino acid, antioxidant activities, and abscisic acid. Gene expression analysis showed enhanced expression of SlCDF3 and SlICS genes and reduced expression of SlACCase, SlAOS, SlGRAS6, SlRBOHD, SlRING1, SlTAF1, and SlZH13 genes following C. bertholletiae application. In conclusion, our study supports the potential of C. bertholletiae as a biofertilizer to reduce plant damage, improve crop endurance and remediation under stress conditions.

Actinomucor elegans and Podospora bulbillosa Positively Improves Endurance to Water Deficit and Salinity Stresses in Tomato Plants.

Fungal strains isolated from the rhizosphere of healthy Solanum lycopersicum were examined to mitigate symptoms of drought and salinity stresses. The fungal strains were identified as Actinomucor elegans and Podospora bulbillosa based on their DNA sequencing and morphological analysis. Additionally, the fungal strains were assayed for a number of plant growth promoting traits and abiotic stresses on solid media. Moreover, a greenhouse experiment was conducted and tomato seedlings were treated with 25% PEG or 1.5% NaCl for 12 days, and the impact of plant growth promoting fungi (PGPF) on tomato seedling performance under these conditions was examined. PGPF application raised the survival of the stressed tomato plants, which was evidenced by higher physiological and biochemical processes. The PGPF-inoculated plants exhibited higher chlorophyll, carotenoid, protein, amino acid, antioxidant activities, salicylic acid, glucose, fructose, and sucrose contents, and showed lower hydrogen peroxide, and lipid metabolism relative to control plants under stress. Analysis using gene expression showed enhanced expression of SlF3H gene and reduced expression of SlNCED1, SlDEAD31, SlbZIP38, and SlGRAS10 genes following PGPFs application. Overall, the outcomes of this study elucidate the function of these fungal strains and present candidates with potential implementation as biofertilizers and in promoting plant stress endurance.

Fabrication of Novel Agrowaste (Banana and Potato Peels)-Based Biochar/TiO2 Nanocomposite for Adsorption of Cr(VI), Statistical Optimization via RSM Approach.

In this research work, a simple, efficient, and eco-friendly procedure for the biosorption of Cr(VI) ions was studied. A detailed comparative study was performed to check the adsorption efficiency of agrowaste (banana and potato peels)-based adsorbents. Firstly, mixed biosorbent was washed, dried and ground into powder, secondly, biosorbent was pyrolyzed into biochar and thirdly TiO2 nanocomposite (TiO2 NC) biosorbent was made by sonicating using prepared biochar and TiO2 NPs. Titanium dioxide nanoparticles (TiO2 NPs) were synthesized by a green method using Psidium guajava leaf extract. The synthesized adsorbents were characterized by SEM, EDX FT-IR, XRD and UV-visible analysis. The effect of four different factors, i.e., pH of the synthetic metallic solution, time, concentration and adsorbent dosage was studied. The optimum conditions were time (120 min), pH (3), concentration (10 ppm) and adsorbent dosage (1.0 g). The kinetic modeling showed that the adsorption of Cr(VI) ion follows a pseudo second-order mechanism and the Langmuir isotherm model was found to fit better for this study. Response surface methodology (RSM)-based optimized parameters provided optimal parameter sets that better represent the adsorption rate models. The uptake capacity of Cr(VI) from aqueous solution was found to be biomass (76.49 mg/L) ˂ biochar (86.51 mg/L) ˂ TiO2 NC (92.89 mg/L). It can be suggested that the produced TiO2 NC could possibly be an efficient biosorbent for the removal of Cr(IV).

Salvianolic Acid Modulates Physiological Responses and Stress-Related Genes That Affect Osmotic Stress Tolerance in Glycine max and Zea mays.

Drought is a serious threat worldwide to soybean and maize production. This study was conducted to discern the impact of salvianolic acid treatment on osmotic-stressed soybean (Glycine max L.) and maize (Zea mays L.) seedlings from the perspective of physiochemical and molecular reactions. Examination of varied salvianolic acid concentrations (0, 0.1, 1, 5, 10, and 25 µM) on soybean and maize seedling growth confirmed that the 0.1 and 1 µM concentrations, respectively, showed an improvement in agronomic traits. Likewise, the investigation ascertained how salvianolic acid application could retrieve osmotic-stressed plants. Soybean and maize seedlings were irrigated with water or 25% PEG for 8 days. The results indicated that salvianolic acid application promoted the survival of the 39-day-old osmotic-stressed soybean and maize plants. The salvianolic acid-treated plants retained high photosynthetic pigments, protein, amino acid, fatty acid, sugar, and antioxidant contents, and demonstrated low hydrogen peroxide and lipid contents under osmotic stress conditions. Gene transcription pattern certified that salvianolic acid application led to an increased expression of GmGOGAT, GmUBC2, ZmpsbA, ZmNAGK, ZmVPP1, and ZmSCE1d genes, and a diminished expression of GmMIPS2, GmSOG1, GmACS, GmCKX, ZmPIS, and ZmNAC48 genes. Together, our results indicate the utility of salvianolic acid to enhance the osmotic endurance of soybean and maize plants.

Ampelopsin Confers Endurance and Rehabilitation Mechanisms in Glycine max cv. Sowonkong under Multiple Abiotic Stresses.

The present investigation aims to perceive the effect of exogenous ampelopsin treatment on salinity and heavy metal damaged soybean seedlings (Glycine max L.) in terms of physiochemical and molecular responses. Screening of numerous ampelopsin concentrations (0, 0.1, 1, 5, 10 and 25 µM) on soybean seedling growth indicated that the 1 µM concentration displayed an increase in agronomic traits. The study also determined how ampelopsin application could recover salinity and heavy metal damaged plants. Soybean seedlings were irrigated with water, 1.5% NaCl or 3 mM chosen heavy metals for 12 days. Our results showed that the application of ampelopsin raised survival of the 45-day old salinity and heavy metal stressed soybean plants. The ampelopsin treated plants sustained high chlorophyll, protein, amino acid, fatty acid, salicylic acid, sugar, antioxidant activities and proline contents, and displayed low hydrogen peroxide, lipid metabolism, and abscisic acid contents under unfavorable status. A gene expression survey revealed that ampelopsin application led to the improved expression of GmNAC109, GmFDL19, GmFAD3, GmAPX, GmWRKY12, GmWRKY142, and GmSAP16 genes, and reduced the expression of the GmERF75 gene. This study suggests irrigation with ampelopsin can alleviate plant damage and improve plant yield under stress conditions, especially those including salinity and heavy metals.

Biocontrol Potential of Bacillus amyloliquefaciens against Botrytis pelargonii and Alternaria alternata on Capsicum annuum.

The aim of this study was to assess the ability of Bacillus amyloliquefaciens, to augment plant growth and suppress gray mold and leaf spot in pepper plants. Morphological modifications in fungal pathogen hyphae that expanded toward the PGPR colonies were detected via scanning electron microscope. Furthermore, preliminary screening showed that PGPR could produce various hydrolytic enzymes in its media. Treatments with B. amyloliquefaciens suppressed Botrytis gray mold and Alternaria leaf spot diseases on pepper caused by Botrytis pelargonii and Alternaria alternata, respectively. The PGPR strain modulated plant physio-biochemical processes. The inoculation of pepper with PGPR decreased protein, amino acid, antioxidant, hydrogen peroxide, lipid peroxidation, and abscisic acid levels but increased salicylic acid and sugar levels compared to those of uninoculated plants, indicating a mitigation of the adverse effects of biotic stress. Moreover, gene expression studies confirmed physio-biochemical findings. PGPR inoculation led to increased expression of the CaXTH genes and decreased expression of CaAMP1, CaPR1, CaDEF1, CaWRKY2, CaBI-1, CaASRF1, CaSBP11, and CaBiP genes. Considering its beneficial effects, the inoculation of B. amyloliquefaciens can be proposed as an eco-friendly alternative to synthetic chemical fungicides.

Rhizospheric Bacillus amyloliquefaciens Protects Capsicum annuum cv. Geumsugangsan From Multiple Abiotic Stresses via Multifarious Plant Growth-Promoting Attributes.

Plant growth-promoting rhizobacteria (PGPR) are beneficial microorganisms that can be utilized to improve plant responses against biotic and abiotic stresses. In this study, we investigated whether PGPR (Bacillus amyloliquefaciens) isolated from the endorhizosphere of Sasamorpha borealis have the potential to sustain pepper growth under drought, salinity, and heavy metal stresses. The bacterial strain was determined based on 16S rDNA and gyrB gene sequencing and characterized based on the following biochemical traits: nitrogen fixation; 1-aminocyclopropane-1-carboxylate deaminase activity; indole acetic acid production; inorganic phosphate, potassium, zinc, and silicon solubilization; and siderophore production. Various abiotic stresses were applied to 28-day-old pepper seedlings, and the influence of the PGPR strain on pepper seedling growth under these stress conditions was evaluated. The application of PGPR improved survival of the inoculated pepper plants under stress conditions, which was reflected by higher seedling growth rate and improved physiochemical traits. The PGPR-treated plants maintained high chlorophyll, salicylic acid, sugar, amino acid, and proline contents and showed low lipid metabolism, abscisic acid, protein, hydrogen peroxide contents, and antioxidant activities under stress conditions. Gene expression studies confirmed our physiological and biochemical findings. PGPR inoculation led to enhanced expression of XTH genes and reduced expression of WRKY2, BI-1, PTI1, and binding immunoglobulin protein (BiP) genes. We conclude that the PGPR strain described in this study has great potential for use in the phytoremediation of heavy metals and for enhancing pepper plant productivity under stress conditions, particularly those involving salinity and drought.

Melatonin Enhances the Tolerance and Recovery Mechanisms in Brassica juncea (L.) Czern. Under Saline Conditions.

Melatonin has been recently known to stimulate plant growth and induce protective responses against different abiotic stresses. However, the mechanisms behind exogenous melatonin pretreatment and restoration of plant vigor from salinity stress remain poorly understood. The present study aimed to understand the effects of exogenous melatonin pretreatment on salinity-damaged green mustard (Brassica juncea L. Czern.) seedlings in terms of oxidative stress regulation and endogenous phytohormone production. Screening of several melatonin concentrations (0, 0.1, 1, 5, and 10 µM) on mustard growth showed that the 1 µM concentration revealed an ameliorative increase of plant height, leaf length, and leaf width. The second study aimed at determining how melatonin application can recover salinity-damaged plants and studying its effects on physiological and biochemical parameters. Under controlled environmental conditions, mustard seedlings were irrigated with distilled water or 150 mM of NaCl for 7 days. This was followed by 1 µM of melatonin application to determine its recovery impact on the damaged plants. Furthermore, several physiological and biochemical parameters were examined in stressed and unstressed seedlings with or without melatonin application. Our results showed that plant height, leaf length/width, and stem diameter were enhanced in 38-day-old salinity-stressed plants under melatonin treatment. Melatonin application obviously attenuated salinity-induced reduction in gas exchange parameters, relative water content, and amino acid and protein levels, as well as antioxidant enzymes, such as superoxide dismutase and catalase. H2O2 accumulation in salinity-damaged plants was reduced by melatonin treatment. A decline in abscisic acid content and an increase in salicylic acid content were observed in salinity-damaged seedlings supplemented with melatonin. Additionally, chlorophyll content decreased during the recovery period in salinity-damaged plants by melatonin treatment. This study highlighted, for the first time, the recovery impact of melatonin on salinity-damaged green mustard seedlings. It demonstrated that exogenous melatonin supplementation significantly improved the physiologic and biochemical parameters in salinity-damaged green mustard seedlings.

Endophytic Enterobacter cloacae exhibits antagonistic activity against Pythium damping-off of cucumber / Enterobacter cloacae endofítica exibe atividade antagonista contra a podridão de pepino causada por Pythium

ABSTRACT: The study was performed to examine the potential presence of biological control agents against Pythium damping-off disease of cucumber.Examining eleven bacterial strains isolated from acid lime roots and rhizosphere soil showed that the bacterial strain RB1 was the most efficient in suppressing mycelial growth of P. aphanidermatum, producing an inhibition zone of 5mm. Scanning electron microscopy study of the mycelia at the interaction zone showed that the pathogen hyphae were deformed and shriveled by the bacterial strain.In pot experiments, pretreatment with the RB1 bacterial strain reduced disease incidence significantly by 63%.The bacterial strain did not exhibit any negative significant effects on cucumber growth (plant height and root dry weight) in comparison with untreated control under growth chamber conditions. Molecular identification of strain RB1 based on the 16S rRNA gene revealed that it is Enterobacter cloacae. Findings from this study suggested that E. cloacae has a potential to be used as a biocontrol agent for suppressingcucumber damping-off disease caused by P. aphanidermatum. This is the first report of the antagonistic activity of E. cloacae against P. aphanidermatum-induced damping-off of cucumber.

RESUMO: O estudo foi realizado para examinar a presença potencial de agentes no controle biológico da doença do apodrecimento do pepino causado por Pythium. Examinando onze cepas bacterianas isoladas de raízes de cal ácida e solo da rizosfera mostraram que a cepa bacteriana RB1 foi a mais eficiente na supressão do crescimento micelial de P. aphanidermatum, produzindo uma zona de inibição de 5 mm. O estudo de microscopia eletrônica de varredura dos micélios na zona de interação mostrou que as hifas do patógeno foram deformadas e enrugadas pela cepa bacteriana. Em experimentos com vasos, o pré-tratamento com a cepa bacteriana RB1 reduziu significativamente a incidência da doença em 63%. A cepa bacteriana não exibiu nenhum efeito negativo. Efeitos significativos no crescimento do pepino (altura da planta e peso seco da raiz), em comparação com o controle não tratado sob condições da câmara de crescimento. A identificação molecular da cepa RB1 com base no gene 16S rRNA revelou que é a Enterobacter cloacae. Os resultados deste estudo sugerem que E. cloacae tem potencial para ser usado como agente de biocontrole para suprimir a doença da podridão de pepino causada por P. aphanidermatum. Este é o primeiro relato da atividade antagônica de E. cloacae contra o amortecimento induzido por P. aphanidermatum de pepino.