Plant Pathogenic Fungus Rhizoctonia Solani Research Articles

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.

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.

Mycoviruses: Antagonistic Potential, Fungal Pathogenesis, and Their Interaction with Rhizoctonia solani.

Mycoviruses, or fungal viruses, are prevalent in all significant fungal kingdoms and genera. These low-virulence viruses can be used as biocontrol agents to manage fungal diseases. These viruses are divided into 19 officially recognized families and 1 unclassified genus. Mycoviruses alter sexual reproduction, pigmentation, and development. Spores and fungal hypha spread mycoviruses. Isometric particles mostly encapsulate dsRNA mycoviruses. The widespread plant-pathogenic fungus Rhizoctonia solani, which has caused a rice sheath blight, has hosted many viruses with different morphologies. It causes significant crop diseases that adversely affect agriculture and the economy. Rice sheath blight threatens the 40% of the global population that relies on rice for food and nutrition. This article reviews mycovirology research on Rhizoctonia solani to demonstrate scientific advances. Mycoviruses control rice sheath blight. Hypovirulence-associated mycoviruses are needed to control R. solani since no cultivars are resistant. Mycoviruses are usually cryptic, but they can benefit the host fungus. Phytopathologists may use hypovirulent viruses as biological control agents. New tools are being developed based on host genome studies to overcome the intellectual challenge of comprehending the interactions between viruses and fungi and the practical challenge of influencing these interactions to develop biocontrol agents against significant plant pathogens.

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.

Improvement of antagonism and fungicides tolerance in Trichoderma harzianum and T.viride local isolates by Ultra-Violet irradiation

Among 22 Iraqi Trichoderma harzianum and T.viride isolates, T3,T5,D5,S1 and S6 related to T. harzianum showed a higher antagonistic activity against the plant pathogenic fungi Fusarium oxysporum and Rhizoctonia solani. These isolates were selected for improvement their biocontrol activity by usi
 ng uv-irradiation. The optimum exposure time to ultraviolet irradiation is 30 min, in which, the percentage of kill was 98-99% .After UV-irradiation treatment (Quartz lamps 30W at 240-280 nm, peak 254nm, distance from the source was 20 cm), antagonistic capability against R. solani and F. oxysporum was improved in mutants MT3,MT5,MS1 and MS6, in which antagonistic reaction score were 3.6,3.9,3.2 and 3.6 against F. oxysporum and 3.7,4,3.4 and 4 against R. solani compared to 3.2, 2.8, 2.6 and 2.8 against F. oxysporum and 2.7, 2.9,3.1 and 3 against R. solani in the wild types T3, T5, S1 and S6,respectively. The growth of T. harzianum isolates (estimated as colony diameter) was significantly increased in all T. harzianum mutants except MT3 and MD5 mutants, CMCase activity was increased from 13.2, 16.2, 10 and 15.8 U/ml / mg protein in the wild isolates T3, T5, S1 and S6 to 18.6, 20.3, 17.7 and 22.5 U/ml / mg protein in the mutants MT3, MT5, MS1 and MS6, respectively. Chitinase activity was also increased in these mutants to 88.4,132.3,86.1 and 136.7 U/ml / mg protein, compared to 82.7, 93.2, 74.5 and 103.2 U/ml / mg protein in their wild, respectively. As comparison between wild types and mutants, the maximum growth was 5.2 and 4.8 cm (as colonies diameters) by MT5 in present of 25% of the recommended field rate of Topsin and Benomyl, compared to 3.3 and 2.2 cm in the wild type T5, respectively, followed by MS6,in which, the maximum growth in this mutant was 4.8 and 4.2 cm in the same concentration of these two fungicides, compared to 3.4 and 2.5 cm in the wild type S6, respectively. According to these results, the mutants MT5 and MS6 developed from UV-irradiation treatment, showed as promising T. harzianum mutants through their antagonistic activity (at mycoparasitism level) , fast growth, higher CMCase and chitinase activity and growth in 25% of the recommended field rate of Elsa , 25 and 50% of the recommended field rate of Diathen and Mizab and all concentrations of Topsin and Benomyl with complete inhibition of pathogenic fungi F. oxysporum and R. solani.

Reduced tillage increases soil biological properties but not suppressiveness against Rhizoctonia solani and Streptomyces scabies

Intensive agricultural crop production can lead to a decline in biological soil characteristics and functions, such as soil microbial biomass and activity, carbon and nutrient cycling and soil suppressiveness, important for the sustainable production of food and feed. There is a need to understand how those soil functions can be improved by agricultural practices. In a long-term field study, we assessed whether reduced tillage could enhance soil biological parameters and soil suppressiveness against the plant-pathogenic fungus Rhizoctonia solani AG2-2IIIB and bacterium Streptomyces scabies. Soil suppressiveness was assessed in bioassays with a susceptible crop while adding pathogen inoculum. Undisturbed soil cores (0–12 cm) were used in these bioassays to include the soil structural aspects, which were likely affected by the tillage treatments. Reduced i.e., non-inversion, tillage was compared with conventional ploughing treatment over 6 years. We found that reduced tillage led to an increase in bacterial and fungal biomass, labile carbon and nitrogen and an increase in the abundance of potential bacterial antagonists, compared to conventional tillage in the upper 12 cm. However, the increase of these microbial parameters did not lead to consistent changes in soil suppressiveness against both R. solani and S. scabies in response to the tillage treatment. Rather, disease suppressiveness varied significantly between field and year of sampling but was not correlated to any of the assessed soil parameters. Thus while reduced tillage can be beneficial for soil biology, other measures will have to be investigated for inducing R. solani and S. scabies disease suppressiveness.

Antifungal activity of Mentha × Piperita L. essential oil

The objective of this study was to investigate the chemical composition and in vitro antifungal activity of Mentha × Piperita L. (peppermint) essential oil (EO) against some plant pathogenic fungi (Alternaria alternata, Penicillium expansum, Rhizoctonia solani, and Rhizopus stolonifer). Antifungal activity of EO against the selected fungi was conducted using the agar diffusion method by adding peppermint EO concentrations (0, 250, 500, 750, 1000, and 2000 ppm). The gas chromatography-mass spectrometry (GC-MS) analysis of peppermint EO showed that the main constituent was menthol (36.4%), followed by menthone (27.7%) and menthyl acetate (11.2%). The mycelium growth of the selected fungi was significantly inhibited by peppermint EO. Light and electron microscopy studies showed that mycelium morphology was seriously changed after treatment with peppermint essential oil. The level of malondialdehyde illustrated that peppermint EO led to lipid peroxidation in the fungal pathogens. Therefore, due to its antifungal properties, peppermint EO can be used as an additive in the food industry and as an active substance in pharmaceuticals.

Functional characterization of the AGL1 aegerolysin in the mycoparasitic fungus Trichoderma atroviride reveals a role in conidiation and antagonism

Aegerolysins are small secreted pore-forming proteins that are found in both prokaryotes and eukaryotes. The role of aegerolysins in sporulation, fruit body formation, and in lysis of cellular membrane is suggested in fungi. The aim of the present study was to characterize the biological function of the aegerolysin gene agl1 in the mycoparasitic fungus Trichoderma atroviride, used for biological control of plant diseases. Gene expression analysis showed higher expression of agl1 during conidiation and during growth in medium supplemented with cell wall material from the plant pathogenic fungus Rhizoctonia solani as the sole carbon source. Expression of agl1 was supressed under iron-limiting condition, while agl1 transcript was not detected during T. atroviride interactions with the prey fungi Botrytis cinerea or R. solani. Phenotypic analysis of agl1 deletion strains (Δagl1) showed reduced conidiation compared to T. atroviride wild type, thus suggesting the involvement of AGL1 in conidiation. Furthermore, the Δagl1 strains display reduced antagonism towards B. cinerea and R. solani based on a secretion assay, although no difference was detected during direct interactions. These data demonstrate the role of AGL1 in conidiation and antagonism in the mycoparasitic fungus T. atroviride.

A novel putative betapartitivirus isolated from the plant-pathogenic fungus Rhizoctonia solani.

In this study, we describe the genome sequence of a novel double-stranded RNA (dsRNA) mycovirus, designated as "Rhizoctonia solani partitivirus 15" (RsPV15), from the phytopathogenic fungus Rhizoctonia solani. RsPV15 consists of two genomic double-stranded RNA segments, dsRNA-1 and dsRNA-2, which are 2433 bp and 2350 bp long, respectively. Each of the dsRNA segments contains a single open reading frame, encoding the putative RNA-dependent RNA polymerase and coat protein, respectively. Homology searches and phylogenetic analysis suggested that RsPV15 is a new member of the genus Betapartitivirus within the family Partitiviridae.

Viruses Infecting the Plant Pathogenic Fungus Rhizoctonia solani.

The cosmopolitan fungus Rhizoctonia solani has a wide host range and is the causal agent of numerous crop diseases, leading to significant economic losses. To date, no cultivars showing complete resistance to R. solani have been identified and it is imperative to develop a strategy to control the spread of the disease. Fungal viruses, or mycoviruses, are widespread in all major groups of fungi and next-generation sequencing (NGS) is currently the most efficient approach for their identification. An increasing number of novel mycoviruses are being reported, including double-stranded (ds) RNA, circular single-stranded (ss) DNA, negative sense (−)ssRNA, and positive sense (+)ssRNA viruses. The majority of mycovirus infections are cryptic with no obvious symptoms on the hosts; however, some mycoviruses may alter fungal host pathogenicity resulting in hypervirulence or hypovirulence and are therefore potential biological control agents that could be used to combat fungal diseases. R. solani harbors a range of dsRNA and ssRNA viruses, either belonging to established families, such as Endornaviridae, Tymoviridae, Partitiviridae, and Narnaviridae, or unclassified, and some of them have been associated with hypervirulence or hypovirulence. Here we discuss in depth the molecular features of known viruses infecting R. solani and their potential as biological control agents.

Trichoderma brevicrassum strain TC967 with capacities of diminishing cucumber disease caused by Rhizoctonia solani and promoting plant growth

Some Trichoderma species have been used as biocontrol agents in agriculture. In order to explore additional useful natural resources, a large scale survey of Trichoderma strains from China was conducted and T. brevicrassum strain TC967 was singled out. In vitro assays indicated that it showed high chitinase activity in agar medium, inhibited growth of the plant-pathogenic fungus Rhizoctonia solani (72.14%) in dual culture, and revealed strong mycoparasitic ability through coiling and penetrating hyphae of the pathogen. In vivo assays indicated that it was able to suppress the cucumber disease caused by R. solani to a low disease index (37.5%). Hyphae of strain TC967 colonized cucumber root surface, induced the expression of pathogenesis-related protein genes PR1, PR4 and PR5. In addition, strain TC967 promoted cucumber growth. This is the first report of T. brevicrassum having biocontrol potential and promoting cucumber growth.

Bio-efficacy of chitinolytic Bacillus thuringiensis isolates native to northwestern Indian Himalayas and their synergistic toxicity with selected insecticides

Pesticidal properties of Bacillus thuringiensis and its associated toxic proteins is an ever-growing science with potential implications in biological pest management. In the present study 80 Bacillus thuringiensis isolates native to Uttarakhand Himalayas were evaluated for chitinolytic activity and potent ones (11 isolates) were further subjected to multiphasic characterization for their antifungal, insecticidal and synergistic properties with selected chemical insecticides. Although all the 11 potent isolates were biologically active, only three isolates (VLBt27, VLBt109 and VLBt238) showed >90% inhibition in radial growth of 3 out of 4 tested plant pathogenic fungi (Rhizoctonia solani, Fusarium oxysporum, Alternaria pori and Pyricularia oryzae). The key antagonism was manifested in the form of disruptions in growing tips and uneven mycelial thickenings. In insect bioassays (against Helicoverpa armigera, Mythimna separata and Thysanoplusia orichalcea), no considerable direct mortality was observed. However, the larval weight reduction was prominent in four isolates (VLBt27, VLBt38, VLBt109 and VLBt135) which accounts to >75% in first instar larvae of H. armigera. Joint action of these four isolates with chemical insecticides showed an overall additive interaction against Brevicoryne brassicae and synergism against H. armigera. All the isolates were compatible with tested insecticides at their field recommended doses except for chlorpyriphos with around 130 kDa protein as chitinase. The study identified VLBt27 and VLBt109, two native isolates of B. thuringiensis with potential antagonistic activity and synergism as well. These isolates have possible implications as single strategy against two diverse pest problems (pathogenic fungi and phytophagous insect) of agriculture with a view of reduced pesticide application.

In-Vitro Inhibition of Pythium ultimum, Fusarium graminearum, and Rhizoctonia solani by a Stabilized Lactoperoxidase System alone and in Combination with Synthetic Fungicides

Advances in enzyme stabilization and immobilization make the use of enzymes for industrial applications increasingly feasible. The lactoperoxidase (LPO) system is a naturally occurring enzyme system with known antimicrobial activity. Stabilized LPO and glucose oxidase (GOx) enzymes were combined with glucose, potassium iodide, and ammonium thiocyanate to create an anti-fungal formulation, which inhibited in-vitro growth of the plant pathogenic oomycete Pythium ultimum, and the plant pathogenic fungi Fusarium graminearum and Rhizoctonia solani. Pythium ultimum was more sensitive than F. graminearum and R. solani, and was killed at LPO and GOx concentrations of 20 nM and 26 nM, respectively. Rhizoctonia solani and F. graminearum were 70% to 80% inhibited by LPO and GOx concentrations of 242 nM and 315 nM, respectively. The enzyme system was tested for compatibility with five commercial fungicides as co-treatments. The majority of enzyme + fungicide co-treatments resulted in additive activity. Synergism ranging from 7% to 36% above the expected additive activity was observed when P. ultimum was exposed to the enzyme system combined with Daconil® (active ingredient (AI): chlorothalonil 29.6%, GardenTech, Lexington, KY, USA), tea tree oil, and mancozeb at select fungicide concentrations. Antagonism was observed when the enzyme system was combined with Tilt® (AI: propiconazole 41.8%, Syngenta, Basel, Switzerland) at one fungicide concentration, resulting in activity 24% below the expected additive activity at that concentration.

Two alphapartitiviruses co-infecting a single isolate of the plant pathogenic fungus Rhizoctonia solani.

Seven dsRNA segments were detected from a single Rhizoctonia solani strain HG81. From the full-length cDNA sequences of four smaller dsRNA segments, the genomes of two related partitiviruses, designated as Rhizoctonia solani partitivirus 3 (RsPV3) and RsPV4, were determined. The genomes of RsPV3 and RsPV4 are both composed of two separate dsRNA segments, with each segment possessing a single open reading frame (ORF). ORF1 from RsPV3 and RsPV4 encodes a putative RNA-dependent RNA polymerase, while ORF2 of RsPV3 and RsPV4 encodes a putative capsid protein. RsPV3 and RsPV4 share high sequence identity with viruses classified within the genus Alphapartitivirus, family Partitiviridae.

Synthesis and characterization of chitosan\u2013copper nanocomposites and their fungicidal activity against two sclerotia-forming plant pathogenic fungi

In this report, the metal-vapor synthesis (MVS) was used for the preparation of copper nanoparticles which was then used for the preparation of chitosan–copper nanocomposite. The antifungal activity of Cu@Chit NCs against two sclerotium-forming plant pathogenic fungi Sclerotium rolfsii (S. rolfsii) and Rhizoctonia solani (R. solani) AG-4 was evaluated in vitro and their effects on hyphal morphology, and sclerotia formation were observed for the first time. The NCs were prepared through impregnation of chitosan with colloid solution of copper nanoparticles in organic solvent (acetone or toluene). Transmission electron microscopy shows that the particles have predominantly spherical form, polydisperse character, the mean diameter about 2–3 nm and a rather uniform distribution in the chitosan matrice. Analysis of the small angle scattering curves suggests that the copper particles in the NCs with the size of ≤2 nm are mostly located in the chitosan pores with the same size. The effect of Cu@Chit NCs on fungal growth reveals some significant inhibitory activity against two tested fungi. The highest level of inhibition against S. rolfsii and R. solani AG-4 was observed using the high concentrations of Cu@Chit NC prepared using acetone as a solvent. A loss of the cytoplasm content, cytoplasmic coagulation, irregular shape of mycelia, or destruction in the hyphae was confirmed. The experiments demonstrate that the Cu@Chit NC synthesized via MVS using acetone was more effective than that of toluene in inhibiting fungal hyphae growth against R. solani AG-4 and S. rolfsii. The results show that the Cu@Chit NCs are fungicidal against both the tested fungus at high concentrations and the fungicidal or fungistatic activity is dependent on the tested fungus species.

English

 Paenibacillus strain, UKCH21, isolated from Uttarakhand State, north western Indian Himalayas was found to produce high levels of extracellular chitinases. The 16s rRNA gene sequence showed 100% homology with P. illinoisensis (Accession No. KR856190) available in the public domain and further phylogenetic study also verified the species identity. The culture supernatants have a maximum chitinase activity of 110.8 U/ml after 3 days of culturing. The isolate showed strong antifungal activity manifested in the form of progressive mycelia degradation in dual culture plates. The pathogenicity was observed as structural deformities like uneven thickening of mycelia as a result of direct degradation of chitin. The optimum pH and temperature of UKCH21 chitinases was found to be 5 and 50°C, respectively. Partial characterization of chitinase gene also confirms the family 18 status of glycosyl hydrolase with substantial variability presented here with. Above all, percent inhibition of growth and the rapid degradation of mycelia of tested plant pathogenic fungi (Rhizoctonia solani, Fusarium solani and Sclerotium rolfsi) in bacteria seeded medium suggest its utilization as potent antifungal biocontrol agent.   Key words: Paenibacillus illinoisensis, UKCH21, chitinase, antifungal, Indian Himalayas.

Plant-mediated green synthesis of silver nanoparticles using Trifolium resupinatum seed exudate and their antifungal efficacy on Neofusicoccum parvum and Rhizoctonia solani.

In recent years, biosynthesis and the utilisation of silver nanoparticles (AgNPs) has become an interesting subject. In this study, the authors investigated the biosynthesis of AgNPs using Trifolium resupinatum (Persian clover) seed exudates. The characterisation of AgNPs were analysed using ultraviolet-visible spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infra-red spectroscopy. Also, antifungal efficacy of biogenic AgNPs against two important plant-pathogenic fungi (Rhizoctonia solani and Neofusicoccum Parvum) in vitro condition was evaluated. The XRD analysis showed that the AgNPs are crystalline in nature and have face-centred cubic geometry. TEM images revealed the spherical shape of the AgNPs with an average size of 17 nm. The synthesised AgNPs were formed at room temperature and kept stable for 4 months. The maximum distributions of the synthesised AgNPs were seen to range in size from 5 to 10 nm. The highest inhibition effect was observed against R. solani at 40 ppm concentration of AgNPs (94.1%) followed by N. parvum (84%). The results showed that the antifungal activity of AgNPs was dependent on the amounts of AgNPs. In conclusion, the AgNPs obtained from T. resupinatum seed exudate exhibit good antifungal activity against the pathogenic fungi R. solani and N. Parvum.

Antifungal Rhizosphere Bacteria Can increase as Response to the Presence of Saprotrophic Fungi

Knowledge on the factors that determine the composition of bacterial communities in the vicinity of roots (rhizosphere) is essential to understand plant-soil interactions. Plant species identity, plant growth stage and soil properties have been indicated as major determinants of rhizosphere bacterial community composition. Here we show that the presence of saprotrophic fungi can be an additional factor steering rhizosphere bacterial community composition and functioning. We studied the impact of presence of two common fungal rhizosphere inhabitants (Mucor hiemalis and Trichoderma harzianum) on the composition of cultivable bacterial communities developing in the rhizosphere of Carex arenaria (sand sedge) in sand microcosms. Identification and phenotypic characterization of bacterial isolates revealed clear shifts in the rhizosphere bacterial community composition by the presence of two fungal strains (M. hiemalis BHB1 and T. harzianum PvdG2), whereas another M. hiemalis strain did not show this effect. Presence of both M. hiemalis BHB1 and T. harzianum PvdG2 resulted in a significant increase of chitinolytic and (in vitro) antifungal bacteria. The latter was most pronounced for M. hiemalis BHB1, an isolate from Carex roots, which stimulated the development of the bacterial genera Achromobacter and Stenotrophomonas. In vitro tests showed that these genera were strongly antagonistic against M. hiemalis but also against the plant-pathogenic fungus Rhizoctonia solani. The most likely explanation for fungal-induced shifts in the composition of rhizosphere bacteria is that bacteria are being selected which are successful in competing with fungi for root exudates. Based on the results we propose that measures increasing saprotrophic fungi in agricultural soils should be explored as an alternative approach to enhance natural biocontrol against soil-borne plant-pathogenic fungi, namely by stimulating indigenous antifungal rhizosphere bacteria.

Transcriptomic changes in the plant pathogenic fungus Rhizoctonia solani AG-3 in response to the antagonistic bacteria Serratia proteamaculans and Serratia plymuthica.

BackgroundImproved understanding of bacterial-fungal interactions in the rhizosphere should assist in the successful application of bacteria as biological control agents against fungal pathogens of plants, providing alternatives to chemicals in sustainable agriculture. Rhizoctonia solani is an important soil-associated fungal pathogen and its chemical treatment is not feasible or economic. The genomes of the plant-associated bacteria Serratia proteamaculans S4 and Serratia plymuthica AS13 have been sequenced, revealing genetic traits that may explain their diverse plant growth promoting activities and antagonistic interactions with R. solani. To understand the functional response of this pathogen to different bacteria and to elucidate whether the molecular mechanisms that the fungus exploits involve general stress or more specific responses, we performed a global transcriptome profiling of R. solani Rhs1AP anastomosis group 3 (AG-3) during interaction with the S4 and AS13 species of Serratia using RNA-seq.ResultsApproximately 104,504 million clean 75-100 bp paired-end reads were obtained from three libraries, each in triplicate (AG3-Control, AG3-S4 and AG3-AS13). Transcriptome analysis revealed that approximately 10 % of the fungal transcriptome was differentially expressed during challenge with Serratia. The numbers of S4- and AS13-specific differentially expressed genes (DEG) were 866 and 292 respectively, while there were 1035 common DEGs in the two treatment groups. Four hundred and sixty and 242 genes respectively had values of log2 fold-change > 3 and for further analyses this cut-off value was used. Functional classification of DEGs based on Gene Ontology enrichment analysis and on KEGG pathway annotations revealed a general shift in fungal gene expression in which genes related to xenobiotic degradation, toxin and antioxidant production, energy, carbohydrate and lipid metabolism and hyphal rearrangements were subjected to transcriptional regulation.ConclusionsThis RNA-seq profiling generated a novel dataset describing the functional response of the phytopathogen R. solani AG3 to the plant-associated Serratia bacteria S4 and AS13. Most genes were regulated in the same way in the presence of both bacterial isolates, but there were also some strain-specific responses. The findings in this study will be beneficial for further research on biological control and in depth exploration of bacterial-fungal interactions in the rhizosphere.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1758-z) contains supplementary material, which is available to authorized users.