Infection of Rice Plants with the Sheath Blight Fungus Causes an Activation of Pentose Phosphate and Glycolytic Pathways

The response of key regulatory enzymes of the pentose phosphate and glycolytic pathways in disease development was assessed in genetically-related rice plants resistant and susceptible to the sheath blight fungus, Rhizoctonia solani. The plants were grown and maintained under greenhouse conditions and inoculated at 50% flowering. Uninoculated healthy plants served as controls. The activities of pentose phosphate pathway enzymes (glucose-6 phosphate dehydrogenase and 6-phosphogluconate dehydrogenase) increased more than two-fold in both the resistant and susceptible plants. Activities of ATP- and pyrophosphate-dependent phosphofructokinase and phosphoenolpyruvate phosphatase increased in infected plants while activity of phosphoenolpyruvate carboxylase in infected plants was lower than in the healthy plants. Furthermore, for enzymes with increased activity, the levels were higher in the resistant line than in the susceptible line. The enhancement of the enzyme activities correlated well with the post infection period. These data suggest that altered carbohydrate metabolism in sheath blight infections may play an important role in modulating the rice plant's response to infection. The isolation of an infection-induced gene encoding a basic enzyme of pentose phosphate and glycolytic pathways could be used to develop plants with more resistance towards sheath blight disease.     

Cytomolecular aspects of rice sheath blight caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Sheath blight, caused by anastomosis group 1-IA of Rhizoctonia solani Kühn (teleomorph Thanatephorus cucumeris (Frank) Donk), is one of the most destructive rice diseases worldwide. The pathogen is able to infect plants belonging to more than 27 families, including many economically important monocots and dicots such as rice, wheat, alfalfa, bean, peanut, soybean, cucumber, papaya, corn, potato, tomato and sugar beet. It is a soil borne necrotrophic fungus that survives in plant debris as sclerotia, which are small brown-to-black, rocklike reproductive structures. The sclerotia can survive in the soil for several years and infect rice plants at the water-plant interface in the flooded field by producing mycelia. Management of rice sheath blight requires an integrated approach based on the knowledge of each stage of the disease and cytomolecular aspects of rice defence responses against R. solani. This review summarizes current knowledge on molecular aspects of R. solani pathogenicity, genetic structure of the pathogen populations, and the rice-R. solani interaction with emphasis on cellular and molecular defence components such as signal transduction pathways, various plant hormones, host defence genes and production of defence-related proteins involved in basal and induced resistance in rice against sheath blight disease.     

Influence of bacteria isolated from rice plants and rhizospheres on antibiotic production by the antagonistic bacterium <Emphasis Type="Italic">Serratia marcescens</Emphasis> strain B2

Serratia marcescens strain B2 is an antagonistic bacterium that produces the red-pigmented antibiotic prodigiosin and suppresses rice sheath blight caused by Rhizoctonia solani AG-1 IA. Rice sheath blight disease was suppressed when plants were inoculated with this bacterium an hour before pathogen inoculation but not when plants were treated 4 weeks before pathogen inoculation. In both cases the bacteria were detected in the rice rhizosphere 4 weeks after inoculation. Bacteria isolated from the rice plant and rhizosphere inhibited biosynthesis of prodigiosin in S. marcescens strain B2. We suggest that bacteria isolated from rice plants and rhizospheres mediate the suppression of antibiotic production of biological control agents and that such suppression is common under field conditions.     

水稻纹枯病生防细菌筛选及其与病原菌侵染垫形成的关系

为筛选对立枯丝核菌Rhizoctonia solani具有拮抗作用的生防细菌,利用稀释涂布平板法,从吉林省水稻纹枯病样中分离筛选出对立枯丝核菌AG-1A具有高拮抗活性的生防菌株,通过gyrB基因序列分析鉴定其分类地位,并采用离体侵染试验和温室防治试验测定筛选生防菌株对水稻纹枯病的防效,在显微镜下观察生防菌株预处理后接种病原菌的水稻叶片表面的菌丝生长情况,分析其生防机理。结果显示,从水稻纹枯病样中共分离获得35株菌株,其中菌株ND11对立枯丝核菌AG-1A的抑制率和抑菌圈直径最大,分别为74.12%和31.50 mm。基于gyrB基因序列分析最终将菌株ND11鉴定为短小芽胞杆菌Bacillus pumilus;该菌株能够减缓立枯丝核菌AG-1A的侵染速度,抑制其侵染垫的形成,对水稻纹枯病的温室防效达70.69%以上。表明短小芽胞杆菌菌株ND11能够通过抑制立枯丝核菌侵染垫的形成来防治水稻纹枯病,且防效较好,具有开发为生防菌剂的潜力。     

Histopathological assessment of wheat seedling tissues infected by Fusarium pseudograminearum

Histopathological assessment of infection by the crown rot pathogen Fusarium pseudograminearum in wheat seedling tissues was performed using fluorescence microscopy. The coleoptiles and leaf sheaths of four host cultivars of differing susceptibility were examined. Leaf sheaths were most frequently penetrated via stomata, indicated by initial lesions forming at the guard cells. Internally, cell wall penetration was facilitated by penetration structures which appeared as hyphal swellings or septate foot‐shaped appressoria. Colonization of leaf sheaths resulted in the re‐emergence of hyphae from stomata on both surfaces of the sheath. These hyphae are hypothesized to have two major roles; first as exploratory hyphae for colonization of new tissues, and secondly as sites of profuse conidial production. The formation of conidia on the leaf sheath surface was only recorded on the most susceptible bread wheat genotype. No other major differences in host–pathogen interactions were observed among these cultivars. Almost all cell types in the leaf sheath tissues were extensively colonized, except for the vascular bundles and silica cells. This investigation provides the first comprehensive assessment of F. pseudograminearum infection structures and growth patterns during the infection of wheat seedlings.     

水稻稻飞虱和纹枯病系统控制研究

在系统分析历史资料、深入研究稻飞虱和纹枯病发生为害规律基础上,组建病虫测报模型,探讨控害丰产配套栽培技术及提高化防水平,定性定量地建立以准确的测报为前提,合理施肥、密植技术为主的生态控制为基础,按复合防治指标进行化学防治,系统控制稻飞虱、纹枯病,经近几年来的应用,收到显著的经济、生态及社会效益。     

水稻与纹枯病菌互作的分子机制研究进展

水稻纹枯病是由死体营养型真菌立枯丝核菌Rhizoctonia solani AG1-IA亚群引起的一种土传病害,在世界水稻种植区均有发生,是限制水稻高产的主要病害之一。虽然国内外学者针对纹枯病菌已开展了大量的研究工作,但由于该菌寄主范围较广、抗性水稻资源缺乏及其田间抗性鉴定的不稳定性等问题,该病害的研究仍没有取得突破性进展。挖掘自然界中存在的纹枯病抗源材料,选育抗病水稻品种是防控该病害、降低水稻产量损失,从而最大限度保障全球水稻产业可持续发展的有效手段。该文对近年来国内外关于水稻纹枯病菌与寄主的互作分子机制、抗性水稻基因资源挖掘及其抗性机制的最新研究进展进行综述,并提出下一步的重点研究方向,以期为推动水稻对纹枯病的抗性机制解析及抗纹枯病水稻育种提供参考。     

Influence of canopy structure on sheath blight epidemics in rice

Sheath blight (caused by Rhizoctonia solani) is one of the most important constraints in achieving high grain yield in intensive rice production systems. Canopy structure can influence the development of sheath blight epidemics. The objective of this study was to determine the effect of canopy structure parameters such as shoot number, leaf area index, biomass production, contact frequency, light transmittance and plant height on the development of sheath blight epidemics in commercial fields. Field experiments were conducted in both early and late seasons of 2009 and 2010 in Wuxue, Hubei province, China. The effects of nitrogen (N) rate and hill density on structure and production parameters and sheath blight severity were investigated. Sheath blight severity was recorded as a sheath blight index or relative lesion height on inoculated and uninoculated plants in each crop. Lesion length was measured on inoculated plants in 2010. The results showed that the sheath blight index increased with an increase of N rate and hill density in uninoculated plots in each trial. Stepwise multiple regression analysis demonstrated that contact frequency was consistently related to sheath blight. Lesion length on inoculated plants was not correlated with canopy structure. These results indicate that canopy structure influences sheath blight epidemics. A ‘healthy’ canopy resulting from appropriate crop management practices can be used to suppress sheath blight epidemics in rice.     

High activities and mRNA expression of pyrophosphate-fructose-6-phosphate-phosphotransferase and 6-phosphofructokinase are induced as a response to Rhizoctonia solani infection in rice leaf sheaths

Rice sheath blight disease caused by Rhizoctonia solani Kuhn results in significant yield and quality losses in rice growing areas worldwide. The glycolytic pathway is important in the resistance response to R. solani infection in rice. This study examined one of the regulatory steps in this pathway catalyzed by pyrophosphate-fructose-6-phosphate-phosphotransferase (PFP) and 6-phosphofructokinase (PFK). PFP and PFK activity in R. solani-infected rice plants increased. The mRNA expression of PFP/PFK isozymes showed that PFK 1, 2, 4 and 5 in the resistant line at 1 dpi were high as compared to the gradual increase observed in the expression of all PFP isozymes. Also PFK 1, PFK 3, PFK 4, PFK 5, PFP 2 and PFP 5 were adaptive to sheath blight disease infection and linked to defence response while, the expressions of PFK 2, PFP 1, PFP 3 and PFP 4 although adaptive, were not specific to R. solani infection. These observations provide evidence that (a) both PFP and PFK have isozymes that play an adaptive role after R. solani infection but while those of PFK are expressed at higher levels within a short time after infection those of PFP are expressed gradually, (b) the adaptive activation of PFP in R. solani-infected rice plants is correlated with the paired expression of its α- and β-subunits as shown by PFP 2 and PFP 5, and (c) the expression of some α-subunits is not specific to R. solani infection as shown by PFP 1, PFP 3 and PFP 4.     

水稻纹枯病生防菌株的筛选、鉴定及其防治效果

为获得对水稻纹枯病有生防潜力的菌株,从湖北省恩施土家族苗族自治州植物根际土壤中分离获得菌株,筛选对水稻纹枯病菌有拮抗作用的菌株,并测定其发酵液的抑菌活性、抑菌谱及生物学特性;基于形态学特征、生理生化特征及分子生物学特征对其进行分类鉴定,并通过离体叶片、室内盆栽和田间试验进一步评价其生防潜力。结果显示,从根际土壤中共分离获得162株菌株,其中生防菌株E12对水稻纹枯病菌的抑制率最高,达94.93%,且该菌株对常见的12种植物病原菌均有抑制作用,抑制率在59.84%~93.14%之间;生防菌株E12能分泌蛋白酶、嗜铁素和淀粉酶,具有固氮及形成生物被膜的能力;结合形态学特征、生理生化特征及分子生物学特征,将菌株E12鉴定为越南伯克霍尔德氏菌Burkholderia vietnamiensis;生防菌株E12处理离体水稻叶片后,其相对病斑长度较对照显著减小,对水稻纹枯病的室内盆栽和田间防治效果分别为62.51%和67.78%。表明越南伯克霍尔德氏菌E12在防控水稻纹枯病方面有较好的潜力。     

Purification of an Elicitor-Inducible Antifungal Chitinase from Suspension-Cultured Rice Cells

Suspension-cultured rice cells showed an appreciable amount of chitinase activity when the cultured cells were treated with an elicitor isolated fromRhizoctonia solani, the rice sheath blight pathogen. A fivefold increase in chitinase activity was observed 24 h after elicitor treatment. The elicitor-inducible chitinase was purified by ammonium sulfate fractionation, chitin affinity chromatography and gel filtration. Its molecular weight is 35 kDa and it has an isoelectric point of 8.3. The 35 kDa basic chitinase inhibited mycelial growth ofR. solani in vitro. Morphological changes appeared within 1 h following exposure of mycelium to the chitinase. The hyphal tip showed marked swelling and subsequently lysis was also observed.     

Rhizoctonia solani: taxonomy,population biology and management of rhizoctonia seedling disease of soybean

Rhizoctonia solani, the most important species within the genus Rhizoctonia, is a soilborne plant pathogen with considerable diversity in cultural morphology, host range and aggressiveness. Despite its history as a destructive pathogen of economically important crops worldwide, our understanding of its taxonomic relationship with other Rhizoctonia‐like fungi, incompatibility systems, and population biology is rather limited. Among the host of diseases it has been associated with, seedling diseases inflicted on soybean are of significant importance, especially in the soybean growing regions of North America. Due to the dearth of resistant soybean genotypes, as well as the paucity of information on the mechanisms of host–pathogen interactions and other molecular aspects of pathogenicity, effective management options have mostly relied upon a combination of cultural and chemical control options. The first section of this review summarizes what is currently known about the taxonomy and systematics, population biology and molecular genetics of R. solani. The second section provides an overview of the pathology and management of rhizoctonia root and hypocotyl rot of soybean, a seedling disease of importance in North America.     

Histopathology of infection and colonization of Quercus ilex fine roots by Phytophthora cinnamomi

Quercus ilex is one of the European forest species most susceptible to root rot caused by the oomycete Phytophthora cinnamomi. This disease contributes to holm oak decline, a particularly serious problem in the ‘dehesas’ ecosystem of the southwestern Iberian Peninsula. This work describes the host–pathogen interaction of Q. ilex and P. cinnamomi, using new infection indices at the tissue level. Fine roots of 6‐month‐old saplings inoculated with P. cinnamomi were examined by light microscopy and a random pool of images was analysed in order to calculate different indices based on the measured area of pathogen structures. In the early stages of invasion, P. cinnamomi colonizes the apoplast and penetrates cortical cells with somatic structures. On reaching the parenchymatous tissues of the central cylinder, the pathogen develops different reproductive and survival structures inside the cells and then expands through the vascular system of the root. Some host responses were identified, such as cell wall thickening, accumulation of phenolic compounds in the middle lamella of sclerenchyma tissues, and mucilage secretion blocking vascular cells. New insights into the behaviour of P. cinnamomi inside fine roots are described. Host responses fail due to rapid expansion of the pathogen and a change in its behaviour from biotrophic to necrotrophic.     

Population genetic evidence that basidiospores play an important role in the disease cycle of rice‐infecting populations of Rhizoctonia solani AG‐1 IA in Iran

The fungus Rhizoctonia solani AG‐1 IA causes sheath blight, one of the most important rice diseases worldwide. The first objective of this study was to analyse the genetic structure of R. solani AG‐1 IA populations from three locations in the Iranian Caspian Sea rice agroecosystem. Three population samples of R. solani AG‐1 IA isolates were obtained in 2006 from infected rice fields separated by 126–263 km. Each field was sampled twice during the season: at the early booting stage and 45 days later at the early mature grain stage. The genetic structure of these three populations was analysed using nine microsatellite loci. While the population genetic structure from Tonekabon and Amol indicated high gene flow, they were both differentiated from Rasht. The high gene flow between Tonekabon and Amol was probably due mainly to human‐mediated movement of infested seeds. The second objective was to determine the importance of recombination. All three populations exhibited a mixed reproductive mode, including both sexual and asexual reproduction. No inbreeding was detected, suggesting that the pathogen is random mating. The third objective was to determine if genetic structure within a field changes over the course of a growing season. A decrease in the proportion of admixed genotypes from the early to the late season was detected. There was also a significant (P = 0·002) increase in the proportion of loci under Hardy–Weinberg equilibrium. These two lines of evidence support the hypothesis that basidiospores can be a source of secondary inoculum.     

A broad diversity survey of Rhizoctonia species from the Brazilian Amazon reveals the prevalence of R. solani AG-1 IA on signal grass and the new record of AG-1 IF on cowpea and soybeans

Leaf blight, sheath blight, and web blight are major diseases caused by Rhizoctonia species on both Fabaceae and Poaceae plant hosts in the Brazilian Amazon agroecosystem. To determine the diversity of Rhizoctonia species associated with foliar diseases on fabaceous (cowpea and soybean) and poaceous (rice and signal grass [Urochloa brizantha]) hosts, a broad survey was conducted in Pará, Rondônia, Roraima, and Mato Grosso, in the Amazon, from 2012 to 2013. We extended our survey to Cerrado areas of Mato Grosso, and the lowlands of Paraíba Valley, in São Paulo, where these Rhizoctonia foliar diseases have not been reported so far. Our findings revealed that these diseases are caused by a diversity of Rhizoctonia solani AG-1 complex. We detected that R. solani AG-1 IA (sexual phase Thanatephorus cucumeris) was the predominant pathogen associated with signal grass leaf blight and collar rot diseases in the Amazon, especially in Rondônia and northern Mato Grosso. In addition, a subgroup of R. solani (AG-1 IF), not previously reported in Brazil, was associated with leaf blight on cowpea and soybean, in Roraima. Another subgroup (AG-1 ID) was also detected in Roraima. In Mato Grosso Cerrados we did not find any of the major Rhizoctonia foliar pathogens. Instead, R. oryzae (Waitea circinata) was the predominant species associated with a collar rot on U. brizantha. In the lowlands of São Paulo, R. oryzae-sativae (Ceratobasidium oryzae-sativae) was the predominant pathogen detected causing the rice sheath spot disease.     

Invertase Activities in Sheath-blight Infected Rice Selection Lines and Fungus Culture Filtrates

Infection of rice plants with Rhizoctonia solani, the sheath blight fungus, at the flowering stage resulted in an increase in acid invertase activity. Activity of the invertase(s) with optimum pH at 3.5 and 4.5 was higher in the susceptible plants compared to resistant plants. Healthy control plants had no change in invertase activity. An invertase of fungal origin produced in sheath blight fungus culture filtrates had an optimum acidic pH of 3.5. The increase in invertase activity observed in the later stages of infection in the susceptible plants compared to the resistant plants demonstrated a relation to fungal growth. These results suggested that the fungus produced an invertase for the hydrolysis of sucrose that resulted in alterations of source-sink relationships in the colonized cells. Thus, the increased invertase activity in the susceptible plants regulated the ratio of hexose to sucrose. This observation is further supported by the lack of a significant invertase increase in the resistant plants. Received 14 July 1999/ Accepted in revised form 9 December 1999     

Effect of various crop establishment methods practised by Asian farmers on epidemics of rice sheath blight caused by Rhizoctonia solani

Establishment methods for rice crops in tropical Asia are very diverse, leading to variation in the structure of rice canopies. Differences in canopy structure can in turn affect the spread of the rice sheath blight pathogen, Rhizoctonia solani . Rice sheath blight epidemics were compared during two seasons in crops established by different methods: direct broadcasting of pregerminated rice seeds, and transplanting of rice seedlings at spacings of 20 × 20 cm, 13 × 25 cm and 25 × 25 cm between hills (i.e. along and between rows, respectively). In both years, the apparent infection rate based on incidence data and the terminal severity of sheath blight were lower in the direct-seeded crops than in any of the transplanted ones, regardless of spacing. The frequency of leaf-to-leaf contacts ( CF ) between hills (or plants) was highest in direct-seeded rice, and lowest in rice transplanted at a spacing of 25 × 25 cm. Larger CF is known to favour rice sheath blight epidemics. The apparent contradiction between higher incidence and lower CF in the transplanted stands than in the direct-seeded stands is interpreted in terms of accessibility of healthy host tissues to the spread of the pathogen in the canopy, and accounts for within-host (rice hill or plant) and between-host (hill or plant) disease spread. The analysis of incidence-severity relationships indicated a less aggregated distribution of the disease in direct-seeded rice, which was related to the spatial distribution of the tillers. These findings have direct implications for the management of the disease.     

Brachypodium distachyon is a pathosystem model for the study of the wheat disease rhizoctonia root rot

Brachypodium distachyon (Bd) is increasingly being used as a model for cereal diseases and to study cereal root architecture. Rhizoctonia solani AG 8 is a necrotrophic root pathogen that infects wheat soon after germination resulting in reduced plant growth and yield loss. Genetic resistance to R. solani AG 8 is not available in commercial wheat cultivars, although some quantitative levels of resistance have previously been found in mutant lines and grass relatives. Resistance mechanisms in cereals remain unknown. The ability to use Bd as a model to study the wheat–R. solani AG 8 pathosystem was investigated. The results presented show that Bd is susceptible to R. solani AG 8 and that the pathogen infects both species to a similar degree, producing comparable disease symptoms. Root length reduction was the primary indicator of disease, with shoots also affected. The second objective was to develop a repeatable phenotyping method to screen Bd populations for resistance to R. solani AG 8. Results of a preliminary experiment provide evidence for variation in resistance between Bd inbred lines. This is the first report showing the potential of Bd as a model plant for discovery of quantitative genetic variation in resistance to a necrotrophic cereal root pathogen.     

Resistance of Sorghum Line CS 621 to Rhizoctonia solani AG1-IA and Other Sorghum Pathogens

Sorghum line CS 621 was evaluated along with other lines and cultivars for resistance to Rhizoctonia sheath blight, tar spot and gray leaf spot for 3 years. CS 621 was consistently resistant to these diseases even under a heavy natural outbreak of tar spot in the breeding nursery. It was also found to be more resistant to Rhizoctonia sheath blight than the resistant lines from Japan during the 1993 to 1994 screening tests. Resistance to Rhizoctonia solani AG1-IA was expressed in the form of reduction in the severity of disease and rate of infection. Evaluation of progenies from crosses involving CS 621 and a susceptible variety, UPL Sg5, indicated that additive and dominant gene effects are important in the expression of quantitative resistance to R. solani. CS 621 could therefore serve as a source of multiple resistance genes in a breeding program for high yield and stability against sorghum diseases. Received 6 August 1999/ Accepted in revised form 18 October 1999     

稻飞虱和纹枯病化控的策略及复合指标研究

采取田间小区和网室盆栽试验,研究稻飞虱和纹枯病混合发生的化控策略及复合防治指标。结果表明,水稻孕穗期(7月上中旬)用高效、长效、安全药剂扑虱灵和粉锈灵混合防治的“治前控后”化防策略保产效果最佳。试验证明,稻飞虱和纹枯病对水稻有明显的交互作用。组建了稻飞虱(白背飞虱、褐飞虱)虫量(X_(1-2))、纹枯病病株率(X_3)的复合为害损失及动态经济阈值模型。经田间小区、大田应用检验,不仅防效高,保产效果好,而且减少施药次数,保护了天敌,三大效益显著。