Interactions between Sarocladium oryzae and stem attacking fungal pathogens of rice

In laboratory tests Sarocladium oryzae, the sheath rot pathogen of rice was found to inhibit the mycelial growth of other stem-attacking rice pathogens. Among those inhibited, Sclerotium oryzae and Gaeumannomyces graminis var. graminis were most sensitive while Pyricularia oryzae and Rhizoctonia solani were less sensitive. Tissue-based tests made with rice culm segments established that Sarocladium oryzae inhibits mycelial growth and delays sclerotium formation in R. solani. Cerulenin, the toxin produced by Sarocladium oryzae showed a toxicity pattern towards rice pathogens similar to that of Sarocladium oryzae. The stem rot pathogen, Sclerotium oryzae was most sensitive to cerulenin. In two greenhouse experiments, IR58 rice plants inoculated with Sarocladium oryzae alone or together with Sclerotium oryzae, G. graminis var. graminis or R. solani were found to have reduced plant height and increased tiller number. Sheath rot severity increased when Sarocladium oryzae was inoculated as a single pathogen or together with others. Sheath rot inoculation reduced stem rot in rice plants by 76 and 58%, respectively, in Experiment 1 and 2. By its known antagonistic interaction towards stem rot and crown sheath rot pathogens which are sensitive to it and by other unknown interactions, sheath rot emerges as the dominant disease.     

Changes in defence-related enzymes in rice responding to challenges by Rhizoctonia solani

Sheath blight disease caused by Rhizoctonia solani Kuhn is becoming a major constraint to rice production, especially in the intensified cultivation system. To know the in rice, it is important to get the knowledge of the activity of defence-related enzymes due to the fungal infection. The pathogen induced superoxide dismutase (SOD) and chitinase activities in rice plants, while suppressing peroxidase (POD) and phenylalanine ammonia-lyase (PAL) activities at 36 and 24 h after inoculation, respectively. Induction of two POD isozymes, POD-3 and -4, up to 48 h after inoculation and disappearance of the said isomers at 72 h onwards in rice–Rhizoctonia interaction implicated the role of these isomers in susceptible host–pathogen interaction. Apart from POD and SOD, the activities of other stress-related enzymes, viz. PAL, polyphenol oxidase (PPO) and β-1,3-glucanase were also studied. From this study, it was found that these defence-related enzymes are most significantly related to host–pathogenic interaction.     

Biological control of rice sheath blight in India: Lack of correlation between chitinase production by bacterial antagonists and sheath blight suppression

Bacterial antagonists of both fluorescent and nonfluorescent groups were screened for in-vitro inhibition of the rice sheath blight (ShB) fungus Rhizoctonai solani Kuhn and chitinase production in the laboratory. Twelve percent of the total 1,757 strains screened inhibited R. solani while 31% of the total 1,366 strains tested were positive for chitinase activity. The efficient strains were then evaluated in the field for ShB suppression. Two strains from each of the three seed-bed experiments were chosen for the field test in a hot-spot location. Additional treatments were a Bacillus and validamycin, the fungicide. There was no correlation between chitinase activity in the antagonists and ShB suppression in the seed-bed or field plots. Two most efficient Pseudomonas putida and P. fluorescens strains afforded 68 and 52% ShB suppression while validamycin afforded 27% disease control.     

Differential expression of defense‐related proteins of rice and sheath blight symptoms in response to active and inactive Rhizoctonia solani toxin

The effects of the phyotoxin from the fungal pathogen Rhizoctonia solani, causing sheath blight on the expression of defense‐related proteins of rice were investigated. The toxin inactivated by chemical treatment and by the toxin‐inactivating enzyme α‐glucosidase produced by Trichoderma viride was used in the study along with the active toxin. Toxin inactivated by T. viride α‐glucosidase and sodium periodate caused significantly less damage and electrolyte leakage to test plants. The active toxin and the pathogen induced chitinase and ß‐1,3‐glucanase synthesis in rice plants, while the inactivated toxin did not have any effect on the expression of these pathogenesis‐related proteins. The toxin was found to suppress the peroxidase activity 72 h after inoculation and the inactivated toxin restored the activity as that of untreated plants. There was no remarkable change in phenylalanine ammonia lyase activity in rice sheath treated with both the forms of the toxin.     

A host-specific toxin of Rhizoctonia solani triggers superoxide dismutase (SOD) activity in rice

Changes in the activity of superoxide dismutase (SOD) in rice in response to treatment with Rhizoctonia solani toxin and/or R. solani elicitor were studied. Treatment of rice leaf sheaths with R. solani-toxin significantly increased the SOD activity within 12?h and the maximum enzyme activity was detected 36?h after treatment at which period a fourfold increase in SOD activity was recorded compared to control plants. Isozyme analysis indicated that five new SOD isozymes (SOD-1, SOD-3, SOD-6, SOD-7 and SOD-8) were induced in rice 1?–?2 days after toxin treatment. In elicitor-treated rice leaf sheaths, SOD-2 increased in activity 1?–?5 days after treatment. Pretreatment of rice leaf sheaths with elicitor suppressed the toxin-induced accumulation of SOD.     

Purification and partial characterization of a phytotoxin produced by sarocladium oryzae,the rice sheath rot pathogen: Reinigung und partielle charakterisierung eines vom reisblattscheidenfäule-pathogen sarocladium oryzae erzeugten phytotoxins

An extracellular, hydrophilic, thermostable phytotoxin was purified to homogeneity from culture fluids of Sarocladium oryzae and sheath rot infected rice plants. The phytotoxin was purified by solvent extraction, gel filtration on Sephadex G-75 and HPLC. Toxicity was evaluated with detached leaf sheath and electrolyte leakage bioassays. Purified phytotoxin induced visible symptoms of the disease, when applied to rice sheath even at a low concentration of 5?μg. The toxin is a glycoprotein with carbohydrate as major component. The importances of the carbohydrate moiety for toxic activity was indicated by inactivation of toxic compound after periodate oxidation. The toxin caused lesions on a number of other monocots and dicots and proved to be non-host specific. This is the first report of the purification and characterization of S. oryzae toxin from in vitro and in vivo and we propose its name SO-toxin.     

Association between sheath blight resistance and chitinase activity in transgenic rice plants expressing McCHIT1 from bitter melon

No usable resources with high-level resistance to sheath blight (SB) have yet been found in rice germplasm resources worldwide. Therefore, creating and breeding new disease-resistant rice resources with sheath blight resistance (SBR) are imperative. In this study, we inoculated rice plants with hyphae of the highly pathogenic strain RH-9 of rice SB fungus Rhizoctonia solani to obtain eight stable transgenic rice lines harbouring the chitinase gene (McCHIT1) of bitter melon with good SBR in the T₅ generation. The mean disease index for SB of wild-type plants was 92% and 37–44% in transgenic lines. From 24 h before until 120 h after inoculation with R. solani, chitinase activity in stable transgenic plants with increased SBR was 2.0–5.5 and 1.8–2.7 times that of wild-type plants and plants of a disease-susceptible stable transgenic line, respectively. The correlation between SBR and chitinase activity in McCHIT1-transgenic rice line plants was significant. This work stresses how McCHIT1 from bitter melon can be used to protect rice plants from SB infection.

     

Preparation and evaluation of Bacillus megaterium-alginate microcapsules for control of rice sheath blight disease

Bacillus megaterium encapsulated in calcium alginate microcapsules was prepared and tested for its efficacy against sheath blight disease of rice. In laboratory conditions, the aqueous suspension (1:100, v/v in potato dextrose agar) of the bacterial microcapsules (10¹⁰ spores/ml) inhibited mycelial growth of Rhizoctonia solani (>99 %) after the microcapsules were produced and stored for 12 months at room temperature (28 ± 2 °C). The survival of the bacterium in the microcapsules in response to ultraviolet (u.v.) irradiation and high temperature was investigated. The survivability of the bacterium in the encapsulated form was greater than that of the fresh cells when it was subjected to u.v. (20-W General electric u.v. lamp from a 25 cm distance for 48 h) and a high temperature treatment (80 °C for 48 h). Cells of the bacterium were detected by scanning electron microscope on both the leaf sheath and the leaf blade (in pot tests in a greenhouse) after spraying encapsulated product. The number of bacteria on the surface of both rice tissues (5 Log. number/g of plant) after spraying with encapsulated product was not significantly different from that after spraying with fresh cells onto the rice seedlings. Spraying the encapsulated B. megaterium on rice plants in the greenhouse was as effective as spraying a chemical fungicide for suppressing rice sheath blight disease.     

Resistance-related physiological response of rice leaves to the compound stress of enhanced UV-B radiation and Magnaporthe oryzae

An enhanced UV-B radiation (5.0?kJ?m^?2) was supplied before, during, and after Magnaporthe oryzae infection. The effects of single and compound stress of the UV-B radiation and M. oryzae on the resistance physiology and gene expression of rice leaves were examined. Results revealed that UV-B radiation given before M. oryzae infection (UV-B?→?M.) significantly increased the pathogenesis-related proteins (PRs) activities of phenylalanine ammonialyase (PAL), lipoxygenase (LOX), chitinase (CHT), and β-1,3-glucanase, the resistance-related substances (flavonoids and total phenols) content, and resistance-related genes (OsPAL and OsCHT) expression, thereby improving the disease resistance of rice leaves. Simultaneous exposure to UV-B radiation and M. oryzae (UV-B/M.) significantly increased the OsLOX2 expression and the PRs activities. Exposure to UV-B radiation after M. oryzae infection (M.?→?UV-B) decreased the flavonoid content, did not improve the PRs activity, and increased OsLOX2 expression. Compound treatments of UV-B?→?M., UV-B/M., and M.?→?UV-B reduced the disease index by 62.3%, 40.2%, and 26.6%, respectively, indicating UV-B radiation inhibited the occurrence of M. oryzae disease, but its inhibitory effect weakened when it was provided after M. oryzae infection. Hence, rice responded to the compound stress of UV-B radiation and M. oryzae through a resistance-related physiological mechanism associated with the sequence of stress occurrence.     

Germin-like proteins (GLPs) in cereal genomes: gene clustering and dynamic roles in plant defence

The recent release of the genome sequences of a number of crop and model plant species has made it possible to define the genome organisation and functional characteristics of specific genes and gene families of agronomic importance. For instance, Sorghum bicolor, maize (Zea mays) and Brachypodium distachyon genome sequences along with the model grass species rice (Oryza sativa) enable the comparative analysis of genes involved in plant defence. Germin-like proteins (GLPs) are a small, functionally and taxonomically diverse class of cupin-domain containing proteins that have recently been shown to cluster in an area of rice chromosome 8. The genomic location of this gene cluster overlaps with a disease resistance QTL that provides defence against two rice fungal pathogens (Magnaporthe oryzae and Rhizoctonia solani). Studies showing the involvement of GLPs in basal host resistance against powdery mildew (Blumeria graminis ssp.) have also been reported in barley and wheat. In this mini-review, we compare the close proximity of GLPs in publicly available cereal crop genomes and discuss the contribution that these proteins, and their genome sequence organisation, play in plant defence.     

Morphological and pathological variations of rice sheath blight inciting south Indian Rhizoctonia solani isolates

Frequent assessment of pathogen diversity is one of the most important criteria in designing disease management programmes. A study on diversity of field isolates of Rhizoctonia solani from sheath blight-infected rice fields of south India has been carried out. A total of 236 R. solani isolates were obtained from 45 locations in the surveyed area. Sclerotial features such as colour, size and shape and distribution pattern were varying among isolates. However, no other morphological features found to differ among isolates. Majority of the R. solani isolates were fast growers as they attained complete mycelial growth within 2 days in a 90-mm Petri plate and the emergence of sclerotial structures was seen even in 4 days of incubation. Selected 10 R. solani isolates exhibited considerable variations in pathogenicity on three different rice cultivars. Vellai ponni was found to be the most susceptible rice cultivar to all the field isolates of R. solani.     

Control of blast and sheath blight diseases of rice using antifungal metabolites produced by Streptomyces sp. PM5

Two antifungal aliphatic compounds, SPM5C-1 and SPM5C-2 with a lactone and ketone carbonyl unit, respectively obtained from Streptomyces sp. PM5 were evaluated under in vitro and in vivo conditions against major rice pathogens, Pyricularia oryzae and Rhizoctonia solani. These compounds were dissolved in distilled water/medium to get the required concentrations. The well diffusion bioassay indicated that the of SPM5C-1 remarkably inhibited the mycelial growth of P. oryzae and R. solani in comparison to SPM5C-2. Though SPM5C-2 showed low antifungal activity against P. oryzae, it was not active against R. solani. Further, SPM5C-1 completely inhibited the growth of P. oryzae and R. solani at concentrations of 25, 50, 75 and 100 μg/ml. Greenhouse experiments revealed that spraying of SPM5C-1 at 500 μg/ml on rice significantly decreased blast and sheath blight development by 76.1% and 82.3%, respectively, as compared to the control with a corresponding increase in rice grain yield.     

Antifungal Hydrolases in Pea Tissue : II. Inhibition of Fungal Growth by Combinations of Chitinase and beta-1,3-Glucanase

Chitinase and β-1,3-glucanase purified from pea pods acted synergistically in the degradation of fungal cell walls. The antifungal potential of the two enzymes was studied directly by adding protein preparations to paper discs placed on agar plates containing germinated fungal spores. Protein extracts from pea pods infected with Fusarium solani f.sp. phaseoli, which contained high activities of chitinase and β-1,3-glucanase, inhibited growth of 15 out of 18 fungi tested. Protein extracts from uninfected pea pods, which contained low activities of chitinase and β-1,3-glucanase, did not inhibit fungal growth. Purified chitinase and β-1,3-glucanase, tested individually, did not inhibit growth of most of the test fungi. Only Trichoderma viride was inhibited by chitinase alone, and only Fusarium solani f.sp. pisi was inhibited by β-1,3-glucanase alone. However, combinations of purified chitinase and β-1,3-glucanase inhibited all fungi tested as effectively as crude protein extracts containing the same enzyme activities. The pea pathogen, Fusarium solani f.sp. pisi, and the nonpathogen of peas, Fusarium solani f.sp. phaseoli, were similarly strongly inhibited by chitinase and β-1,3-glucanase, indicating that the differential pathogenicity of the two fungi is not due to differential sensitivity to the pea enzymes. Inhibition of fungal growth was caused by the lysis of the hyphal tips.     

Rice oxalate oxidase gene driven by green tissue‐specific promoter increases tolerance to sheath blight pathogen (Rhizoctonia solani) in transgenic rice

Rice sheath blight, caused by the necrotrophic fungus Rhizoctonia solani, is one of the most devastating and intractable diseases of rice, leading to a significant reduction in rice productivity worldwide. In this article, in order to examine sheath blight resistance, we report the generation of transgenic rice lines overexpressing the rice oxalate oxidase 4 (Osoxo4) gene in a green tissue‐specific manner which breaks down oxalic acid (OA), the pathogenesis factor secreted by R. solani. Transgenic plants showed higher enzyme activity of oxalate oxidase (OxO) than nontransgenic control plants, which was visualized by histochemical assays and sodium dodecylsulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Transgenic rice leaves were more tolerant than control rice leaves to exogenous OA. Transgenic plants showed a higher level of expression of other defence‐related genes in response to pathogen infection. More importantly, transgenic plants exhibited significantly enhanced durable resistance to R. solani. The overexpression of Osoxo4 in rice did not show any detrimental phenotypic or agronomic effect. Our findings indicate that rice OxO can be utilized effectively in plant genetic manipulation for sheath blight resistance, and possibly for resistance to other diseases caused by necrotrophic fungi, especially those that secrete OA. This is the first report of the expression of defence genes in rice in a green tissue‐specific manner for sheath blight resistance.     

Suppression of Rhizoctonia solani and induction of host plant resistance by Paenibacillus kribbensis PS04 towards controlling of rice sheath blight

We have identified a promising antagonistic micro-organism for suppressing Rhizoctonia solani and triggering induced resistance in rice. The isolate PS04 was identified as Paenibacillus kribbensis. PS04 and its crude metabolites showed inhibitory effects against R. solani both in vitro and in vivo, and with high stability against temperature and pH. Strain PS04 is a great biocontrol agent.     

Chitosan-induced defence responses in tomato plants against early blight disease caused by Alternaria solani (Ellis and Martin) Sorauer

The in vitro antifungal properties of chitosan and its role in protection of tomato from early blight disease were evaluated. Chitosan inhibited the radial and submerged growth of Alternaria solani at 1 mg/ml, and controls tomato plants from blight pathogen. Chitosan induces the level of chitinase activity and new isoforms of chitinase resulting in the reduction of early blight disease severity in tomato leaves. These results suggested the role of chitosan in activation of defence responses as well as protecting tomato plants from A. solani infection.     

Influence of soil factors and cultural practice on biological control of sheath blight of rice with antagonistic bacteria

Fluorescent and nonfluorescent strains of bacteria isolated from rice rhizospheres on the International Rice Research Institute (IRRI) farm were evaluated for in vitro antibiosis towards the sheath blight (ShB) pathogen Rhizoctonia solani, and for suppression of ShB in detached rice leaves. Efficient strains were located on the basis of consistent performance in two laboratory tests. Among nine efficient strains, 3 strains were identified as Pseudomonas fluorescens, 5 strains were tentatively identified as Bacillus spp. and one strain was identified as Enterobacter. In three greenhouse tests lowland rice soils with optimum pH for rice growth (pH 5.5–6.5), acidic pH (pH 5.0) and boron toxicity were found more suitable for biological control of ShB and, less frequently, also yield increases than were alkaline (pH 6.9) and zinc-deficient soils. Bacterial treatments afforded significant ShB reductions in 3 field experiments, but no significant yield increases resulted. In direct-seeded rice best performances by bacterial treatments in terms of ShB suppression were 66 and 98% during DS 1988 and WS 1988, respectively, which were comparable to or better than the performance of validamycin (a fungicide routinely used for ShB control) which afforded 42 and 63% disease suppression, respectively, in the same experiments. Although bacterial treatments caused ShB reductions both in direct-seeded and transplanted rice crops, disease control was more pronounced in direct-seeded than in transplanted crops. These results indicate that carefully selected strains of bacterial antagonists have the potential for ShB suppression in rice at least in areas where direct-seeding is practised.     

Involvement of secondary metabolites and extracellular lytic enzymes produced by Pseudomonas fluorescens in inhibition of Rhizoctonia solani, the rice sheath blight pathogen

Fourteen strains of Pseudomonas fluorescens isolated from rhizosphere soil of rice were tested for their antagonistic effect towards Rhizoctonia solani, the rice sheath blight fungus. Among them, PfMDU2 was the most effective in inhibiting mycelial growth of R. solani in vitro. Production of chitinase, beta-1,3-glucanase, siderophores, salicylic acid (SA) and hydrogen cyanide (HCN) by P. fluorescens strains was evaluated. The highest beta-1,3-glucanase activity, siderophore production, SA production and HCN production were recorded with PfMDU2. A significant relationship between the antagonistic potential of P. fluorescens against R. solani and its level of beta-1,3-glucanase, SA and HCN was observed.