Morphological,pathological and molecular characterisation of rice sheath blight disease causal organism Rhizoctonia solani AG-1 IA in Egypt

Rice sheath blight disease caused by Rhizoctonia solani is considered a distractive soil-borne disease of rice production worldwide. The study aimed to determine the causal organism of sheath blight symptoms in Egyptian rice fields. Sheath blight symptoms were first observed in a small area during 2013, 2014 and 2016 seasons, later in a wide area of rice fields in 2016 to 2018 seasons. Pathogen identification was carried out based on morphological traits and internal transcribed spacer sequencing. Thirty-six isolates were identified as R. solani fungus. The isolates exhibited a wide range of variability in their morphological traits and virulence patterns. Five isolates were sequenced and aligned with Chinese isolates with 75–100% identity. This is the first report of R. solani AG-1 IA that associated with rice sheath blight in Egypt. Initiate a breeding program for disease resistance and integrated disease management procedures are important to keep the disease under control.     

Morphological variability in Rhizoctonia solani isolates from different agro-ecological zones of West Bengal,India

Sheath blight caused by Rhizoctonia solani Kühn is one of the most important diseases of rice, resulting in significant yield loss in rice every year. The rice-based intensified cropping system, edapho-climatological and host variations make the disease problem more complicated. However, the incidence and severity of the disease differ from one location to other, one geographical area to other and even differs from country and region wise. The reasons for this disease severity have been attributed to the variation in host genotype, virulence of the pathogen, prevalence of congenial soil physico-chemical and plants’ surrounding environment and cultural practices. Sixty-seven number of isolates of R. solani from rice, 12 no. of R. solani isolates associated with maize, sugarcane, weeds, cabbage, pointed gourd, watermelon, potato, dolichos bean and aparajita were isolated from different agro-ecological region of West Bengal and three no. of isolates of R. oryzae-sativae obtained from Department of Plant Pathology, BCKV, were used in the present study. Cultural and morphological characteristics revealed considerable diversity among the R. solani isolates. Cultural and morphological analysis of WB isolates of rice has indicated that the diversity among the isolates does not correlated with their origin. On the basis of morphological characteristics, R. solani isolates could be easily separated from R. oryzae-sativae isolates. The no. of sclerotia, hyphal length, wt. of sclerotia and mycelial growth rate are the important morphological markers for differentiation of R. oryzae-sativae from R. solani isolates.     

Pathogenic and Genetic Variation among the Isolates of Rhizoctonia solani (AG 1‐IA), the Rice Sheath Blight Pathogen

Sheath blight disease of rice caused by Rhizoctonia solani is one of the most dreaded plant diseases faced by the rice farmers all over the world. None of the commercially cultivated rice varieties have sufficient level of field resistance, and the disease is presently being managed by chemical pesticides. In this study, 40 isolates of rice sheath blight pathogen, collected from diverse rice ecosystems from 12 different states of India, were characterized for their morphological, pathological and genetic variation. The isolates showed wide morphological variation in terms of size of sclerotia and abundance of sclerotia production. The virulence of each pathogen isolate was studied on four rice varieties, that is TN1, IR 64, Tetep and Swarnadhan in glasshouse, and observations were taken by measuring the relative lesion height. The relative lesion heights produced by these isolates on four different rice varieties varied widely. Genetic variation of the isolates was analysed using ISSR markers. The primers based on AG, GA, AC and CA repeats were informative and revealed polymorphism among the isolates. The polymorphism information content (PIC) of the primers ranged from 0.80 to 0.96, while the resolving power (Rp) ranged from 3.7 to 15.35. Largely, grouping of the isolates happened based on their geographical origin. One isolate from Titabar, Assam, and another from Adialabad, Telangana, were quite distinct from rest of the isolates.     

Biological control of rice sheath blight using hyphae-associated bacteria: development of an <Emphasis Type="Italic">in planta</Emphasis> screening assay to predict biological control agent performance under field conditions

A pathogenicity assay for in planta screening of biological control agents (BCAs) towards sheath blight in rice was developed, and used to evaluate a panel of Rhizoctonia solani Kühn hyphae-associated bacteria for their ability to control sheath blight under screenhouse conditions. The best performing BCA, Burkholderia sp. strain A-7.3, was selected for field trials. Disease incidence and disease severity were significantly reduced leading to a significant grain yield increase of 7%. Furthermore, R. solani sclerotia formation and sclerotia viability were significantly reduced, thus lowering the reservoir for primary infections in the next crop cycle. The paper presents a reliable pathogenicity assay that can be used to test BCA performance under different scenarios e.g. plant variety and soil conditions, and help predict translation of results obtained under screenhouse conditions to field performance. Furthermore, it supports the hypothesis that hyphae-associated bacteria are a promising source of niche-specific BCAs towards fungal pathogens.     

Comparison ofRhizoctonia solani isolates from web blight and basal canker of cowpea and from soil

Summary Isolates ofRhizoctonia solani from web blight and stem basal canker of cowpea and those obtained from soil had similar linear growth rates on potato dextrose agar (PDA) at various temperatures but differed in other features. The web blight isolates differed from the basal canker and the soil isolates in cultural appearance on PDA and on potato marmite agar (PMA). The web blight isolates readily formed discrete sclerotia on PDA, PMA and soil but the other isolates did not. In greenhouse tests, the former were generally the most virulent in inciting foliage and stem basal necrosis and damping-off of seven crop species. Of the plants tested, the legumes were the most susceptible toR. solani.     

Genetic Structure and Aggressiveness of Rhizoctonia solani AG1‐IA,the Cause of Sheath Blight of Rice in Southern China

One hundred and eighty isolates of Rhizoctonia solani AG1‐IA, the causal agent of rice sheath blight, were obtained from six locations in southern China. The genetic structure of R. solani isolates was investigated using random amplified polymorphic DNA (RAPD) markers, and a considerable genetic variation among R. solani isolates was observed. Most of the genetic diversity was distributed within populations, rather than among them. The distribution pattern of the genetic variation of R. solani appears to be the result of high gene flow (Nm) and low‐genetic differentiation among populations. The aggressiveness of R. solani was visually assessed by rice seedlings of five different cultivars in the glasshouse. All isolates tested were found to induce significantly different levels of disease severity, reflecting considerable variation in aggressiveness. The isolates were divided into highly virulent, moderately virulent and weakly virulent groups, and the moderately virulent isolates were dominant in R. solani population. No significant correlation was observed among the genetic similarity, pathogenic aggressiveness and geographical origins of the isolates. Information obtained from this study may be useful for breeding for improved resistance to sheath blight.     

Baseline sensitivity and efficacy of thifluzamide in Rhizoctonia solani

Thifluzamide is a SDHI (succinate dehydrogenase inhibitor) fungicide, which interferes with succinate ubiquinone reductase in the mitochondrial electron transport chain of fungi. Presently, jinggangmycin is the major fungicide extensively used for the control of rice sheath blight caused by Rhizoctonia solani and resistance to jinggangmycin was first reported to occur in China. A total of 128 isolates of R. solani from Anhui Province of China were characterised for the baseline sensitivity to thifluzamide. The isolates were very sensitive to thifluzamide and the baseline sensitivity curve was unimodal with an average EC₅₀ value of 0.058 ± 0.012 µg mL^?1. However, EC₅₀ values of boscalid (another SDHI fungicide) for inhibition of mycelial growth of 22 arbitrarily selected R. solani isolates ranged from 1.89 to 2.68 µg mL^?1. Thifluzamide applied at 110 µg mL^?1 exhibited excellent protective and curative activity against rice sheath blight and provided 81.1–91.0% protective or curative control efficacy. In field trials in 2010 and 2011, control efficacies of thifluzamide at 82 g.a.i ha^?1 15 and 30 days after second application were 84.2% and 86.7%, respectively, suggesting excellent activity against sheath blight. There was a statistically significant difference in the efficacy between thifluzamide and boscalid or jinggangmycin. These results suggested that thifluzamide should be a good alternative fungicide to jinggangmycin for the control of rice sheath blight.     

Comparison among four triazole fungicides on growth and development of sheath blight of rice pathogen Rhizoctonia solani Kühn AG1-1A

The sheath blight of rice pathogen Rhizoctonia solani Kühn AG1-1A has long been known as a major crisis in global rice production. The present study aimed to compare the efficacy of four triazole fungicides at different doses on the growth and development of R. solani Kühn AG1-1A. Obtained results demonstrated inhibitory effects of all four fungicides on mycelial growth, sclerotia formation and biomass production of R. solani Kühn AG1-1A with significant variation among the treatment doses as well as fungicide molecules (p ≤ 0.05). At the respective EC₅₀ doses all four fungicides inhibited cell wall degrading enzymes viz. invertase, cellulase, polygalacturonase and pectin lyase, of the sheath blight pathogen. The extent of inhibition of the enzymes significantly varied among the fungicides. It is important to note that in spite of having common mode of action, all four triazole fungicides demonstrated significant variation in their fungicidal efficacy on R. solani Kühn AG1-1A.     

Dispersal of rice sheath blight fungus,Rhizoctonia solani AG-1(IA), and subsequent disease development in paddy fields, from survey of vegetative compatibility groups

The rice sheath blight fungus,Rhizoctonia solani AG-1(IA), was isolated from surface soil and stubble in the pretrans-planting season (early March) and from sheath blight lesions on plants at the maturing stage (late September) in two paddy fields (TW and TM:8 a). Isolates were classified into vegetative compatibility groups (VCG) based on the formation of a barrage zone (or interaction zone) at the colony interface between isolates on paired cultures. In field TW surveyed in 1991 and 1992, 128–142 isolates obtained in the pretransplanting season and 82–85 isolates from lesions on mature plants were classified into 27–29 and 28–30 VCGs, respectively. Seven to 10 VCGs (26–35%) were common between both samples, and of 30 VCGs from mature plants 12 (40%) survived the winter 1991/1992 and recovered from stubble and/or soil. In field TM in 1992, 111 isolates in the pretransplanting season and 99 from mature plants were divided into 18 and 14 VCGs, respectively. Seven (39%) out of 18 VCGs in the pretransplanting season were recovered again from mature plants. Occurence of common VCGs throughout the year indicated that ca. one third of VCGs found in the pretransplanting season caused sheath blight on rice plants about 6 mo later in the same field. Further, it was shown that about the same rate of VCGs which had caused disease overwintered on stubble and/or surface soil until the following pretransplanting season.     

Endophytic fungus Piriformospora indica induced systemic resistance against rice sheath blight via affecting hydrogen peroxide and antioxidants

In this study, the effect of endophytic fungus Piriformospora indica on Rhizoctonia solani AG1-IA, causal agent of sheath blight disease, was investigated. In addition, plant defence responses activated in P. indica-inoculated rice plants were analysed. Two-week-old seedlings were inoculated by dipping their roots in P. indica chlamydospore suspension and transferred to pots containing sterilized soil. After two weeks, the seedlings pre-inoculated with P. indica were inoculated with R. solani. Statistical analysis of biological indicators showed that application of P. indica increased both fresh and dry weight of rice shoots and roots, compared to those of uninoculated healthy controls and the samples only inoculated with R. solani. Accumulation of hydrogen peroxide (H₂O₂) and activity of antioxidants such as superoxide dismutase (SOD) and guaiacol peroxidase (GPX) in plants inoculated with P. indica, R. solani, and P. indica-R. solani were investigated. The obtained results revealed that P. indica not only increased the plant biomass, but also delayed the infection process of R. solani and decreased sheath blight severity. Decreased severity of the disease was associated with decreased levels of H₂O₂ and increased SOD activity. Considering the necessity of reducing fungicide application, using P. indica in seedling bed before transplantation to the field could be a novel and effective method to increase rice production and decrease sheath blight progress.     

Survival of Rhizoctonia solani AG‐1 IA,the Causal Agent of Rice Sheath Blight,under Different Environmental Conditions

The ability of Rhizoctonia solani AG‐1 IA, the causal agent of rice sheath blight, to survive in diseased rice straw and as sclerotia and mycelia was investigated. After storage for 10 months at 4°C, 25°C and non‐air‐conditioned natural room temperature (NRT, temperature range from 6°C to 35°C), sclerotia placed inside a desiccator, soaked in sterile water or immersed in wet paddy soil were viable. In contrast, only 15% of sclerotia in dry paddy soil survived. Survival of mycelia was severely affected by temperature and humidity. After 10 months in a desiccator at 4°C, 55% of mycelia samples could survive, whereas at 25°C and NRT, mycelial samples survived for only 7 and 5 months, respectively. However, mycelia stored in sterile water at constant temperatures (4°C or 25°C) survived for 10 months. A certain amount of UV radiation had no obvious effect on the survival of sclerotia or mycelia. The survival rate of the fungus in diseased rice straw stored for 16 months could reach 100% at 4°C, 50% at 25°C and 35% at NRT. The survival rates of the pathogen in diseased rice straw buried in dry, wet and flooded paddy soils after 10‐month storage at NRT were 75, 100 and 100%, respectively, indicating that soil humidity is a crucial factor for the survival of this fungus.     

利用纤维二糖差向异构酶制备乳果糖的研究进展

乳果糖是由D-半乳糖和D-果糖两个基团通过β-1,4糖苷键连接而成的还原型二糖;乳果糖口服液具有治疗慢性便秘和肝性脑病的功效,在100多个国家作为常见非处方药(OTC)使用,需求量十分巨大;乳果糖还可以作为益生元改善人体肠道菌群关系。乳果糖的生产依赖化学法,其催化剂对人体有害,下游分离难度大。近年来,纤维二糖差向异构酶被发现能够高效催化乳糖制备乳果糖,该技术绿色环保、步骤简单,具有很强的产业化前景。本文结合自身研究经历对纤维二糖差向异构酶的研发情况进行总结,并综述了乳果糖酶法制备技术的现状。     

Association of the Hydrolytic Enzyme Chitinase against Rhizoctonia solani in Rhizobacteria-treated Rice Plants

Twenty‐two strains of Pseudomonas fluorescens isolated from the rhizosphere soil of nine plant species were screened in vitro for their inhibitory effect on the mycelial growth of the rice sheath blight fungus, Rhizoctonia solani. Of the 22 strains, two promising strains (Pf1 and FP7) were assessed for their effect on seedling vigour and their ability to promote growth in vitro of four cultivars of rice. Both bacterial strains induced systemic resistance in rice cv. IR 50, which is susceptible to sheath blight. After inoculation of the sheaths with the pathogen, Pseudomonas‐treated plants showed an increase in chitinase activity significantly higher than that of untreated control plants. A twofold increase in chitinase activity occurred 2 days after inoculation of plants with the pathogen. Western blot analysis of chitinase indicated the expression of 28 and 38 kDa proteins in rice sheaths against R. solani. Increased induction of the pathogenesis‐related chitinase isoform in Pseudomonas‐treated rice in response to R. solani infection indicates that the induced chitinase has a definite role in suppressing disease development.     

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.     

Integration of soil solarization with chemical, biological and cultural control for the management of soilborne diseases of vegetables

The long-term effectiveness of soil solarization integrated with (integration of pest management [IPM]) a biological control agent (Trichoderma virens), chemical fungicide (pentachloronitrobenzene [PCNB]), organic amendment (chicken litter) or physical method (black agriplastic mulch) to reduce southern blight (Sclerotium rolfsii) and southern root-knot diseases (Meloidogyne incognita) were evaluated on vegetable production. Results showed that the long-term effectiveness of IPM plus soil solarization reduced soilborne diseases of vegetables more than two years following the termination of solarization. These disease management strategies in 1991 and 1992, following soil solarization in 1990, reduced the numbers of sclerotia in the soil, and the number of plants killed by southern blight and root-knot of tomatoes, compared to nonsolarized bare soil treatment. The integration of a reduced dosage level of PCNB or T. virens in field plots, reduced southern blight of tomatoes by 100% and 71%, respectively, in solarized soil, compared to nonsolarized bare soil two years following soil solarization. PCNB effectively controlled southern blight in nonsolarized bare soil both years. All solarized treatments, except PCNB plus solarized soil increased tomato yields compared to nonsolarized bare soil plots. In the second study (1992) following soil solarization in 1991, the effectiveness of solarized bare soil, and nonsolarized bare soil mulched with black agriplastic film, with or without Reemay spunbounded polyester row cover, were effective in reducing root-knot of tomatoes as indicated by the root-knot gall index. Following a one year fallow period in 1994 three years following soil solarization, the root-knot gall index for severity of tomato roots grown in solarized bare soil, nonsolarized bare soil, black agriplastic mulched bare nonsolarized soil and black agriplastic mulched solarized bare soil, were 1.0, 3.0, 3.0 and 2.0, respectively, on a 0–5 scale, where 0=0% and 5=100% root-knot galled. In the third study 1992 and 1993, different dosage levels of chicken litter were used to amend soil artificially infested with sclerotia of S. rolfsii at different depths following solarization, decreased the number of viable sclerotia by 85–100%. All solarized treatments and nonsolarized bare soil amended with 18.8 MT/ha of chicken litter, were effective in controlling southern root-knot damage, and postharvest storage root rots of sweetpotato storage roots (Fusarium root rot [Fusarium solani] and Java black rot [Diplodia tubericola]). Our study showed that all soil solarization treatments, and soils amended with chicken litter, stimulated a shift in the soil microbial population dynamics. Rhizobacteria of Bacillus spp. and fluorescent pseudomonads increased significantly in the rhizosphere, rhizoplane, and interior root tissues of tomatoes and sweetpoatoes, grown in solarized soil compared to nonsolarized soil. These microorganisms may have contributed to the increased growth response of vegetables and some were probably suppressive to soilborne diseases     

Targeted disruption of a G protein α subunit gene results in reduced growth and pathogenicity in <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Cloning and disruption of Rga1, the gene encoding the G protein α subunit in the rice sheath blight fungus Rhizoctonia solani, was investigated. The deduced primary structure of the Rga1-encoded protein showed high identity to those of Gα subunits from other filamentous fungi. Disruption of Rga1 led to decreased vegetative growth and pathogenicity. The Rga1 disruptant showed altered colony morphology. In addition, the sclerotia formation ability of the disruptant was completely lost. These results suggest that the Gα subunit encoded by Rga1 is involved in a signal transduction pathway in R. solani that controls growth, development and pathogenicity.     

Identification of major quantitative trait loci qSBR11-1 for sheath blight resistance in rice

Sheath blight caused by Rhizoctonia solani Kühn is one of the important diseases of rice, resulting in heavy yield loss in rice every year. No rice line resistant to sheath blight has been identified till date. However, in some rice lines a high degree of resistance to R. solani has been observed. An indica rice line, Tetep, is a well documented source of durable and broad spectrum resistance to rice blast as well as quantitative resistance to sheath blight. The present study identified genetic loci for quantitative resistance to sheath blight in rice line Tetep. A mapping population consisting of 127 recombinant inbred lines derived from a cross between rice cultivars HP2216 (susceptible) and Tetep (resistant to sheath blight) was evaluated for sheath blight resistance and other agronomic traits for 4 years across three locations. Based on sheath blight phenotypes and genetic map with 126 evenly distributed molecular markers, a quantitative trait loci (QTLs) contributing to sheath blight resistance was identified on long arm of chromosome 11. Two QTL mapping approaches i.e., single marker analysis and composite interval mapping in multi environments were used to identify QTLs for sheath blight resistance and agronomical traits. The QTL qSBR11-1 for sheath blight resistance was identified between the marker interval RM1233 (26.45 Mb) to sbq33 (28.35 Mb) on chromosome 11. This region was further narrowed down to marker interval K39516 to sbq33 (~0.85 Mb) and a total of 154 genes were predicted including 11 tandem repeats of chitinase genes which may be responsible for sheath blight resistance in rice line Tetep. A set of 96 varieties and a F₂ population were used for validation of markers linked to the QTL region. The results indicate that there is very high genetic variation among varieties at this locus, which can serve as a starting point for allele mining of sheath blight resistance.     

Identification and characterization of rhizosphere fungal strain MF‐91 antagonistic to rice blast and sheath blight pathogens

Aim

To examine the inhibition effects of rhizosphere fungal strain MF‐91 on the rice blast pathogen Magnaporthe grisea and sheath blight pathogen Rhizoctonia solani.

Methods and Results

Rhizosphere fungal strain MF‐91 and its metabolites suppressed the in vitro mycelial growth of R. solani. The inhibitory effect of the metabolites was affected by incubation temperature, lighting time, initial pH and incubation time of rhizosphere fungal strain MF‐91. The in vitro mycelial growth of M. grisea was insignificantly inhibited by rhizosphere fungal strain MF‐91 and its metabolites. The metabolites of rhizosphere fungal strain MF‐91 significantly inhibited the conidial germination and appressorium formation of M. grisea. Moreover, the metabolites reduced the disease index of rice sheath blight by 35·02% in a greenhouse and 57·81% in a field as well as reduced the disease index of rice blast by 66·07% in a field. Rhizosphere fungal strain MF‐91 was identified as Chaetomium aureum based on the morphological observation, the analysis of 18S ribosomal DNA internal transcribed spacer sequence and its physiological characteristics, such as the optimal medium, temperature and initial pH for mycelial growth and sporulation production.

Conclusions

Rhizosphere fungus C. aureum is effective in the biocontrolling of rice blast pathogen M. grisea and sheath blight pathogen R. solani both in in vitro and in vivo conditions.

Significance and Impact of the Study

This study is the first to show that rhizosphere fungus C. aureum is a potential fungicide against rice blast and sheath blight pathogens.