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.     

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.     

Rice siR109944 suppresses plant immunity to sheath blight and impacts multiple agronomic traits by affecting auxin homeostasis

Plant small RNAs (sRNAs) play significant roles in regulating various developmental processes and hormone signalling pathways involved in plant responses to a wide range of biotic and abiotic stresses. However, the functions of sRNAs in response to rice sheath blight remain unclear. We screened rice (Oryza sativa) sRNA expression patterns against Rhizoctonia solani and found that Tourist‐miniature inverted‐repeat transposable element (MITE)‐derived small interfering RNA (siRNA) (here referred to as siR109944) expression was clearly suppressed upon R. solani infection. One potential target of siR109944 is the F‐Box domain and LRR‐containing protein 55 (FBL55), which encode the transport inhibitor response 1 (TIR1)‐like protein. We found that rice had significantly enhanced susceptibility when siR109944 was overexpressed, while FBL55 OE plants showed resistance to R. solani challenge. Additionally, multiple agronomic traits of rice, including root length and flag leaf inclination, were affected by siR109944 expression. Auxin metabolism‐related and signalling pathway‐related genes were differentially expressed in the siR109944 OE and FBL55 OE plants. Importantly, pre‐treatment with auxin enhanced sheath blight resistance by affecting endogenous auxin homeostasis in rice. Furthermore, transgenic Arabidopsis overexpressing siR109944 exhibited early flowering, increased tiller numbers, and increased susceptibility to R. solani. Our results demonstrate that siR109944 has a conserved function in interfering with plant immunity, growth, and development by affecting auxin homeostasis in planta. Thus, siR109944 provides a genetic target for plant breeding in the future. Furthermore, exogenous application of indole‐3‐acetic acid (IAA) or auxin analogues might effectively protect field crops against diseases.     

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.     

水稻纹枯病菌的矿质营养生态位

运用生态位原理与方法,研究了水稻纹枯病菌的矿质元素营养生态位.结果表明,在分蘖盛期、孕穗期、抽穗期和蜡熟期,水稻纹枯病菌矿质营养生态位宽度指数分别为0.2710、0.3865、0.4252和0.4817,即随着水稻的生长而逐渐增大,但各生育期的生态位宽度总体上仍偏小.表明在水稻生长的各时期水稻纹枯病菌只占有了矿质营养类型中的较少部分.水稻纹枯病菌总是优先占有低Mg、低Zn、低Si的营养位,说明Mg、Zn、Si的含量与水稻对纹枯病的抗性紧密相关.     

云南30个水稻品种对水稻白叶枯病的抗性评价

【目的】水稻白叶枯病是一种严重危害水稻的细菌性病害,培育抗性品种是治理该病害的重要途径。因此,明确云南水稻材料对检疫性病害水稻白叶枯病的抗性,可以为该病害的防治与监测、水稻栽培的合理布局和良好抗性资源的获取提供依据。【方法】采用剪叶接种法测定云南稻区30个品种对7个不同致病型白叶枯病菌的抗性。【结果】在供试的30个云南水稻品种中,2个品种(玉粳16和JS42糯稻)对7个不同致病型菌株均表现为抗性;15个品种对7个致病型菌株均表现感病;对HEN11、SCYC-6、YN7、YN11、FUJ、YN241和PX099等7个致病型菌株表现抗性的水稻品种分别占26.67%、16.67%、23.33%、13.33%、6.67%、10.00%和20.00%。此外,区试材料的抗性比例高于主栽品种,地方稻未发现抗性品种。【结论】现在生产上的大部分水稻品种对优势致病型病原菌入侵的抵抗能力降低甚至丧失。针对云南地区的优势致病小种FUJ筛选得到2个抗性品种:玉粳16和JS42糯稻。     

Pyramiding transgenic resistance in elite indica rice cultivars against the sheath blight and bacterial blight

Elite indica rice cultivars were cotransformed with genes expressing a rice chitinase (chi11) and a thaumatin-like protein (tlp) conferring resistance to fungal pathogens and a serine-threonine kinase (Xa21) conferring bacterial blight resistance, through particle bombardment, with a view to pyramiding sheath blight and bacterial blight resistance. Molecular analyses of putative transgenic lines by polymerase chain reaction, Southern Blot hybridization, and Western Blotting revealed stable integration and expression of the transgenes in a few independent transgenic lines. Progeny analyses showed the stable inheritance of transgenes to their progeny. Coexpression of chitinase and thaumatin-like protein in the progenies of a transgenic Pusa Basmati1 line revealed an enhanced resistance to the sheath blight pathogen, Rhizoctonia solani, as compared to that in the lines expressing the individual genes. A transgenic Pusa Basmati1 line pyramided with chi11, tlp, and Xa21 showed an enhanced resistance to both sheath blight and bacterial blight. S. Maruthasalam and K. Kalpana have contributed to this article equally.     

Development and evaluation of Trichoderma asperellum preparation for control of sheath blight of rice (Oryza sativa L.)

Sheath blight, which is caused by Rhizoctonia solani, is a disease that majorly impacts rice production. A biocontrol agent used for control rice sheath blight must be sprayed on the stem at specific times during rice growth, a process that is labour-intensive and renders the antagonist vulnerable to environmental factors. In this study, Trichoderma asperellum T12 was used to produce preparation by solid-state fermentation using a surface-response method. Rice hull was selected as a carrier based on its ability to sustain the T12 floating in the water and protect T12 from ultraviolet irradiation. The production of a T12-based preparation required 32% wheat bran, 7% inoculum, 2.3 g kg^?1 (NH₄)₂SO₄ and 65% water content, with fermentation at 27.5°C for 30 days and agitation every six days. The preparation demonstrated 90% biocontrol efficacy and significantly (P > 0.05) increased the seed-set rate and 1000-grain weight as compared with the pathogen treatment. The population of Trichoderma on the surface of rice leaf sheath in the treatment applied with T12 preparation increased from 232 cfu (colony forming units) g^?1 fw (fresh weight) to 436 cfu g^?1 fw during rice growth stage, which was significantly (P > 0.05) higher than pathogen treatment. The population of R. solani on the leaf sheath increased from 41 cfu g^?1 fw to 271 cfu g^?1 fw in the pathogen treatment, while remained stable (P > 0.05) at level of 10–23 cfu g^?1 fw in T12 preparation applied treatment. Biocontrol of sheath blight by the addition of the preparation to the soil is effective and decreases the costs of agro-industrial waste disposal.     

Rapid suppression of defence enzymes and compounds by sheath blight (Rhizoctonia solani) and sheath rot (Sarocladium oryzae) toxins in rice cell suspension cultures

To understand the suppression mechanisms against disease resistance in rice, we took advantage of the fact that suspension cultured cells exhibit many of the defence responses that are characteristic of intact tissues. In this study we constitutively measured the Rhizoctonia solani and Sarocladium oryzae toxins, induced and suppressed levels of phenylalanine ammonia lyase, peroxidase, superoxide dismutase, phenols, catalase, β-1,3-glucanase and chitinase in rice suspension cultured cells. The addition of Rhizoctonia solani and Sarocladium oryzae toxins separately in suspension cultured cells shows the suppression of defence enzymes and compounds at 24 h and 48 h respectively except SOD. The rice cultivar IR50 delays the disease suppression effect when compared to the other cultivars viz., Pusa Basmati and Co 43. The PR proteins (namely β-1,3-glucanase and chitinase) activities in rice suspension cultured cells were reduced during 48 h and 72 h after the addition of Rhizoctonia solani toxin, whereas the activities were suppressed only after 72 h when inoculated with Sarocladium oryzae toxin. Selective suppression of these defence enzymes and compounds by Rhizoctonia solani and Sarocladium oryzae toxin shows that toxins play a major role during pathogenesis in rice cells.     

Potential of Serratia marcescens strain B2 for biological control of rice sheath blight

Serratia marcescens strain B2 inhibited mycelial growth of the rice sheath blight pathogen Rhizoctonia solani AG-1 IA. Rice plants were treated with bacterial suspension and then challenge inoculated with the pathogen. Application of S. marcescens effectively reduced the incidence of sheath blight. S. marcescens survived in soil under glasshouse conditions at ca. 10⁸ colony forming units g^-1 of soil for 4 weeks after application. These results suggest that S. marcescens has potential as an effective and persistent biological control agent for rice sheath blight.     

Marker assisted selection of bacterial blight resistance genes in rice

Bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae is one of the most important diseases affecting rice production in Asia. We were interested in surveying rice genotypes that are popularly used in the Indian breeding program for conferring resistance to bacterial blight, using 11 STMS and 6 STS markers. The basis of selection of these DNA markers was their close linkage to xa5, xa13, and Xa21 genes and their positions on the rice genetic map relative to bacterial blight resistance genes. Eight lines were found to contain the xa5 gene while two lines contained Xa21 gene and none of the lines contained the xa13 gene with the exception of its near-isogenic line. Using the polymorphic markers obtained in the initial survey, marker-assisted selection was performed in the F3 population of a cross between IR-64 and IET-14444 to detect lines containing multiple resistance genes. Of the 59 progeny lines analyzed, eight lines contained both the resistance genes, xa5 and Xa4.     

Greenhouse and field trials of the bacterial antagonists in pelletformulations to suppress sheath blight of rice caused by Rhizoctoniasolani

Bacterial formulations, produced using both Bacillus megaterium andB. pumilus individually with pharmaceutical technology, were testedunder both greenhouse and field conditions. In the greenhouse testing,some bacterial formulations, for instance For 7 minus Lac and For 16 minusLac, performed as well as freshly prepared bacterial antagonists insuppress sheath blight disease. In the field testing, For 16 minus Lac wasnot effective in suppressing sheath blight development. Failure of the For16 minus Lac to suppress sheath blight disease in the field trial may be dueto the dilution and inactivation of antibiotics produced by B.megaterium in the aquatic environment in the rice field and climaticconditions during the formulation application.     

Isozyme polymorphism and virulence of Indian isolates of the rice sheath blight fungus

Inadequate information about the genetic structure of the polyphagous Rhizoctonia solani has made sheath blight resistance breeding a difficult task. To assess the variability in the Indian populations of sheath blight fungus, 18 isolates were collected from different rice growing regions of India and analyzed for virulence and electrophoretic profiles of 13 isozymes. The virulence spectrum of all 18 isolates was examined on susceptible IR50 and tolerant Swarnadhan varieties, based on which the isolates could be grouped as highly virulent, moderately virulent or a virulent. A total of 11 enzyme systems with 153 electrophoretic phenotypes were applied to characterize the genetic variation among the isolates. Cluster analyses based on isozyme patterns resulted in one major cluster comprising 16 virulent isolates, with two a virulent isolates loosely linked to this at 0.13 similarity. Isozyme systems of esterases (both and ) and 6-phosphogluconic dehydrogenase could be used to fingerprint the individual isolates.This revised version was published online in October 2005 with corrections to the Cover Date.     

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.     

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.     

Prospect of the QTL-qSB-9^Tq utilized in molecular breeding program of japonica rice against sheath blight

The major QTL-qSB-9Tq conferring partial resistance to rice (Oryza sativa L.) sheath blight (Rhizoctonia solani Kühn) has been verified on chromosome 9 of the indica rice cultivar, Teqing. In this study, the prospect of this QTL utilized in molecular breeding program of japonica rice for sheath blight resistance was investigated. Most of the japonica rice cultivars showed lower level of sheath blight resistance than the indica rice cultivars. At the corresponding site of qSB-9Tq, nine typical japonica rice culfivars from different ecological regions or countries proved to possess the susceptible allele(s). Introgression of qSB-9Tq into these cultivars enhanced their resistance level by decreasing sheath blight score of 1.0 (0.5-1.3), which indicated that qSB-9Tq had a large potential in strengthening the resistance of japonica rice to sheath blight. The use of the three molecular markers, which were polymorphic between Teqing and many japonica rice cultivars, promotes the application of qSB-9Tq in a concrete molecular breeding program.     

Functional validation of pathogenicity genes in rice sheath blight pathogen Rhizoctonia solani by a novel host-induced gene silencing system

Rice sheath blight, caused by the soilborne fungus Rhizoctonia solani, causes severe yield losses worldwide. Elucidation of the pathogenic mechanism of R. solani is highly desired. However, the lack of a stable genetic transformation system has made it challenging to examine genes' functions in this fungus. Here, we present functional validation of pathogenicity genes in the rice sheath blight pathogen R. solani by a newly established tobacco rattle virus (TRV)–host-induced gene silencing (HIGS) system using the virulent R. solani AG-1 IA strain GD-118. RNA interference constructs of 33 candidate pathogenicity genes were infiltrated into Nicotiana benthamiana leaves with the TRV-HIGS system. Of these constructs, 29 resulted in a significant reduction in necrosis caused by GD-118 infection. For further validation of one of the positive genes, trehalose-6-phosphate phosphatase (Rstps2), stable rice transformants harbouring the double-stranded RNA (dsRNA) construct for Rstps2 were created. The transformants exhibited reduced gene expression of Rstps2, virulence, and trehalose accumulation in GD-118. We showed that the dsRNA for Rstps2 was taken up by GD-118 mycelia and sclerotial differentiation of GD-118 was inhibited. These findings offer gene identification opportunities for the rice sheath blight pathogen and a theoretical basis for controlling this disease by spray-induced gene silencing.     

湿地稻-鸭复合系统中水稻纹枯病的变化规律

为了探明稻田养鸭对水稻纹枯病的发生、发展的影响 ,为稻 -鸭复合系统中水稻纹枯病的防治提供依据 ,笔者在中稻、晚稻田进行了稻田养鸭试验。试验结果表明 ,在中稻田每 6 6 6 .7m2 放养体重 15 0 g左右鸭子 15～ 2 0只 ,能使纹枯病病蔸率减少5 6 .0 % ,病株率减少 5 7.74 % ,同时比用井岗霉素防治的小区病蔸率下降 9.0 % ,病株率下降 15 .2 5 % ,病情指数比空白对照下降2 6 .4 6 ,比施用井岗霉素的施药区减少 0 .95 ;防治效果显著 ,基本可控制纹枯病的危害     

水稻白叶枯病抗性基因Xa21和稻瘟病抗性基因Pi-d2激酶蛋白质在毕赤酵母中的表达及条件优化

【目的】白叶枯病和稻瘟病是最主要的水稻病害,Xa21是水稻白叶枯病抗性基因,Pi-d2是稻瘟病抗性基因,二者都编码类受体激酶蛋白质。本研究旨在毕赤酵母系统中表达XA21和PI-D2激酶蛋白质。【方法】用Xa21和Pi-d2的激酶区PCR产物,构建了pPICZαA-Xa21K、pPICZαA-Pi-d2K重组质粒,酶切及测序验证后,将重组质粒线性化,转化到毕赤酵母菌株中,系统地比较了不同酵母菌株(KM71、GS115、X33),不同甲醇浓度(1%、2%、3%),不同pH(pH5、pH6、pH7、pH8)值,不同诱导时间(24h、48h、72h)条件下激酶蛋白质的表达情况。【结果】XA21和PI-D2激酶蛋白质可以在毕赤酵母中表达,但表达的蛋白质不能分泌到培养基上清中,而只能在菌体中检测到,对表达条件的系统比较发现,毕赤酵母菌株KM71和X33、2%的甲醇诱导浓度、pH5和48h以上的诱导时间有利于激酶蛋白质的表达,最后我们在酵母裂解物上清中获得了纯化的考染可见的激酶蛋白质。【结论】在毕赤酵母中表达了XA21和PI-D2激酶蛋白质,为下一步生化特性研究奠定了基础。