Fungal Pathogen Rhizoctonia Solani Research Articles

Evaluation of Sheath Blight Resistance in Diverse Rice Varieties Under Artificial Inoculation and Field Conditions

Rice (Oryza sativa L.) a vital food crop for nearly half of the global population, facing significant threats from various biotic and abiotic stresses. Among biotic stresses, diseases pose the most common challenge. Sheath blight, caused by soil-borne fungal pathogen Rhizoctonia Solani Kühn (R. solani), is a major concern, leading to substantial damage to rice and cereal crops worldwide. This study evaluated sheath blight resistance across diverse rice varieties, including short grain, long grain, aromatic, hybrid, and wild types, under both artificial inoculation and field conditions. Ten host plant species were assessed for resistance to sheath blight caused by Rhizoctonia Solani, using physiological screening one-week post-inoculation. The highly virulent R. solani isolate (AG-1 IA anastomosis group) was cultured on PDA media, covering the entire plate 3 to 5 days. Screening of cultivated rice varieties, was conducted using field conditions and humidified chamber methods, with results confirmed through molecular characterization. Among the ten cultivated rice varieties, Arize 6444 and Dhanya 748 exhibited the lowest disease index, classifying them as moderately resistant. In contrast, Kalanamak and Pusa Basmati were highly susceptible, displaying high disease indices. Among wild rice accessions, Oryza rufipogon showed a disease index of five or less. Visual ratings indicated the highest susceptibility in Kalanamak, followed by Pusa Basmati and Swarna sub-1, while the lowest visual ratings were observed in wild rice accessions Oryza australiensis and Oryza rufipogon across all tested hosts. The findings underscore the importance of identifying resistant rice varieties to manage sheath blight effectively. The moderately resistant cultivars and wild rice accessions identified in this study offer valuable genetic resources for breeding programs aimed at enhancing sheath blight resistance in rice.

Order of arrival and nutrient supply alter outcomes of co-infection with two fungal pathogens.

A pathogen arriving on a host typically encounters a diverse community of microbes that can shape priority effects, other within-host interactions and infection outcomes. In plants, environmental nutrients can drive trade-offs between host growth and defence and can mediate interactions between co-infecting pathogens. Nutrients may thus alter the outcome of pathogen priority effects for the host, but this possibility has received little experimental investigation. To disentangle the relationship between nutrient availability and co-infection dynamics, we factorially manipulated the nutrient availability and order of arrival of two foliar fungal pathogens (Rhizoctonia solani and Colletotrichum cereale) on the grass tall fescue (Lolium arundinaceum) and tracked disease outcomes. Nutrient addition did not influence infection rates, infection severity or plant biomass. Colletotrichum cereale facilitated R. solani, increasing its infection rate regardless of their order of inoculation. Additionally, simultaneous and C. cereale-first inoculations decreased plant growth and-in plants that did not receive nutrient addition-increased leaf nitrogen concentrations compared to uninoculated plants. These effects were partially, but not completely, explained by the duration and severity of pathogen infections. This study highlights the importance of understanding the intricate associations between the order of pathogen arrival, host nutrient availability and host defence to better predict infection outcomes.

Research Progress on Diseases Caused by the Soil-Borne Fungal Pathogen Rhizoctonia solani in Alfalfa

Rhizoctonia solani is a soil-borne fungal species with worldwide distribution and poses serious threats to a wide range of economically important crops such as grain and forage crops. This pathogen has survival capabilities within plants and soil, giving rise to sclerotia and persisting for several months or years. Alfalfa (Medicago sativa) is the most widely grown and important forage crop in China and worldwide. The unique characteristics of alfalfa, such as excellent forage quality, ruminant desirability, and substantial biomass output, distinguish it from other fodder crops, and it is also known as the “Queen of Forages”. However, the production of alfalfa is seriously affected by R. solani, with yield losses of 20% to 60% globally. This review firstly summarizes diseases such as seedling damping-off, root rot, crown rot, root cankers, stem cankers, blight, and stem rot caused by R. solani in alfalfa and the survival mechanism of this pathogen. The techniques for R. solani detection and quantification from plants and soils, as well as management through host resistance, cultural practices, fungicides, and biological control, were then overviewed. This review provides scientific knowledge to enable researchers to efficiently manage R. solani in alfalfa production.

In vitro and in vivo antifungal efficacy of individual and consortium Bacillus strains in controlling potato black scurf and possible development of spore-based fungicide

Potato is a major crop whose yield and quality are impacted by the pathogenic fungus Rhizoctonia solani. Here, a control approach based on the use of endophytic bacillus spores and bacterial consortia was assessed as an innovative solution to improve tuber productivity and health. Endophytic potato bacteria extracted from several organs (leaves, stems and roots) evaluated for their antagonistic activity against R. solani by the double culture technique and their ability to rot potato slices. Additionally, the ability of Bacillus strains to generate hydrolytic enzymes and produce lipopeptides (using ESI-MS/MS mass spectrometry) was assessed. Then, the effectiveness of selected bacterial strains, either individually or in consortium (after carrying out compatibility tests) in the suppression of R. solani was evaluated in the greenhouse. Six endophytes not affecting potato tubers and having an antagonistic effect against R. solani were identified: B. halotolerans SpS5, B. velezensis KnL15, B. aryabhattai FaR1, B. amyloliquefaciens LiR9, B. haynesii ReR10 which produce several types and combinations of lipopeptides belonging to different families (iturins, fengycins and surfactins) as well as hydrolytic enzymes (chitinase, protease and cellulase). The vegetative and sporulating strain SpS5, as well as the 5 other Bacillus and their Sps5-based consortia, were found to be very effective in increasing the yield of healthy, high-quality tubers and reducing disease severity. Based on the results mentioned above, a microbial fungicide from the spores of (i) B. halotolerans SpS5 for which we sequenced the whole genome or (ii) bacterial consortia could be developed to combat black scurf potato and other fungal infections.

OsEIL2 balances rice immune responses against (hemi)biotrophic and necrotrophic pathogens via the salicylic acid and jasmonic acid synergism.

Plants typically activate distinct defense pathways against various pathogens. Heightened resistance to one pathogen often coincides with increased susceptibility to another pathogen. However, the underlying molecular basis of this antagonistic response remains unclear. Here, we demonstrate that mutants defective in the transcription factor ETHYLENE-INSENSITIVE 3-LIKE 2 (OsEIL2) exhibited enhanced resistance to the biotrophic bacterial pathogen Xanthomonas oryzae pv oryzae and to the hemibiotrophic fungal pathogen Magnaporthe oryzae, but enhanced susceptibility to the necrotrophic fungal pathogen Rhizoctonia solani. Furthermore, necrotroph-induced OsEIL2 binds to the promoter of OsWRKY67 with high affinity, leading to the upregulation of salicylic acid (SA)/jasmonic acid (JA) pathway genes and increased SA/JA levels, ultimately resulting in enhanced resistance. However, biotroph- and hemibiotroph-induced OsEIL2 targets OsERF083, resulting in the inhibition of SA/JA pathway genes and decreased SA/JA levels, ultimately leading to reduced resistance. Our findings unveil a previously uncharacterized defense mechanism wherein two distinct transcriptional regulatory modules differentially mediate immunity against pathogens with different lifestyles through the transcriptional reprogramming of phytohormone pathway genes.

Development of activation-tagged gain-of-functional mutants in indica rice line (BPT 5204) for sheath blight resistance.

The development of sheath blight (ShB) resistance varieties has been a challenge for scientists for long time in rice. Activation tagging is an efficient gain-of-function mutation approach to create novel phenotypes and to identify their underlying genes. In this study, a mutant population was developed employing activation tagging in the recalcitrant indica rice (Oryza sativa L.) cv. BPT 5204 (Samba Mahsuri) through activation tagging. In this study, we have generated more than 1000 activation tagged lines in indica rice, from these mutant population 38 (GFP- RFP+) stable Ds plants were generated through germinal transposition at T2 generation based on molecular analysis and seeds selected on hygromycin (50mg/L) containing medium segregation analyses confirmed that the transgene inherited as mendelian segregation ratio of 3:1 (3 resistant: 1 susceptible). Of them, five stable activation tagged Ds lines (M-Ds-1, M-Ds-2, M-Ds-3, M-Ds-4 and M-Ds-5) were selected based on phenotypic observation through screening for sheath blight (ShB) resistance caused by fungal pathogen Rhizoctonia solani (R. solani),. Among them, M-Ds-3 and M-Ds-5 lines showed significant resistance for ShB over other tagged lines and wild type (WT) plants. Furthermore, analysed for launch pad insertion through TAIL-PCR results and mapped on corresponding rice chromosomes. Flanking sequence and gene expression analysis revealed that the upregulation of glycoside hydrolase-OsGH or similar to Class III chitinase homologue (LOC_Os08g40680) in M-Ds-3 and a hypothetical protein gene (LOC_Os01g55000) in M-Ds-5 are potential candidate genes for sheath blight resistance in rice. In the present study, we developed Ac-Ds based ShB resistance gain-of-functional mutants through activation tagging in rice. These activation tagged mutant lines can be excellent sources for the development of ShB resistant cultivars in rice.

Genome analysis and hyphal movement characterization of the hitchhiker endohyphal Enterobacter sp. from Rhizoctonia solani.

Bacterial-fungal interactions are pervasive in the rhizosphere. While an increasing number of endohyphal bacteria have been identified, little is known about their ecology and impact on the associated fungal hosts and the surrounding environment. In this study, we characterized the genome of an Enterobacter sp. Crenshaw (En-Cren), which was isolated from the generalist fungal pathogen Rhizoctonia solani, and examined the genetic potential of the bacterium with regard to the phenotypic traits associated with the fungus. Overall, the En-Cren genome size was typical for members of the genus and was capable of free-living growth. The genome was 4.6 MB in size, and no plasmids were detected. Several prophage regions and genomic islands were identified that harbor unique genes in comparison with phylogenetically closely related Enterobacter spp. Type VI secretion system and cyanate assimilation genes were identified from the bacterium, while some common heavy metal resistance genes were absent. En-Cren contains the key genes for indole-3-acetic acid (IAA) and phenylacetic acid (PAA) biosynthesis, and produces IAA and PAA in vitro, which may impact the ecology or pathogenicity of the fungal pathogen in vivo. En-Cren was observed to move along hyphae of R. solani and on other basidiomycetes and ascomycetes in culture. The bacterial flagellum is essential for hyphal movement, while other pathways and genes may also be involved.IMPORTANCEThe genome characterization and comparative genomics analysis of Enterobacter sp. Crenshaw provided the foundation and resources for a better understanding of the ecology and evolution of this endohyphal bacteria in the rhizosphere. The ability to produce indole-3-acetic acid and phenylacetic acid may provide new angles to study the impact of phytohormones during the plant-pathogen interactions. The hitchhiking behavior of the bacterium on a diverse group of fungi, while inhibiting the growth of some others, revealed new areas of bacterial-fungal signaling and interaction, which have yet to be explored.

Isolation, Characterization, and Screening of Endospore Forming Endophytic Bacteria from Cowpea [Vigna unguiculata (L.) Walp.] for Plant Growth Promotion and Abiotic Stress Mitigation

We performed isolation and characterization of endospore forming endophytic bacteria (EEB) from endorhizospheric region of healthy bush cowpea var. Bhagyalakshmi. All the EEB isolates were evaluated for their ability to produce plant growth promoting traits such as production of indole acetic acid (IAA), gibberellic acid, ammonia, ACC deaminase, volatile organic compounds and siderophore, and capacity for nitrogen fixation, and solubilization of phosphorus and potassium. Roll towel assay was used for assessing the seedling vigour index of cowpea after seed priming with the EEB isolates. Based on growth promotion and seedling vigour index, seven superior bacterial isolates were selected, and antagonistic activity against the fungal pathogens Fusarium oxysporum, Rhizoctonia solani, Colletotrichum sp., Phytophthora sp., and Pythium sp., was assayed. The isolates were pooled together to form a consortium and biopriming of cowpea seeds were done. Bioprimed seeds were tested for plant growth promotion and abiotic stress management in cowpea under protray condition. The seedlings of the bioprimed cowpea seeds showed better seedling vigour index and stress tolerance compared to hydro-primed seeds and the unprimed control. EEB isolated from the host plant could be developed as plant growth promoting bioagents.

ANTAGONISTIC ACTIVITY OF ENDOPHYTIC FUNGI ISOLATED FROM ALOE VERA LEAVES AGAINST SOME PLANT PATHOGENIC FUNGI

The present study was aimed to test the antagonistic activity of endophytic fungi isolated from Aloe vera leaves against a number of plant-pathogenic fungi. The results showed that the highest antagonistic activity was exerted by Penicillium chermesinum against the pathogenic fungus Rhizoctonia solani, with an inhibition rate of 78.57%. Also, Talaromyces verruculosus and P. chermesinum showed their highest antagonistic activities against pathogenic fungus Fusarium 1 with inhibition rates of 61.79% and 61.53%, respectively, while T. verruculosus showed an inhibition rate of 62.91% against the pathogenic fungus Fusarium 2. Most of the endophytic fungi showed the least percentage of inhibition against Macrophomina phaseolina compared to the rest of the pathogenic fungi. The fungal filtrates showed a concentration-dependent inhibition rate against the plant pathogens, with a range of 0.00-48.13%. The highest inhibitory activity of Aspergillus niger was recorded at a concentration of 60% against Fusarium 2, R. solani, and Fusarium 1, with rates of 48.13, 37.39, and 36.66%, respectively. The results also showed the ineffectiveness of all fungal filtrates against the pathogenic fungus M. phaseolina. Chemical analysis of the filtrates of the endophytic fungi which showed antagonistic activity against pathogenic fungi was also conducted by using GC-MS. The results showed the presence of effective compounds with biological effects, such as Pentadecanoic acid, Oleic Acid, Limonene, cis-Vaccenic acid, Hexanedioic acid, bis(2-ethylhexyl) ester, Cycloheptasiloxane, and tetracamethyl-.

Pathogen-induced methylglyoxal negatively regulates rice bacterial blight resistance by inhibiting OsCDR1 protease activity

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB), a globally devastating disease of rice (Oryza sativa) that is responsible for significant crop loss. Sugars and sugar metabolites are important for pathogen infection, providing energy and regulating events associated with defense responses; however, the mechanisms by which they regulate such events in BB are unclear. As an inevitable sugar metabolite, methylglyoxal (MG) is involved in plant growth and responses to various abiotic stresses, but the underlying mechanisms remain enigmatic. Whether and how MG functions in plant biotic stress responses is almost completely unknown. Here, we report that the Xoo strain PXO99 induces OsWRKY62.1 to repress transcription of OsGLY II genes by directly binding to their promoters, resulting in overaccumulation of MG. MG negatively regulates rice resistance against PXO99: osglyII2 mutants with higher MG levels are more susceptible to the pathogen, whereas OsGLYII2-overexpressing plants with lower MG content show greater resistance than the wild type. Overexpression of OsGLYII2 to prevent excessive MG accumulation confers broad-spectrum resistance against the biotrophic bacterial pathogens Xoo and Xanthomonas oryzae pv. oryzicola and the necrotrophic fungal pathogen Rhizoctonia solani, which causes rice sheath blight. Further evidence shows that MG reduces rice resistance against PXO99 through CONSTITUTIVE DISEASE RESISTANCE 1 (OsCDR1). MG modifies the Arg97 residue of OsCDR1 to inhibit its aspartic protease activity, which is essential for OsCDR1-enhanced immunity. Taken together, these findings illustrate how Xoo promotes infection by hijacking a sugar metabolite in the host plant.

Bacillus subtilis and Bacillus amyloliquefaciens Mix Suppresses Rhizoctonia Disease and Improves Rhizosphere Microbiome, Growth and Yield of Potato (Solanum tuberosum L.).

Black scurf and stem canker caused by Rhizoctonia solani is a significant disease problem of potatoes. Currently, chemical methods are the primary means of controlling this pathogen. This study sought to explore an alternative approach by harnessing the biocontrol potential of a bacterial mix of Bacillus subtilis and Bacillus amyloliquefaciens against black scurf, and to determine their effect on rhizosphere microorganisms of soil microbiota. This study showed that these bacteria demonstrate antagonistic activity against Rhizoctonia solani. Reduced damage to potato plants during the growing season in Siberia was observed. The index of disease development decreased from 40.9% to 12.0%. The treatment of tubers with this mix of bacteria also led to a change in the composition of the rhizosphere microbiota (according to CFU, 16S and ITS sequencing). This effect was accompanied by a positive change in plant physiological parameters (spectrophotometric analysis). The concentration of chlorophyll in potatoes with the bacterial mix treatment increased by 1.3 fold (p ≤ 0.001), and of carotenoids by 1.2 fold (p ≤ 0.01) compared with the control. After bacterial mix treatment, the length of the aerial parts of plants was 1.3 fold higher (p ≤ 0.001), and the number of stems 1.4 fold higher (p ≤ 0.05). The yield of potatoes was increased by 8.2 t/ha, while the large tuber fraction was increased by 16% (p ≤ 0.05). The bacteria mix of Bacillus subtilis and Bacillus amyloliquefaciens suppressed the plant pathogenic fungus Rhizoctonia solani, and simultaneously enhanced the physiological parameters of potato plants. This treatment can be used to enhance the yield/quality of potato tubers under field conditions.

Mycoparasitism related targets of Tmk1 indicate stimulating regulatory functions of this MAP kinase in Trichoderma atroviride

Mycoparasitism is a key feature of Trichoderma (Hypocreales, Ascomycota) biocontrol agents. Recent studies of intracellular signal transduction pathways of the potent mycoparasite Trichoderma atroviride revealed the involvement of Tmk1, a mitogen-activated protein kinase (MAPK), in triggering the mycoparasitic response. We previously showed that mutants missing Tmk1 exhibit reduced mycoparasitic activity against several plant pathogenic fungi. In this study, we identified the most robustly regulated targets that were governed by Tmk1 during mycoparasitism using transcriptome and proteome profiling. Tmk1 mainly exerts a stimulating function for T. atroviride during its mycoparasitic interaction with the fungal plant pathogen Rhizoctonia solani, as reflected by 89% of strongly differently responding genes in the ∆tmk1 mutant compared to the wild type. Specifically, 54% of these genes showed strong downregulation in the response with a deletion of the tmk1 gene, whereas in the wild type the same genes were strongly upregulated during the interaction with the fungal host. These included the gene encoding the mycoparasitism-related proteinase Prb1; genes involved in signal transduction pathways such as a candidate coding for a conserved 14-3-3 protein, and a gene coding for Tmk2, the T. atroviride cell-wall integrity MAP kinase; genes encoding a specific siderophore synthetase, and multiple FAD-dependent oxidoreductases and aminotransferases. Due to the phosphorylating activity of Tmk1, different (phospho-)proteomics approaches were applied and identified proteins associated with cellular metabolism, energy production, protein synthesis and fate, and cell organization. Members of FAD- and NAD/NADP-binding-domain proteins, vesicular trafficking of molecules between cellular organelles, fungal translational, as well as protein folding apparatus were among others found to be phosphorylated by Tmk1 during mycoparasitism. Outstanding downregulation in the response of the ∆tmk1 mutant to the fungal host compared to the wild type at both the transcriptome and the proteome levels was observed for nitrilase, indicating that its defense and detoxification functions might be greatly dependent on Tmk1 during T. atroviride mycoparasitism. An intersection network analysis between the identified transcripts and proteins revealed a strong involvement of Tmk1 in molecular functions with GTPase and oxidoreductase activity. These data suggest that during T. atroviride mycoparasitism this MAPK mainly governs processes regulating cell responses to extracellular signals and those involved in reactive oxygen stress.

Dissecting genomic regions and underlying sheath blight resistance traits in rice (Oryza sativa L.) using a genome-wide association study.

The productivity of rice is greatly affected by the infection of the plant pathogenic fungus Rhizoctonia solani, which causes a significant grain yield reduction globally. There exist a limited number of rice accessions that are available to develop sheath blight resistance (ShB). Our objective was to identify a good source of the ShB resistance, understand the heritability, and trait interactions, and identify the genomic regions for ShB resistance traits by genome-wide association studies (GWAS). In the present study, a set of 330 traditional landraces and improved rice varieties were evaluated for ShB resistance and created a core panel of 192 accessions used in the GWAS. This panel provides a more considerable amount of genetic variance and found a significant phenotypic variation among the panel of rice accessions for all the agro-morphological and disease-resistance traits over the seasons. The infection rate of ShB and disease reaction were calculated as percent disease index (PDI) and area under the disease progress curve (AUDPC). The correlation analysis showed a significant positive association between PDIs and AUPDC and a negative association between PDI and plant height, flag leaf length, and grain yield. The panel was genotyped with 133 SSR microsatellite markers, resulting in a genome coverage of 314.83 Mb, and the average distance between markers is 2.53 Mb. By employing GLM and MLM (Q + K) models, 30 marker-trait associations (MTAs) were identified with targeted traits over the seasons. Among these QTLs, eight were found to be novel and located on 2, 4, 8, 10, and 12 chromosomes, which explained the phenotypic variation ranging from 5% to 15%. With the GWAS approach, six candidate genes were identified. Os05t0566400, Os08t0155900, and Os09t0567300 were found to be associated with defense mechanisms against ShB. These findings provided insights into the novel donors of IC283139, IC 277248, Sivappuchithirai Kar, and Bowalia. The promising genomic regions on 10 of 12 chromosomes associated with ShB would be useful in developing rice varieties with durable disease resistance.

Evaluation of some bioactive compounds of <i>Azadirachta indica</i> extracts and its application as safe fungicide against different plant pathogenic fungi

Extracts of leaves, seeds and twigs of Azadirachta indica was carried out using Soxhlet extraction technique. These extracts were subjected to percent yield, in vitro phytochemical screening and antifungal activities (minimum fungicidal concentration (MFC), minimum inhibitory concentration (MIC), zone of inhibition and spore germination assay) against plant pathogenic fungi Cladosporium fulvum, Colletotrichum coccodes, Fusarium oxysporum and Rhizoctonia solani. The Soxhlet extractive solvents were methanol, ethanol, chloroform, ethyl acetate and n–hexane. The maximum extractive yield were calculated in ethanol leaves (26±1.5%), methanol leaves (22±01%), n–hexane seeds (32±0.8%) and ethanol seeds (24±01%), while twigs calculated minimum extractive yield. The phytochemicals screening findings confirmed the occurrence of terpenoids, phenols, flavonoids, glycosides, alkaloids, tannins and saponins. The antifungal activities observed that leaves and seeds extracts of neem might cause the inhibition of mycelium and spore germination of tested fungi, though the antifungal activities rate of under study fungal strains diverse with dissimilar concentration and nature of extracts. However the entire concentrations of extract and crude extract especially the methanol and ethanol extracts of neem seeds and leaves reduce the plant pathogenic fungi growth at noteworthy rate. It was concluded from our results that neem leaves and seeds extracts were effectual as antifungal agents against the entire under study fungal strains however the most sensitive fungi were Rhizoctonia solani and Fusarium oxysporum. The findings acquired from this study revealed that chemical constituents from neem seeds and leaves may be formulated as latent agrochemical fungicides.

In vitro inhibition mechanism of Trichoderma asperellum isolates from corn against Rhizoctonia solani causing banded leaf and sheath blight disease and its role in improving the growth of corn seedlings

BackgroundOne of the primary corn diseases, banded leaf and sheath blight, is carried on by the pathogenic fungus Rhizoctonia solani. Efforts to control R. solani are more directed at environmentally friendly control using a biological control agent, such as Trichoderma spp. as antagonistic agents and plant growth promoter. This study aimed to identify T. asperellum isolates based on molecular characteristics and to determine the in vitro inhibition mechanism against R. solani and its role in enhancing the growth of corn seedlings.ResultsThe HMRP7, HMRF7A, HMEDF1B, HMEDF6A, and CHM01 isolates were identified as T. asperellum with a genetic distance coefficient value of 0.000 and a very high similarity of 100%. Meanwhile, the RsHM isolate was identified as R. solani with a homology level of > 90% and genetic distance coefficient values ranging from 0.000 to 0.032. The antagonistic ability showed that the five T. asperellum isolates were able to inhibit the growth of R. solani in vitro on PDA medium with an inhibition percentage of ≥ 50%, so they were categorized as antagonist agents. T. asperellum showed the antagonistic mechanism in inhibiting the R. solani growth through the action of parasitism. The five T. asperellum isolates tested on corn seedlings showed a significantly high difference from the control treatment on the observational variables of maximum growth potential (MGP), growth rate (GtR), growth simultaneity (GS), vigor index (VI), germination rate (GR), and median germination time (T50).ConclusionCHM01 isolate showed better potential than other isolates in inhibiting the growth of R. solani in vitro on PDA medium with a parasitism mechanism and enhancing the growth of corn seedlings.

Biocontrol of Rhizoctonia Solani in Cucumber Plants Using Endobacteria: Effects On Disease Severity and Chlorophyll Content

This study aimed to investigate the potential of endobacteria isolated from different parts of cucumber plants (roots, stems, and leaves) to inhibit the growth of the pathogenic fungus Rhizoctonia solani. Molecular identification confirmed the presence of R. solani in cucumber plants displaying symptoms of wilting and yellowing. Laboratory experiments revealed variations in citrate consumption and gram stain results among different endobacterial isolates. In field experiments, treatments involving Bacillus subtilis with Tricozone and Pseudomonas fluorescens with Tricozone demonstrated significant reductions in disease severity and increased chlorophyll content in cucumber plants compared to the pathogen control. Additionally, Acinetobacter baumannii with Hymazole at the first concentration showed superior effects on leaf area. These findings highlight the potential of endobacteria as biocontrol agents against R. solani, with implications for improving cucumber plant health and productivity.

Jasmonate biosynthesis enzyme allene oxide cyclase 2 mediates cold tolerance and pathogen resistance.

Allene oxide cyclase (AOC) is a key enzyme in the biosynthesis of jasmonic acid (JA), which is involved in plant growth and development as well as adaptation to environmental stresses. We identified the cold- and pathogen-responsive AOC2 gene from Medicago sativa subsp. falcata (MfAOC2) and its homolog MtAOC2 from Medicago truncatula. Heterologous expression of MfAOC2 in M. truncatula enhanced cold tolerance and resistance to the fungal pathogen Rhizoctonia solani, with greater accumulation of JA and higher transcript levels of JA downstream genes than in wild-type plants. In contrast, mutation of MtAOC2 reduced cold tolerance and pathogen resistance, with less accumulation of JA and lower transcript levels of JA downstream genes in the aoc2 mutant than in wild-type plants. The aoc2 phenotype and low levels of cold-responsive C-repeat-binding factor (CBF) transcripts could be rescued by expressing MfAOC2 in aoc2 plants or exogenous application of methyl jasmonate. Compared with wild-type plants, higher levels of CBF transcripts were observed in lines expressing MfAOC2 but lower levels of CBF transcripts were observed in the aoc2 mutant under cold conditions; superoxide dismutase, catalase, and ascorbate-peroxidase activities as well as proline concentrations were higher in MfAOC2-expressing lines but lower in the aoc2 mutant. These results suggest that expression of MfAOC2 or MtAOC2 promotes biosynthesis of JA, which positively regulates expression of CBF genes and antioxidant defense under cold conditions and expression of JA downstream genes after pathogen infection, leading to greater cold tolerance and pathogen resistance.

Comparative bioactive compounds and antifungal potential evaluation of Mentha longifolia (L.) Hudson (Lamiaceae) extracts against plant pathogenic fungi

The present study was design to evaluate the Mentha longifolia stem, flower and leaves infusion, decoction and hydroalcoholic extracts to determine the bioactive molecules and antifungal activities by zone of inhibition technique, Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were assay by broth dilution technique and spore germination assay using Light Compound Microscope against plant pathogenic fungi (Fusarium oxysporum, Aspergillus parasiticus, Rhizoctonia solani and Alternaria alternata). The Mentha longifolia phytochemicals were found in the range flavonoids 3.8 - 2.4%, saponins were 5.6-3.4%, alkaloids 0.7-0.1% and total phenolic were 40-15 mg Gallic acid/g. The maximum zone of inhibition 23±1 mm and 22±1 mm were found by hydroalcoholic leaves extract against Fusarium oxysporum and Aspergillus parasiticus respectively. The MIC range of hydroalcoholic were 12.5-100 mg/mL, infusion were 25-100 mg/mL and decoction were 25-200 mg/mL. While MFC range of hydroalcoholic were 25-200 mg/mL, infusion were 50- 200 mg/mL and decoction were 50-200 mg/mL. Spore germination assay reaveled that increse in the extract concenetration decrese the spore germination. The results revealed that Mentha longifolia chemical composition might be build up as latent bio- fungicides against plant pathogenic fungi.

Antifungal Activity of Bioactive Compounds Produced by the Endophytic Fungus Paecilomyces sp. (JN227071.1) against Rhizoctonia solani.

Biologically active natural compounds are molecules produced by plants or plant-related microbes, such as endophytes. Many of these metabolites have a wide range of antimicrobial activities and other pharmaceutical properties. This study aimed to evaluate (in vitro) the antifungal activities of the secondary metabolites obtained from Paecilomyces sp. against the pathogenic fungus Rhizoctonia solani. The endophytic fungus Paecilomyces was isolated from Moringa oleifera leaves and cultured on potato dextrose broth for the production of the fungal metabolites. The activity of Paecilomyces filtrate against the radial growth of Rhizoctonia solani was tested by mixing the filtrate with potato dextrose agar medium at concentrations of 15%, 30%, 45%, and 60%, for which the percentages of inhibition of the radial growth were 37.5, 50, 52.5, and 56.25%, respectively. The dual culture method was conducted on PDA medium to observe the antagonistic nature of the antibiotic impacts of Paecilomyces sp. towards the pathogenic fungus. The strength of the antagonistic impacts was manifested by a 76.25% inhibition rate, on a scale of 4 antagonistic levels. Ethyl acetate extract of Paecilomyces sp. was obtained by liquid-liquid partition of the broth containing the fungus. Gas chromatography-mass spectrometry (GC-MS) analysis identified the presence of important chemical components e.g., (E) 9, cis-13-Octadecenoic acid, methyl ester (48.607), 1-Heptacosanol, 1-Nonadecene, Cyclotetracosane (5.979), 1,2-Benzenedicarboxylic acid, butyl 2-methylpropyl ester, di-sec-butyl phthalate (3.829), 1-Nonadecene, n-Nonadecanol-1, Behenic alcohol (3.298), n-Heptadecanol-1, 1-hexadecanol, n-Pentadecanol (2.962), Dodecanoic acid (2.849), 2,3-Dihydroxypropyl ester, oleic acid, 9-Octadecenal, and (Z)-(2.730). These results suggest that secondary metabolites of the endophytic Paecilomyces possess antifungal properties and could potentially be utilized in various applications, such as environmental protection and medicine.

New insight on tomato seed priming with Anabaena minutissima phycobiliproteins in relation to Rhizoctonia solani root rot resistance and seedling growth promotion

AbstractCyanobacteria phycobiliproteins (PBPs) are already exploited in the food industries and for biotechnological applications but not in the agricultural field. Different concentrations (0.6 – 4.8 mg/mL) of Anabaena minutissima PBPs were applied to tomato seed to study their priming effect against the soil-borne fungal pathogen Rhizoctonia solani and in promoting plant growth. PBPs increased seedling emergence and vigour, showed activity against root rot disease (67%), and enhanced plant dry weight, length, and height. Generally, no dose effect has been observed except for dry weight (55% at 4.8 mg/mL). Seed treatment primed seeds and seedlings by leading to the activation of defence responses raising phenol (26% in hypocotyls) and flavonoid (26 and 45% in hypocotyls and epicotyls, respectively) contents and chitinase (4-fold at 2.4 and 4.8 mg/mL in hypocotyls) and β-1,3-D-glucanase (up to about 2-fold at all doses in epicotyls) activities. Micro-Attenuated Total Reflection Fourier Transform Infrared revealed changes in functional groups of primed seeds, hypocotyls and exudates released into the agar because of treatment. Protein extract from PBP-primed seedlings inhibited mycelial growth (67% for epicotyl proteins) and caused morphological alterations in hyphae. This research emphasizes the potential priming role of PBPs applied by seed treatment against soil-borne pathogens.