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1.
Strains of the pathogen causing bacterial blight of kiwifruit in New Zealand, previously identified as Pseudomonas viridiflava, were examined using phenotypic and genotypic methods. Percentage DNA–DNA reassociation values for strains of the pathogen, with the type strains representing P. viridiflava and P. savastanoi, and representative strains within P. syringae , were obtained using the S1 endonuclease method. Strains of the pathogen were most similar to the type strain of P. savastanoi. This similarity was supported by examination of the Δ T m between representative strains. It is concluded that the pathogen can be considered as a member of the P. savastanoi genomic species. The pathogen from kiwifruit in New Zealand was also differentiated in genomic terms from P. syringae pv. actinidiae . Strains of the kiwifruit pathogen compared using the Biomerieux API Biotype 100 system exhibited consistent determinative tests which distinguished the pathogen from P. viridiflava and P. syringae pv. actinidiae . The origins of the pathogen in New Zealand are discussed.  相似文献   

2.
中国猕猴桃细菌性花腐病菌的鉴定   总被引:4,自引:0,他引:4  
 从福建、湖南和湖北猕猴桃病花上分离到能引起花腐病的32个细菌菌株,经细菌学和BiologGN测试板测定,可以看出中国的猕猴桃细菌性花腐病菌与新西兰的猕猴桃花腐病菌、丁香假单胞菌丁香致病变种Pseudomonas syringae pv.syringae和绿黄假单胞菌P.viridiflava相似,与萨氏假单胞菌P.savastanoi和猕猴桃溃疡病菌P.syringae pv.actinidiae有更多的不同,但是DNA/DNA同源性测定结果却显示出中国的菌株可分为2个类型:第1个类型与新西兰猕猴桃花腐病菌和萨氏假单胞菌有很高的同源性,第2类型与绿黄假单胞菌有很高的同源性,说明中国菌株分别属于这2个种。第1类型来自于福建和湖北,第2类型来自于湖南。  相似文献   

3.
The bacterial canker of kiwifruit caused by Pseudomonas syringae pv. actinidiae is a severe threat to kiwifruit production worldwide. Many aspects of P. syringae pv. actinidiae biology and epidemiology still require in-depth investigation. The infection by and spread of P. syringae pv. actinidiae in xylem and phloem was investigated by carrying out artificial inoculation experiments with histological and dendrochronological analyses of naturally diseased plants in Italy. We found that the bacterium can infect host plants by entering natural openings and lesions. In naturally infected kiwifruit plants, P. syringae pv. actinidiae is present in the lenticels as well as in the dead phloem tissue beneath the lenticels, surrounded by a lesion in the periderm which appears to indicate the importance of lenticels to kiwifruit infection. Biofilm formation was observed outside and inside plants. In cases of advanced stages of P. syringae pv. actinidiae infection, neuroses of the phloem occur, which are followed by cambial dieback and most likely by infection of the xylem. Anatomical changes in wood such as reduced ring width, a drastic reduction in vessel size, and the presence of tyloses were observed within several infected sites. In the field, these changes occur only a year after the first leaf symptoms are observed suggesting a significant time lapse between primary and secondary symptoms. It was possible to study the temporal development of P. syringae pv. actinidiae-induced cambial dieback by applying dendrochronology methods which revealed that cambial dieback occurs only during the growing season.  相似文献   

4.
Pseudomonas syringae pv. actinidiae (Psa) is responsible for bacterial canker of kiwifruit. Biovar 3 of Psa (Psa3) has been causing widespread damage to yellow‐ and green‐fleshed kiwifruit (Actinidia spp.) cultivars in all the major kiwifruit‐producing countries in the world. In some areas, including New Zealand, P. syringae pv. actinidifoliorum (Pfm), another bacterial pathogen of kiwifruit, was initially classified as a low virulence biovar of Psa. Ability to rapidly distinguish between these pathovars is vital to the management of bacterial canker. Whole genome sequencing (WGS) data were used to develop PCR assays to specifically detect Psa3 and Pfm from field‐collected material without the need to culture bacteria. Genomic data from 36 strains of Psa, Pfm or related isolates enabled identification of areas of genomic variation suitable for primer design. The developed assays were tested on 147 non‐target bacterial species including strains likely to be found in kiwifruit orchards. A number of assays did not proceed because although they were able to discriminate between the different Psa biovars and Pfm, they also produced amplicons from other unrelated bacteria. This could have resulted in false positives from environmental samples, and demonstrates the care that is required when applying assays devised for pure cultures to field‐collected samples. The strategy described here for developing assays for distinguishing strains of closely related pathogens could be applied to other diseases with characteristics similar to Psa.  相似文献   

5.
Since 2008, bacterial canker of kiwifruit (Actinidia deliciosa and A. chinensis) caused by Pseudomonas syringae pv. actinidiae (Psa) has resulted in severe economic losses worldwide. Four biovars of Psa can be distinguished based on their biochemical, pathogenicity and molecular characteristics. Using a range of biochemical, molecular and pathogenicity assays, strains collected in France since the beginning of the outbreak in 2010 were found to be genotypically and phenotypically diverse, and to belong to biovar 3 or biovar 4. This is the first time that strains of biovar 4 have been isolated outside New Zealand or Australia. A multilocus sequence analysis based on four housekeeping genes (gapA, gltA, gyrB and rpoD) was performed on 72 strains representative of the French outbreak. All the strains fell into two phylogenetic groups: one clonal corresponding to biovar 3, and the other corresponding to biovar 4. This second phylogenetic group was polymorphic and could be divided into four lineages. A clonal genealogy performed with a coalescent approach did not reveal any common ancestor for the 72 Psa strains. Strains of biovar 4 are substantially different from those of the other biovars: they are less aggressive and cause only leaf spots whereas Psa biovars 1, 2 and 3 also cause canker and shoot die‐back. Because of these pathogenic differences, which were supported by phenotypic, genetic and phylogenetic differences, it is proposed that Psa biovar 4 be renamed Pseudomonas syringae pv. actinidifoliorum pv. nov. Strain CFBP 8039 is designated as the pathotype strain.  相似文献   

6.
Pseudomonas syringae pv. actinidiae (Psa) is a Gram‐negative bacterium that causes the bacterial canker of both green (Actinidia deliciosa) and yellow (Actinidia chinensis) fleshed kiwifruit. Since the emergence of an economically devastating Psa outbreak in Japan in the 1980s, the disease took a contagious turn causing severe economic loss to kiwifruit industries in Italy, South Korea, Spain, New Zealand and other countries. Research shows that the pathogenic strains isolated from different infected orchards vary in their virulence characteristics and have distinct genes coding for the production of different toxins. The global Psa outbreak has activated research around the world on developing efficient strategies to contain the pandemic and minimize loss to the kiwifruit industry. Chemical and biological control options, orchard management and breeding programmes are being employed in this global effort. Synergy between different disease control strategies has been recognized as important. Phytotoxicity, resistance development and regulatory measures in certain countries restrict the use of copper compounds and antibiotics, which are otherwise the mainstay chemicals against bacterial plant diseases. Therefore, because of the limitations of existing chemicals, it is important to develop novel chemical controls against Psa. Antimicrobial peptides, which are attractive alternatives to conventional antibiotics, have found promising applications in plant disease control and could contribute to expanding the chemical control tool box against Psa. This review summarizes all chemical compounds trialled so far against Psa and provides thoughts on the development of antimicrobial peptides as potential solutions for the future.  相似文献   

7.
Pseudomonas syringae pv. actinidiae (Psa) was identified as the causal agent of severe epidemics of bacterial canker on Actinidia chinensis (yellow kiwifruit) in central Italy occurring during 2008–9. A total of 101 strains were obtained from infected leaves, twigs, branches and trunks of cvs Hort16A, Jin Tao and CK3. Outbreaks were also found on A. deliciosa cv. Hayward. A representative set of 21 strains were compared with other Psa strains isolated from previous outbreaks in Japan and Italy as well as with P. s. pv. syringae strains obtained from A. chinensis and with strains of genomospecies 8. Repetitive‐sequence PCR (rep‐PCR) typing using BOX and ERIC primer sets revealed that all Psa strains obtained during 2008–9 showed the same fingerprinting profile. This profile, however, was different from those of strains previously isolated in Japan and Italy. Multilocus sequence typing (MLST) of gapA, gltA, gyrB and rpoD revealed a higher genetic variability among the strains than rep‐PCR, with some of them showing the same sequence pattern although isolated from different areas, cultivars and years. None of the recently obtained strains possessed genes coding for phaseolotoxin or coronatine, and all had an effector protein, namely hopA1, differentiating them from the strains causing past outbreaks in Japan and Italy. All isolates were inhibited in vitro by copper‐based compounds, antibiotics, geraniol, citronellol and by a chitin‐based organic compound. The recent epidemics found in central Italy on yellow kiwifruit appear to have been caused by a different Psa population than those previously recorded in Japan, South Korea and Italy.  相似文献   

8.
Journal of General Plant Pathology - Loop-mediated isothermal amplification (LAMP) was designed to rapidly detect biovars 1 and 3 of Pseudomonas syringae pv. actinidiae (Psa), a serious, global...  相似文献   

9.
猕猴桃溃疡病菌在中国的适生性分析   总被引:2,自引:0,他引:2  
通过分析猕猴桃溃疡病菌在中国的适生性,为科学制定有效的检疫监管措施,防范其入侵和扩散,确保猕猴桃产业健康发展提供理论依据。本研究根据前人研究结果,采用模糊数学综合评判的原理和方法,定量分析猕猴桃细菌性溃疡病菌(Pseudomonas syringae pv.actinidiae)在我国各个地区的适生性。猕猴桃溃疡病菌在我国最适宜的省份主要分布在四川、云南、贵州、福建、安徽、湖南、湖北、河南、江西、陕西、浙江、重庆、西藏。鉴于该病具有发生发展迅速,危害性强,防治难度大等特点,应当加强猕猴桃种苗等繁殖材料的检疫,加强对果园的管理和病害监测,积极采取有效的防治措施并加强抗病育种方面的研究。  相似文献   

10.
猕猴桃溃疡病菌的分子检测技术研究   总被引:3,自引:0,他引:3  
 猕猴桃溃疡病是猕猴桃生产上的主要病害,为建立该病的快速诊断技术,本实验通过RAPD分析获得一条1 300 bp左右的致病菌的特异片段,对该片段进行克隆测序,在测序的基础上设计并合成一对特异引物F7/R7,优化特异引物扩增条件,并验证引物的特异性和灵敏性。利用该特异引物对包括猕猴桃溃疡病菌在内的14个菌株基因组DNA进行PCR扩增表明,只有猕猴桃溃疡病菌能扩增出1条约为950 bp的特异条带,其他菌株及对照均未扩增出特异条带。对采自果园的染病枝干组织和接种致病菌的枝干组织的检测表明,该特异引物能特异性地检测到猕猴桃溃疡病菌的存在,其在组织中的检测灵敏度为100 fg/μL。因此,利用设计合成的特异引物F7/R7,参考优化的体系和程序,结合简单的试剂盒法提取猕猴桃溃疡病菌或植物组织DNA,可以在短时间内完成对该病原菌的分子检测。  相似文献   

11.
Occurrence of Pseudomonas syringae pv. actinidiae on kiwifruit in Italy   总被引:2,自引:0,他引:2  
Pseudomonas syringae pv. actinidiae has been isolated from kiwifruit plants for the first time in Italy. Biochemical tests were consistent with those characterizing the type-strain; pathogenicity tests yielded severe blights in the inoculated kiwifruit plants and no symptoms on lilac, pear and peach. Nutritional tests as well as whole-cell protein profiles revealed slight differences between the strains isolated in Japan and those of the present study. The main symptoms observed in the field are a red-rusty exudation covering the bark of twigs and trunks, blight of young canes and plants, angular leaf spots surrounded by chlorotic haloes and tiny cankers along the twigs.  相似文献   

12.
Y. J. Koh 《Plant pathology》2018,67(5):1208-1219
Bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae (Psa), is a severe threat to kiwifruit production in Korea. An existing infection risk model from New Zealand was adopted to respond to this epidemic. Disease incidence (proportion of diseased leaves on each vine) and weather (hourly temperature, rainfall and relative humidity) data required to develop the model were collected and analysed in the study. Disease incidence data were used to modify and validate the existing model. Because the Psa risk model was originally developed in a region where the characteristic climatic conditions are completely different from those in Korea, the temperature and rainfall functions of the existing model were modified. Analyses using statistical correlation and prediction–realization tables revealed that the modified model is valid with high agreement (a correlation coefficient of 0.85 and an accuracy of 85.7%, respectively) between the observed disease incidence and simulated disease risk from the model. The model was also found to be more highly sensitive to the presence or absence of rainfall than any other weather variable inputs. Uncertainty in simulated disease risk was measured based on the level of uncertainty in temperature input from weather forecasts. Overall, these results indicate that the modified Psa risk model can be used to provide practical and applicable information for timely disease control to the kiwifruit growers in Korea.  相似文献   

13.
由丁香假单胞菌猕猴桃致病变种Pseudomonas syringae pv. actinidiae (Psa)侵染引起的猕猴桃细菌性溃疡病(kiwifruit bacterial canker)是全球猕猴桃生产上最具毁灭性的细菌病害。为探明福建、安徽、四川和陕西4省Psa菌株的生物型和遗传多样性,用5对PCR特异性引物PsaJ-F/-R、PsaK-F/-R、Tac-F/-R、Con002-F/-R和avrRps4-F1/-R2检测Psa菌株的生物型;用4对PCR引物27F/1492R、PsaF1/PsaR2、gapA-Fps/Rps和rpoD+364s/-1222ps分别扩增16S rRNA、ITS、gapA和rpoD基因,进行多基因联合分析Psa菌株的遗传多样性。结果表明,特异性引物Tac-F/-R从47株Psa菌株中均能扩增出一条545 bp的特异条带,其他4对引物未扩增出任何条带,说明供试Psa菌株的生物型均为biovar 3。多基因联合分析表明,4省Psa存在丰富的遗传多样性,4个群体共检测出27个单倍型,单倍型多样性为0.955。安徽、福建、四川和陕西群体的单倍型数差异较大,分别为1、8、12个和12个。4个群体的多态性位点数、核苷酸多样性和平均核苷酸差异数差异极显著(P<0.01),其中福建群体的多态性最丰富,而安徽群体的多态性最低。AMOVA分析表明,3.6%的遗传变异来源于种群间,而96.4%的遗传变异来源于种群内,说明种群内变异是遗传变异的主要来源。遗传分化分析表明,安徽省Psa群体与其他3个群体间的遗传分化极高(Fst>0.175),福建、四川和陕西群体间的遗传分化水平较低(Fst<0.017)。研究结果有利于了解福建省Psa的来源,为阻断Psa的传播和猕猴桃细菌性溃疡病的长期可持续控制提供了理论参考。  相似文献   

14.
Since 2002, severe leaf spotting on parsley (Petroselinum crispum) has occurred in Monterey County, CA. Either of two different pathovars of Pseudomonas syringae sensu lato were isolated from diseased leaves from eight distinct outbreaks and once from the same outbreak. Fragment analysis of DNA amplified between repetitive sequence polymerase chain reaction; 16S rDNA sequence analysis; and biochemical, physiological, and host range tests identified the pathogens as Pseudomonas syringae pv. apii and P. syringae pv. coriandricola. Koch's postulates were completed for the isolates from parsley, and host range tests with parsley isolates and pathotype strains demonstrated that P. syringae pv. apii and P. syringae pv. coriandricola cause leaf spot diseases on parsley, celery, and coriander or cilantro. In a multilocus sequence typing (MLST) approach, four housekeeping gene fragments were sequenced from 10 strains isolated from parsley and 56 pathotype strains of P. syringae. Allele sequences were uploaded to the Plant-Associated Microbes Database and a phylogenetic tree was built based on concatenated sequences. Tree topology directly corresponded to P. syringae genomospecies and P. syringae pv. apii was allocated appropriately to genomospecies 3. This is the first demonstration that MLST can accurately allocate new pathogens directly to P. syringae sensu lato genomospecies. According to MLST, P. syringae pv. coriandricola is a member of genomospecies 9, P. cannabina. In a blind test, both P. syringae pv. coriandricola and P. syringae pv. apii isolates from parsley were correctly identified to pathovar. In both cases, MLST described diversity within each pathovar that was previously unknown.  相似文献   

15.
利用叠氮溴乙锭(ethidium monoazide bromide,EMA)与实时荧光定量PCR技术相结合(EMA-qPCR),建立了一种有效快速检测猕猴桃溃疡病菌活菌的方法。以猕猴桃溃疡病菌ITS序列为检测靶标,菌体经EMA渗透处理,再进行qPCR特异性扩增。结果显示,qPCR检测灵敏度为2cfu;当EMA的浓度为2.0μg/mL时,能有效抑制1.0×10~7 cfu/mL经高温灭活的死菌的扩增,对活菌的扩增没有影响。当活菌数在1.0×10~1~1.0×10~5 cfu范围内,每个qPCR反应体系中活菌数与Ct值呈线性相关(R~2=0.988)。不同温度处理活菌菌悬液后用EMA-qPCR检测猕猴桃溃疡病菌的存活情况并与平板计数法进行比较,结果表明待检样品可在4℃和20℃短期保存。对疑似带病猕猴桃材料进行EMA-qPCR检测,结果表明能减少猕猴桃溃疡病菌PCR的假阳性结果。本研究建立的EMAqPCR方法是一种有效检测猕猴桃溃疡病菌活菌的方法,能有效避免PCR检测实际样品可能造成的假阳性结果。  相似文献   

16.
A virulent strain of Pseudomonas syringae pv. actinidiae biovar 3 (Psa), which causes bacterial canker in kiwifruit, was first recorded in New Zealand in November 2010. This strain has severely affected Actinidia chinensis var. chinensis ‘Hort16A’ kiwifruit productivity but its effect on green Actinidia chinensis var. deliciosa ‘Hayward’ kiwifruit productivity has been variable. An observational study design was used to develop explanatory models to quantify the impacts of Psa infection on productivity (tray equivalents per hectare) of Hayward kiwifruit harvested in 2012, using data captured by industry from 2599 orchards. A total of 934 orchards were Psa positive at the end of the study period. Multivariable linear regression was used to model 2012 productivity in the presence of Psa, while controlling for regional differences, elevation, 2011 productivity, harvest dates and application of agrichemicals. The model showed productivity was initially higher in the presence of Psa, and was not reduced until after 1 year of infection. The relationship between protective spray use and productivity was also quantified. It is likely that improved disease management has offset the impact of the disease and future research should consider a reassessment of the effects of disease after longer term exposure to Psa in New Zealand. The use of an observational cohort study to assess disease impacts using multivariable analysis could have wider application in the field of plant epidemiology.  相似文献   

17.
Knowing the population structure of a pathogen is fundamental for developing reliable phytosanitary legislation, detection techniques, and control strategies based on the actual aggressiveness and distribution of the pathogen. Currently, four populations of Pseudomonas syringae pv. actinidiae (Psa) have been described: Psa 1, Psa 2, Psa 3 and Psa 4. However, diagnostic assays specific for Psa populations do not detect Psa 4, the less virulent (LV) strains isolated in New Zealand. Similarly, multilocus sequence typing (MLST) of housekeeping genes, or broad Psa strain genome comparisons, revealed that Psa 4‐LV strains clustered separately from other Psa populations. In order to examine whether the placement of Psa 4 in the pathovar actinidiae was appropriate, various tests were carried out. It was shown that the Psa 4‐LV strains induced leaf and shoot wilting in Prunus cerasus, extensive necrotic lesions in Capsicum annuum fruits, and no significant symptoms in Actinidia deliciosa. Moreover, repetitive‐sequence PCR fingerprinting, type III secretion system effector protein genes detection and colony morphology clearly indicated the distinctiveness of Psa 4‐LV strains from the other three Psa populations. Rep‐PCR molecular typing revealed a high similarity of the Psa 4‐LV strains with members of Pseudomonas avellanae species. The Psa 4‐LV strains, most probably, belong to a new, still unnamed pathovar. It was concluded that the Psa 4‐LV strains isolated in New Zealand do not belong to the pathovar actinidiae, and, consequently, three Psa populations pathogenic to Actinidia spp. should currently include Psa 1, Psa 2 and Psa 3.  相似文献   

18.
 利用MT选择性培养基从加拿大进境油菜籽样品中分离到2株细菌分离物2305-1和5309-1,对分离物进行致病性测定、LOPAT测试、Biolog测试、hrpZ和cfl基因序列分析,以及多位点序列分析。结果表明:2株分离物人工接种油菜、花椰菜和番茄幼苗都能引起典型黑斑症状;LOPAT测试和Biolog测试结果与丁香假单胞菌(Pseudomonas syringae)的各项生理指标一致;hrpZ基因序列与十字花科黑斑病菌(P. syringae pv. maculicola)和番茄细菌性叶斑病菌(P. syringae pv.tomato)的序列相似性均为99.18%~100%;cfl基因序列分析表明分离物2305-1和5309-1基因组中存在冠毒素合成基因;选择gyrBropDgltAgap1、 acnBpgi 6个看家基因进行多位点序列分析,系统发育树显示分离物2305-1和5309-1均与P. syringae pv. ma-culicola聚在一起。根据试验结果将分离物2305-1和5309-1鉴定为十字花科黑斑病菌P. syringae pv. maculicola。  相似文献   

19.
广东南瓜细菌性叶枯病及其病原鉴定   总被引:1,自引:0,他引:1  
 在广东省雷州市发生一种南瓜(Cucurbita moschata)叶枯病,病株叶片边缘开始出现水渍状病斑,逐步发展成大病斑,后期病斑焦枯;在叶片上也可形成近圆形水渍状病斑,伴有黄色晕圈,后期病斑联合形成不规则大枯斑;叶柄和匍匐茎被侵染后呈水渍状腐烂。从病斑上分离到一种细菌,在KB培养基上,菌落为椭圆形,乳白色,半透明,边缘参差不齐,紫外灯照射下产生荧光反应。致病性测定结果表明,该病原细菌可侵染6个南瓜品种引起与田间症状相同的叶枯病。生理生化试验结果表明,该病原细菌与丁香假单胞丁香致病变种(Pseudomonas syringae pv. syringae)的特性一致。应用假单胞菌属特异引物Ps-for/Ps-rev和丁香假单胞丁香致病变种组群特异性引物Group III-F/Group III-R,可从该病原细菌中扩增出预期大小分别为1 018 bp和750 bp的目的片段。应用丁香致病变种syrB基因特异性引物B1/B2,可从该病原菌中扩增出预期大小为750 bp的丁香霉素基因片段。基于16S rDNA与gyrB基因序列系统进化分析均表明,南瓜叶枯病菌株与已报道的P. syringae pv. syringae菌株HS191(CP006256)亲缘关系最近,二者聚类在一起形成一个小分支。人工接种条件下,该病原细菌还可侵染西葫芦、丝瓜、茄子、番茄、菜豆、扁豆等植物。这些结果表明,引起广东省南瓜叶枯病的病原为丁香假单胞丁香致病变种(Pseudomonas syringae pv. syringae)。这是首次在中国发现丁香假单胞丁香致病变种引起南瓜叶枯病。  相似文献   

20.
Pseudomonas syringae pv. maculicola causes bacterial leaf spot on cruciferous plants in Australia. This is the first record confirming the identity of seven isolates currently stored as P. syringae pv. maculicola in the herbarium of the Department of Agriculture, Orange, Australia (Herb. DAR). The isolates were identified using pathogenicity testing on cauliflower and fatty acid methyl ester analysis. They clustered together using repetitive sequence-based polymerase chain reaction (rep-PCR) and PCR-restriction fragment-length polymorphism (RFLP) of the 16S−23S internal transcribed spacer region (ITS) using seven restriction enzymes. Identification was confirmed by comparison of these isolates with known overseas isolates of P. syringae pv. maculicola , but there was significant variation in their pathogenicity and genetic structure.  相似文献   

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