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.     

球孢白僵菌不同世代菌株胞外蛋白酶与毒力的关系

以蝉蜕、蝇明壳两种培养基培养球孢白僵菌不同世代菌株分析其类枯草杆菌蛋白（Ｐｒ１）和总胞外蛋白酶的产生水平及其与菌株毒力的相关性。结果表明,菌株的Ｐｒ１产生水平达到最高的时间较总胞外蛋白酶早１～２天,在单蜕培养基中,菌株Ｐｒ１产生水平较高,而总胞外蛋白产生水平大大低于蝇明壳培养基中的产生水平。随民代数的增加,Ｐｒ１和总胞外蛋白酶产生水平与其毒力的相关系数分别为０．９２和０．８９,高于 包外蛋白酶与其     

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.     

Foliar calcium concentration of potato and its relation to genotype lateness and tolerance of cyst nematodes

In field trials during three years respectively 18, 22 and 57 potato genotypes were grown on soils moderately or heavily infested with potato cyst nematodes (Globodera pallida) after soils were fumigated or not. Nematode infection increased leaf calcium contents but genotypes that were relatively tolerant of potato cyst nematodes (Globodera pallida) had lower leaf calcium concentrations on a particular sampling date. Tolerance of potato cyst nematodes was closely linked to genotype lateness and may be related to tolerance of drought. When using the method in plant breeding and screening for genotypes with tolerance, plants may be planted in infected or in uninfected soils, without influencing the outcome. The relationship between tolerance and calcium contents was clearest when sampling took place towards the end of the growing season when the variability was at it highest with a good distinction between newly formed leaves in late maturing genotypes and aged leaves in early maturing cultivars.     

Anastomosis Groups, Pathogenicity and Sensitivity to Fungicides of Rhizoctonia solani Isolates Collected on Potato Crops in France

A collection of 241 isolates of Rhizoctonia solani obtained from potato plants grown in different areas in France was characterized for anastomosis grouping, symptomatology on tubers of different cultivars and sensitivity to three fungicides. Most isolates collected belonged to (anastomosis groups (AGs)) AG 3, but 2% and 4% of the isolates were AG 5 and AG 2-1. AG 3 and AG 2-1 isolates were mostly obtained from sclerotia on tubers, but all AG 5, some AG 3 and some AG 2-1 isolates were recovered from superficial tuber alterations, like deformations, corky or scabby lesions. Sclerotia were formed on tubers produced by healthy stem cuttings grown in soil artificially infested with AG 3, but not on tubers grown in soil infested with either AG 5 or AG 2-1. No variation in susceptibility to sclerotial formation was observed among five potato cultivars. In all cases, a large proportion of tubers showed superficial corky lesions, often associated with deformations. The proportion of tubers with lesions and deformations was highest in soil infested with AG 2-1 and significantly lower on cv. Samba in all treatments. All isolates were highly sensitive to flutolanil, iprodione and pencycuron, except the AG 5 isolates, moderately sensitive to pencycuron. These results show that, although AG 3 is the most common R. solani group on potato in France, AG 5 and AG 2-1 may be present. Isolates differed for pathogenicity. In vitro sensitivity to fungicides varied among AGs.     

Multiple Selection of Potato Cyst Nematode Globodera pallida Virulence on a Range of Potato Species. I. Serial Selection on Solanum-hybrids

A series of selection experiments on potato cyst nematode Globodera pallida, pathotype Pa1, tested the virulence response of the nematode to a range of resistant potato Solanum genotypes. Alleles conferring virulence against all four Solanum sources used in the study (i.e. Solanum vernei, S. multidissectum, S. sanctae-rosae and S. tuberosum ssp. andigena) were detected. Selection for multiple virulence against a combination of resistant sources resulted in the originally-selected virulence genes being retained or lost. The mechanism, or basis, of potato cyst nematode resistance differs between the Solanum species. The appropriate use of resistance cultivars produced from a range of Solanum-resistant species offers a management tool for controlling potato cyst nematode levels in infested land.     

The influence of cyst nematodes and drought on potato growth. 3. Effects on carbon isotope fractionation

A good agreement was found between drought induced water-use efficiencies and carbon isotope fractionation of potato plants of the same age. Discrimination of¹³Carbon isotope increased with plant age and with the rank order of the distinguished plant organs. In young plants, nematode infection (contrary to drought) reduced¹³C discrimination but also reduced the water-use efficiency, leading to the conclusion that nematode infection, by slowing down the plant metabolism may have reduced the extrusion of¹³C. Towards senescence both nematode infection and drought reduced¹³C discrimination and increased water-use efficiencies indicating that then nematode caused water stress in the plants.     

The influence of cyst nematodes and drought on potato growth. 1. Effects on plant growth under semi-controlled conditions

Potatoes were grown under a permanent rain shelter in mobile containers in soil with or without potato cyst nematodes (Globodera pallida). The plants were subjected to an early drought stress period starting at planting until 43 days after planting, to a late drought stress period starting at 43 days until senescence at 92 days and to a drought control. Dry matter weight and characteristics of leaves, stems, stolons and roots were determined at periodic harvests. The early drought stress and nematode infection affected many plant organ characteristics in similar ways. Numbers of leaves, specific leaf area, plant height, specific stem weight, leaf area ratio, mean tuber weight and harvest index were reduced by both stress factors at early stages of growth.Later on, interactions between both stress factors which influence the development rate of the plants led to more diverse plant reactions. Plants of all treatments rapidly senesced at about 90 days after planting. Uninfected plants had then depleted the soil nutrient supply whereas the plants grown in the inoculated soil senesced as a result of the potato cyst nematode infection.     

高毒力金龟子绿僵菌的筛选及其胞外蛋白酶产量测定

以大蜡螟Galleria mellonella L.幼虫为供试昆虫,对从果园土壤中筛选出的金龟子绿僵菌Metarhizium anisopliae的生物活性进行了测定,共获得4株高毒力菌株,其LT50值在1.85～2.56 d之间,LD50值在6.11×102～1.55×104 spores之间。利用明胶-琼脂平板分析法对筛选出的高毒力绿僵菌菌株产胞外蛋白酶的水平进行了测定,结果显示,各菌株间产酶水平差异较大,且菌株产酶水平的高低与其对大蜡螟幼虫的毒力之间存在明显的线性相关性。     

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.     

氮磷钾及有机肥对马铃薯生长发育和干物质积累的影响

采取田间试验,探讨不同施肥处理对马铃薯生长发育及干物质积累的影响.结果表明,N1PKM处理(氮300 kg·hm^-2,磷200 kg· hm^-2,钾200 kg·hm^-2,有机肥17.5 t·hm^-2)的马铃薯叶片叶绿素含量最高,比对照处理的叶片叶绿素a、b、总量分别高出15.12％、18.18％和36.37％。施用磷肥有效促进了叶面积的增加,N₁PKM、N₁P、N₁PK、P处理的叶面积比对照分别高出27.78％,24.57％,20.26％和20.16％。其中施用磷肥促进马铃薯根系生长的效果最明显,而单施钾肥、有机肥对根长发育的影响不明显。氮、磷、钾、有机肥配比施用可以促进马铃薯地上及地下部的生长,采用氮磷钾与有机肥配合施用时,马铃薯干物质累积总量N₁PKM＞ N1PK＞ N₁K＞N₁P。N₁PKM处理的马铃薯生物量较对照显著提高了117.1％,可以作为旱地马铃薯的施肥方案。     

Influence of environmental factors on field concentrations of<Emphasis Type="Italic">Alternaria solani</Emphasis> conidia above a South African potato crop

Trends in weather variables and concentrations of airborne conidia ofAlternaria solani were monitored in a potato field in South Africa during three potato-growing seasons in 2001 and 2002. Distinct seasonal variation was noted, with a drop in spore numbers during winter. Peaks in spore concentration coincided with periods favorable for spore formation and dispersal; most notable was the effect of interrupted wetting periods. Diurnal periodicity of spore dispersal was also observed, with the peak of spore concentrations between 9h00 and 18h00. Few spores were sampled at night, when wind velocity and temperature are lowest and relative humidity is highest. Increased numbers of spores were sampled during days of harvesting or when other ground-operated farm equipment was used. The results obtained in this study will be useful in establishing decision support systems to control early blight on potatoes in southern Africa. http://www.phytoparasitica.org posting July 10, 2003.     

Assessment of recent outbreaks of <Emphasis Type="Italic">Dickeya</Emphasis> sp. (syn. <Emphasis Type="Italic">Erwinia chrysanthemi</Emphasis>) slow wilt in potato crops in Israel

Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification. All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation, and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA.     

Infestation by the cyst forming nematodeGlobodera pallida of potato tubers in Southern Italy

A case of heavy infestation on tubers of three potato (Solanum tuberosum L) cultivars (Nicola, Spunta and Sieglinde) by the cyst nematodeGlobodera pallida Stone, 1973 was observed in Southern Italy on early producing potato cultivars in spring. Tubers were covered by white females and cysts but no other external symptoms were detectible on their surfaces. Detailed observations were directed to study the response of phellem and secondary cortex tissues induced by the expansion of syncytia during nematode feeding activity. The micro details of histological changes observed on serially sectioned infected tissues are described and illustrated. The nematode's feeding activity was confined to the outer part of the tuber while the host-parasite relationships were similar to those induced by the same nematode in the feeder roots.     

Evaluation of biocontrol preparations and plant extracts for the control of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on potato leaves

The potential of biocontrol products and plant extracts for control of late blight on potato plants, caused by Phytophthora infestans was evaluated in detached leaf assays and on potted plants. Based on an initial screening of 22 preparations and plant extracts, the 10 most active treatments were selected for further investigation. In the detached leaf assays the commercial preparations Elot-Vis, Serenade and Trichodex, and plant extracts of Rheum rhabarbarum and Solidago canadensis showed a significant effect on the level of infestation by P. infestans. However, none of the treatments was as effective as copper. In the case of Serenade, the metabolites produced by its active micro-organism, Bacillus subtilis, were demonstrated to be the effective component of the formulation, and not the micro-organism itself. In order to take curative and protective modes of action into account, the test substances were applied 24 h before, or 90 min after inoculation with P. infestans. Generally, better effects were obtained when the applications were made 24 h before inoculation. For defining the optimum time of application, potted plants were treated 72 or 24 h before, and 1 and 24 h after inoculation with P. infestans. In these tests, Trichodex showed no activity, while Elot-Vis gave best results when applied 1 day before inoculation. Serenade and the extracts of R. rhabarbarum and S. canadensis (all at 5% concentration) however, were effective when applied up to 3 days before and just after inoculation with P. infestans. The results of the experiments on potted plants indicated direct effects on the pathogen for all agents except the extract of S. canadensis, but other mode of actions, e.g. induced resistance, could not be ruled out. None of the treatments had a curative effect.     

Quantification of microsclerotia ofVerticillium dahliae in plant material by image analysis

A procedure for the quantification of microsclerotia ofVerticillium dahliae with an image analysis system was compared with counting by eye. Colonised potato plant material was used from plants grown in pathogen-free soil in a greenhouse and from twelve crops (including four potato cultivars) grown outdoors in pots filled with pathogen-free soil under natural conditions. The values obtained from the potato material from the greenhouse were comparable for both methods. Variation in the results mainly resulted from sampling errors. The numbers of microsclerotia in plants grown outdoors were overestimated by image analysis for most crops. The source of the error was related to the presence of plant and soil particles that did not discolour during boiling of the samples in sodium hydroxide. Image analysis was a suitable and reliable method for assessing the number of microsclerotia only in potato haulm samples from plants grown in pathogen-free soil in the greenhouse.Abbreviations cv coefficient of variation - IA image analysis - MS microsclerotia, microsclerotium     

History of potato wart disease in Europe – a proposal for harmonisation in defining pathotypes

Potato wart disease, caused by the chytridiomycete Synchytrium endobioticum, was first introduced into Europe in the late 19th century. It spread quickly, and today is reported in 15 European countries. Initially, only one pathotype was found, and the disease was efficiently controlled using resistant cultivars. In 1941, however, formerly resistant cultivars showed wart formation in the field simultaneously in Germany and South Bohemia (Czech Republic), indicating the occurrence of new pathotypes. New pathotypes have since been reported from Germany, The Netherlands, Czech Republic, Ukraine and Canada. Today the pathogen is present in The Netherlands (only in fields for ware and starch potatoes) but restricted to two demarcated areas and subject to official control. Outside these areas, the pathogen is absent. For pathotyping, different countries have used different sets of differential cultivars, and the usual system of numerical coding of pathotypes has not been consistently followed. In this review we propose a new standardised code to be used for the 43 pathotypes currently known and described in Europe. The code is a combination of a numerical and letter code, combining the two terminologies used by former West and East Germany, respectively. We also plead for harmonisation in the choice of differential cultivars used for pathotype identification. The set of differentials described in the international standard for diagnosis of S. endobioticum issued by the European and Mediterranean Plant Protection Organisation (EPPO), should serve as a basis. Through close collaboration of European countries dealing with new pathotypes of potato wart disease, a final agreed upon set of differentials, combined with a set of reference isolates, should ultimately be established, allowing a clear distinction between the most important pathotypes occurring in Europe.     

Modes of Action of Pantoea agglomerans CPA-2, an Antagonist of Postharvest Pathogens on Fruits

Pantoea agglomerans CPA-2 is an effective antagonist against the postharvest pathogens Penicillium digitatum and Penicillium italicum on citrus fruits but its mode of action is unknown. Possible mechanisms studied in this work were antibiosis, induced resistance, competition and production of chitinolytic enzymes. P. agglomerans CPA-2 was unable to produce antibiotics or chitinolytic enzymes under the conditions tested. Induction of resistance by P. agglomerans CPA-2 was studied in oranges by measuring phenylalanine ammonia lyase and peroxidase enzyme activity in the orange peel at different time points after inoculation with the antagonist and/or the pathogen. No significant augmentation of enzyme activity after inoculation of oranges with P. agglomerans CPA-2 in the presence or absence of the pathogen was observed. P. agglomerans was effective only when it is in close contact with the pathogens. Competition for nutrients was studied using tissue culture plates with cylinder inserts, which allowed competition for nutrients to be studied without competition for space since physical contact between pathogen and antagonist was avoided. The presence of P. agglomerans in the tissue culture wells clearly decreased the germination of Penicillium conidia present in the cylinder when diluted orange peel extract or diluted potato dextrose broth was the nutrient source. Germination of Penicillium conidia, however, was almost completely inhibited when pathogen and antagonist were in physical contact. These results indicate that competition for nutrients is one of the modes of action of P. agglomerans CPA-2, but that physical contact between pathogen and antagonist is important for effective control.     

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.