Homothallism in Peronospora viciae f.sp. pisi and the effect of temperature on oospore production

Monoconidial cultures derived from seven P. viciae f.sp. pisi isolates, obtained from different countries, were able lo produce oospores, Apparently, these isolates were homothallic. Oospore production of one isolate was studied at 5, 10, 15 and 20°C in systematically colonized shoots, and in local lesions on leaflets, stem parts and pods of the pisum sativum cv. Kelvedon Wonder. The number of oospores produced per gram systemically colonized tissue increased with temperature. In lesions of leaflets and of stem parts, including tendrils, petioles and main stem, most oospores were produced at 20°C. At 10°C, a few oospores were found in stem parts but none in leaflet lesions. At 5° C, no oospores were formed at all. In pods, moe oospores were produced at 15 and 20°C than at 10°C, but the numbers of oospores was smaller than in the other plant parts. Oospores formed at lower temperatures were larger than those formed at higher temperatures. At 20°C, similar oospore densities were found in leaflet lesions of three cultivars widely differing in resistance to downy mildew.     

Factors affecting germination of oospores of Peronospora viciae f.sp. pisi in vitro

The effects of host plant exudates, light and temperature on germination of oospores of Peronospora viciae f.sp. pisi in vitro were investigated. Seed and root exudates did not increase percentage germination, whereas light inhibited germination. The first germ tubes appeared after 4, 7, and 14 days of incubation at 15, 10 and 5 °C, respectively. The eventual level of germination was highest and had similar values at 5 and 10 °C. At 20 °C germination was poor and at 25 °C no germination was observed. Oospores placed on membrane filters were incubated on soil. When oospores were retrieved from the membrane filters after six days and placed in water at 10 °C, they germinated within 2 days. On soil significantly less oospores germinated than in water. Germinability of oospores stored in the dark at 5 or 20 °C at 30 or 76% RH was studied over a two-year period. Germinability generally increased over time, but fluctuations were observed indicating the occurrence of secondary dormancy. Time courses of germinability were generally similar for oospores stored at several temperatures and humidities. No effect of light on time course of germinability was found when oospores were exposed to alternating light-dark periods or stored in continuous dark for 140 days. Percentage germination observed in a germination assay was correlated with percentage infection determined in a bioassay.     

Production of oospores by Peronospora viciae f.sp. fabae

Oospore production inVicia faba, cv. Metissa, was quantified in the field after plants had been inoculated with a sporangium suspension of a homothallic isolate ofPeronospora viciae f.sp.fabae. Oospores were produced abundantly during the hole growing season from 3 weeks after inoculation on. Oospores were found in all plant parts above soil level, except in the seeds. Most oospores were found in the leaves. Less oospores were formed in leaves inoculated in an older stage than in those inoculated in a younger stage. Towards the end of the season, in August, numbers of oospores in pods strongly increased.Oospore production in leaves of three cultivars, Metissa, Toret and Maris Bead, was studied in growth chambers at 5, 10, 15 and 20 °C at 16 h light. Oospores were formed earlier at higher temperatures than at lower temperatures. The ultimate numbers of oospores produced in leaves were highest at 10 and 15 °C. Similar numbers of oospores were formed in leaves of cultivars Metissa and Toret. In leaves of cv. Maris Bead significant less oospores were produced than in leaves of cv. Metissa and cv. Toret. Total numbers of oospores produced were not related to the level of host plant resistance to downy mildew. The percentage of asexually sporulating leaf area, assessed in a resistance test, was largest in cv. Metissa and smallest in cv. Toret.     

Factors governing oospore production by Peronospora farinosa f. sp. spinaciae in cotyledons of spinach

Oospore production inPeronospora farinosa f. sp.spinaciae was induced in spinach seedlings. This was done in detached cotyledons as wel as in undetached cotyledons on living plants, and in growth chambers as well as in the field. In young leaf tissue the fungus, produced oospores more often than in old tissue. The four spinach cultivars tested, viz. Breedblad Scherpzaad, Symphony, Novires and Huro, did not show significant differences in numbers of cotyledons with oospores, nor in numbers of oospores per cotyledon, though cv. Breedblad Scherpzaad was less susceptible to oospore formation in the field. Comparison of various inoculum densities showed a tendency to a smaller number of cotyledons with oospores with a lower inoculum density. Oospores were abundantly formed, when the plants were exposed to stress conditions in the second half of the latent period.Samenvatting Methoden werden ontwikkeld om oosporenvorming doorPeronospora farinosa f. sp.spinaciae in kiemplanten van spinazie te induceren. Dit gebeurde met afgeplukte kiembladeren en in kiembladeren aan de levende plant, zowel in klimaatcellen als tevelde.In jonge kiembladeren werden vaker en meer oosporen gevormd, dan in oudere kiembladeren. De vier getoetste cultivars Breedblad Scherpzaad, Symphony, Novires en Huro vertoonden geen opvallende verschillen in aantallen kiembladeren met oosporen en aantallen oosporen per kiemblad, hoewel cultivar Breedblad Scherpzaad in het veld als minst vatbaar voor oosporenvorming naar voren kwam.Vergelijking van verschillende inoculum dichtheden toonde een tendens aan voor een kleiner aantal cotylen met oosporen bij lagere inoculumdichtheden. Er werden overvloedig oosporen gevormd wanneer de planten in de tweede helft van de latentieperiode in een stress situatie werden gebracht.     

Variation for virulence among Scandinavian isolates of Peronospora viciae f.sp. pisi (pea downy mildew) and responses of pea genotypes

Variation for virulence among Scandinavian isolates of Peronospora viciae f.sp. pisi (downy mildew) on different pea genotypes was investigated. Variation for virulence was found within and between mass-conidial isolates which were originally obtained from oospore populations. Virulence towards pea cultivars that have not been commerically cultivated in Scandinavia was observed. The specific resistance of the pea cultivars Puget, Cobri, Gastro, Starcovert and Starnain was confirmed. One pea breeding line showed a stable partial resistance to different isolates of the fungus and it was also highly resistant to race 8'from The Netherlands which is considered to be virulent on most pea genotypes carrying known specific resistance factors.     

Extraction of oospores of Peronospora viciae from soil

A method for the extraction of oospores of Peronospora viciae from soil is described. Approximately 75% of the oospores added to silty clay loam and loamy sand soil samples were recovered. Percentage recovery was independent of oospore density. This extraction method did not affect viability as determined by the tetrazolium bromide test and a germination assay and may therefore be used to study survival of oospores. Numbers of oospores extracted from soil samples taken from seven fields naturally infested with oospores of P. viciae f.sp. pisi ranged from 2 to 21 oospores per g soil. Oospore density was not significantly correlated with disease incidence as determined by a bioassay.     

Production of conidia by Peronospora farinosa f. sp. spinaciae

Conidium production byPeronospora farinosa f. sp.spinaciae (pathotype 3) was measured daily on colonies induced on leaves of spinach cvs Breedblad Scherpzaad (BS) and Huro. Distinction was made between colonies grown at temperatures of 10 and 15°C. Spore production was expressed as number of spores produced per stoma. There was a significant difference between total spore production on lesions on BS at 10 °C and that on lesions on the three other cultivartemperature combinations tested. Also a significant difference in the average sporulation period was observed between lesions produced at 10 and those at 15 °C. The same significant difference in response to temperature was found in the sporulation per stoma in the course of time.Samenvatting De conidiënproduktie vanPeronospora farinosa f. sp. spinaciae (fysio 3) werd dagelijks bepaald aan kolonies, welke op bladeren van de spinaziecultivars Breedblad Scherpzaad (BS) en Huro werden geïnduceerd. Daarbij werd onderscheid gemaakt tussen kolonies welke zich bij 10 en bij 15 °C hadden ontwikkeld. De sporenproduktie werd omgerekend naar het aantal sporen per huidmondje. Er was een significant verschil tussen de totale sporenproduktie op lesies op BS bij 10 °C en die op lesies op de drie andere getoetste cultivar-temperatuur combinaties. Er kon ook een significant verschil in de gemiddelde sporulatieperiode worden waargenomen tussen de lesies ontstaan bij 10 en bij 15 °C. Een dergelimk significant verschil in temperatuurreactie kon ook worden gevonden in het verloop van de soorulatie per huidmondje in de tijd.     

The occurrence of a new genetic variant of Fusarium oxysporum f.sp. pisi

During a survey of root diseases of pea in Denmark, a new genetic variant of Fusarium oxysporum f.sp. pisi was isolated from vining peas in two widely separated geographical regions. In terms of pathogenicity on a set of differential pea lines, the Danish isolated closely resembled a race 6 isolate from the United States, DNA extracts of the isolates, restricted with the endonuclease Hind III, then probed with a homologous repetitive genomic fragment from the plasmid pDG106 by the Southern hybridization technique, gave a unique'fingerprint'pattern distinctly different from the American race 6 and all other known races. When probed with pDG312, containing a homologous ribosomal repeat unit, the pattern obtained for the Danish isolates was indistinguishable from races 1, 5 and 6 but distinctly different from 2A and 2B. The Danish isolates represent a separate vegetative compatibility group because they are compatible with each other but incompatible with the other known races. In pigmentation the new variant resembled races 1, 5 and 6 for the first 8-12 days, after which it began to secrete a dark purple pigment resembling that of race 2A and 2B. Until an additional line in the host differentials can separate the new genetic variant it should be considered a subgroup of F. oxysporum f. sp. pisi race 6.     

Germination of Peronospora farinosa f. sp. spinaciae conidia: a two-topped temperature curve

Samenvatting Bij verschillende herkomsten vanP. farinosa f. sp.spinaciae werden twee optima geconstateerd in de curve welke het verband aangeeft tussen conidiënkiemingspercentage en temperatuur.     

Soil receptivity toFusarium solani f. sp.pisi and biological control of root rot of pea

Potential antagonists ofFusarium solani f. sp.pisi (Fsp) were selected from soil samples with varying degrees of receptivity to this pathogen. They were tested against Fsp isolate 48 (Fs48), in increasingly complex systems. Most species testedin vitro were able to antagonize Fs48. No relation could be establishedin vitro between the receptivity of the soil from which an isolate originated and its antagonism to Fs48. In soils naturally infested with pea root rot pathogens, which were stored humid at 4°C for a period longer than a year, various isolates ofFusarium, Gliocladium andPenicillium spp. were able to reduce root rot. After sterilization of these soils, onlyGliocladium roseum isolates, added at 10⁵ conidia g^–1 dry soil, significantly reduced disease severity and prevented root weight losses caused by Fs48 at 10⁴ conidia g^–1 dry soil. In soils in which the biota were activated by growing peas before the assays, doses of 10⁶ and 10⁷ ofG. roseum were required to reduce root rot. In these soils, the antagonistic effects of fluorescent pseudomonad strains from soil of low receptivity to Fsp were variable. Some strains of fluorescent pseudomonads, from soil moderately receptive to Fsp and from highly infested soils, were also able to reduce root rot. Disease suppression by pseudomonad strains was more evident in the absence than in the presence ofAphanomyces euteiches in the root rot pathogen complex. The role of receptiveness of the soil with regard to potential antagonists is discussed.     

Colonization and sporulation of Peronospora viciae on cultivars of Pisum sativum

The behaviour of Peronospora viciae was examined on four pea ( Pisum sativum ) cultivars (Maro, Superb, Victory Freezer and Surprise) which appeared to exhibit varying degrees of resistance to this pathogen. Penetration into all cultivars was similar and mainly by direct entry through the cuticle. The pathogen grew in a similar manner and at a similar rate in all the cultivars during the early stages (<96 h) of tissue colonization. As it approached the onset of sporulation its growth slowed down in the more resistant cultivars but in no instance were any hypersensitive or other host cell reactions observed. Greater differences between the cultivars were seen in the extent of sporulation, with the least resistant cultivar (Superb) bearing the largest number of sporangiospores. There was much less sporulation on the most resistant cultivar (Maro) and the process was more dependent on temperature than it was on Superb. The development of mature plant resistance occurred in a similar way and on a similar time-scale in all cultivars. The youngest leaflets on plants which were less than 8 weeks old at inoculation showed the least resistance to infection and colonization.     

Toxicity of Glucosinolate Degradation Products from Brassica napus Seed Meal Toward Aphanomyces euteiches f. sp. pisi

ABSTRACT Brassica tissues are potentially useful in the control of Aphanomyces root rot of peas (Pisum sativum), but identity of the responsible compounds and specific impacts of those compounds on the pathogen's infection potential remain uncertain. Brassica napus seed meals and water extracts from these meals were used to determine the effect of glucosinolate hydrolysis products on Aphanomyces euteiches f. sp. pisi. B. napus meal ('Dwarf Essex') containing glucosinolates and intact myrosinase, the enzyme responsible for glucosinolate hydrolysis, completely inhibited infection by A. euteiches f. sp. pisi oospores. Water extracts from this meal, likewise, severely inhibited infection by oospores, as well as mycelial growth. Extracts from autoclaved 'Dwarf Essex' meal, in which myrosinase was denatured, and a low glucosinolate B. napus variety ('Stonewall') produced little disease reduction and had less impact on mycelial growth. Gas chromatographic analysis of Brassica tissues and water extracts confirmed that glucosinolates remained in autoclaved 'Dwarf Essex' meal and that 'Stonewall' meal contained low glucosinolate concentrations. 5-Vinyloxazolidine-2-thione was identified by mass spectrometry as a dominant glucosinolate hydrolysis product in aqueous extracts of the inhibitory meal. Bioassays conducted with aqueous solutions of this compound reduced mycelial growth, but not to the extent of those from intact 'Dwarf Essex' meal. Water-soluble compounds produced from the hydrolysis of glucosinolates in B. napus tissues reduced A. euteiches oospore infection and inhibited mycelial growth, thus, demonstrating potential utility of Brassica species in the control of A. euteiches.     

Resistance to Peronospora viciae expressed as differential colony growth in two cultivars of Pisum sativum

Peronospora viciae sporulated frequently following localized inoculation of leaflets of Pisum sativum cv. Krupp Pelushka but rarely on cv. Early Onward (60 and < 1% of plants respectively). Similar results were obtained following inoculation of seedling shoot apices. The proportions of sporangia that germinated, formed appressoria, invaded via stomata or cuticle, and established colonies, were similar on leaflets of both cultivars. After 2 days, total hyphal lengths of most colonies were greater than 100 μm (94% in Krupp Pelushka, 64% in Early Onward), but in Krupp Pelushka there were five times as many haustoria per unit length of hypha. Subsequently, only occasional colonies in Early Onward grew to the same extent as in Krupp Pelushka, with 8% and 88% respectively attaining lengths > 800 μm after 4 days. Reduced growth of hyphae in Early Onward was associated with increased frequency of host cells stained by trypan blue.     

香蕉枯萎病菌群体多样性研究进展

香蕉枯萎病严重威胁世界香蕉产业,而目前尚无有效防治药剂。开发快速诊断技术以加强检疫,控制其传播速度,同时加快选育抗病品种是有效控制该病的根本策略。然而,无论是研发快速诊断的分子技术还是进行抗病品种的选育,都需要深入了解病原菌的群体结构及其基因多样性背景。香蕉枯萎病菌经过一个多世纪的变异与进化,已分化出4个生理小种,23个营养亲合群和多个基因多样性类群。本文从香蕉枯萎病的起源及其病原菌培养性状、生理小种、致病性、营养亲合群及基因多样性等方面研究进展进行梳理和分析,以期为下一步的研究提供思路和启发。     

Appearance of race Pfs:5 of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, in Japan

The races for the causal agent of spinach downy mildew Peronospora farinosa f. sp. spinaciae were identified by inoculation of race-differential cultivars. One isolate was identified as Pfs:5s and the others belonged to a new race. This is the first report of race Pfs:5 and another new race in Japan.     

Temperature adaptation in Australasian populations of Puccinia striiformis f. sp. tritici

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the major fungal pathogens of wheat. A new pathotype was introduced to Australia in 2002 and several derivative pathotypes were detected in subsequent seasons. It has been suggested that the severity of stripe rust outbreaks in Australia since 2002 could be as a result of traits other than virulence in the pathogen population. This study was conducted to investigate the hypothesis that the stripe rust pathogen population dominant in Australia since 2002 was better adapted to warm temperature conditions compared to previous pathogen populations. Sixteen pathotypes were selected to examine the influence of two contrasting temperature regimes during the 24 h incubation (10°C and 15°C) and the subsequent post‐inoculation (17°C and 23°C) periods on latent period and infection efficiency on four susceptible wheat cultivars. In addition, the effect of two contrasting incubation temperatures on urediniospore germination was examined. The results indicated that pathotypes of P. striiformis f. sp. tritici detected after 2002 did not show evidence of adaptation to high temperatures, which suggests that other factors contributed to the observed increased aggressiveness.     

Virulence patterns in populations of Erysiphe graminis f.sp. hordei in Europe in 1986

A total of 3166 single colony progeny of Erysiphe graminis f.sp. hordei , constituting random samples of the aerial inoculum from 68 regions of Western Europe, were analysed in the laboratory for virulence to barley resistance genes Mlra, Mlg, Mla6+Mlg. MILa, Mla7+Mlk, Mlall, Mla9, MlaI3 + MlRu3. Mlal, Mla3 and mlo. No isolate virulent to mlo was detected. Distinct regional patterns were observed for the frequencies of most of the other virulence genes. They stressed the role of selection exerted by the cultivars grown, as well as the effect of wind dissemination of mildew populations. These two phenomena are particularly well exemplified by the data on early stages in the evolution of the newly emerging Val3 virulence in Denmark. In contrast with the preceding evolution, there was a considerable decrease of the frequency of a broadly distributed virulence ( Va6); the most probable reasons for this are outlined.     

Pisatin involvement in the variation of inhibition of Fusarium oxysporum f. sp. pisi spore germination by root exudates of Pisum spp. germplasm

Fusarium wilt caused by Fusarium oxysporum f. sp. pisi (Fop) is one of the major constraints of pea worldwide. Its control is difficult and is mainly based on the use of resistant cultivars. This study aimed to identify and characterize resistance mechanisms interfering with Fop spore germination, as an additional pre‐penetration resistance mechanism little explored so far. For this, root exudates were collected from 12 pea accessions with differential responses to the disease, from resistant to susceptible, and their effects on Fop germination and growth were determined. While root exudates from most accessions stimulated Fop germination, the root exudates of three accessions, JI 1412, JI 2480 and P42, did not stimulate, or even inhibited, Fop germination. Although some additional compounds might be involved, the analysis showed that the most active metabolite was the pea phytoalexin pisatin. Pisatin was identified in the active fraction of pea root exudate extracts and its amount in the root exudates was negatively correlated with the extent of Fop germination. This suggests an important role of pisatin in the constitutive defence of pea against F. oxysporum.     

小麦白粉病菌有性生殖与自然群体交配型检测

 本研究根据MAT1-1-1(GenBank登录号HQ171902)和MAT1-2-1(GenBank登录号HQ171899)的部分基因序列设计和筛选其特异性引物,使用PCR方法对2012年度采自四川、陕西、云南、贵州、河南、安徽、江苏、河北、山东和甘肃10个省30个采集地点共516个小麦白粉病菌菌株的交配型基因进行检测。结果显示30个采集地点两种交配型基因都能检测到,其中28个采集地的两种交配型菌株比例(MAT1-1-1 :MAT1-2-1)符合1:1。选择具不同交配型基因的小麦白粉病菌株配对接种,16个随机杂交组合都能产生闭囊壳。对获得的闭囊壳进行子囊孢子释放,均可获得子囊孢子。该研究结果为深入研究有性生殖对小麦白粉病菌株毒性多样性的作用奠定了基础。