水稻基部叶鞘封泥对水稻纹枯病的影响

采用先封泥于水稻基部叶鞘再人工接种水稻纹枯病菌,调查水稻纹枯病发生情况,同时测定水稻叶鞘过氧化物酶活性和可溶性糖含量变化。连续3年试验的调查结果表明：先封泥于水稻基部叶鞘再人工接种水稻纹枯病菌的发病率、病情指数明显低于井冈霉素处理和对照,其控病效果为72.80%~81.21%。先封泥再接病菌处理的过氧化物酶活性比对照高,但低于井冈霉素处理;先封泥再接病菌使可溶性糖含量降低,但不同处理之间差异不显著。     

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.     

内生真菌紫杉木霉ZJUF0986菌株及其活性代谢产物防治水稻纹枯病的效果

从南方红豆杉分离到的一种新内生真菌紫杉木霉Trichoderma taxi菌株ZJUF0986,与水稻纹枯病菌对峙培养。结果发现,该内生真菌通过菌丝缠绕附着等重寄生方式,导致纹枯病菌菌丝断裂或其内含物降解直至死亡。其产生的活性代谢产物也能强烈抑制纹枯病菌菌丝的生长,显著降低病原菌的菌核萌发率;对病原菌菌丝生长和菌核萌发的有效中浓度EC50分别为1．08和3．59μg／ml。盆栽试验结果表明,菌株ZJUF0986浓度80μg／ml的活性代谢产物对水稻纹枯病的防效达63．82％,与50μg／ml井冈霉素A的防治效果相当。     

伯克霍尔德氏菌JP2-270抗水稻纹枯病菌机理的初步研究

水稻纹枯病是由立枯丝核菌Rhizoctonia solani引起的,是水稻的三大病害之一.本研究采用基因敲除、荧光定量PCR和靶向代谢物差异分析等方法,初步探索了伯克霍尔德氏菌Burkholderia sp.JP2-270抑制水稻纹枯病菌的作用机制.首先,选取了水稻稻瘟病菌Magnaporthe oryza、香蕉枯萎病...     

秸秆还田对水稻根际微生物多样性和水稻纹枯病发生的影响

为探究稻麦轮作模式下秸秆还田对水稻根际微生物多样性与水稻纹枯病发生的影响,本研究采用Illumina测序技术结合生物信息学比较分析了秸秆还田与不还田处理的水稻根际微生物群落多样性差异,并采用实时荧光定量PCR （real-time fluorescent quantitative PCR,RT-qPCR）对这2种稻田土壤中水稻纹枯病菌Rhizoctonia solani含量进行测定,并采用随机五点取样法调查稻田中水稻纹枯病发病率和病情指数。结果表明,秸秆还田后水稻根际细菌群落Chao1丰富度指数和Shannon多样性指数均有所增加,分别增加14.42%和4.39%,其中前者增幅显著;根际真菌群落Chao1丰富度指数降低29.60%,而Shannon多样性指数略有增加,增加5.96%;在门水平上,秸秆还田和不还田处理的水稻根际土壤优势细菌门均为变形菌门、放线菌门、绿弯菌门和酸杆菌门,优势真菌门均为子囊菌门、接合菌门和担子菌门;在属水平上,秸秆还田处理的硝化螺旋菌属和芽胞杆菌属相对丰度分别比不还田处理的显著增加96.11%和106.19%;在水稻孕穗期秸秆还田土壤中水稻纹枯病菌含量比不还田处理的显著增加120.20%;秸秆还田后水稻纹枯病发生加重,病情指数比不还田处理的增加18.10%~45.41%。     

广西水稻内生细菌的动态分布及其对水稻纹枯病菌的拮抗作用

从广西稻区74种水稻品种中采集了222个水稻标样,分离出1903株内生细菌。经形态和生理生化测定,确定这些菌株分属于芽孢杆菌属Bacillus、鞘胺醇单孢菌属Sphingomonas、土壤杆菌属Agrobacterium、伯克霍尔德氏菌属Burkholderia、肠杆菌属Enterobacter和成团泛菌属Pantoea。其中,Bacillus和Enterobacter为主要类群,Bacillus为优势类群。水稻内生细菌的类群随不同的水稻品种和栽培地区而变化。水稻内生细菌在植株器官中的分布以根部数量最多,其次是叶鞘和叶。内生细菌的总量在水稻生育期的变化趋势是从苗期到孕穗期上升,从灌浆期到乳熟期下降。分离到的1903株内生细菌中,有412个菌株对水稻纹枯病菌Rhizoctoniasolani有拮抗作用。     

微生物杀菌剂及其混配剂防治水稻纹枯病的田间药效试验

对微生物杀菌剂及其混配剂对水稻纹枯病进行田间防治试验,结果表明枯草芽孢杆菌10亿/g可湿性粉剂、井冈霉素·蜡质芽孢杆菌12.5%可湿性粉剂对水稻纹枯病有较好的防治效果。在发病初期施药,枯草芽孢杆菌10亿/g可湿性粉剂1500g/hm2防效为75.4%,产量增幅21.5%;井冈霉素·蜡质芽孢杆菌12.5%可湿性粉剂2880g/hm2防效为83.89%。可以作为井冈霉素的替代产品,进行推广应用。     

氯苯醚酰胺(Y13149)生物活性初步研究及水稻纹枯病菌对其敏感性基线的建立

氯苯醚酰胺[N-(2-(2,4-二氯苯氧基)苯基)-3-(二氟甲基)-1-甲基-1H-吡唑-4-酰胺,Y13149]是由华中师范大学基于药效团连接碎片虚拟筛选策略(PFVS)快速发现的新型琥珀酸脱氢酶抑制剂类(SDHIs)化合物。采用菌丝生长速率法,测定了该创制化合物的农药生物活性。结果表明:氯苯醚酰胺对真菌具有较广的抗菌谱,对供试10种植物病原真菌均具有较好的抑制活性,EC50值在0.008~15.25 mg/L之间,其对立枯丝核菌的抑制活性最强,但对辣椒疫霉和链状腐霉活性较弱。田间试验表明,氯苯醚酰胺对水稻纹枯病具有较好的防治效果,有效剂量75 g/hm2时的平均防效为79.6%,其防效总体略好于噻呋酰胺(75 g/hm2的平均防效为77.3%)。此外,氯苯醚酰胺对采自浙江和江苏两省的125株水稻纹枯病菌的EC50值分布在0.002~0.259 mg/L之间,且呈近似正态单峰曲线,平均EC50值为(0.021 ±0.008) mg/L,因此可将其作为水稻纹枯病菌对氯苯醚酰胺的敏感性基线,用于田间抗药性监测。     

Characterization of Rhizoctonia Species Associated with Foliar Necrosis and Leaf Scorch of Clonally-propagated Eucalyptus in Brazil

The objective was to identify and characterize the causal agent of foliar necrosis and leaf scorch of Eucalyptus spp. in Brazil. Nineteen putative isolates of Rhizoctonia obtained from Eucalyptus plants during clonal propagation were compared with isolates from other hosts and with tester strains of anastomosis groups of Rhizoctonia solani. Features compared were morphological characteristics of anamorphs and teleomorphs, numbers of nuclei per cell in the vegetative hyphae, anastomosis of hyphae, and ability to produce necrotic lesions on cuttings and damping-off of E. grandis×E. urophylla hybrid seedlings. Rhizoctonia solani AG1 (AG1-IB like) was the most frequent causal agent isolated from Eucalyptus plants and cuttings with symptoms of leaf scorch and foliar necrosis respectively. These isolates were highly virulent on Eucalyptus cuttings and presented naturally epiphytic growth on Eucalyptus shoots. Binucleate isolates and isolates of R. solani AG4 were also virulent on cuttings and were most virulent on Eucalyptus seedlings causing pre- and post-emergence damping-off. Virulence on Eucalyptus cuttings and seedlings was not restricted to a single species or anastomosis group of Rhizoctonia.     

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。     

禾谷丝核菌环介导等温扩增检测技术体系的建立

为建立简便、快速和灵敏的小麦纹枯病菌——禾谷丝核菌Rhizoctonia cerealis的分子检测技术体系,基于环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术,以核糖体DNA内转录间隔区(internal transcribed spacer,ITS)为靶标序列,设计并筛选特异性引物,建立禾谷丝核菌的LAMP检测方法,并对其特异性、灵敏度和应用效果进行测定。结果表明,基于筛选获得的4条禾谷丝核菌特异性引物建立的LAMP方法能够从15种植物病原菌中特异性地检出禾谷丝核菌,近缘种立枯丝核菌Rhizoctonia solani和引致早期症状易混淆病害的病原菌禾顶囊壳菌Gaeumannomyces graminis、麦根腐平脐蠕孢菌Bipolaris sorokiniana和禾谷镰孢菌Fusarium graminearum均未检出;该LAMP方法在日光下的灵敏度为10^-1 ng/μL,在蓝光下的灵敏度为10^-3 ng/μL;对发病组织的检测显示该LAMP方法能从人工接种的小麦发病组织中...     

Investigations on the effect of haulm destruction and additional root cutting on black scurf on potato tubers

In greenhouse experiments factors which are involved in the stimulation of sclerotia formation byRhizoctonia solani Kühn on potato tubers after haulm destruction were investigated. Cutting off the shoot stimulated the production of sclerotia as much as chemical haulm destruction. This was also observed when potato tubers were grown away from the roots in a separate compartment filled with steamed perlite. Fewer sclerotia were produced when roots were cut in addition to haulm destruction compared to haulm destruction alone. Cutting off the roots alone often stimulated sclerotia production. The data indicate that sclerotia production was directly affected by the tuber and probably due to physiological changes in the tuber caused by elimination of the shoot.Samenvatting In kasproeven werd onderzocht welke factoren betrokken zijn bij de stimulering van de produktie van sclerotiën op aardappel doorRhizoctonia solani Kühn als gevolg van loofvernietiging. Afknippen van het loof stimuleerde de vorming van sclerotiën evenzeer als chemische vernietiging. Na afknippen van het loof trad de stimulering ook op bij knollen, die zich ruimtelijk gescheiden van de wortels in gestoomd vochtig perliet konden ontwikkelen. Doodspuiten plus doorsnijden van de wortels resulteerde in minder lakschurft dan doodspuiten alleen. Wanneer alleen de wortels werden doorgesneden resulteerde dat vaak in meer lakschurft. De vorming van sclerotiën wordt dus direct door de knol beïnvloed. Dit effect lijkt voort te komen uit fysiologische veranderingen in de knol.     

噻呋酰胺与嘧菌酯混配对马铃薯黑痣病菌增效作用及其田间防效

为明确噻呋酰胺与嘧菌酯混配对马铃薯黑痣病菌Rhizoctonia solani的增效作用及对马铃薯黑痣病的田间防效,采用菌丝生长速率法和田间药效试验测定二者混配组合对其的联合毒力,分别用Wadley法和Abbott法评价混配组合的增效作用,并比较拌种、沟施及苗期茎基部喷淋3种施药方式对黑痣病的田间防效。结果表明,噻呋酰胺与嘧菌酯以质量比3∶1、2∶1、1∶1和1∶2进行混配时抑制马铃薯黑痣病菌菌丝生长的增效系数分别为1.7、2.7、2.5和1.9,其中2∶1和1∶1的混配组合增效作用较为明显。240 g/L噻呋酰胺悬浮剂与250 g/L嘧菌酯悬浮剂1∶1桶混以240 g(a.i.)/hm~2沟施对马铃薯黑痣病的防效优于拌种和苗期茎基部喷淋的防效,沟施方式下对植株黑痣病的防效为65.3%,对薯块黑痣病的防效为71.6%,增效系数为1.1,表现增效作用。表明噻呋酰胺与嘧菌酯有效成分质量比1∶1的混配组合以240 g(a.i.)/hm~2为推荐剂量沟施可有效防治马铃薯黑痣病。     

Phylogenetic analysis of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> subgroups associated with web blight symptoms on common bean based on ITS-5.8S rDNA

Sixty-eight Rhizoctonia solani isolates (31 AG-1, 37 of AG-2-2) associated with web blight (WB) of common bean, Phaseolus vulgaris, were examined for sequence variations in the ITS-5.8S rDNA region. The isolates were collected in bean-growing lowland and mountainous regions in Central and South America. Sequences of these isolates were aligned with other known R. solani sequences from the NCBI GenBank and distance and parsimony analysis were used to obtain phylogenetic trees. WB isolates of AG-1 formed two clades separated from known AG-1 subgroups. WB isolates of AG-2-2 formed one clade separated from known AG-2-2 subgroups. Other isolates belonged to AG-1 IA and AG-1 IB. Based on phylogenetic analysis, we confirmed that at least five genetically different subgroups incite WB of common beans. Three new subgroups of R. solani have been identified and designated as AG-1 IE, AG-1 IF and AG-2-2 WB. DNA sequences of these isolates provided needed information to design taxon-specific primers that can be employed in ecological/epidemiological studies and seed health tests.     

Molecular and Pathotype Analysis of the Rice Blast Fungus in North Vietnam

Rice blast caused by Pyricularia oryzae is a devastating disease worldwide. In Vietnam, rice blast is especially severe in the Red River Delta in the North. The genetic diversity of 114 P. oryzae isolates collected from rice in 2001 in the Red River Delta and nine additional Vietnamese P. oryzae isolates was analysed using Amplified Fragment Length Polymorphism (AFLP). DNA similarity and cluster analysis based on 160 polymorphic AFLP markers showed twelve different AFLP genetic groups among the 123 field isolates. Isolates collected from japonica hosts clustered separately from indica host isolates with at least 60% dissimilarity with little evidence for gene flow between the two populations. In the 2001 population originating from indica hosts, three genetic groups were predominant and represented 99% of the isolates sampled. One predominant clonal lineage represented 59% of the 2001 indica host population and was found in eleven provinces in the Red River Delta of North Vietnam. Significant genotype flow could be demonstrated between the indica population south of Red river and the indica population north of Red river. There was significant linkage disequilibrium between the AFLP loci within the indica population, indicating that this is not a random mating population. Pathogenicity tests of 25 isolates selected from the 12 AFLP groups on a set of 29 differential rice lines revealed two avirulent isolates and 23 pathotypes. Different combinations of known resistance genes were found to have potential for blast resistance breeding for North Vietnam. First two authors contributed equally     

An Aspartic Protease With Antimicrobial Activity is Induced after Infection and Wounding in Intercellular Fluids of Potato Tubers

Aspartic proteases (APs) one of the main proteinase classes, have different physiological functions in animals, fungi and viruses. In plants, knowledge of the biological roles of APs is less well developed. An AP has been purified from potato tuber and leaves (Guevara et al., 1999, 2001). In this paper, the changes in the level of AP in response to infection by Phytophthora infestans (P. infestans) and wounding were studied in intercellular washing fluids (IWFs) from tuber disks of two potato cultivars differing in their susceptibility to P. infestans. A differential induction was observed between both cultivars: in the resistant cultivar, induction was higher and faster in infected tissues than in wounded ones. In the susceptible cultivar, a lower and later accumulation was observed than in the resistant cultivar. In addition, AP had a direct inhibitory effect on the germination of cysts of P. infestans and conidia of Fusarium solani. The pattern of accumulation and in vitro activity of AP suggest that this enzyme may have a role in the defense response of potato.     

Development and Evaluation of an in vitro Detached Leaf Assay forc Pre-Screening Resistance to Fusarium Head Blight in Wheat

An in vitro detached leaf assay, involving the inoculation of detached leaves with Microdochium nivale, was further developed and used to compare with whole plant resistance ratings to Fusarium head blight (FHB) of 22 commercial cultivars and published information on 21 wheat genotypes, identified as potential sources for FHB resistance. An incubation temperature of 10 °C and isolates of M. nivale var. majus of intermediate pathogenicity were found to be the most suitable for the differential expression of several components of partial disease resistance (PDR), namely incubation period, latent period and lesion length, in wheat genotypes used in the detached leaf assay. There were highly significant differences (P < 0.001) for each component of PDR within commercial cultivars and CIMMYT genotypes. Positive correlations were found between incubation period and latent period (r = 0.606; P < 0.001 and r = 0.498; P < 0.001, respectively, for commercial cultivars and CIMMYT genotypes), inverse correlations between incubation period and lesion length (r = -0.466; P < 0.01 and r = –0.685; P < 0.001, respectively) and latent period and lesion length (r = –0.825; P < 0.001 and r = –0.848; P < 0.001, respectively). Spearman rank correlations between individual PDR components and UK 2003 recommended list ratings were significant for incubation period (r_s = 0.53; P < 0.05) and latent period (r_s = 0.70; P < 0.01) but not for lesion length (r _s = –0.26). Commercial cultivars identified with high resistances across all three PDR components in the detached leaf assay also had high whole plant FHB resistance ratings, with the exception of cv. Tanker which is more susceptible than the results of the detached leaf assay suggested, indicating an additional susceptibility factor could be present. Agreement between resistances found in the detached leaf assay and resistance to FHB suggests resistances detected in detached leaves are under the same genetic control as much of the resistances expressed in the wheat head of the commercial cultivars evaluated. In contrast, high resistances in each of the PDR components were associated with higher susceptibility across 19 CIMMYT genotypes previously evaluated as potential breeding sources of FHB resistance (incubation period: r = 0.52; P < 0.01, latent period: r = 0.53; P < 0.01, lesion length: r = –0.49; P < 0.01). In particular, the CIMMYT genotypes E2 and E12 together with Summai #3, known to have high levels of whole plant FHB resistance, showed low levels of resistance in each PDR component in the detached leaf assay. Such whole plant resistances, which are highly effective and not detected by the detached leaf assay, do not appear to be present in Irish and UK commercial cultivars. The most resistant Irish and UK commercial cultivars were comparable to the genotype Frontana and the most resistant CIMMYT germplasm evaluated in the leaf assay.     

Effect of haulm destruction supplemented by cutting off roots on the incidence of black scurf and skin damage,flexibility of harvest period and yield of seed potatoes in field experiments

In field experiments, supplementing chemical haulm destruction (CHD) with cutting off roots resulted in a lower incidence of black scurf and skin damage (ripping off the skin) at harvest date than CHD alone. The lower susceptibility to skin damage at harvest allowed harvesting to beging on an earlier date, when only a few sclerotia ofRhizoctonia solani had developed. Furthermore, black scurf often developed more slowly after haulm destruction if roots had been severed and this enabled harvesting to be postponed.At harvest, gross yield was highest if roots had not been cut through prior to CHD: the extra weight existed merely of water. Weight loss during storage, however, as well as grading losses resulting from black scurf were greater after CHD alone. This resulted in an equal or even lower net yield after CHD alone than after CHD supplemented by root severing. The favourable effects of supplementing CHD with cutting off roots almost equaled those of the mechanical removal, often called haulm pulling or plant pulling. Factors that may affect the development of black scurf are discussed.Samenvatting Pootaardappelen raakten minder bezet met lakschurft en werden minder ontveld bij de oogst na looftrekken of na wortelsnijden plus doodspuiten dan na loofklappen plus spuiten. Al vanaf de derde dag na trekken of wortelsnijden plus doodspuiten was de mate van ontvelling zo gering, dat met het oogsten kon worden begonnen. Op zo'n vroeg tijdstip na loofdoding konden knollen worden geoogst met nog weinig lakschurft, want de stimulering van lakschurft werd pas zichtbaar vanaf 7 à 10 dagen na loofdoding. Lakschurft ontwikkelde zich het traagst na looftrekken. Ook wortelsnijden bij het doodspuiten gaf meestal een tragere ontwikkeling van lakschurft dan klappen plus spuiten. Daardoor konden na wortelsnijden plus doodspuiten en vooral na looftrekken op nog latere tijdstippen knollen met weinig lakschurft worden geoogst.Na klappen plus spuiten was het bruto gewicht bij de oogst het hoogst. Dat meergewicht bleek louter uit water te bestaan en het verloop in knolvochtgehalte leek erop te wijzen dat de wortels nog gedurende een week na doodspuiten blijven functioneren. Dat gaf wel knollen met een hoger vochtgehalte die meer ontvelden en meer vocht en dus gewicht verloren. Dit groter gewichtsverlies bij bewaren en de hogere leesverliezen door lakschurft deden de meeropbrengst teniet: zo werd het netto knolgewicht na klappen plus spuiten, al naar gelang het moment van oogsten, meestal gelijk aan, of lager dan het netto knolgewicht na het trekken of na wortelsnijden plus doospuiten. In dit artikel worden de factoren besproken, die mogelijk van invloed zijn op de ontwikkeling van lakschurft.     

Interactions Between the Potato Cyst Nematode Globodera rostochiensis and Diseases Caused by Rhizoctonia solani AG3 in Potatoes Under Field Conditions

Findings from 2 years of field experiments investigating the relationship between Globodera rostochiensis and Rhizoctonia solani on unique field sites are reported. In 2000, a field experiment was positioned on land that had previously been used for experimental work investigating integrated potato cyst nematode (PCN) management methods. This study had produced an ‘untypical’ mosaic of PCN population densities ranging from 5 to 221 eggs g⁻¹ soil. In 2001, the field experiment was conducted on a different field site and overlaid on a focus of G. rostochiensis population densities ranging from 11 to 108 eggs g⁻¹ soil. In each experiment, potatoes (cv. Désirée) were grown in plots with similar population densities of G. rostochiensis that were either uninoculated or inoculated with R. solani. A series of potato plant harvests were undertaken to investigate the effects of nematode infestation on the incidence and severity of R. solani diseases and the associated development of plants. In both experiments, a clear relationship was found between the density of G. rostochiensis juveniles present in potato roots and the incidence of stolons infected by R. solani, 6 weeks after planting. For the first time this interaction has been determined under field conditions. The results of the study suggest that the interaction between nematode and fungus is indirect and possible mechanisms are discussed.     

Comparison of sequences for the internal transcribed spacer region in <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG 1-ID and other subgroups of AG 1

The rDNA-ITS sequence of Rhizoctonia solani AG 1-ID was determined and compared to those of R. solani AG 1-IA, AG 1-IB, and AG 1-IC. The similarity of the isolates from each AG 1 subgroup was almost identical (99%–100%), whereas it was lower between subgroups (91%–95%) than within subgroups. Phylogenetic analysis indicated that isolates of AG 1-ID and other subgroups were separately clustered. Isolates of R. solani AG 1 were clearly separated from R. solani AG 2-1, AG 4, and binucleate Rhizoctonia AG-Bb and AG-K. These results showed that analysis of the rDNA-ITS sequence is an optimal criterion for differentiating R. solani AG 1-ID from other subgroups of R. solani AG 1.