Inactivation of sclerotia of Rhizoctonia solani on potato tubers by Verticillium biguttatum,a soil-borne mycoparasite

Experiments in the laboratory and on farms with potato tubers in storage are described in which sclerotia ofRhizoctonia solani were inactivated after inoculation of infected tubers with a suspension of conidia and hyphal fragments ofVerticillium biguttatum. Sclerotia on freshly harvested tubers can be killed in a period of six to eight weeks, provided that (1) a direct contact between sclerotia and conidia ofV. biguttatum is obtained, (2) the temperature during the storage period is at least 15 °C, but preferably closer to 20 °C during the first weeks, and (3) the relative humidity of the air between the tubers is at least 99%. Seed tubers are only certified as export quality if the infection withR. solani, visible as sclerotia on the tubers, is assessed as below a specified incidence. To restore the economical value of tubers with many sclerotia, living sclerotia can be inactivated byV. biguttatum. However, also dead sclerotia have to be removed, as dead and living sclerotia cannot be distinguished visually by inspectors. A satisfactory way to remove dead sclerotia from tubers has not yet been found.     

Suppression of Rhizoctonia solani in potato fields. II. Effect of origin and degree of infection with Rhizoctonia solani of seed potatoes on subsequent infestation and on formation of sclerotia

The seed potatoes used in these experiments had been grown in a slightly acid pleistocene sandy soil or in a marine, holocene sandy loam. They were free of sclerotia ofR. solani or lightly or moderately speckled with them. Seed potatoes from the sandy soil produced plants that suffered less fromRhizoctonia than plants from seed potatoes that had been grown on the marine sandy loam. Similarly harvested tubers had, in a non-conducive soil and in conducive soils with a (very) low inoculum density ofR. solani, fewer sclerotia when they came from seed potatoes grown in an acid sand. In each soil, the degree of infestation of the crop not only depended on the severity of infection of the seed potatoes, but also on their origin. With regard to sclerotia production on tubers, three types of soil were distinguished: suppressive, conducive with a high, and conducive with a very low inoculum density ofR. solani. The differences in infestation and in the amounts of sclerotia on tubers between the crop grown from seed potatoes from the sandy soil and that from seed potatoes from the marine sandy loam soil, is attributed to a richer load of antagonists on the former and possibly to a larger proportion of saprophyticRhizoctonia strains among their sclerotia. The antagonists seem to be inhabitants of the subterranean parts of the plant and to function independently of the soil. This implies possibilities for their use in biological control.     

Postharvest dark skin spots in potato tubers are an oversuberization response to Rhizoctonia solani infection

Israeli farmers export 250,000 tons of potato tubers annually, ≈40,000 tons of which are harvested early, before skin set. In recent years, there has been an increase in the occurrence of dark skin spots on early-harvested potato tubers ('Nicola') packed in large bags containing peat to retain moisture. The irregular necrotic spots form during storage and overseas transport. Characterization of the conditions required for symptom development indicated that bag temperature after packing is 11 to 13°C and it reaches the target temperature (8°C) only 25 days postharvest. This slow decrease in temperature may promote the establishment of pathogen infection. Isolates from typical lesions were identified as Rhizoctonia spp., and Koch's postulates were completed with 25 isolates by artificial inoculation performed at 13 to 14°C. Phylogenetic analysis, using the internal transcribed spacer sequences (ITS1 and ITS2) of rDNA genes, assigned three isolates to anastomosis group 3 of Rhizoctonia solani. Inoculation of wounded tubers with mycelium of these R. solani isolates resulted in an oversuberization response in the infected area. With isolate Rh17 of R. solani, expression of the suberin biosynthesis-related genes StKCS6 and CYP86A33 increased 6.8- and 3.4-fold, respectively, 24 h postinoculation, followed by a 2.9-fold increase in POP_A, a gene associated with wound-induced suberization, expression 48 h postinoculation, compared with the noninoculated tubers. We suggest that postharvest dark spot disease is an oversuberization response to R. solani of AG-3 infection that occurs prior to tuber skin set.     

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.     

马铃薯立枯丝核菌生物学特性研究

采用生长速率法研究了黑龙江省马铃薯立枯丝核菌(Rhizoctonia solani Kühn)菌丝生长的条件。结果表明病菌菌丝在10~35℃均能生长,适宜温度是25~30℃,30℃菌丝生长最快。在pH为3~12的范围内病菌菌丝均能生长,以pH 5~7菌丝生长速度较快。黑暗对菌丝生长有促进作用,光照可促进菌核的形成。不同碳源、氮源对立枯丝核菌菌丝生长的影响均较明显,可溶性淀粉和酵母粉对菌丝生长有显著的促进作用。立枯丝核菌菌丝体及菌核的致死温度分别为49℃和51℃。明确了黑龙江省马铃薯立枯丝核菌生长的环境条件,可为病害防治提供理论依据。     

Infection of potato by Rhizoctonia solani: effect of anastomosis group

Glasshouse and field experiments showed that the pathogenicity and disease type on potato varied between different anastomosis groups (AGs) of Rhizoctonia solani. For example, severe stem and stolon disease developed in plants inoculated with a single isolate of AG3PT and AG5. Severe root disease was observed with single isolates of AG8 and to a lesser extent AG3PT, but rarely with single isolates of the other AGs tested. In both field and glasshouse experiments the AG2‐1 isolate (X81) produced only small lesions (<5 mm). However, this was not representative of two other AG2‐1 isolates. When AG2‐1 isolates of the three different rDNA IGS1 types were tested in a glasshouse trial, one caused more severe stem and stolon infection than AG3PT. In the field experiment, the yield of tubers, by weight, was significantly less (P < 0·05) in all inoculated plants than for uninoculated (control) plants. Yield losses were greatest and tuber numbers smallest in plots inoculated with an AG8 isolate, suggesting that root infection is important in determining quantitative yield loss. The incidence of black scurf was greatest in the progeny tubers in plots inoculated with AG3PT (83·9%), whereas only very small amounts of black scurf developed on tubers from plants infected with AG2‐1 (510 bp) or AG5 isolates. This is supported by laboratory tests, where isolates of AG3PT produced significantly more sclerotia on potato dextrose agar than isolates of AGs 2‐1, 4, 5 and 8.     

水稻纹枯病菌菌核上的真菌和细菌种类及其对水稻纹枯病菌生长的影响

 A total of 136 fungal and 86 bacterial isolates were isolated from the sclerotia of Rhizoctonia solani,collected from three different sources.These fungal isolates were identified as the following 12 genera:Alternaria,Aspergillus,Cladosporium,Coniothyrium,Curvularia,Gliocladium,Fusarium,Metarhizium,Peni-cillium,Phoma,Phytophthora and Trichoderma.Some isolates of Trichoderma and Gliocladium were found to inhibit the growth of R.solani strongly in vitro.Among the 86 bacterial isolates,20 of them could inhibit the growth of R.solani,whereas 5 isolates were found to promote the growth of mycelia and the formation of sclerotia of R.solani.These bacterial isolates with antagonistic actives to R.solani were identified as Bacillus and Pseudomonas.     

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.     

甘肃省定西市马铃薯黑痣病菌菌丝融合群的鉴定及药剂筛选

马铃薯黑痣病立枯丝核菌(Rhizoctonia solani)菌丝存在遗传分化现象。从定西市马铃薯产区采集茎部和薯块病样80份进行融合群测定。结果表明,80株菌株中与标准菌株AG3融合的占总数的70.0%、与AG4-HG-Ⅱ融合的占15.0%、与AG2-1融合的占7.5%。为了有效控制马铃薯黑痣病,采用生长速率法在室内选取9种杀菌剂进行药效评价。结果表明:30%噻呋酰胺悬浮剂EC50最低,为0.015 2μg/mL;其次是40%氟胺·异菌脲悬浮剂、3%多抗霉素可湿性粉剂和250g/L嘧菌酯悬浮剂,其EC50分别为0.064 1、0.129 3和0.176 7μg/mL;EC50最大的是25.75%多抗·福美双可湿性粉剂,为2.930 4μg/mL。田间拌种,30%噻呋酰胺悬浮剂和250g/L嘧菌酯悬浮剂防效较好,达70%以上,值得在生产中推广。     

Isolation of mycoparasite Verticillium biguttatum from sclerotia of Rhizoctonia solani in the United Kingdom

Verticillium biguttatum was isolated from sclerotia of Rhizoctonia solani removed from black scurf-infected potato tubers which had been field-grown in the UK. Two isolates were identified by means of morphological and physiological characteristics, together with their ability to overgrow cultures of R. solani. A Dutch isolate M73 was used for comparison.     

Infection of potato and wheat by isolates of Rhizoctonia solani and R. cerealis

Isolates of Rhizoctonia were obtained from potato crops with stem canker or black scurf and from cereal crops with sharp eyespot. Those with many nuclei per cell, wide cells, darkening colonies and fast growth were assigned to R. solani ; those with two nuclei per cell, narrow cells, pale colonies and slow growth were assigned to R. cerealis . Only R. solani was obtained from potatoes and only R. cerealis from cereals. On young plants in the glasshouse, the isolates of R. solani infected potato substantially but not wheat; R. cerealis infected wheat substantially and potato slightly. This host preference was shown at temperatures between 10 and 25°C in growth rooms. R. solani on potato caused more disease with increasing temperature; no trend with temperature was observed for R. cerealis on wheat.     

宁夏马铃薯主栽品种对黑痣病的抗性鉴定

为明确宁夏马铃薯主栽品种对黑痣病的抗性水平,利用田间和室内接种相结合的方法对供试马铃薯品种的抗病性进行评价。室内接种试验表明,供试的20份马铃薯材料中没有免疫及高抗品种,接种量与抗性水平呈负相关,其中‘陇薯7号’,‘黑美人’,‘青薯9号’在接种量为5~15g水平上表现为中抗。通过聚类分析,接种量为5g和10g时,以欧式距离13作为最佳聚类分割点,供试品种可分为3个类群。田间接种综合评价分析表明,不同品种间抗性存在显著差异,通过聚类分析,以欧式距离13作为最佳聚类分割点,试验品种可分为4个高感品种、11个中感品种和5个中抗品种。     

Dynamics of soilborne Rhizoctonia solani in the presence of Trichoderma harzianum: effects on stem canker, black scurf and progeny tubers of potato

Stem canker and black scurf are diseases of potato caused by the fungus Rhizoctonia solani . Spatiotemporal experimentation and empirical modelling were applied for the first time to investigate the effect of antagonistic Trichoderma harzianum on the dynamics of soilborne R. solani on individual potato plants. Trichoderma harzianum reduced the severity of symptoms, expressed as 'rhizoctonia stem lesion index' (RSI), during the first 7 days post-inoculation when the inoculum of R. solani was placed at certain distances (30–60 mm) from the host. For example, with inoculum at 40 mm from the host, RSI was 6 and 40 with and without T. harzianum , respectively. At later observation times, the antagonistic effect was overcome. Trichoderma harzianum reduced the severity of black scurf on progeny tubers. Furthermore, the mean number of progeny tubers per potato plant was reduced by the biocontrol treatment (means of 6·5 ± 1·1 and 9·9 ± 2·7 tubers per plant with and without T. harzianum , respectively), as was the proportion of small (0·1–20·0 g) tubers (48% and 66% with and without T. harzianum , respectively). Additionally, there were fewer malformed and green-coloured tubers in pots treated with T. harzianum than in those without T. harzianum .     

Suppression of Rhizoctonia solani in potato fields. 1. Occurrence

A search was made forRhizoctonia solani-suppressive soils by establishing many small experimental plots, half of which were planted withRhizoctonia-infected seed potatoes and the other half with disinfected seed stock. The sclerotium index of the harvested tubers was compared witht that of the seed potatoes. In suppressive soils, the sclerotium index of the harvest is much lower than that of the seed potatoes. None of the plots on holocene marine soils (loamy sand, sandy loam, clay loam and clay) proved to be suppressive in 1978 and 1979. Only on pleistocene, slightly acid sandy soil suppressiveness was observed. In 1978, four out of twelve plots showed suppressiveness when the plots were planted with seed potatoes produced on a sandy soil. In 1979, only two out of thirtyone plots were slightly suppressive when planted with seed potatoes produced on a young clay loam from a new polder. A higher percentage of sclerotia on tubers from sandy soils proved to be infected with antagonistic fungi (73%) than of those on tubers from marine clay or loam soils (25%). Factors that influence suppressiveness are suggested.     

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.     

碳酸氢盐对马铃薯黑痣病菌毒力及田间防效

为明确碳酸氢盐对马铃薯黑痣病菌的毒力及田间防效,采用菌丝生长速率法,测定了碳酸氢盐对马铃薯黑痣病菌的毒力;并通过盆栽防治试验及田间防治试验,研究了碳酸氢盐对马铃薯黑痣病的防治效果。结果表明:碳酸氢铵抑制马铃薯黑痣病菌菌丝生长的有效中浓度EC50仅为0.71g/L,碳酸氢钾EC50为3.8g/L;在PSA培养基中同时加入0.79g/L碳酸氢铵、1g/L碳酸氢钾与在PSA培养基中含1.58g/L碳酸氢铵的抑菌效果一样;尿素对马铃薯黑痣病没有防治效果。碳酸氢铵与碳酸氢钾混配对该病有良好的防治作用,相对尿素最高防治效果可达到63.4%。覆膜栽培能够提高碳酸氢盐的防效和马铃薯产量。碳酸氢盐可用于田间马铃薯黑痣病的防治。     

The occurrence of Rhizoctonia solani on subterranean parts of wild plants in potato fields

The subterranean parts of 1585 wild plants (weeds) belonging to 52 species, found in potato fields in the northern part of the Netherlands were examined for the presence ofRhizoctonia solani. The fungus could be isolated from 30 plants belonging to 12 species. Of these isolates 62% proved to be pathogenic to potato sprouts.Some species seemed more frequently colonized byR. solani than others, particularlySolanum nigrum, Elytrichia repens andMatricaria recutita. On some common weeds, viz.Chenopodium album andPoa annua, noR. solani was observed, while about 1% of allPolygonum persicaria were infected.Disease symptoms were only observed once onElytrichia repens as sharp eyespots, presumably caused byR. cerealis.The incidence ofR. Solani on weeds increased markedly towards the end of the growing season.

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Samenvatting Wortels, wortelstokken en ondergrondse stengeldelen van onkruiden voorkomend in aardappelakkers in Groningen, Friesland en Drenthe zijn onderzocht op de aanwezigheid vanRhizoctonia solani. Van 1585 planten, behorend tot 52 soorten, herbergde bijna 2%, d.w.z. 29 planten behorend tot 12 soorten,R. solani op de ondergrondse delen. Van dezeRhizoctonia-isolaten bleek 62% aardappelspruiten aan te tasten.Bepaalde onkruidsoorten bleken vaker drager te zijn vanR. solani dan andere. Dit was het geval met kweek, zwarte nachtschade en echte kamille. Op enkele algemene onkruiden als witte ganzevoet en straatgras is geenRhizoctonia waargenomen. Van het algemeen op zandgronden voorkomende perzikkruid bleek ongeveer 1%Rhizoctonia op de wortels te herbergen.Typische ziektesymptomen — scherpe oogvlekken — zijn alleen waargenomen op stengels van kweek. Het voorkomen vanR. solani op onkruiden neemt sterk toe tegen het eind van het groeiseizoen (eind augustus en september).