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.     

Rice Resistance to Sheath Blight Mediated by Potassium

This study investigated the effect of potassium (K) on sheath blight (Rhizoctonia solani) development on rice plants from cultivars BR‐IRGA 409 and Labelle grown in nutrient solution containing 0, 50 and 100 mm of K. Sheath blight progress on inoculated sheaths was evaluated by measuring the relative lesion length at 48, 72, 96 and 120 h after inoculation (hai). Data were used to calculate the area under relative lesion length progress curve (AURLLPC). The foliar K concentration on leaf sheaths tissue increased by 61.48 and 116.05% to cultivars BR‐IRGA 409 and Labelle, respectively, as the K rates increased from 0 to 100 mm . A linear model best described the relationship between the AURLLPC and the K rates. The AURLLPC decreased by 29.2 and 21.3% for cultivars BR‐IRGA 409 and Labelle, respectively, as the K rates in the nutrient solution increased. It can be concluded that high K concentration on leaf sheaths tissue was important to decrease sheath blight symptoms on rice leaf sheaths.     

Understanding sheath blight resistance in rice: the road behind and the road ahead

Rice sheath blight disease, caused by the basidiomycetous necrotroph Rhizoctonia solani, became one of the major threats to the rice cultivation worldwide, especially after the adoption of high‐yielding varieties. The pathogen is challenging to manage because of its extensively broad host range and high genetic variability and also due to the inability to find any satisfactory level of natural resistance from the available rice germplasm. It is high time to find remedies to combat the pathogen for reducing rice yield losses and subsequently to minimize the threat to global food security. The development of genetic resistance is one of the alternative means to avoid the use of hazardous chemical fungicides. This review mainly focuses on the effort of better understanding the host–pathogen relationship, finding the gene loci/markers imparting resistance response and modifying the host genome through transgenic development. The latest development and trend in the R. solani–rice pathosystem research with gap analysis are provided.     

Activation of Systemic Resistance to Magnaporthe oryzae in Rice by Substances Produced by Fusarium solani Isolated from Soil

Of 70 micro‐organisms (fungi, bacteria and actinomycetes) isolated from soil using vegetable tissue baits, 16 produced substances in culture fluids capable of preventing the development of blast caused by Magnaporthe oryzae on rice leaves with little or no inhibitory effect on the conidial germination of the pathogen. Isolate KS‐F14, which secreted substances capable of activating resistance in untreated leaves, was selected and identified as Fusarium solani. The resistance‐inducing substances were effective at pH values ranging from 5 to 10 and were stable under high temperatures, maintaining approximately the same level of activity even after autoclaving for 20 min. After application, the activated resistance in rice leaves persisted for 14 days. The polar solvent extracts of freeze‐dried KS‐F14 secretions were effective in activating resistance against M. oryzae in rice plants. The non‐polar solvent extracts were also effective, albeit not as effective as the polar solvent extracts, indicating that although the majority of the secreted resistance‐inducing compounds are hydrophilic, some of the compounds are hydrophobic. Treating secretions with cation or anion exchange resins only partially reduced their resistance‐inducing ability, suggesting that the resistance‐inducing components include both charged and non‐charged compounds. The resistance‐inducing compounds produced by F. solani have the potential to be developed into a commercial product for the control of rice blast and possibly other plant diseases.     

Effect of Magnesium on the Development of Sheath blight in Rice

This study investigated the effect of magnesium (Mg) on sheath blight, caused by Rhizoctonia solani, development on rice plants from cultivars BR‐IRGA 409 and Labelle grown in nutrient solution containing 0.062, 0.125, 0.25 and 0.50 mm of Mg. Sheath blight progress on inoculated sheaths was evaluated by measuring lesions expansion (mm) at 24, 48, 72 and 96 h after inoculation. Data were used to calculate the area under lesion expansion progress curve (AULEPC). The relationship between the foliar Mg concentration and the Mg rates was quadratic. The Mg concentration on leaf sheaths tissue was highest at the Mg rates of 0.389 and 0.400 mm , respectively, for cultivars BR‐IRGA 409 and Labelle. A linear model best described the relationship between the AULEPC and the Mg rates. The AULEPC decreased by 48.7 and 26.2% for plants of cultivars BR‐IRGA 409 and Labelle, respectively, as the Mg rates in the nutrient solution increased. The results permitted to conclude that high foliar Mg concentration played a pivotal role to decrease sheath blight lesions expansion.     

Biocontrol potential of phylloplane bacterium Ochrobactrum anthropi BMO‐111 against blister blight disease of tea

Aims

The present study was carried out to screen the phylloplane bacteria from tea for antagonism against grey blight caused by Pestalotiopsis theae and blister bight caused by Exobasidium vexans and to further evaluate the efficient isolates for disease control potential under field condition.

Methods and Results

A total of 316 morphologically different phylloplane bacteria were isolated. Among the antagonists, the isolates designated as BMO‐075, BMO‐111 and BMO‐147 exhibited maximum inhibitory activity against both the pathogens under in vitro conditions and hence were selected for further evaluation under microplot field trial. Foliar application of 36‐h‐old culture of BMO‐111 (1 × 10⁸ colony‐forming units ml^?1) significantly reduced the blister blight disease incidence than the other isolates. The culture of BMO‐111 as well as its culture filtrate effectively inhibited the mycelial growth of various fungal plant pathogens. The isolate BMO‐111 was identified as Ochrobactrum anthropi based on the morphological and 16S rDNA sequence analyses.

Conclusions

It could be concluded that the biocontrol agent O. anthropi BMO‐111 was effective against blister blight disease of tea.

Significance and Impact of the Study

Further study is required to demonstrate the mechanism of its action and formulation for the biocontrol potential against blister blight disease of tea.     

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.     

Identification of rice Allene Oxide Cyclase mutants and the function of jasmonate for defence against Magnaporthe oryzae

Two photomorphogenic mutants of rice, coleoptile photomorphogenesis 2 (cpm2) and hebiba, were found to be defective in the gene encoding allene oxide cyclase (OsAOC) by map‐based cloning and complementation assays. Examination of the enzymatic activity of recombinant GST–OsAOC indicated that OsAOC is a functional enzyme that is involved in the biosynthesis of jasmonic acid and related compounds. The level of jasmonate was extremely low in both mutants, in agreement with the fact that rice has only one gene encoding allene oxide cyclase. Several flower‐related mutant phenotypes were observed, including morphological abnormalities of the flower and early flowering. We used these mutants to investigate the function of jasmonate in the defence response to the blast fungus Magnaporthe oryzae. Inoculation assays with fungal spores revealed that both mutants are more susceptible than wild‐type to an incompatible strain of M. oryzae, in such a way that hyphal growth was enhanced in mutant tissues. The level of jasmonate isoleucine, a bioactive form of jasmonate, increased in response to blast infection. Furthermore, blast‐induced accumulation of phytoalexins, especially that of the flavonoid sakuranetin, was found to be severely impaired in cpm2 and hebiba. Together, the present study demonstrates that, in rice, jasmonate mediates the defence response against blast fungus.     

Somatic Hybrids Between Potato and S. berthaultii Show Partial Resistance to Soil‐Borne Fungi and Potato Virus Y

Three tetraploid somatic hybrid lines produced by protoplast fusion between a dihaploid potato, Solanum tuberosum, cultivar BF15 and the wild potato species Solanum berthaultii were evaluated here for their response to different soil‐borne pathogens, that is Fusarium solani, Pythium aphanidermatum and Rhizoctonia solani as well as to infection by potato virus Y (PVY). Both hybrid and BF15 plants grown in vitro were inoculated with the tested pathogen strains, that is R. solani, P. aphanidermatum, or F. solani. The growth level and disease severity index of these plants were compared to the susceptible commercial cultivar Spunta. A better growth of inoculated hybrid plants and restricted disease symptoms were observed in comparison with the commercial plants. Under glasshouse conditions and after inoculation with R. solani and P. aphanidermatum, improved resistance of the hybrid plants to these pathogens was confirmed. Indeed, these plants showed no significant damage following inoculation and a better development in R. solani‐infected plants. The susceptibility of the hybrid tubers to R. solani, P. aphanidermatum, and to F. solani infection was also determined. A significant reduction of tissue colonisation was observed in all the hybrid lines compared to the cultivated cultivars. The STBc and STBd hybrids also showed improved resistance to the PVY ordinary strain (PVY^o) under glasshouse conditions.     

Inhibition of OsSWEET11 function in mesophyll cells improves resistance of rice to sheath blight disease

Pathogen–host interaction is a complicated process; pathogens mainly infect host plants to acquire nutrients, especially sugars. Rhizoctonia solani, the causative agent of sheath blight disease, is a major pathogen of rice. However, it is not known how this pathogen obtains sugar from rice plants. In this study, we found that the rice sugar transporter OsSWEET11 is involved in the pathogenesis of sheath blight disease. Quantitative real‐time polymerase chain reaction (qRT‐PCR) and β‐d ‐glucuronidase expression analyses showed that R. solani infection significantly enhanced OsSWEET11 expression in leaves amongst the clade III SWEET members. The analyses of transgenic plants revealed that Ossweet11 mutants were less susceptible, whereas plants overexpressing OsSWEET11 were more susceptible, to sheath blight compared with wild‐type controls, but the yield of OsSWEET11 mutants and overexpressors was reduced. SWEETs become active on oligomerization. Split‐ubiquitin yeast two‐hybrid, bimolecular fluorescence complementation and co‐immunoprecipitation assays showed that mutated OsSWEET11 interacted with normal OsSWEET11. In addition, expression of conserved residue mutated AtSWEET1 inhibited normal AtSWEET1 activity. To analyse whether inhibition of OsSWEET11 function in mesophyll cells is related to defence against this disease, mutated OsSWEET11 was expressed under the control of the Rubisco promoter, which is specific for green tissues. The resistance of transgenic plants to sheath blight disease, but not other disease, was improved, whereas yield production was not obviously affected. Overall, these results suggest that R. solani might acquire sugar from rice leaves by the activation of OsSWEET11 expression. The plants can be protected from infection by manipulation of the expression of OsSWEET11 without affecting the crop yield.     

Isolation and Identification of Bacillus amyloliquefaciens IBFCBF‐1 with Potential for Biological Control of Phytophthora Blight and Growth Promotion of Pepper

In this study, 76 bacterial strains were isolated from the rhizosphere soil of pepper. Of these, 23 bacterial isolates capable of inhibiting Phytophthora capsici growth were selected. Among the antagonistic bacteria, one strain, IBFCBF‐1 showed the strongest antagonistic activity, and was identified as Bacillus amyloliquefaciens based on the results of 16S rRNA gene sequence analysis, physiological and biochemical testing, and morphological characteristics. When tested with a dual‐culture method and with laboratory greenhouse studies, the strain IBFCBF‐1 was found to be a potential biocontrol agent for controlling the plant pathogen, P. capsici. Moreover, it showed high efficiency and broad‐spectrum antifungal properties in vitro. Under greenhouse conditions, IBFCBF‐1 could significantly promote the growth of pepper seedlings, and was able to solubilize phosphate, and produce indole acetic acid (IAA) and ammonia. This study clearly demonstrated that IBFCBF‐1 is a potential candidate exhibiting phytophthora blight‐suppressive and plant growth‐promoting effects on pepper.     

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.     

Greek indigenous streptomycetes as biocontrol agents against the soil‐borne fungal plant pathogen Rhizoctonia solani

Aims

To examine the biocontrol potential of multiactive Greek indigenous Streptomyces isolates carrying antifungal activity against Rhizoctonia solani that causes damping‐off symptoms on beans.

Methods and Results

A total of 605 Streptomyces isolates originated from 12 diverse Greek habitats were screened for antifungal activity against R. solani DSM843. Almost one‐third of the isolates proved to be antagonistic against the fungus. From the above isolates, six were selected due to their higher antifungal activity, identified by analysing their 16S rRNA gene sequence and studied further. The obtained data showed the following: firstly, the isolates ACTA1383 and ACTA1557 exhibited the highest antagonistic activity, and therefore, they were selected for in vivo experiments using bean seeds as target; secondly, in solid and liquid culture experiments under optimum antagonistic conditions, the medium extracts from the isolates OL80, ACTA1523, ACTA1551 and ACTA1522 suppressed the growth of the fungal mycelium, while extracts from ACTA 1383 and ACTA1557 did not show any activity.

Conclusions

These results corresponded important indications for the utility of two Greek indigenous Streptomyces isolates (ACTA1557 and ACTA1383) for the protection of the bean crops from R. solani damping‐off symptoms, while four of them (isolates OL80, ACTA1523, ACTA1551 and ACTA1522) seem to be promising producers of antifungal metabolites.

Significance and Impact of the Study

This is the first study on the biocontrol of R. solani using multiactive Streptomyces isolates originated from ecophysiologically special Greek habitats. Our study provides basic information to further explore managing strategies to control this critical disease.     

Antagonistic potential of native strain Streptomyces aurantiogriseus VSMGT1014 against sheath blight of rice disease

A total of 132 actinomycetes was isolated from different rice rhizosphere soils of Tamil Nadu, India, among which 57 showed antagonistic activity towards Rhizoctonia solani, which is sheath blight (ShB) pathogen of rice and other fungal pathogens such as Macrophomina phaseolina, Fusarium oxysporum, Fusarium udum and Alternaria alternata with a variable zone of inhibition. Potential actinomycete strain VSMGT1014 was identified as Streptomyces aurantiogriseus VSMGT1014 based on the morphological, physiological, biochemical and 16S rRNA sequence analysis. The strain VSMGT1014 produced lytic enzymes, secondary metabolites, siderophore, volatile substance and indole acetic acid. Crude metabolites of VSMGT1014 showed activity against R. solani at 5 µg ml^?1; however, the prominent inhibition zone was observed from 40 to 100 µg ml^?1. Reduced lesion heights observed in culture, cells-free filtrate, crude metabolites and carbendazim on challenge with pathogen in the detached leaf assay. The high content screening test clearly indicated denucleation of R. solani at 5 µg ml^?1 treatment of crude metabolite and carbendazim respectively. The results conclude that strain VSMGT1014 was found to be a potential candidate for the control of ShB of rice as a bio fungicide.     

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.     

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.

     

Phenotyping of Partial Physiological Resistance to Rice Sheath Blight

Sheath blight, caused by Rhizoctonia solani, is one of the most important rice diseases worldwide especially under irrigated agro‐ecosystems. To date, no rice accession with complete resistance to sheath blight has been reported. However, a number of genotypes with varying levels of resistance have been reported. Twelve genotypes (including mega varieties) viz. Tetep, Jasmine 85, Te‐Qing, Duduruchi, Betichikon, Khatochalani, D‐6766, D‐256, Swarna, Sarju‐52, MTU‐1010 and Samba Mashuri were evaluated for quantitative measurement of partial physiological resistance to sheath blight under controlled conditions using detached tiller method. Three independent experiments, each involving three replications, were conducted. Seven days after inoculation, the following disease variables were measured: number of lesions, lesion length, vertical sheath colonization (VSC) on the tiller, disease severity, relative vertical sheath colonization (RVSC) and survival of the leaf blade. Variation between rice genotypes was observed for all the disease variables. Disease severity and VSC were the two most correlated variables, whereas the number of lesions and mean lesion length were the least correlated variables. The ranking of varieties often differed depending on the disease variable considered. Amongst the genotypes tested, D‐256, Tetep and Jasmin‐85 had the lowest number of lesions and disease severity. Similarly, Tetep and D‐256 showed the lowest levels of RVSC, whilst Jasmine‐85 was found to be intermediate. D‐6766, Samba Mashuri and Betichikon showed the highest levels of disease variables. The fraction of dead leaves ranged from 0.00 to 0.38. No dead leaves were observed in Te‐Qing, Swarna and MTU‐1010. The highest fraction of dead leaves was observed for Betichikon (0.38) followed by Duduruchi and D‐6766 (0.33). Our results suggest that this method in combination with other phenotyping methods could be used to quantify partial resistance to rice sheath blight.