PF7-5对烟草角斑病的室内及大田防治效果

为验证荧光假单胞杆菌PF7-5对烟草角斑病的生物防治效果,设计了荧光假单胞杆菌PF7-5对烟草角斑病菌的室内和大田抑菌试验。结果表明,室内试验中荧光假单胞杆菌PF7-5代谢产物对烟草角斑病的抑菌效果达到71%,大田试验中PF7-5活菌的50倍稀释液田间防治效果达87.34%,说明荧光假单胞杆菌PF7-5对烟草角斑病有一定的防治效果。     

Screening wild cherry (Prunus avium) for resistance to bacterial canker by laboratory and field tests

Currently, bacterial canker caused by Pseudomonas syringae is a major cause of dieback and tree death in wild cherry (Prunus avium) plantations. The evaluation of breeding collections is needed to produce less susceptible clones or cultivars. Resistance tests were performed using excised shoots (1 and 2 years old) from 79 clones in the laboratory. A subset of 10 clones was also tested in the field. The clones were inoculated with four to seven isolates of a set of 15 isolates of P. s. pv. morsprunorum, P. s. pv. syringae, P. s. pv. persicae, P. syringae pv. avii and P. fluorescens. In the laboratory tests, older and larger shoots were more susceptible. In the field test, size and age of the shoots were not related to girdling by the bacterial canker. Two‐year‐old shoots were best for clonal discrimination. Correlations between 1 and 2‐year‐old shoots were significant but not high. The isolates varied a lot between experiments, but as the clone × isolate interactions were always low, breeding could thus be facilitated. The ranking of clones was conserved quite well between two laboratory tests, but not between two others. Good agreement was found for the best clones in the laboratory tests and in the field test. However, the two worst clones in the latter were among the best in one laboratory test. At least two independent tests in the laboratory are needed to evaluate resistance/susceptibility of clones. Broad sense heritability for resistance varied from 0.27 to 0.51. Although moderate, such heritability clearly encourages a breeding approach to reduce the problem of bacterial canker.     

Pseudomonas syringae pv. aesculi: foliar infection of Aesculus species and temperature–growth relationships

Since 2001, the incidence of bleeding canker of horse chestnut (Aesculus hippocastanum) has increased markedly in western Europe. The causal agent, the bacterium Pseudomonas syringae pv. aesculi, originally isolated from foliar lesions on Indian horse chestnut (Aesculus indica) in India, is a bark killing pathogen on A. hippocastanum. In this study, P. syringae pv. aesculi was found as a foliar epiphyte on both A. hippocastanum and A. indica trees growing in the UK. When Aesculus leaves were challenged with cell suspensions (10⁹ CFU ml^?1) of Pseudomonas syringae pv. aesculi, a high level of asymptomatic infection occurred in all the species tested. The degree of re‐isolation of the bacterium after surface sterilization of leaves ranged from 33% (A. pavia) to 84 and 97% for A. hippocastanum and A. chinensis, respectively. The studies suggest both epiphytic and intrafoliar populations of P. syringae pv. aesculi could play a role in the incidence and spread of bleeding canker of horse chestnut. Growth–temperature responses of P. syringae pv. aesculi indicated a minimum of approximately ?4°C and a maximum of approximately 35°C, with an optimum of approximately 25°C. These findings show that P. syringae pv. aesculi is not restricted to bark lesions but is likely to be widespread in the environment. It is also capable of causing foliar infection of several Aesculus species and could persist under extremes of weather in the UK.     

Characterization of endophytic bacteria with plant growth promotion and biological control potential isolated from walnut trees

In this study, a total of 68 endophytic bacteria were isolated from different tissues of walnut trees. About 55% and 22% of the strains had the ability to produce indole acetic acid and gibberellic acid, respectively. The capability of isolates to solubilize phosphate, growth on N‐free medium, siderophore, protease and lipase production was varied. Based on phenotypic grouping and plant growth promotion properties, twelve isolates were selected and 16S rRNA gene‐based phylogenetic analysis revealed that strains showed 99%–100% similarity to Pseudomonas, Bacillus, Arthrobacter, Roseomonas and Streptomyces genera. Amongst the selected strains, PS12, KS54, JS66 and KS74 showed root and shoot growth enhancement of poplar cutting. NS70, KS54 and PL36 strains showed antagonistic activity against Pseudomonas syringae pv. syringae;RR47, KS74 and NR69 strains had inhibition effects against Brenneria nigrifluens; and JS66 and RR26 strains had antagonistic activity against both phytopathogens under in vitro conditions. This is the first reported study to elucidate the endophytic bacterial diversity associated with walnut trees with beneficial attributes.     

Fast molecular detection of Pseudomonas syringae pv. aesculi in diseased horse chestnut trees

A molecular technique was used to detect the bacterium Pseudomonas syringae pv. aesculi in horse chestnut trees (Aesculus hippocastanum), affected by the recently recognized European ‘Pseudomonas horse chestnut bark disease’. The technique helped identify the pathogen within 6 h of sample preparation including DNA extraction, polymerase chain reaction (PCR) and electrophoresis until gel documentation. PCR primer pairs derived from the gyrase B gene sequence were used. Because of the great similarity in the gyrase B gene sequences of the numerous closely related P. syringae pathovars, the primers were not only totally specific to the pathovar aesculi, but also detected a few other pathovars. The assumption that other bacteria should not occur at least near to a necrotic lesion of a horse chestnut tree was corroborated by sequence identity of the PCR products obtained with the gyrase B gene sequence of P. syringae pv. aesculi. Koch’s postulates were fulfilled for an isolate of P. syringae pv. aesculi obtained from a diseased horse chestnut tree sampled in Hamburg in 2007.     

Pseudomonas syringae pv. aesculi associated with horse chestnut bleeding canker in Germany

Bacteria were isolated from necrotic lesions on a horse chestnut tree (Aesculus hippocastanum) with bleeding canker in Hamburg, Germany. Sequencing of the rDNA-ITS region revealed great similarity to pathovars of Pseudomonas syringae. Pseudomonas syringae pv. aesculi was identified by sequence homology of the gyrase B gene. This is the first report of P. syringae pv. aesculi in Germany. Phytophthora was not detected.     

Variation in pathogenicity among the three subspecies of Phytophthora alni on detached leaves,twigs and branches of Alnus glutinosa

Pathogenicity tests were carried out on leaves, twigs and branches of Alnus glutinosa using several isolates of Phytophthora alni ssp. alni, P. alni ssp. multiformis and P. alni ssp. uniformis in vitro. Healthy fresh leaves were collected from disease‐free areas and inoculated with mycelium on agar discs or by dipping in zoospore suspensions. In addition, twigs and branches were collected from both disease‐free and disease‐affected areas, inoculated with mycelium on agar discs and incubated at four temperatures (15, 20, 25, 30°C). All subspecies tested were pathogenic but with varied level of virulence. In inoculation tests on foliage, wounding was a key factor in causing infections: lesions on inoculated wounded leaves were larger than on non‐wounded leaves. In the twig and branch inoculation tests, no differences in virulence were observed among the P. alni subspecies in terms of sampling locations, but lesions differed in size according to incubation temperature, with the largest lesions occurring on tissues incubated at 25°C. The work is the first to report foliar necrosis caused by P. alni on A. glutinosa. P. alni ssp. uniformis was the least virulent of the subspecies in branch inoculations. These findings demonstrate that various tissues of A. glutinosa could act as sources of pathogen inoculum and may disseminate alder Phytophthora in natural ecosystems.     

Characterisation of bacteria from poplars and willows exhibiting leaf spotting and stem cankering in New Zealand

During the unusually moist 1985–86 growing season, field grown poplars and willows exhibited on the current seasons growth, terminal and side shoot blackening, dieback, leaf spots and stem cankering. Bacteria isolated from infected tissues were: Pseudomonas syringae pv. syringae, P. fluorescens, Erwinia herbicola and Xanthomonas campestris pv. populi (poplar only). Isolates of P. syringae pv. syringae alone were pathogenic to the original hosts and evoked hypersensitive responses in tobacco leaves. The remaining bacteria were non-pathogenic and accordingly were considered to be components of the epiphytic microflora.     

Susceptibility of silver birch and black alder to several Phytophthora species isolated from soils in declining broadleaf forests in western Ukraine

In declining broadleaf forests in western Ukraine, several Phytophthora species including P. plurivora, P. bilorbang, P. polonica, P. gonapodyides and P. cactorum were recovered using soil baiting assays and identified using morphological and molecular methods. Pathogenicity tests of selected isolates were performed on black alder (Alnus glutinosa (L.) Gaerth.) and silver birch (Betula pendula Roth.) to assess susceptibility of these two tree species to the newly detected Phytophthora species. Phytophthora plurivora, P. bilorbang and P. polonica showed higher pathogenicity in both alder and birch compared to the other tested Phytophthora species.     

The bacterial disease of ash (Fraxinus excelsior), caused by Pseudomonas syringae subsp. savastanoi pv. Fraxini II. Etiology and taxonomic considerations

The results of studies of various bacteria isolated from characteristic excrescences of the so-called bacterial canker of common ash are reported. A fungus (formerly known as Plenodomus rabenhorstii Preuss) associated with this disease, is described and renamed as Phoma riggenbachii Boerema et Janse. Based on biochemical, serological and pathological work the name Pseudomonas syringae subsp. savastanoi (ex Smith) subsp. nov., nom. rev. pv. fraxini is proposed for the ash bacterium. The closely related causal organisms of olive and oleander have been compared and named pv. oleae and pv. nerii.     

Investigations on Heterobasidion annosum s.lat. in central and eastern Asia with the aid of mating tests and DNA fingerprinting

To investigate the taxonomy of Heterobasidion in Eurasia, 49 specimens belonging to H. annosum sensu lato from Asia were identified with the aid of mating tests. Most of the specimens originated from north‐eastern and south‐western China and from the Altai region in southern Siberia, but a few isolates from Kirghizia, Japan and India were also tested. In addition to mating tests, the material from China was investigated with DNA fingerprinting. Heterobasidion annosum sensu stricto was identified only from the Altai region. Homokaryotic isolates from other specimens, except the Indian ones, were sexually compatible with H. parviporum, but they also showed a high degree of compatibility with H. abietinum and with the North American S group. The isolates from SW China (eastern Himalayas) mated with about equal frequency with the European strains of H. parviporum and H. abietinum. However, the DNA fingerprinting showed that these isolates were more closely related to H. parviporum, and hence they were tentatively included in this species. The North American S group was more distant from these Eurasian taxa. Four old isolates from India mated only weakly with the members of the H. annosum s.lat. According to the species concept presented, the distribution of H. parviporum extends from western Europe through southern Siberia to northern China, Japan and the eastern Himalayas. H. annosum s.str. is so far identified only from the Altai region outside Europe, and H. abietinum only from Europe.     

Occurrence of mycoplasma-like organisms in diseased and non-symptomatic alder trees (Alnus spp.)

Mycoplasma-like organisms (MLOs) were detected in almost all examined trees of Alnus glutinosa and A. incana older than approximately 5 years. The minority of trees showed symptoms of decline while the others appeared healthy. MLOs were also found in A. glutinosa var. barbata, A. hirsuta, A. rubra, A. rugosa, A. subcordata, and A. tenuifolia.     

猕猴桃体内酚含量、保护酶活性与抗溃疡病关系的初步研究

分别在2005年的夏季(6月)、秋季(10月)、冬季(1月)和2006的春季(3月),对感、抗性不同的猕猴桃品种的1年生枝条韧皮部酚含量、SOD、POD活性与抗病性表现进行了相关性研究,结果表明:夏、秋、春季酚含量与相对感病指数都存在线性相关,相关系数分别为-0.873,-0.863,-0.801,线性方程分别为Y=14.224-0.429X,Y=6.293-0.133X,Y=5.808-0.120X,且达到显著水平;而SOD、POD活性与抗病性之间均仅有一次存在相关性。说明,酚可以作为猕猴桃溃疡病抗病鉴定的指标。     

Occurrence of horse chestnut bleeding canker caused by Pseudomonas syringae pv. aesculi in the Czech Republic

Between 2008 and 2010, horse chestnut (Aesculus hippocastanum) trees growing at 216 locations in the Czech Republic were surveyed for bleeding canker disease. Typical symptoms of bleeding canker were found at 16 locations, and samples were collected from five of these locations. The bacterium Pseudomonas syringae was isolated from five locations, and Pseudomonas syringae pathovar aesculi, which is the causal agent of bleeding canker disease, was isolated at one location. This is the first report of P. syringae pv. aesculi in the Czech Republic.     

Assessment of intrapathovar variability of Xanthomonas axonopodis pv. commiphorae through phenotypic and molecular markers

Gummosis of guggal (Commiphora wightii) caused by Xanthomonas axonopodis pv. commiphorae (Xac) is one of the major reasons for drastic reduction in guggal population under natural plant stand. The pathogen spreads mainly through human activities (tapping). We isolated 43 Xac strains from 14 locations representing four different regions spread over three districts of Gujarat state, India. A polyphasic approach was followed to characterize these strains and to measure phenotypic and genetic variations among them. All strains were identical in colony morphology and were virulent on guggal. Some of the strains showed differential reactions towards certain biochemical tests viz., acid production from carbon sources. Whole cell protein profiles were apparently uniform for all strains with similarity coefficients ranging between 0.75 and 1.00. Clustering based on sodium dodecyl sulphate–polyacrylamide gel electrophoresis banding patterns showed heterogeneity among the isolates, but strains originating from same zone had similar protein profiles. Inter simple sequence repeats (ISSR)‐based and repetitive elements (rep)‐based PCR analyses revealed genetic heterogeneity among the strains. Both these methods were equally effective in deciphering variability among the strains and indicated similar types of variability with highly significant correlation (r = 0.91) on the similarity matrices. The variation matrix analysis on combined ISSR‐ and rep‐PCR data suggested 66.1% variability among Xac strains. The present study established that Xac strains from different geographical locations had profound genetic heterogeneity.     

Identification of Kenyan Armillaria isolates by cultural morphology,intersterility tests and analysis of isozyme profiles

Three groups of morphologically distinct isolates were observed among nine Kenyan Armillaria isolates based on their mycelium and rhizomorphs characteristics. Seven of the isolates were interfertile with testers of North American biological species III, VII and IX. However, tests with benomyl segregants BEN 433, BEN 157-10 and BEN AVK were intersterile with the same testers and also with the European A. mellea, A. lutea and A. ostoyae. The analysis of isozyme profiles showed that morphologically similar isolates had similar isozyme profiles of esterases. Their profiles however differed from those of the European A. mellea, A. lutea and A. ostoyae. On the basis of intersterility tests and analysis of isozyme profiles, the Kenyan isolates should be considered different.     

Susceptibility of species of Juglans to pathovars of Xanthomonas campestris

Bacterial blight is considered one of the most serious diseases affecting the genus Juglans. Artificial inoculations with Xanthomonas campestris pv. Juglandis were performed in the field by spraying seedlings of Juglans cinerea, Juglans hindsii, Juglans mandshurica, Juglans nigra, Juglans regia and Juglans sieboldiana. Juglans nigra, J. cinerea ana J. sieboldiana proved to be the most resistant. The multiplication ability of Xanthomonas campestris pv. corylina, X. c. pv. juglandis, and X. c. pv. campestris was evaluated by injecting 2-year-old seedling leaves of J. mandshurica, J. nigra and J. regia. The bacterial growth was monitored in the infected tissue over 12 days. Xanthomonas c. pv. juglandis exhibited a high growth rate and induced black greasy spots in J. mandshurica and J. regia; X. c. pv. campestris grew in leaf tissues without inducing symptoms, X. c. pv. corylina showed a very low growth rate. None of the tested bacterial strains multiplied in the leaves of J. nigra. This confirms the results obtained in the field tests. Juglans regia was the most susceptible among the walnut species tested.     

Decay and canker caused by Inonotus rickii spreading on more urban tree species

Inonotus rickii was detected for the first time causing cankers and decay in Acer negundo and Celtis australis in Italy. In a boxelder boulevard, declining trees showed sparse foliage, exudations and cracks in the bark; in some cases, chlamydospore masses were present. Five isolates were collected and compared by growth tests in vitro and electrophoretic analyses; three isolates from the same boulevard showed very similar physiological characters. The increasing importance of the pathogen in urban areas is underlined and discussed.     

Bacterial canker of Maackia amurensis var. buergeri caused by a putative Pseudomonas syringae

A new disease of Maackia amurensis var. buergeri was recently found on the northern island of Hokkaido, Japan. Affected trees were heavily damaged and had cankers on both trunks and branches. After natural infection, a series of swellings on the bark surface developed longitudinally. These swellings burst and coalesced to become long cankers. It is proposed that the disease be designated ‘bacterial canker of Maackia’. The causal pathogen was isolated and characterized tentatively as Pseudomonas syringae on the basis of laboratory tests. Pathogenicity of the bacterium was confirmed by inoculation into the host.     

Virulence and double‐stranded RNA in Sphaeropsis sapinea

Forty wildtype isolates of Sphaeropsis sapinea were grouped into the morphotypes A and B based on previously defined differences in cultural and morphological criteria as well as restriction sites for Dde I and Bst UI endonucleases in nuclear ribosomal DNA amplicons. Thirteen of 20 type A isolates and nine of 20 type B isolates contained detectable dsRNA (55%) of different molecular weight and size. dsRNA was transmitted into conidia at a frequency of 71–100%. By selecting single conidia, dsRNA‐free subcultures were obtained from six of 22 isolates containing dsRNA. Pathogenicity tests on expanding buds of landscape trees of three species of Pinus showed highly significant statistical interactions between isolate virulence, Pinus species, and year. Pine species‐year had a profound impact on virulence. The pattern in the interactions was revealed by principal component analysis of the interaction sums of squares of the anova (Additive Main Effects and Multiplicative Interaction; AMMI). Pinus sylvestris was highly interactive in its susceptibility to S. sapinea with seasonal effects. P. nigra and P. resinosa were more stable. The interactivity analysis was used to apportion interaction to specific isolates to improve the accuracy of the estimates of virulence. Estimates of the relative virulence of isolates were predicted over five different Pinus species‐years. Isolates were ranked in virulence and interactivity using the AMMI model. This model permitted mean separation tests of the relative virulence among isolates over the combined Pinus species‐years. One isolate was identified as potentially having dsRNA‐mediated hypovirulence based on the significantly greater virulence of its isogenic, dsRNA‐free subculture, as expressed over the three Pinus species and 2 years. Type A isolates containing dsRNA ranged from stable to highly interactive and from low to high in virulence. Type B isolates containing dsRNA were similar in interactivity but virulence ranged from avirulent to moderate, seldom exceeding the mean for S. sapinea. dsRNA‐free isogenic subcultures tended not to express higher virulence than their dsRNA‐containing parent strains but often changed in interactivity. Therefore, in one year a dsRNA‐free subculture might be more virulent than its dsRNA‐containing parent. In another year the dsRNA‐free subculture might be less virulent.