立枯丝核菌的生物防治

本文扼要叙述了立枯丝核菌生防因子的种类生防几生防因子在根部的定殖,并讨论了有关问题。     

Resistance in Rhizoctonia solani to tolclofos-methyl

Isolates ofRhizoctonia solani were adapted in vitro to grow on a medium amended with tolclofos-methyl at a concentration 500 times that which initially almost completely inhibited growth.Acquired resistance was retained after five transfers on a fungicide-free medium. Pathogenicity of resistant isolates was not reduced, but their growth rates on PDA were significantly lower than those of the original isolates. Recovery of the resistant isolates was not improved on a selective medium amended with tolclofos-methyl.Samenvatting Na overenting op een medium dat tolchlofos-methyl bevatte, raakten enkele isolaten vanRhizoctonia solani gewend aan 500 maal de dosis die oorspronkelijk bijna alle groei verhinderde.Resistente isolaten vanR. solani bleven minder gevoelig voor tolchlofos-methyl na vijf overentingen op een medium zonder het fungicide. De pathogeniteit van resistente isolaten was niet verminderd, maar hun groeisnelheid op PDA was significant vertraagd vergeleken bij die van de oorspronkelijke isolaten. Isolatie van de resistente stammen werd niet verbeterd op een selectief medium waaraan tolchlofos-methyl was toegevoegd.     

Degradation of Nitralin by Rhizoctonia solani

Degradation studies of nitralin [4-(methylsulfonyl)-2.6-dinitro-N.N-dipropylaniline] were performed with pure cultures of Rhizactoma solani Kühn. Czapek Dos Broth was selected as the growth medium and contained no components which interfered with extraction and subsequent TLC analysis. Ten to I I degradation products were detected after 14 to 21 days incubation at 33°C, The nonpolar fraction (ethyl acetate exlraclahkl contained at least nine compounds visible on TLC plales. Only one product other than the parent nitralin was identified the monoalkylated derivate. 4-(meihylsulfonyl)-2.6-dinitro-N-pro-pylanilinc. After 21 days approximately 10% of the original nitralin could be detected. The aqueous phase (after ethyl ace-late extraction) was concentrated, placed on a Sephadex G10 column and separated into at leasi two products. No ¹⁴CO₂ evolution was detected from ¹⁴C-ring labeled nitralin. Dégradation de la nilralinepar Rhizoclonia solani Dcs ètudes de dégradation de la nitraline [4-(méthylsulfonyl)-2,6-dinitro-N,N-dipropilanihne) ont été effectuées avec des cultures pures de Rhizurtvnia solani Kühn. Le milieu de culture de Czapek. Dox, Broth, a été choisi; il ne contenait pas de composés qui auraient pu interférer a vet I'extraction el avec l'analyse chromalographique en couchc mince ulténeure. Dtx à 11 produits de dégradation ont été décelés après 14 à 21 jours d'incubation à 33 C, La fraction non polaire (extructible par l'acétate d'éthyle) contenait iu minimum 9 composés visibles sur les chromatogrammes Un produit seulement. autre que la nitralinc apparentée. a été identific. le dèrivé monoalkylé. 4-(methylsulfonyl)-2.6-dinilro-N-propylflniline, Aprés 2l jours. 10% environ de la nitralinc origmale avait pu ètre décelée. La phase aqueuse (aprés extraction à l'acétate d'ethyle) a été con-centrée. placée sur une colonne de Scphadex G10 el séparée en deux produits au moins. II n'a pas été décelé d'èevolution du ¹⁴CO₂ provenant du cycle marqué au ¹⁴C de la nitraline. Abhau van Nitrulirt durch Rhizoctonia solani Es wurden Abbaustudien mit Nitralin unter Verwendung von Rcinkulturen von Rhizoctonia solani Kühn durchgeführt. Also Wuchsmedmm wurde Czapek Dox Broth verwendet. das keine Substanzen enthielt. die die Extraktion und die daraffogende dünnschichtchromatographische Analyse störten. Nach einer Inkuhationszeit von 14 bis 21 Tagen nei einer Temperatur von 13°C. fturden 10 bis 11 Abbauprodukte nachgewiesen. Die nicht-polare Fraktion (extrahierbar mil Äthylacetal) enthielt mindestens 9 auf Dünnschichtplatten sichtbare Verbindungen Eine Substanz wurde identifiziert—das monoalkylierte Derivat 4-(Methylsulfonyl)-2.6-dinitro-N-propylanilin Nach 21 Tagen konnten noch ungefähr 10% des ursprünglichen Nitralins nach. gewiesen werden. Die wässrige Phase (nach Extraktion mit Äthylacetat) wurde eingeengt und auf einer Sephadex G 10 Säule in mindestens 2 Verbindungen aufgetrennt. Aus ^l4C-ring-markiertem Nitralin konnte keine ¹⁴CO₂ -Freisetzung nachge-wiesen werden.     

Hyperparasites of Rhizoctonia solani in Dutch potato fields

Gliocladium roseum was found to be the most common and probably the most effective mycoparasite in potato fields in the northern parts of the Netherlands. It is able to parasitize and kill living hyphae at temperatures of 12°C and higher. Sclerotia ofR. solani are often infected and killed by this fungus under suitable conditions, i.e. at temperatures of 16°C and more. Killing of sclerotia by other antagonistic organisms was also observed. It is also shown by not parasitic fungi and is caused by toxins produced by the antagonist.The development of theG. roseum population was studied during the growth of a potato crop in two soils. In both soils its initial level was very low. In both a slightly acid sandy soil and a neutral sandy loam, suppression ofR. solani can occur;G. roseum accumulated in the former mainly under continuous potato crops,Colletotrichum coccodes was the main antagonist in the latter.Samenvatting In de meeste Nederlandse aardappelakkers komen schimmels voor dieRhizoctonia solani kunnen aantasten en doden. De meest algemene, en waarschijnlijk ook de meest belangrijke, die we tot nu toe vonden, isGliocladium roseum (Tabel 1). Het is bekend, dat deze schimmel stoffen produceert die voorR. solani giftig zijn. Met behulp hiervan kanG. roseum, evenals andere antibiotisch actieve micro-organismen, ook de sclerotiën doden (Tabel 2). Voor doding doorG. roseum is de temperatuur een factor van belang. Hyfen worden nog gedood bij een temperatuur van 12°C, waarbij de sclerotiën niet meer aangetast kunnen worden. Gedurende het winterseizoen worden sclerotiën door deze schimmel naar alle waarschijnlijkheid niet gedood.De ontwikkeling van de populatie vanG. roseum en andere antagonisten vanR. solani werd gevolgd in aardappelvelden op een licht zure zandgrond en op een neutrale zware zavel. Op de zandgrond werden twee proefplekken bemonsterd: één waarop voor het vierde achtereenvolgende jaar aardappelen werden geteeld en één met een vruchtwisselingsschema van graan, bieten en aardappelen.In de zandgrond nam in het groeiseizoen de populatie vanG. roseum toe. Op de proefplek waar voor het vierde jaar achtereen aardappelen stonden werdR. solani vanaf half augustus onderdrukt, evenwel niet volledig. Ook in het vruchtwisselingsstuk breiddeG. roseum zich flink uit, doch een onderdrukking vanR. solani werd niet bereikt.In de zware zavel nam de populatie vanG. roseum niet toe. Hier werdR. solani — uit besmet pootgoed — onderdrukt doorColletotrichum coccodes (zelf een pathogeen van stolonen) en antagonistische bacteriën. De resultaten zijn vermeld in Tabel 3.De besmetting van de geoogste knollen met sclerotiën, zoals die voorkwam op de zandgrond, is in Tabel 4 vermeld. Op de zavel leverde schoon pootgoed een bijna schone oogst (2% van de knollen was zeer licht bezet met sclerotiën). Besmet pootgoed leverde een oogst met 58% schone knollen, 35% met een zeer lichte en 7% met een iets zwaardere sclerotiënbezetting. Hoewel uit 100% besmet pootgoed een veel schonere oogst werd verkregen, was eerder toch een beschadiging van het gewas opgetreden. Pas tegen het eind van het groeiseizoen werdR. solani flink onderdrukt.     

水稻纹枯病的发生流行与控制技术

水稻纹枯病自20世纪60年代在大冶地区流行以来，发病程度逐年加重，成为大冶水稻3大病害之首，对水稻生产构成严重威胁。近几年，通过试验研究，逐步形成了一套行之有效的综合防治方法，能有效地控制水稻纹枯病的发生与危害。     

利用cDNA-AFLP分析纹枯病菌诱导的玉米差异表达基因

 以纹枯病菌AG1-IA(Rhizoctonia solani)诱导玉米高耐纹枯病自交系R15,采用cDNA-AFLP技术分析其基因差异表达谱。在拔节期对R15幼苗进行接菌处理,12、24、36、48、60 h分别取材,以不接菌为对照。用56对AFLP选扩增引物对处理和对照的cDNA进行AFLP分析,得到87个差异片段,回收并剔除假阳性,克隆获得18条阳性差异条带(TDFs)。BLASTn比对结果表明,其中可以找到同源序列的有13个TDFs,按其功能可分为信号传导(2个)、抗病与防御基因(2个)、转录调控(2个)、能量代谢(2个)等。对13个TDFs基因进行了半定量RT-PCR分析,结果表明13个差异片段在对照与处理,或是处理的不同时段存在着表达量的差异。     

Characterization of Rhizoctonia solani Associated with Soybean in Brazil

Rhizoctonia solani causes pre- and post-emergence damping-off, root and hypocotyl rot and foliar blight in soybean. Foliar blight has resulted in yield losses of 31–60% in north and northeast Brazil. The aim of this study was to characterize isolates of R. solani associated with soybean in Brazil. Among 73 Rhizoctonia isolates examined, six were binucleate and 67 were multinucleate. The multinucleate iso1ates were characterized according to hyphal anastomosis reaction, mycelial growth rate, thiamine requirement, sclerotia production, and RAPD molecular markers. Four isolates that caused hypocotyl rot belonged to AG-4 and using RAPD analysis they grouped together with the HGI subgroup. Another isolate that caused root and hypocotyl rots was thiamine auxotrophic, grew at 35°C, and belonged to AG-2-2 IIIB. All 62 isolates that caused foliar blight belonged to AG-1 IA. RAPD analysis of R. solani AG-1 IA soybean isolates showed high genetic similarity to a tester strain of AG-1 IA, confirming their classification. The teleomorph of R. solani, Thanatephorus cucumeris was produced in vitro by one AG-1 IA isolate from soybean. The AG-4 and AG-2-2 IIIB isolates caused damping-off and root and hypocotyl rots of soybean seedlings cv. FT-Cristalina, under greenhouse conditions. The AG-2-2 IIIB isolate caused large lesions on the cortex tissue, that was distinct from the symptoms caused by AG-4 isolates. The AG-1 IA isolates caused foliar blight in adult soybean plants cv. Xingu under the greenhouse and also in a detached-leaf assay.     

Rhizoctonia Blight of Yacon Caused by Rhizoctonia solani AG-1 (IB)

Severe blight of stems and leaves caused by Rhizoctonia solani AG-1 (IB) was found on yacon (Smallanthus sonchifolius), a Compositae tuber crop, grown in Ehime Prefecture, Japan, from August to September 1996 and 1999. We named it “Rhizoctonia blight of yacon” as a new disease. Received 15 October 2001/ Accepted in revised form 25 November 2001     

Characterization of Rhizoctonia solani from potato in Great Britain

One hundred and thirty five isolates of Rhizoctonia solani were obtained from British potato crops between 2001 and 2003. Isolates were assigned to anastomosis group (AG) using conventional PCR assays for AG2-1 or AG3 or through the observation of hyphal interactions, where appropriate. A previously published primer set was modified in this study to enhance specificity for AG3PT. Most of the isolates (92·6%) belonged to AG3PT whilst some (6·7%) belonged to AG2-1. Only one isolate recovered (0·7%) belonged to AG5. Isolates of AG2-1 were diverse, with variation in both the length of the rDNA intergenic spacer 1 (IGS1) region and the categories of hyphal interaction observed between pairings of AG2-1 isolates. No variation in the length of the rDNA IGS1 region was observed amongst the AG3 isolates collected. Tests carried out on potato stems with a sub-set of the isolates revealed a wide range of aggressiveness amongst AG2-1 isolates. Sequencing of the rDNA internal transcribed spacer (ITS) region of the AG2-1 isolates and construction of a neighbour joining tree with other AG2-1 sequences available indicated that AG2-1 isolates with the short IGS1 region were closely related. This is the first investigation which provides evidence of the relative AG composition of R. solani populations causing disease in potato crops in Great Britain.     

Association of Binucleate Rhizoctonia with Soybean and Mechanism of Biocontrol of Rhizoctonia solani

ABSTRACT The association of binucleate Rhizoctonia (BNR) AG-K with soybean and the interaction of BNR, R. solani AG-4, and soybean seedlings were investigated to elucidate the mechanism of biocontrol of R. solani by BNR. Sixty-hour-old seedlings were inoculated and incubated in a growth chamber at 24 degrees C; plants were examined with light microscopy and with scanning and transmission electron microscopy at various times following inoculation. BNR grew over hypocotyls, roots, and root hairs, but only colonized epidermal cells. Hyphae of BNR appeared to attach to the epidermis and, 5.5 h following inoculation, began penetrating cells by means of penetration pegs without forming distinct appressoria or infection cushions. There was evidence of cuticle degradation at the point of penetration. Infection hyphae moved to adjacent epidermal cells by direct penetration of epidermal radial walls. There were epidermal and cortical cell necrosis, beginning with the fragmentation of the tonoplast and followed by the disintegration of cytoplasm, organelles, and plasma membranes. Cell necrosis was also observed in adjacent cells where there was no evidence of BNR hyphae. Cell walls were not destroyed. After 144 h, there was noevidence of BNR hyphae in cortical cells. Attempted penetrations were observed, but papillae formed on the inside of cortical cell walls. Pre-inoculation of soybean seedlings with BNR 24 or 48 h before inoculation with R. solani (1 cm between inocula) affected the growth of R. solani on soybean tissue. There were fewer hyphae of R. solani, the hyphae branched sparingly, and infection cushions were rare when compared with hyphal growth on soybean inoculated only with R. solani. These effects were observed before the BNR hyphae began to intermingle with the hyphae of R. solani on the surface of the inoculated host. Preinoculation of soybean seedlings 24 h before inoculation with R. solani significantly (P = 0.05) reduced disease incidence and severity caused by R. solani AG-4. The lesions caused by R. solani always appeared distally, not proximally, to the BNR inoculum. The interactions of intermingling hyphae of BNR and R. solani were examined in vitro and on the surface of the host. There was no evidence of lysis, mycoparasitism, inhibition of growth, or any other form of antagonism between hyphae. The results of these studies strongly suggest that induced resistance is the mechanism of biocontrol of R. solani on soybean by BNR. The inhibition of hyphal growth of R. solani on the surface of soybean tissue preinoculated with BNR appears to be a novel characteristic of induced resistance.     

毒素在稻纹枯病菌致病中的作用分析

用水稻胚根生长抑制法测定了10个稻纹枯病菌菌株在寄主体外产生毒素的能力,发现不同菌株的产毒能力各不相同,且产毒量差异极显著。用水稻离体叶接种法测定了这10个菌株的致病力,结果表明：不同菌株的致病力也各不相同,且差异极显著。相关分析发现,菌株的体外产毒能力与致病力高度正相关（R =0.915 2）。这些结果表明：稻纹枯病菌毒素的分泌可能在其致病过程中起重要作用。     

Molecular tools to investigate Rhizoctonia solani distribution in soil

Real-time PCR protocols were developed to detect and discriminate 11 anastomosis groups (AGs) of Rhizoctonia solani using ribosomal internal transcribed spacer (ITS) regions (AG-1-IA, AG-1-IC, AG-2-1, AG-2-2, AG-4HGI+II, AG-4HGIII, AG-8) or β-tubulin (AG-3, AG-4HGII, AG-5 and AG-9) sequences. All real-time assays were target group specific, except AG-2-2, which showed a weak cross-reaction with AG-2^tabac. In addition, methods were developed for the high throughput extraction of DNA from soil and compost samples. The DNA extraction method was used with the AG-2-1 assay and shown to be quantitative with a detection threshold of 10⁻⁷ g of R. solani per g of soil. A similar DNA extraction efficiency was observed for samples from three contrasting soil types. The developed methods were then used to investigate the spatial distribution of R. solani AG-2-1 in field soils. Soil from shallow depths of a field planted with Brassica oleracea tested positive for R. solani AG-2-1 more frequently than soil collected from greater depths. Quantification of R. solani inoculum in field samples proved challenging due to low levels of inoculum in naturally occurring soils. The potential uses of real-time PCR and DNA extraction protocols to investigate the epidemiology of R. solani are discussed.     

稻曲病菌和水稻纹枯病菌真菌病毒的研究进展

由稻绿核菌(Ustilaginoidea virens)和立枯丝核菌(Rhizoctonia solani)引起的稻曲病和水稻纹枯病是水稻上的2种重要真菌病害.真菌病毒是一类以真菌和卵菌为寄主的病毒,一些引起寄主真菌低毒力的真菌病毒具有生防潜力,可用来防治植物真菌病害.评述了从稻曲病菌和水稻纹枯病菌中分别发现并完成测序...     

Antagonism of Azotobacter chroococcum isolates to Rhizoctonia solani

Antagonism of isolates ofAzotobacter chroococcum toRhizoctonia solani on agar plates was studied, and isolates were tested for their ability to controlR. solani infection of potato sprouts in sterilized and unsterilized soil. The degree of antagonism exhibited varied strongly among the isolates and was also found to be temperature-dependent. At 25, 20 and 15°C, all but one strongly antagonisticAzobacter isolate effectively prevented infection of sprouts of potatoes planted in a soil heavily infected with a pathogenic isolate ofR. solani. At 10°C none was effective.     

黄绿木霉发酵液对水稻纹枯病菌作用的研究

利用黄绿木霉菌发酵液处理水稻纹枯病丝核菌,初步探讨了黄绿木霉菌发酵液对水稻纹枯病丝核菌的抑菌能力及抑菌机理。试验结果表明：黄绿木霉菌发酵液抑菌作用稳定,经121 ℃处理25 min后,抑菌率为100%; pH3～8的范围内抑菌率均为100%。经黄绿木霉菌发酵液处理的水稻纹枯病丝核菌菌丝电导率与呼吸强度几乎下降为0,光学显微镜观察到菌体形态发生变化,透射电镜下观察到细胞壁出现孔洞。用处理后的水稻纹枯病丝核菌菌丝接种水稻植株,发病率明显降低。     

应用稻苗离体叶片法筛选水稻纹枯病新农药活性的研究

采用水稻离体叶片法测定了农药对水稻纹枯病的活性。结果表明,该法与“植株法”比较有很好的相关性,具有占用空间小、温湿度容易控制、快速、准确等优点,适合于大量化合物的筛选试验。