Phytotoxicity on cotton ex-plants of an 18.5 kDa protein from culture filtrates of Verticillium dahliae

A phytotoxic protein that evokes the typical symptoms of Verticillium wilt disease in seedlings of Gossypium hirsutum L. (Upland cotton) was isolated from culture filtrates of Verticillium dahliae. The protein was purified by ammonium sulfate precipitation, Sephadex-G100 fractionation, and native PAGE. The 18.5 kDa protein, designated VD18.5, appears to be a single subunit protein with an isoelectric point between 3 and 5. VD18.5 induces symptoms of leaf dehydration, chlorosis, necrosis and stem discoloration in seedlings of the disease susceptible cotton cultivar Siokra 1–4. The LD₅₀ of VD18.5 on protoplasts of Siokra 1–4 was 18 μg mL⁻¹. VD18.5 had no noticeable effect on Pima S-7, which is a disease resistant cultivar. Phytotoxic activity was partially destroyed at high temperature and was abolished by digestion with proteinase K. Mass spectrometry fingerprinting and protein sequence data from VD18.5 yielded no significant matches when submitted to the Mascot search engine and NCBI non-redundant protein databases, respectively. These results suggest that VD18.5 is a novel protein that may be involved in the development of some of the symptoms associated with Verticillium wilt disease in the cotton plant.     

几种生化物质与西瓜抗蚜性的相关性

为了初步探讨西瓜抗蚜性的营养与次生代谢物质防御效应,采用分光光度法测定并分析了几种生化物质与西瓜抗蚜性的相关性。结果表明,抗性品种黑皮、绿美人、黑美人和惠兰的游离氨基酸、脯氨酸和可溶性氮含量分别为2.22~2.27、0.23~0.26和1.88~3.03 mg/g,显著低于感性品种金美人、小玲、甜美人和花绿的3.24~3.83、0.67~0.87和5.26~5.82 mg/g;抗性品种的糖/氮比为5.38~7.94,显著高于感性品种;抗、感品种间可溶性糖和维生素C(Vc)含量无显著差异;瓜蚜为害4 d后的抗性品种叶组织中的总酚和单宁酸含量显著增加,感性品种显著降低;丙二醛含量增加幅度显著高于抗性品种。研究表明,西瓜品种抗蚜性与糖/氮比及总酚、单宁酸含量均呈显著正相关,与可溶性氮、游离氨基酸、脯氨酸及丙二醛含量均呈显著负相关,但与可溶性糖和Vc含量无显著相关性。     

Nutritional requirements of the mycoparasitic fungus Verticillium biguttatum

Verticillium biguttatum was able to grow axenically in a synthetic liquid medium with a compound containing ammonium or amino group as nitrogen source, glucose as carbon source and biotin as growth factor. Among various carbon and nitrogen compounds tested, highest mycelial production was achieved with mannitol and with two ammonium salts and glutamine; sporulation reached highest values with galactose and glutamine. Highest yields of mycelium and conidia were obtained at pH 4.3 and 5.1, respectively. Although neutral and alkaline conditions were growth-limiting in the synthetic medium some growth ofV.biguttatum occurred on solid media at pH 7.0 and on sclerotia ofRhizoctonia solani in natural soil at pH 7.2–7.3.Samenvatting Een synthetisch vloeibaar medium met glucose als koolstofbron, een ammonium- of aminogroep bevattende verbinding als stikstofbron en biotine als groeifactor voldeed aan de voedingseisen vanVerticillium biguttatum. Van de verschillende koolstof- en stikstofbronnen leverden mannitol en twee ammoniumzouten en glutamine de hoogste opbrengsten aan mycelium; de opbrengst aan conidiën was het hoogst met galactose en glutamine. De hoogste opbrengsten aan mycelium en conidiën werden bereikt bij respectievelijk pH 4,3 en 5.1. Ofschoon neutrale en alkalische omstandigheden de groei vanV. biguttatum in het synthetische medium beperkten, werd enige groei vanV. biguttatum waargenomen op vaste voedingsbodems bij pH 7,0 en op sclerotia vanR. solani in natuurlijke grond bij pH 7,2–7,3.     

Sequence Analysis Shows that a Dwarfing Disease on Rice, Wheat and Maize in China is Caused by Rice Black-streaked Dwarf Virus

Isolates of plant reoviruses causing severe stunting and dark leaf symptoms on wheat in Hebei province, on maize in Hubei province and on rice in Zhejiang province, China have been characterized. Four of the ten genome segments corresponding to Rice black-streaked dwarf virus (RBSDV) S7, S8, S9, S10 were amplified by RT-PCR from the Hebei and Zhejiang isolates and sequenced. Sequences of S9 and S10 were also obtained from the Hubei isolate. Sequences of corresponding segments of the Chinese isolates were very similar to each other (94.0–99.0% identical nucleotides and 96.3–100% identical amino acids) and were closer to those of a previously reported Japanese RBSDV isolate (90.0–94.9% identical nucleotides and 91.1–98.6% identical amino acids) than to those of an Italian Maize rough dwarf virus isolate (85.1–88.1% identical nucleotides and 85.5–94.3% identical amino acids). The Chinese isolates should be classified as RBSDV.     

Purification of polygalacturonases produced by the pear scab pathogens, Venturia nashicola and Venturia pirina

An exopolygalacturonase and three endopolygalacturonases were purified from mycelia of pear scab pathogens, Venturia pirina and Venturia nashicola. The molecular weight of the isolated exoPG from V. pirina was 43 kDa, and the endoPGs from V. nashicola were 42 kDa as estimated by SDS–polyacrylamide gel electrophoresis. The pH optimum of the exoPG activity from V. pirina was 5.0. TheK_m and V_maxvalues of the exoPG were 0.08 mg ml⁻¹and 4.44 × 10⁻³ mmol reducing group min⁻¹ mg protein⁻¹. The N-terminal amino acid sequence of the exoPG from V. pirina was similar to that of the exoPG from Fusarium oxysporum f. sp. melonis, and the N-terminal amino acid sequences of the three endoPGs fromV. nashicola races 1, 2 and 3 were similar to other fungal endoPGs with a conserved motif of ASxxxTFTxAAAxxxG.     

Effect of benomyl on soil fungi associated with rye. 1. Effect on the incidence of sharp eyespot caused by Rhizoctonia cerealis

Effects of benomyl on incidence of pathogens affecting the culm base of rye were studied in field trials and growth chamber experiments. Spraying of the crop with the fungicide at a high dosage (2.4 kg.ha^–1) resulted in a tenfold increase of sharp eyespot caused byRhizoctonia cerealis and reduced foot rot symptoms caused by fusaria by 50%. In a field trial at a low dosage (0.24 kg.ha^–1) a slight increase of sharp eyespot was observed. In one year, probably because of wet conditions during the infection period, sharp eyespot did not occur in either benomyl-treated or untreated plots, but eyespot caused byPseudocercosporella herpotrichoides was abundant. Its occurrence was reduced from 74% affected culm bases in untreated plots to 8% and 1% in plots that received 0.24 and 2.4 kg.ha^–1 of the fungicide, respectively.In growth chambers seedlings were grown in two sandy soils inoculated withR. cerealis. The soil was kept dry at about 35% of the moisture holding capacity. In plots with benomyl (1 mg.kg^–1; moisture content 11% of fresh weight), fewer seedlings emerged than in plots without the fungicide. This result was highly significant (P<0.01) for one soil but not for the other. The number of seedlings that remained free of disease symptoms was higher (P<0.01) in untreated than in fungicide-treated plots of both soils.Isolates of pathogens obtained from diseased culms were tested for their sensitivity to benomyl. Growth of all of them includingR. cerealis was inhibited, although not always completely suppressed, at 10 g.ml^–1 on potato-dextrose agar. ED₅₀ values of most isolates ofR. cerealis were between 2.2 and 3.1 g.ml^–1. The fungus was slightly but consistently less sensitive thanF. culmorum. Mycelial growth ofF. nivale was appreciably more sensitive than that of the otherFusarium spp. from cereals.P. herpotrichoides andF. nivale were the most sensitive pathogens tested with ED₅₀ values of <1 g.ml^–1. Accordingly,F. nivale was absent on culms from treated plots. In a growth chamber experiment, seedlings were protected from infection by supplying the fungicide (1 mg.kg^–1) to previously inoculated soil.In a laboratory assay the effect of benomyl on microbial antagonism toR. cerealis was estimated for rhizosphere soil. Enhanced incidence of sharp eyespot in treated crops was associated with adverese effects of the fungicide on microbial antagonism. There is presumptive evidence thatR. cerealis is suppressed by bacteria after wet periods during the vegetation period of the crop and by fungi after dry periods. Only fungal antagonism, which may be less effective, is affected by benomyl. The response to benomyl of the microflora in different soils varied. Reasons for this inconsistency are suggested.Samenvatting In veldproeven en in een klimaatkamer werd de invloed van benomyl op het optreden van voetziekten in rogge onderzocht. In veldjes die bespoten waren met een hoge dosis van het fungicide (in totaal 2.4 kg.ha^–1) bleken tienmaal zoveel halmen met scherpe oogvlekken, veroorzaakt doorRhizoctonia cerealis, voor te komen dan in onbespoten veldjes. Daarentegen was voetrot veroorzaakt doorFusarium-soorten met 50% verminderd. In een volgende veldproef, waarbij een voor de praktijk geadviseerde dosis (0.24 kg.ha^–1) was toegepast, werd een lichte toename van scherpe oogvlekken waargenomen.In een ander jaar trad scherpe oogvlekkenziekte in het geheel niet op, ook niet in met benomyl behandelde veldjes. De vochtige omstandigheden tijdens de infectieperiode zijn daarvan waarschijnlijk de oorzaak. Daarentegen werd de oogvlekkenziekte, welke doorPseudocercosporella herpotrichoides werd veroorzaakt, veel aangetroffen. In de onbehandelde veldjes waren 74% van de halmen aangetast tegen 8 en 1% in de veldjes die met het fungicide waren behandeld in doseringen van 0.24 en 2.4 kg.ha^–1.De invloed van het fungicide op de aantasting van kiemplanten werd in klimaatkamerproeven onderzocht. Daartoe werden twee zandgronden metR. cerealis geënt. De grond werd droog gehouden (op 35% van het waterhoudend vermogen). In grond met fungicide (1 mg.kg^–1) was de opkomst minder dan in grond zonder fungicide. Dit was zeer significant (P<0.01) voor één van de beide zandgronden, maar niet voor de andere. Het aantal gezonde kiemplanten was in beide gevallen duidelijk hoger (P<0.01) voor de onbehandelde grond.De isolaten van ziekteverwekkers uit aangetaste halmen werden op hun gevoeligheid voor het fungicide getoetst. Op aardappel-glucoseagar werden alle isolaten in hun groei geremd bij een benomyl-concentratie van 10 g.ml^–1.R. cerealis was iets minder gevoelig danF. culmorum. Voor het overgrote deel van de isolaten vanR. cerealis lag de ED₅₀ waarde tussen 2,2 en 3,1 g.ml^–1. De myceliumgroei vanF. nivale werd meer geremd dan die van de andereFusarium-soorten.P. herpotrichoides enF. nivale waren met een ED₅₀ waarde van <1 g.m.^–1 de gevoeligste pathogenen die uit de halmvoeten werden geïsoleerd. Dat de populatie vanF. nivale in benomylhoudende grond wordt onderdrukt, blijkt uit (1) het feit dat de schimmel niet voorkwam op halmen uit behandelde veldjes en (2) de bescherming tegen infectie van kiemplanten als aan de besmette grond fungicide (1 mg.kg^–1) was toegevoegd.In laboratoriumproeven werd de invloed van benomyl op het microbiële antagonisme in rhizosfeergrond tegenR. cerealis bepaald. Een toename in het optreden van scherpe oogvlekkenziekte in behandelde gewassen bleek gepaard te gaan met een remming van het antagonisme tegen de ziekteverwekker. Er zijn sterke aanwijzingen datR. cerealis na vochtige perioden tijdens de vegetatieperiode door bacteriën wordt onderdrukt en na droge perioden door schimmels. Het antagonisme van de laatste groep lijkt minder effectief te zijn en alleen dit antagonisme wordt door benomyl verlaagd. Tenslotte wordt een mogelijke oorzaak aangegeven voor de ongelijke respons op het fungicide van het microbieel antagonisme in verschillende gronden.     

Resistance Reaction of Conifer Species (European Larch, Norway Spruce, Scots Pine) to Infection by Selected Necrotrophic Damping-off Pathogens

Three conifer species (European larch, Norway spruce, Scots pine) were investigated for their resistance to five damping-off pathogens (Rhizoctonia solani, Fusarium solani, F. oxysporum, F. culmorum, F. avenaceum). Inoculation of the primary roots of seedlings with these pathogens caused host cell death which did not prevent the invasive growth of these fungi; seedlings that had formed secondary and tertiary roots could overcome the infection to a significant degree. Infections with R. solani caused significant mortality to all the conifer species. In contrast, the tree species expressed different levels of resistance when challenged with the Fusarium isolates, with Norway spruce being the most resistant compared to uninoculated controls. Some of the Fusarium isolates were more pathogenic to certain hosts than others; F. oxysporum for European larch, F. avenaceum for Scots pine, F. solani for European larch; only F. culmorum was significantly pathogenic to Norway spruce. No significant differences in disease severity were observed at different soil pH (4.3–7.5). Disease progression was delayed at lower (10–15°C) rather than higher temperatures (20–25°C).     

First report of a <Emphasis Type="Italic">Grapevine Algerian latent virus</Emphasis> disease on statice plants (<Emphasis Type="Italic">Limonium sinuatum</Emphasis>) in Japan

A viral disease was found in Nagano Prefecture, Japan, on statice (Limonium sinuatum) with chlorotic leaf spot, necrotic stunt, and dwarfing. Spherical virus particles 30 nm in diameter were isolated from infected plants and statice seedlings and caused identical symptoms 4 weeks after mechanical inoculation. Nucleotide and deduced amino acid sequences of the coat protein showed 98% and 98.7% identities with those of Grapevine Algerian latent virus (GALV) nipplefruit strain. This is the first report in Japan of a viral disease on statice caused by GALV. The nucleotide sequence data reported here are available in the DDBJ/EMBL/GenBank databases under accession AB461854.     

Identity and pathogenicity of Rhizoctonia spp. associated with bare patch disease of cereals at a field site in Western Australia

Roots of seedlings of wheat and barley affected by bare patch disease at a field site in Western Australia were assessed for root damage and plated to isolate fungi. The patches were variable in shape and size and had the most severely affected plants in the centre. Of the 165 isolates ofRhizoctonia spp. obtained, 90% were multinucleate and 10% binucleate, the former being predominant in the plants at the centre of the patch. The relative frequency of binucleate isolates increased with proximity to the periphery. The increase in activity of avirulent binucleate isolates towards the periphery of the patch may be related to the sharp and abrupt edging of the patch. A variety of other species of fungi such asFusarium spp.,Mortierella spp.,Bipolaris sorokiniana, Pythium sp. andTrichoderma sp. were encountered within the patches. The multinucleate isolates belonging to anastomosis groups (Ag) 2–1, 2–2 and 8 (Thanatephorus cucumeris) were most pathogenic to wheat. The binucleate isolates of Ag C, D, E, and K (Ceratobasidium sp.) were less pathogenic. It is suggested that the bare patch disease is caused by a complex of root rot fungi composed of one or more anastomosis groups ofRhizoctonia spp. and other associated fungi.Samenvatting Van kiemplanten van tarwe en gerst, afkomstig van een met kale-plekkenziekte besmet perceel in West Australië werd de mate van wortelbeschadiging bepaald en werden schimmels uit de wortels geïsoleerd. De plekken waren verschillend van vorm en afmeting; de zwaarst aangetaste planten werden in het centrum ervan aangetroffen. Van de 165 verkregen isolaten vanRhizoctonia spp. was 90% meerkernig en 10% tweekernig. De meerkernige overheersten in de centra van de plekken. Relatief gezien nam het aantal tweekernige isolaten toe naarmate de herkomst dichter bij de periferie van de plekken was. De scherpe begrenzing van de ziekte aan de randen van de plekken zou in verband kunnen staan met het toenemen van de activiteit van de avirulente tweekernige isolaten in de nabijheid van de periferie van de plekken. Een aantal andere schimmels, zoalsFusarium spp.,Mortierella spp.,Bipolaris sorokiniana, Pythium sp. enTrichoderma sp. werd eveneens in de plekken aangetroffen. De meerkernige isolaten die tot de anastomosegroepen Ag 2–1, 2–2 en 8 (Thanatephorus cucumeris) behoren, waren voor tarwe het meest pathogeen. De tweekernige isolaten van de anastomosegroepen Ag C, D, E en K (Ceratobasidium sp.) waren minder pathogeen. Gesuggereerd wordt, dat de kale-plekkenziekte veroorzaakt wordt door een complex van verschillende wortelschimmels, die behoren tot een of meer anastomosegroepen vanR. solani en andere daarmee geassocieerde schimmels.     

华北大黑鳃金龟Holotrichia oblita中肠羧酸酯酶基因 HoCL1的克隆与序列分析

利用粉纹夜蛾(Trichoplusia ni)围食膜蛋白多克隆抗体,从已构建的华北大黑鳃金龟 Holotrichia oblita 中肠cDNA表达文库中筛选得到1个编码羧酸酯酶的cDNA克隆 HoCL1 ,其开放阅读框(ORF)长1 599 bp,编码532个氨基酸,推导的蛋白质分子质量为59.5 kDa,等电点(p I)为4.5。 HoCL1蛋白具有羧酸酯酶的保守结构域:1个二硫键形成的位点和1个丝氨酸活性中心,三联体催化活性中心位于Ser207、Asp333和His422上,不含有氮联糖基化位点和氧联糖基化位点,只含有3 个半胱氨酸残基。依据氨基酸序列同源性分析和保守结构域分析,HoCL1属于B类酯酶,与赤拟谷盗 Tribolium castaneum 羧酸酯酶相似性最高,为35.2%。通过与其他昆虫羧酸酯酶序列比对及构建系统发育树,发现HoCL1羧基端的氨基酸序列保守性低,但靠近N端的活性中心处的氨基酸序列则高度保守,可与赤拟谷盗、异色瓢虫 Harmonia axyridis 羧酸酯酶聚类在一起。羧酸酯酶 HoCL1 基因的克隆鉴定为进一步研究该基因在华北大黑鳃金龟体内的表达及功能奠定了基础。     

Studies on in vitro Growth and Pathogenicity of European Fusarium Fungi

The effect of temperature on the in vitro growth rates and pathogenicity of a European Fusarium collection consisting of isolates of Fusarium graminearum, F. culmorum, F. avenaceum, F. poae and Microdochium nivale was examined. Irrespective of geographic origin, the optimum temperature for the growth of F. graminearum, F. culmorum and F. poae was 25 °C, while that for F. avenaceum and M. nivale was 20 °C. In general, the growth rates of F. graminearum, F. culmorum and F. poae increased between 10 and 25 °C and those of F. avenaceum and M. nivale increased between 10 and 20 °C. Pathogenicity tests were carried out by examining the effect of the five species on the in vitro coleoptile growth rate of wheat seedlings (cv. Falstaff). Irrespective of geographic origin, the temperature at which F. avenaceum, F. culmorum and F. graminearum caused the greatest retardation in coleoptile growth ranges 20–25 °C (>89.3% reduction), whilst for F. poae and M. nivale it was 10–15 °C (>45.6% retardation), relative to uninoculated control seedlings. In general, F. culmorum and F. graminearum were the most pathogenic of the five species, causing at least a 69% reduction in coleoptile growth at 10, 15, 20 and 25 °C. General linear model analysis (GLIM) showed that species accounted for 51.3–63.4% of the variation in isolate growth and from 19.5% to 44.3% of the variation in in vitro pathogenicity. Country of origin contributed from 22.6% to 51.9% to growth rate variation and from 0.73% to 7.61% to pathogenicity variation. The only significant correlation between in vitro growth and pathogenicity was that observed for M. nivale at 15 °C (r = -0.803, P < 0.05).     

Induction of Systemic Acquired Resistance in Pepper Plants by Acibenzolar-S-methyl against Bacterial Spot Disease

The ability of acibenzolar-S-methyl to induce resistance in pepper plants against Xanthomonas campestris pv. vesicatoria was investigated in both growth chamber and open field conditions. Growth chamber experiments showed that acibenzolar-S-methyl (300M) treatment protects pepper plants systemically and locally against X. campestris pv. vesicatoria. Evidence for this was a reduction in the number and diameter of bacterial spots and bacterial growth in planta. Systemic protection was also exerted by the acibenzolar-S-methyl acid derivative, CGA 210007, which may be produced by hydrolysis in the plant. The efficacy of acibenzolar-S-methyl was also found in open field conditions, where both leaves and fruit were protected from the disease. The highest efficacy (about 67%) was obtained by spraying the plants 6–7 times every 8–12 days with a mixture of acibenzolar-S-methyl and copper hydroxide (2.5 + 40ghl^–1 active ingredient). Persistence and translocation data obtained from the growth chamber experiments revealed a persistence of acibenzolar-S-methyl lasting five days after treatment with rapid translocation and negligible levels of acid derivative formation. Since the protection exerted by acibenzolar-S-methyl against bacterial spot disease was observed when the inducer was completely degraded, it would appear to be due to SAR activation.     

Studies on the Inherent Resistance Risk to Fenhexamid in Botrytis cinerea

After chemical mutagenesis with N-methyl-N-nitrosoguanidine (MNNG) two phenotypes that were highly or moderately resistant to fenhexamid, were isolated from a wild-type strain of Botrytis cinerea, at a mutation frequency of 0.9 × 10^–5. Resistance factors, based on EC⁵⁰ values, were 460–570 and 10–15, respectively. The mutation(s) for resistance to fenhexamid did not affect the sensitivity of mutant strains to the benzimidazole benomyl, the phenylpyridinamine fluazinam, the anilinopyrimidine cyprodinil, the guanidine iminoctadine or to the sterol-biosynthesis-inhibiting fungicides fenarimol, fenpropimorph and tridemorph. On the contrary, an increased sensitivity (EC⁵⁰ ratios of 0.2–0.6) of fenhexamid-resistant strains to the phenylpyrrole fludioxonil and the dicarboximide iprodione was observed. Study of fitness parameters of fenhexamid-resistant isolates of both phenotypic classes showed that these mutation(s) had no effect on mycelial growth and sensitivity to high osmolarity, but they did affect one or more of some other characteristics, such as sporulation, conidial germination and sclerotia production. In tests on cucumber seedlings under greenhouse conditions, all highly fenhexamid-resistant isolates tested presented decreased infection ability compared with the wild-type. Preventive applications of a commercial formulation of fenhexamid, Teldor 50 WP, were effective against lesion development on cotyledons by the wild-type, but ineffective, even in high concentrations, against disease caused by the fenhexamid-resistant isolates. The risk of resistance problems arising during commercial use of fenhexamid is discussed.     

Siderophore-mediated competition for iron and induced resistance in the suppression of fusarium wilt of carnation by fluorescent Pseudomonas spp

The mechanisms of suppression of fusarium wilt of carnation by two fluorescentPseudomonas strains were studied.Treatments of carnation roots withPseudomonas sp. WCS417r significantly reduced fusarium wilt caused byFusarium oxysporum f. sp.dianthi (Fod). Mutants of WCS417r defective in siderophore biosynthesis (sid^–) were less effective in disease suppression compared with their wild-type. Treatments of carnation roots withPseudomonas putida WCS358r tended to reduce fusarium wilt, whereas a sid^– mutant of WCS358 did not.Inhibition of conidial germination of Fod in vitro by purified siderophores (pseudobactins) of bothPseudomonas strains was based on competition for iron. The ferrated pseudobactins inhibited germination significantly less than the unferrated pseudobactins. Inhibition of mycelial growth of Fod by bothPseudomonas strains on agar plates was also based on competition for iron: with increasing iron content of the medium, inhibition of Fod by thePseudomonas strains decreased. The sid^– mutant of WCS358 did not inhibit Fod on agar plates, whereas the sid^– mutants of WCS417r still did. This suggests that inhibition of Fod by WCS358r in vitro was only based on siderophore-mediated competition for iron, whereas also a non-siderophore antifungal factor was involved in the inhibition of Fod by strain WCS417r.The ability of thePseudomonas strains to induce resistance against Fod in carnation grown in soil was studied by spatially separating the bacteria (on the roots) and the pathogen (in the stem). Both WCS417r and its sid^– mutant reduced disease incidence significantly in the moderately resistant carnation cultivar Pallas, WCS358r did not.It is concluded that the effective and consistent suppression of fusarium wilt of carnation by strain WCS417r involves multiple mechanisms: induced resistance, siderophore-mediated competition for iron and possibly antibiosis. The less effective suppression of fusarium wilt by WCS358r only depends on siderophore-mediated competition for iron.     

Rootstock resistance to Fusarium wilt and effect on watermelon fruit yield and quality

The potential of grafted watermelon for resistance toFusarium oxysporum f.sp.niveum on some Curcurbitaceae,Lagenaria, Luffa, Benincasa and commercial rootstocks was evaluated. Effects of grafting on yield and quality of diseased plants were evaluated. All grafted plants and rootstocks were resistant to the three known races (0, 1, and 2) ofF. oxysporum f.sp.niveum except watermelon cv. ‘Crimson Tide’, which was susceptible to race 2. Fruit yield was positively (21–112%) affected byLagenaria rootstocks but negatively affected (200–267%) byCucurbita rootstocks when compared with the control. While only minor differences in fruit quality were determined in control and grafted plants onLagenaria rootstocks, the quality parameters for watermelon grafted ontoCucurbita rootstocks were lower than in the control. The reasons for low yield and quality might be due to an incompatibility betweenCucurbita rootstocks and watermelon. These results showed that rootstock influence on disease resistance as well as yield and quality of scion fruit is important in determining the potential use of grafting applications in watermelon. http://www.phytoparasitica.org posting Feb. 2, 2003.     

First report of Rhizomonas sp. causing corky root of lettuce in Europe

Typical symptoms of corky root were observed on iceberg lettuce (Lactuca sativa L.) in the Netherlands and England, on prickly lettuce (Lactuca serriola L.) in Spain, and on sowthistle (Sonchus oleraceus L.) in Greece. Slow-growing bacteria with similar colonies as strains ofRhizomonas suberifaciens or otherRhizomonas species were isolated from soil surrounding plants with corky root symptoms using lettuce seedlings as bait. Crude lysate from all strains was tested for DNA homology with DNA fromR. suberifaciens strain CA1 (R. sub. CA1) andRhizomonas sp. strain WI4 (R. sp. WI4). Strains that had homology values higher than that ofR. sp. WI4 orR. sub. CA1 were tested for pathogenicity on 1-wk-old lettuce seedlings, cv. Salinas, Two strains from the Netherlands induced typical symptoms of corky root on lettuce. These strains tested negative with monoclonal antibody MAb-Rs1 specific forRhizomonas suberifaciens in an enzyme-linked immunosorbent assay (ELISA). They had low DNA homology withR. sub. CA1 (4–9%) and low to moderate DNA homology withR. sp. WI4 (4–17%). Some nonpathogenic strains had moderate to high levels of DNA homology withR. sub. CA1 orR. sp. WI4 (19–84%). All strains had fatty acid profiles similar to those ofRhizomonas species. This is the first report ofRhizomonas sp. causing corky root of lettuce in Europe.     

Genetic Variation in Melampsora Larici-epitea on Biomass willows Assessed using AFLP

Five pathotypes belonging to formae speciales larici-epitea typica (LET), larici-retusae (LR) and larici-daphnoides (LD) of Melampsora larici-epitea were examined using amplified fragment length polymorphism (AFLP). Of 213 AFLP markers scored, several were found to be exclusive to different formae speciales. The dendrogram placed the five pathotypes into distinct groups. Within pathotypes, average Nei & Li's similarity coefficients were calculated as 0.71–0.85. The similarities were 0.66–0.72 among the three pathotypes within LET and 0.34–0.44 between pathotypes belonging to different formae speciales. When assessed using the Shannon index, the diversity within locations was estimated as 0.55–0.59, greater than that found within pathotypes (0.24–0.42). The average per-locus diversity was 0.37 among the pathotypes and 0.11 among the locations. When the data from both LET and LR isolates were examined using AMOVA, the majority of the variation (70.85%) was attributed to among pathotypes within location. When only LET types were included, approximately half of the variation was partitioned to among pathotypes within location and the other half to among the isolates within collection. It appears that the degree of differentiation of LET4 on S. × mollissima between Loughgall and Long Ashton sites has decreased markedly since 1992, when it was first detected.     

Line pattern in Limonium latifolium caused by tobacco rattle virus

Tobacco rattle virus (TRV) was isolated from plants ofLimonium latifolium showing bright yellow or red line patterns and ringspots on the leaves. It was proved that this virus, designated TRV-Lim, was the causal agent of the disease. In its reactions onNicotiana clevelandii it resembled a yellow strain of TRV from Oregon (USA), but the symptoms inN. glutinosa, N. megalosiphon, N. tabacum andPetunia hybrida were more comparable to those caused by socalled unstable variants of TRV. Dilution end-point was 10^–6–10^–7, thermal inactivation at 76–80°C, and ageing in vitro 55–60 days. The purified virus suspension contained particles of three normal lengths, 70, 102, and 194 nm. The virus sedimented as three components with average sedimentation coefficients of 129, 161 and 206 S, respectively. In purified suspensions TRV-Lim had two different buoyant densities. A serological relationship was found with TRV isolated from Europe and Brazil.Samenvatting Tabaksratelvirus (TRV) werd geïsoleerd uitLimonium latifolium planten die heldergeel of rood figuurbont op de bladeren vertoonden. Er werd aangetoond dat dit virus, aangeduid als TRV-Lim, de ziekteverwekker was. De reacties van dit virus opNicotiana clevelandii deden denken aan die van een gele stam van TRV afkomstig uit Oregon (VS), maar de symptomen opN. glutinosa, N. megalosiphon, N. tabacum enPetunia hybrida vertoonden meer gelijkenis met die welke veroorzaakt worden door de zogenaamde onstabiele varianten van TRV. De verdunningsgrens was 10^–6–10^–7, de inactiveringstemperatuur 75–80°C en de houdbaarheid in vitro 55–60 dagen. De gezuiverde virussuspensie bevatte deeltjes met drie normale lengtes, nl. 70, 102 en 194 nm. Het virus sedimenteerde, als drie componenten met gemiddelde sedimentatiecoëfficiënten van respectievelijk 129, 161 en 206 S. In gezuiverde suspensie vertoonde TRV-Lim twee verschillende zweefdichtheden. Het virus was serologisch verwant aan TRV-isolaten uit Europa en Brazilië.     

Etiology of asparagus replant-bound early decline

Asparagus replant-bound early decline (ARED) was characterized and its etiology was elucidated in experiments under greenhouse and field conditions. Selective soil treatments were used to differentiate between autotoxic compounds and soil-borne pathogens as causal agents. In greenhouse experiments, there were symptoms of ARED within 12—15 weeks. Asparagus plants grown in soil formerly used for asparagus (asparagus soil) showed brown lesions on primary and secondary roots, and many secondary roots had rotted. Root weights of plants grown in asparagus soil were lower than those of plants grown in fresh soil.Fusarium oxysporum f. sp.asparagi (Foa) was by far the most common species among the fungi isolated from roots with lesions. Under greenhouse and field conditions, there were similar symptoms, which indicates that the results obtained under greenhouse conditions are similar to those in the field. The vertical distribution of the ARED-causing factor(s) was studied in a greenhouse experiment in which plants were grown in soil from three layers: 0–30, 30–60, and 60–90 cm. For all four asparagus soils tested, there were ARED symptoms and similar disease severity in samples from all three depths. The causal factor persisted at least 11 years after soil was no longer used for asparagus. When asparagus soil was diluted with fresh soil to give mixtures with 100%, 80%, 50%, 20% and 0% asparagus soil, disease severity did not decrease with increasing dilution of the asparagus soil from 100% to 20%. Disease severity of all mixtures with asparagus soil was significantly higher than that for fresh soil. The results imply that ARED is caused by a pathogen colonizing the soil rather than inhibition by autotoxins released from residues of the preceding asparagus crop. This conclusion is supported by the results of greenhouse and outdoor experiments with heat and fungicide treatments of soil. ARED was nullified by heat treatments of 30 min at 55 or 60 °C but not 45 and 50 °C, eliminating autotoxins as an important cause of ARED because they are heat-stable. Foa is eliminated by a 30-min soil treatment at 55–60 °C but not 50 °C. Prochloraz, known for its toxicity toF. oxysporum, also nullified ARED. Disease severity level was related to the density of Foa in soil. The results provide conclusive evidence thatF. oxysporum f. sp.asparagi is the main cause of ARED in the Netherlands, which largely removes the need to discriminate between early decline and replant-bound early decline, because Foa is the main cause of both diseases.