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1.
利用MT选择性培养基从进境加拿大豌豆样品上分离到一株细菌分离物(编号1314),对该分离物进行PCR检测、16S和23S rRNA序列扩增、多位点序列分析、Biolog测定、烟草过敏性反应和致病性测试。豌豆细菌性疫病菌(Pseudomonas syringae pv. pisi, Ppi)特异引物AN7F/AN7R扩增分离物1314和Ppi菌株ATCC 11043得到预期272 bp的条带,二者的PCR产物序列一致,与GenBank中Ppi (X97405)的序列相似性为99.57%。分离物1314的部分16S rRNA、23S rRNA序列以及16S-23S序列均与Ppi菌株ATCC 11043一致。选择gap1、gltA、gyrB和ropD 4个看家基因进行多位点序列分析,系统发育树显示分离物1314与Ppi菌株聚在同一组内。Biolog鉴定结果表明:分离物1314为Ppi,PROB值为0.898,SIM为0.72。该分离物人工接种豌豆茎秆能引起水渍状症斑,接种烟草叶片产生过敏性反应。基于上述试验结果,将分离物1314鉴定为豌豆细菌性疫病菌。 相似文献
2.
利用MT选择性培养基从来自日本的菜豆种子样品上分离到1株细菌分离物（编号1160）,对该分离物进行培养性状观察、特异引物PCR检测、多位点序列分析、烟草过敏性反应和致病性测试。结果表明：该分离物菌落在MT上呈白色、略扁平、无水解圈,经菜豆晕疫病菌（Pseudomonas savastanoi pv. phaseolicola,Psp）特异引物PHA19/PHA95及P5.1/P3.1扩增,获到437 bp及500 bp目的条带,并与Psp菌株LMG2245及GenBank中Psp（基因登录号：AB237164、CP000058）的序列相似性均为100%。选择gap1、gltA、gyrB、rpoD 4个看家基因进行多位点序列分析（MLSA）,系统发育树显示分离物1160与Psp菌株LMG2245、1448A聚在同一分支。该分离物人工接种菜豆叶片能引起典型的黄色晕疫病害症状,接种菜豆荚果引起水渍及褐变病害症状;接种烟草叶片可产生过敏性反应。根据上述试验结果,将分离物1160鉴定为菜豆晕疫病菌。这也是全国首次从进境商品中截获菜豆晕疫病菌。 相似文献
3.
《植物病理学报》2016,(1)
γ-变形菌纲(γ-Proteobacteria)的黄单胞菌属(Xanthomonas)的大多数种类可引起植物病害,多数是我国检疫对象。与其他革兰氏阴性植物病原细菌一样,植物病原黄单胞菌可通过高度保守的Ⅲ型分泌系统(type-Ⅲsecretion system,T3SS)分泌效应蛋白(T3SS-secreted effectors,T3SEs)进入植物细胞,在非寄主植物和抗病寄主植物上产生过敏反应(hypersensitive response,HR)以及在感病寄主植物上具有致病性。尚不清楚哪些种类的黄单胞菌具有T3SS和缺少哪些T3SE是否可作为检疫的依据。搜集7种检疫性植物病原黄单胞菌,通过PCR和Southern杂交试验结果发现:香蕉细菌性青枯病菌(X.campestris pv.musacearum)的ICMP287和ATCC49084菌株、甘蔗流胶病菌(X.axonopodis pv.vasculorum)ATCC13901菌株、洋葱细菌性叶枯病菌(X.axonopodis pv.allii)的LMG576和LMG578菌株中不含有tale基因,并且ATCC13901菌株既不含有T3SS基因也不含有hpal和xopQ基因;菜豆细菌性疫病菌(X.campestris pv.phaseoli)ATCC49119菌株不含有hpal基因。相应地,推测含有2~12个tale基因的黄单胞菌有:大豆斑疹病菌(X.axonopodis pv.glycines)ICMP5732和ATCC43911菌株、豌豆细菌性疫病菌(X.axonopodis pv.vignicola)ATCC11648菌株、棉花细菌性角斑病菌(X.campestris pv.malvacearum)ATCC12131和(X.campestris pv.phaseoli)ATCC49119菌株。大豆细菌性斑疹病菌ATCC43911菌株尽管含有hpal、xopQ和hrcC基因,但在非寄主烟草上不能激发HR反应;而甘蔗流胶病菌ATCC13901菌株不含有hpal、xopQ和hrcC基因,却激发烟草产生HR反应。这些结果对于分析比较不同植物病原黄单胞菌的致病性因子和设计特定的植物检疫靶点提供了科学线索。 相似文献
4.
进境柠檬样品上柑桔溃疡病菌的检测 总被引：1，自引：0，他引：1
从进境柠檬样品上分离到一株疑似柑桔溃疡病菌的分离物X206,对分离物进行了菌落形态学特征观察、16S r RNA序列测定、PCR检测、Biolog测定及致病性测试。试验结果表明分离物X206在NA培养基上形成黄色,有光泽,圆形,全缘,微隆起,粘稠状的菌落。特异引物Xac01/Xac02扩增分离物X206的DNA得到582bp的预期产物,产物序列与柑桔溃疡病菌序列的相似性为100%。其16S r RNA序列与柑桔溃疡病菌的序列完全一致。Biolog测定将分离物X206和柑桔溃疡病菌阳性菌株FZ01均鉴定为Xanthomonas campestris pv.dieffenbachiae。致病性测试显示分离物X206能导致柑桔叶片产生明显的溃疡病斑。根据试验结果将分离物X206鉴定为柑桔溃疡病菌(Xanthomonas axonopodis pv.citri)。 相似文献
5.
7种检疫性植物病原黄单胞菌III型分泌系统和效应蛋白编码基因的差异性分析 总被引：1，自引：0，他引：1
γ-变形菌纲(γ-Proteobacteria)的黄单胞菌属(Xanthomonas)的大多数种类可引起植物病害,多数是我国检疫对象。与其他革兰氏阴性植物病原细菌一样,植物病原黄单胞菌可通过高度保守的III型分泌系统(type-III secretion system, T3SS)分泌效应蛋白(T3SS-secreted effectors, T3SEs)进入植物细胞,在非寄主植物和抗病寄主植物上产生过敏反应(hypersensitive response, HR)以及在感病寄主植物上具有致病性。尚不清楚哪些种类的黄单胞菌具有T3SS和缺少哪些T3SE是否可作为检疫的依据。搜集7种检疫性植物病原黄单胞菌,通过PCR和Southern杂交试验结果发现:香蕉细菌性青枯病菌(X. campestris pv. musacearum)的ICMP287和ATCC49084菌株、甘蔗流胶病菌(X. axonopodis pv. vasculorum)ATCC13901菌株、洋葱细菌性叶枯病菌(X. axonopodis pv. allii)的LMG576和LMG578菌株中不含有tale基因,并且ATCC13901菌株既不含有T3SS基因也不含有hpa1和xopQ基因;菜豆细菌性疫病菌(X. campestris pv. phaseoli)ATCC49119菌株不含有hpa1基因。相应地,推测含有2~12个tale基因的黄单胞菌有:大豆斑疹病菌(X. axonopodis pv. glycines)ICMP5732和ATCC43911菌株、豌豆细菌性疫病菌(X. axonopodis pv. vignicola)ATCC11648菌株、棉花细菌性角斑病菌(X. campestris pv. malvacearum)ATCC12131和(X. campestris pv. phaseoli)ATCC49119菌株。大豆细菌性斑疹病菌ATCC43911菌株尽管含有hpa1、xopQ和hrcC基因,但在非寄主烟草上不能激发HR反应;而甘蔗流胶病菌ATCC13901菌株不含有hpa1、xopQ和hrcC基因,却激发烟草产生HR反应。这些结果对于分析比较不同植物病原黄单胞菌的致病性因子和设计特定的植物检疫靶点提供了科学线索。 相似文献
6.
7.
对经甘肃口岸进境的30批菜豆Phaseolus vulgaris种子进行了普通细菌性疫病菌的检测,利用选择性培养基MT从波兰进境菜豆种子上分离到1株细菌597,对该分离物进行菌落形态特征观察、致病性测定、16S rDNA及16S-23S rDNA ITS序列分析和特异性PCR检测。结果表明,该分离物在MT培养基上菌落呈黄色、圆形、黏稠、表面光滑向外隆起、菌落周围有水解圈。分离物597接种菜豆幼苗后导致叶片枯萎,接种点干枯。结合菌落形态、16S-23S rDNA ITS序列、特异性PCR检测结果,将分离物597鉴定为地毯草黄单胞杆菌菜豆致病变种Xanthomonas axonopodis pv.phaseoli。 相似文献
8.
《中国植保导刊》2021,(3)
在口岸送检的鲜食马铃薯上分离到一株细菌分离物0297,对该分离物在薯块上的为害症状和菌落形态特征进行观察,并经特异引物PCR检测、多位点序列分析、致病性测定。结果表明:患病薯块切面维管束变褐,按压分泌乳白色菌脓,分离物菌落在NA培养基上呈有光泽的乳白色;青枯病菌特异引物RS32/RS37扩增分离物0297得到预期583 bp的条带,与GenBank中青枯病菌(JQ323519)的序列相似性为100%;选择gyrB、gapA、adk和gdhA 4个看家基因进行多位点序列分析,系统发育树显示分离物0297与青枯病菌株Ralstonia solanacearum Phylotype II race3(UW448、BR113、UW504、UW224、BS025、UW551)聚集在同一分支上;该分离物人工接种马铃薯与番茄茎能引起茎叶褐变、萎蔫等典型病害症状。根据上述试验结果,将分离物0297鉴定为青枯病菌(Ralstonia solanacearum Phylotype II race3)。 相似文献
9.
菜豆普通细菌性疫病是黑龙江省近年来发生面积较广的菜豆病害之一,严重时可导致菜豆大面积减产,并影响籽粒品质。为发掘菜豆普通细菌性疫病的生防菌资源,本研究从健康菜豆根际土壤中分离筛选到一株对菜豆普通细菌性疫病菌具有显著抑制效果的拮抗菌株B2。经菌落形态观察、生理生化测试、16SrDNA和gyr B基因序列分析,将其鉴定为贝莱斯芽胞杆菌Bacillusvelezensis,该菌株在平板试验中对菜豆普通细菌性疫病病原菌Xanthomonas axonopodis pv. phaseoli(Xap)和Xanthomonas fuscans subsp.fuscans(Xff)的抑菌圈直径分别为19.3和21.8mm。在室内盆栽试验中,菌株B2对Xap和Xff的防效分别为55.42%和67.77%。通过菜豆叶片防御酶活性及丙二醛含量测定显示,菌株B2能显著提高菜豆叶片中SOD、POD、CAT、PPO酶活性,降低丙二醛含量,表明菌株B2能诱导植物防御反应起到抵抗病原菌的作用。综上,菌株B2在由Xap和Xff两种病原菌引起的菜豆普通细菌性疫病生物防治中具有良好的开发应用潜力。 相似文献
10.
大豆灰斑病菌毒素生物活性分析 总被引：11，自引：1，他引：10
大豆灰斑病菌[Cercosporium sojina(Hara) Liu&Guo]可以产生有毒代谢物。滤液透析及热稳定性测试结果表明:该毒素可以穿过半透膜,为较小分子化合物并具有很强的热稳定性。该菌培养物浸提液经浓缩,硅胶柱层析可得粗毒素。生物测定结果表明:该毒素对大豆幼苗及叶片具有致萎作用。针刺叶片可产生类似真菌感染的病斑。浸渍处理后,可使叶组织失绿、萎蔫最后坏死,同时毒素对抗性不同的品种有鉴别作用。 相似文献
11.
引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种 总被引：3，自引：0，他引：3
1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。 相似文献
12.
利用BIOLOG鉴定系统快速鉴定菜豆萎蔫病菌的研究 总被引：3，自引：1，他引：3
本研究利用美国Biolog公司生产的MicroStationTM V3.5系统对我国一类危险性病害菜豆萎蔫病菌及其相关菌进行了快速鉴定研究。研究结果表明,来自不同国家和寄主的24株菜豆萎蔫病菌及其相关致病变种,23株准确鉴定至种水平,其中13株鉴定至致病变种水平,种水平的鉴定准确率为95.8%,另外1株至属水平。同时2株苜蓿萎蔫病菌和2株番茄溃疡病菌均鉴定至致病变种水平。经聚类分析研究,结果支持了对格兰氏阳性植病细菌在属、种水平的分类。本研究是首次使用该系统对格兰氏阳性植病细菌进行鉴定研究,试验证明Biolog鉴定系统用于快速鉴定菜豆萎蔫病菌是一个很有用的工具,且由于其标准化程度高、快速准确,符合我国口岸植物检疫的要求。 相似文献
13.
落葵上发现短小杆菌属(Curtobacterium)一个新的致病变种 总被引：4，自引：1，他引：3
1994年在江苏省南京及镇江地区新发现了落葵细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析和DNAG+C mol%测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛,亚极生或侧生,结合其生理生化特性、细胞化学成分和DNAG+C mol%测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur.flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下一个新的致病变种,定名为Curtobacterium flaccumfaciens pv.basellae pv.nov.(萎蔫短小杆菌落葵致病变种)。 相似文献
14.
采用ERIC-PCR,BOX-PCR和ITS的分析方法,对分离自我国内蒙古自治区5个市的21个糖甜菜叶斑病菌菌株进行多样性分析,并与其他12种病原细菌进行比较。ERIC-PCR揭示,在相似性80%上所有参试菌株分为26簇,而BOX-PCR只得到20个簇,暗示这两种短重复序列在基因组中的分布不同;将两者电泳图谱结合,得到介于上述两者间的结果,分为23个簇;在相似率达87%时,ITS分析将21个糖甜菜叶斑病菌菌株分成7簇。3种分析方法相互验证,均说明内蒙古糖甜菜叶斑病菌基因组存在显著多样性。ERIC和BOX聚类证明了糖甜菜叶斑病菌与短小杆菌属(Curtobacterium)亲缘关系较近,与其他属细菌亲缘关系较远。研究证明,ERIC和BOX扩增基因组DNA指纹比ITS图谱具有更强的多样性。 相似文献
15.
《EPPO Bulletin》2011,41(3):320-328
Specific scope
This standard describes a diagnostic protocol for Curtobacterium flaccumfaciens pv. flaccumfaciens. 1 1 Use of names of chemicals or equipment in these EPPO Standards implies no approval of them to the exclusion of others that may also be suitable.Specific approval and amendment
Approved in 2011‐09.16.
Daniele M. Nascimento Letícia R. Oliveira Luana L. Melo João C. Silva José M. Soman Karine T. Girotto Renan P. Eburneo Marcos R. Ribeiro-Junior Maria M. P. Sartori Tadeu A. F. Silva Júnior Antonio C. Maringoni 《Plant pathology》2020,69(7):1357-1367
Weeds are important alternative hosts of pathogens, responsible for the survival and spread of phytopathogenic bacteria. Our study evaluated the potential of weeds as hosts of Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), causal agent of bacterial wilt, one of the main diseases of common beans. Cff survival was evaluated in the phyllosphere and in the rhizosphere of 21 weeds, in four experiments under field conditions, during the years 2018 and 2019. The aerial part of the plant was inoculated by spraying bacterial suspension (107 cfu/ml) of Cff, while the soil of the growing pots was infested with the same suspension. Cff survival was evaluated every 7 days, for 70 days. The identity of the bacterium was confirmed by PCR with the specific primers CffFOR2 and CffREV4, from strains recovered from all samples. Principal component analysis (PCA) showed that high temperatures and rainfall reduced Cff survival in the phyllosphere, while high temperatures reduced the survival of the bacterium in the rhizosphere. Our results demonstrated that Amaranthus viridis (family Amaranthaceae), Conyza bonariensis, Emilia fosbergii, Galinsoga parviflora, Gnaphalium purpureum (Asteraceae), Raphanus sativus, Lepidium virginicum (Brassicaceae), Commelina benghalensis (Commelinaceae), Ipomoea triloba (Convolvulaceae), Cyperus rotundus (Cyperaceae), Senna obtusifolia (Fabaceae), Digitaria insularis (Poaceae), Nicandra physalodes, and Solanum americanum (Solanaceae) are potential hosts for Cff. Their eradication in common bean fields is recommended, especially in fields with a history of bacterial wilt occurrence. 相似文献
17.
Ricardo M. Gonçalves Carlos A. Schipanski Lincom Koguishi José M. Soman Renate K. Sakate Tadeu A. F. Silva Júnior Antonio C. Maringoni 《European journal of plant pathology / European Foundation for Plant Pathology》2017,148(2):357-365
Alternative hosts are an important way of phytopathogenic bacteria survival between crop seasons, constituting a source of inoculum for the following crops. Bacterial wilt, caused by Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), is one of the most important diseases for common bean, and little information is available about the host range of the bacterium. In this study, we assessed possible alternative hosts for Cff, especially those cultivated during winter, in rotation systems with common bean. Plants of barley, black oat, canola, radish, ryegrass, wheat and white oat, were assessed under field and greenhouse conditions. Cff colonized epiphytically all plant species and endophytically black oat, ryegrass, wheat and white oat plants assessed in the greenhouse assays. Under field conditions, Cff colonized all plant species by except radish. All bacterial strains re-isolated from the plants were pathogenic to common bean and identified as Cff by PCR with specific primers. Based on our results, the cultivation of bean crop in succession with barley, black oat, canola, ryegrass, wheat and white oat should not be recommended, mainly in areas with a history of bacterial wilt occurrence. In these cases, the better option for crop rotation during the winter is radish, a non-alternative host for Cff. 相似文献
18.
Mixtures of plant growth-promoting rhizobacteria enhance biological control of multiple cucumber pathogens 总被引：1，自引：0，他引：1
ABSTRACT Plant growth-promoting rhizobacteria (PGPR) strains INR7 (Bacillus pumilus), GB03 (Bacillus subtilis), and ME1 (Curtobacterium flaccumfaciens) were tested singly and in combinations for biological control against multiple cucumber pathogens. Investigations under greenhouse conditions were conducted with three cucumber pathogens-Colletotrichum orbiculare (causing anthracnose), Pseudomonas syringae pv. lachrymans (causing angular leaf spot), and Erwinia tracheiphila(causing cucurbit wilt disease)-inoculated singly and in all possible combinations. There was a general trend across all experiments toward greater suppression and enhanced consistency against multiple cucumber pathogens using strain mixtures. The same three PGPR strains were evaluated as seed treatments in two field trials over two seasons, and two strains, IN26 (Burkholderia gladioli) and INR7 also were tested as foliar sprays in one of the trials. In the field trials, the efficacy of induced systemic resistance activity was determined against introduced cucumber pathogens naturally spread within plots through placement of infected plants into the field to provide the pathogen inoculum. PGPR-mediated disease suppression was observed against angular leaf spot in 1996 and against a mixed infection of angular leaf spot and anthracnose in 1997. The three-way mixture of PGPR strains (INR7 plus ME1 plus GB03) as a seed treatment showed intensive plant growth promotion and disease reduction to a level statistically equivalent to the synthetic elicitor Actigard applied as a spray. 相似文献
19.
Isolates of Pseudomonas syringae pv. phaseolicola from Africa and other bean growing areas were categorized into nine races on the basis of their reactions to eight differential cultivars following artificial inoculation. Eight hundred and ninety-three isolates representing 303 disease occurrences were initially identified as P.s. pv. phaseolicola by their pathogenicity to bean, cultural and serological characteristics and phage sensitivity. These tests also served to distinguish P.s. pv. phaseolicola from the closely related pathovars P.s . pv. glycinea and P.s. pv. syringae . Detailed race determinations were carried out on 175 selected isolates of p.s. pv. phaseolicola representative of the different geographical regions and hosts in which the pathogen was found and nine races were identified. A number of races (1,2,5,6 and 7) were distributed worldwide with race 6 predominant. Other races were found mainly in Africa; races 3 and 4 in East/Central Africa and races 8 and 9 in Southern Africa. Most isolates were obtained from the major host, Phaseolus vulgaris . Alternative natural hosts included 10 legume species representative of seven different genera ( Cajanus cajan, Desmodium sp., Lablab purpureus, Macroptilium atropurpureum, Neonotonia wightii, Phaseolus acutifolius, P. coccineus, P. lunatus, Vigna angularis and V. radiata ). Of these, Desmodium sp. constitutes a new host record. 相似文献