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1.
2015年,在广东省广州市番茄种植区发生番茄髓部坏死病,从番茄病样中分离到一种细菌。随机选取5株细菌进行致病性测定,结果显示,这5株细菌均可侵染番茄植株产生典型的髓部坏死症状,与田间病株的症状相同。5株细菌的16S rDNA序列间同源性为99.9%~100%,且与菊苣假单胞菌(Pseudomonas cichori)MAFF211996、TIs01和IP1-05菌株的16S rDNA序列同源性最高,为99%。利用菊苣假单胞菌hrcR基因的特异引物Hrp1a/Hrp2a进行PCR,从这5株细菌的DNA中均能扩增出大小约897 bp的特异片段;这些片段序列间同源性为99%~99.9%,且与P.cichori 83-1菌株hrcR基因的序列同源性为99.9%。这些结果表明,引起广东番茄髓部坏死病的病原为菊苣假单胞菌(P.cichorii)。生理生化试验显示,该病原菌与文献报道的菊苣假单胞菌的生理生化特征均相吻合,进一步证实引起广东番茄髓部坏死病的病原为菊苣假单胞菌。 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.
马铃薯内生细菌的分离及环腐病拮抗菌的筛选鉴定 总被引：27，自引：1，他引：26
本研究从大同、太原和内蒙古等地采集马铃薯块茎,分离到240株内生细菌,通过离体抑菌作用测定,共得到55株对环腐病菌有拮抗作用的菌株,占菌株总数的22.9%,抑菌圈半径最大的可达13 mm。按抑菌圈半径大小将拮抗菌分为强、中、弱三类。从中筛选出9个对环腐病等病菌具有较强拮抗作用的内生菌株进行了细菌学鉴定,结果表明:118为荧光假单胞生物型V (Pseudomonas fluorescens biovar V);110为短小芽孢杆菌(Bacillus pumilus);085为嗜热脂肪芽孢杆菌(Bacillus stearothermophilus);069为草生欧文氏菌(Erwinia herbicola);043为草莓黄单胞菌(Xanthomonas fragariae);A-10'、T3为芽孢杆菌属(Bacillus);H1-6为荧光假单胞菌(Pseudomonas fluorescens);116为短小杆菌属(Curtobacterium)。 相似文献
4.
2013年12月,甘肃省白银市水川镇日光温室中的西葫芦发生了严重的根腐和茎基腐,部分棚室病株率达50%,从病根和病茎上分离得到拟漆斑菌属真菌3株,病株分出率27.3%。采用胚根和茎基部接种法测定了菌株FG-62对西葫芦的致病性:茎基部接种后27 d,植株开始出现凋萎;接种后40 d,两种接种法的西葫芦苗均呈现严重根腐和茎基腐症状,茎基部接种的西葫芦凋萎株率达30%;从病根和病茎上均可再分离出原接种菌。菌株FG-62在PDA平板上25℃培养14 d,产生大量墨绿色至黑色分生孢子座,分生孢子无色至淡榄黑色,单胞,杆状或腰鼓状,两端钝圆,大小为(7.04~9.15)μm ×(1.97~2.46)μm,聚集的分生孢子呈黑色。BLASTn分析结果显示,菌株FG-62(GenBank 登录号 MK252098)的rDNA-ITS序列与露湿拟漆斑菌Paramyrothecium roridum分离物E-469 (GenBank 登录号KY582183.1)和CBS 357.89(源自模式材料,GenBank 登录号NR_145077.1)的序列相似性分别达99.65%和96.83%。依据病原菌形态学和rDNA-ITS序列,将其鉴定为露湿拟漆斑菌P. roridum (Basionym:Myrothecium roridum)。这是露湿拟漆斑菌引起西葫芦根腐和茎基腐的首次报道。 相似文献
5.
烟草疫霉拮抗菌株P 72 10的鉴定及其拮抗代谢 产物 初步分析 总被引：1，自引：0，他引：1
为探讨烟草根际生防细菌的生防机制,从重庆地区连作烟田健康烟株根际土壤中分离筛选到1株对烟草疫霉具有较强拮抗作用和对黑胫病具有良好防效的细菌菌株P-72-10。根据培养性状、形态特征、生理生化特性、基因组DNA的(G+C)mol%含量测定以及16S rDNA基因序列分析确定该菌株的分类地位。该菌株菌落乳白色,能产生水溶性荧光色素,革兰氏染色反应阴性,菌体杆状、大小(8.1~16.2)μm×(1.8~4.8)μm,单端生鞭毛,不形成芽孢。The BIOLOG GN2 结果显示该菌株属于假单胞菌属Pseudomonas。该菌株基因组DNA的(G+C)mol%含量为 60.72 mol%。16S rDNA基因序列分析显示该菌株与假单胞菌属荧光假单胞杆菌多个菌株的序列同源性达到99%,GenBank上的登录号为：HQ888871。结合其形态特征和生理生化特性,将菌株P-72-10鉴定为荧光假单胞杆菌P. fluorescens。平板检测拮抗代谢产物结果表明：该菌株具有分解纤维素、蛋白质和结合Fe 3+的能力,但不能分解几丁质。 相似文献
6.
甘肃省定西市当归“水烂病”病原鉴定及致病性测定 总被引：1，自引：0，他引：1
水烂病是甘肃省当归上发生的一种病害.本试验对水烂病病原菌的形态特征、培养特性、生理生化指标进行了测定,同时结合16SrDNA序列测定和相似性比对,发现该病原菌与GenBank数据库已知荧光假单胞杆菌第2生物型(Pseudornonas fluorescens生物型Ⅱ)的序列相似性达99％,其片段大小为1445 bp,编号为ZB1.致病性测定结果表明,该病原菌对‘岷归2号’(新繁品种)致病力最强,‘岷归1号’(主栽品种)次之,对‘朝鲜当归’(引进品种)的致病力最弱.本研究首次报道了荧光假单胞杆菌第2生物型能引起当归水烂病. 相似文献
7.
拮抗细菌FD6分离自福建闽侯青口青菜根围土壤,采用凹玻片法和离体叶片接种法测定菌株FD6的抑菌能力,经16S rDNA序列比对和相关生理生化性状分析,对生防细菌FD6进行了鉴定,并通过PCR扩增、薄层层析、薄层色谱生物自显影等探讨FD6产生抗生素的种类及其抑菌效果。结果表明,FD6的细菌悬浮液可显著抑制灰霉病菌分生孢子萌发,抑制率达99%,FD6培养滤液的抑制率仅为31%;细菌悬浮液对番茄灰霉病防效达59.7%。FD6的16S rDNA序列与假单胞菌属Pseudomonas的相似性达99%,系统进化树显示与荧光假单胞菌P.fluorescens遗传距离最近,结合生理生化表型特征将FD6菌株鉴定为荧光假单胞菌P.fluorescens。菌株FD6可产生硝吡咯菌素、2,4-二乙酰基间苯三酚、藤黄绿脓菌素、嗜铁素、氢氰酸和蛋白酶等抗菌物质,不产生吩嗪-1-羧酸,其中,硝吡咯菌素可直接抑制番茄灰霉病菌孢子萌发和菌丝的生长。 相似文献
8.
云南灯盏花根腐病的2种新病原鉴定及防治药剂的室内筛选 总被引：1，自引：0，他引：1
引起云南省灯盏花根腐病的病原为茄病镰刀菌(Fusarium solani)、尖孢镰刀菌(F.oxysporum)和半裸镰刀菌(F.semitectum)3种真菌,茄病镰刀菌和尖孢镰刀菌为灯盏花根腐病的2种新病原,其中茄病镰刀菌为主要致病菌,其分离率为42.86%,茄病镰刀菌有伤接种发病率为70%,无伤接种发病率为56.7%。采用室内生长速率法测定了50%多菌灵可湿性粉剂、70%甲基硫菌灵可湿性粉剂、64%恶霜·锰锌可湿性粉剂、10%苯醚甲环唑水分散粒剂、10亿活芽胞/g 枯草芽胞杆菌·荧光假单胞杆菌可湿性粉剂5种供试杀菌剂对灯盏花根腐病主要病原茄病镰刀菌的毒力。结果表明：5种供试药剂对茄病镰刀菌都有抑制作用。对茄病镰刀菌的有效中浓度（EC50）分别为0.000 4、0.050 7、0.053 5、0.081 3和8.624 0 mg/mL,其中50%多菌灵可湿性粉剂有效中浓度最低,相对抑制效果最好。5种药剂的毒力相关系数均在0.89以上,表明药剂质量浓度与抑制作用呈较高相关性。 相似文献
9.
边缘假单胞菌的分离鉴定及其特性 总被引：1，自引：0，他引：1
边缘假单胞菌Pseudomonas marginalis是假单胞菌科假单胞菌属植物病原细菌,能引发百合软腐病、草莓花疫病、番茄细菌性髓部坏死病、马蹄莲块茎软腐病等,还可引发洋葱、莴苣、中国白菜、大蒜、姜、花椰菜、水稻等软腐病。边缘假单胞菌具有如此广泛的宿主致病性取决于其生理特点,尤其是在0 ℃生长良好、5 ℃能大量繁殖等特点。2007年在我国甘肃发现由边缘假单胞菌边缘致病变种引发的马铃薯腐烂病,而有关边缘假单胞菌分离鉴定和特性的研究则鲜有报道。本试验从苗木中分离得到边缘假单胞菌,对其形态特征、生理生化特性和致病性进行研究,并通过16S rRNA序列分析进一步验证其分类地位。 相似文献
10.
《中国生物防治学报》2020,(2)
为筛选防治水稻稻瘟病的假单胞菌Pseudomonas,通过平板对峙试验从291株细菌中筛选获得对稻瘟病菌具有明显拮抗作用的菌株S149,对稻瘟病菌P131菌丝抑制率达到83.33%,发酵液和无菌上清液的抑制作用分别为67.39%和56.52%。16S rDNA序列分析显示,菌株S149与荧光假单胞菌P. fluorescens基因序列的同源性最高,结合形态观察,鉴定为荧光假单胞菌。对镰刀菌等10种植物病原菌有较明显拮抗作用,发酵液对稻瘟病菌分生孢子萌发和附着胞形成的抑制效果达到89%以上,无菌上清液对孢子萌发和附着胞形成的抑制作用达到81%以上。温室条件下对叶瘟的预防效果达到79.98%;田间试验结果证实对水稻幼苗有明显的促生效果,提高种子发芽7.15%、促进根长10.36%和增加株高10.33%;对叶瘟和穗颈瘟的田间防效分别达到73.32%和70.21%,与75%三环唑可湿性粉剂没有显著差异,与绿地康有显著差异。研究表明,荧光假单胞菌S149不仅可以用于防治水稻稻瘟病,而且可以促进水稻生长,具有较好的防治稻瘟病等多种真菌病害的应用潜力。 相似文献
11.
ABSTRACT Pectolytic strains of Pseudomonas fluorescens are opportunistic pathogens of broccoli, causing head rot in temperate regions of the world. In this study, we investigated the potential of two bacterial isolates, P. fluorescens m6418 and Bacillus sp. A24, for biological control of broccoli head rot caused by P. fluorescens 5064, isolated from diseased broccoli in Scotland, UK. P. fluorescens m6418, a Tn5 mutant of wild-type 5064, is nonpathogenic and overproduces an extracellular metabolite with strong antimicrobial activity. In this study, we identified the anti-microbial metabolite produced by strain m6418 as pyrrolnitrin. P. fluorescens m6418 had significant inhibitory effects against strain 5064 both in culture and on broccoli leaves. In an excised broccoli head pathogenicity test, strain m6418, when coinoculated with P. fluorescens 5064, reduced disease by 41%. Bacillus sp. A24 produces an enzyme that can degrade N-acyl homoserine lactones, signaling molecules employed by bacteria for quorum sensing. Bacillus sp. A24 was capable of out-competing P. fluorescens 5064 when grown together in culture, and could degrade the quorum sensing signal of P. fluorescens 5064 (and thereby attenuate its virulence gene production). However, Bacillus sp. A24 had only a limited biocontrol effect on P. fluorescens 5064 in the excised broccoli head assay. 相似文献
12.
海南一品红细菌性叶斑病病原菌的分离与鉴定研究 总被引：1，自引：0，他引：1
为明确海南省近几年发生的一品红细菌性叶部病害的病原菌特别是该病原菌的分子特征,为该病害的治理提供可行的依据,本文描述了2005年海南省部分花圃一品红植株上发生的细菌性叶斑病症状,并通过致病性测定、BIOLOG分析和16SrDNA序列比较将分离的病原菌鉴定为黄单胞菌属(Xanthomonas);部分碳源利用测定进一步显示2005年海南分离的菌株与2004年海南报道的细菌性疫病病原菌存在差异,但与杭州地区报道的一品红细菌性叶斑病菌基本一致。利用黄单胞菌模式种典型菌株的核糖体DNA内转录间隔区(internal transcribed spacer,ITS)序列构建系统发育树,结果显示3个海南菌株与巴豆黄单胞菌(Xanthomonas codiaei)和地毯草黄单胞菌(X.axonopodis)单独聚合成群,其中与巴豆黄单胞菌亲缘关系最近。 相似文献
13.
Md. Aktaruzzaman Young-Gyu Lee Tania Afroz Byung-Sup Kim 《European journal of plant pathology / European Foundation for Plant Pathology》2018,150(1):245-251
In this study, we identified the causative agent of postharvest gray-mold rot in sweet persimmon fruit that were collected from Gangneung, Gangwon Province, Korea in October 2016. Symptoms included extensive growth of mycelia on post harvested fruit. The fungus was isolated from infected fruit and cultured on potato dextrose agar (PDA). For identification of the fungus, we examined morphology characteristics and rDNA sequencing analysis of the fungus and confirmed its pathogenicity according to Koch’s postulates. The results of morphological examinations, pathogenicity tests, 5.8S rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4) and the five nuclear protein-coding genes G3PDH, HSP60, RPB2, MS547 and TUB revealed that the causal agent of postharvest gray-mold rot on sweet persimmon fruit in Korea was Botrytis cinerea. 相似文献
14.
新疆加工番茄腐霉根腐病病原鉴定及其rDNA的ITS区段分析 总被引：2，自引：1，他引：1
为了明确新疆加工番茄根腐病的病原种类,从成苗期到果实成熟期对各主栽区的病原物进行分离,并进行形态学鉴定和ITS序列分析.结果表明,从162个根腐病样中获得120个分离物,其中腐霉菌93株,占总分离物的77.5%.腐霉菌分离物鉴定为3个种,瓜果腐霉Pythium aphani-dermatum(Edson)Fitzp,56株,占腐霉分离物的60.22%;简囊腐霉P.monospermum Pringsheim,6株,占6.45%;棘腐霉P.acanthicum Drechsler,2株,占2.15%;另有29个分离物因未诱发出雄器和藏卵器而无法鉴定到种. 相似文献
15.
2015年春季在昆明市区绿化带种植的中华常春藤上发现一种由细菌侵染而引起的病害,称为中华常春藤细菌性叶斑病。通过发病症状、菌落形态观察、致病性测定、Biolog分析,16S rDNA序列和核糖体DNA内转录间隔区(internal trans-cribed spacer, ITS )序列分析比较,对昆明地区的常春藤叶斑病病原菌及其系统进化关系进行研究。分离病原菌接种中华常春藤叶片完成科赫法则验证,发病初期在叶片表面形成带有黄色叶晕的不规则褐色斑点,后期叶片边缘形成倒V字型坏死并起皱。将菌株CCT1和CCT6测序结果与现有的黄单胞菌菌株的16S rDNA序列和核糖体DNA的ITS序列构建进化树,结果均显示病原菌与野油菜黄单胞菌的序列相似度最大,属于同一支。研究确定该病原菌为野油菜黄单胞菌(Xanthomonas campestris)。这是中国首次报道由X. campestris 引起的中华常春藤叶斑病。 相似文献
16.
油菜菌核病生防芽孢杆菌的分离鉴定及其脂肽化合物分析 总被引：8，自引：4，他引：4
采用平板拮抗筛选,分别从西藏日喀则地区和拉萨地区杂草根围土壤中筛选到2个对油菜菌核病菌有显著拮抗活性的芽孢杆菌菌株RJGP16和YBWC43。通过生理生化鉴定、16S rDNA序列分析和BOX-PCR指纹图谱分析,鉴定菌株RJGP16为萎缩芽孢杆菌Bacillus atrophaeus, 菌株YBWC43为解淀粉芽孢杆菌Bacillus amyloliquefaciens。离体叶片试验结果显示,菌株RJGP16和YBWC43对油菜菌核病菌防治效果分别为50.24%和100.00%。脂肽化合物种类分析显示,菌株RJGP16产生脂肽化合物表面活性素和芬枯草菌素,菌株YBWC43产生杆菌霉素D和芬枯草菌素。表明菌株RJGP16和YBWC43对油菜菌核病的防治效果与其产生的脂肽化合物有关。 相似文献
17.
Ching-Hong YANG David E CROWLEY Anthony G HAY Noel T KEEN 《Journal of General Plant Pathology》2000,66(3):225-233
Specific Pseudomonas strains were detected by PCR amplification of the 16S-23S rDNA spacer region followed by denaturing gradient gel electrophoresis
(DGGE) to generate DNA banding profiles. In initial studies, two diverse sequence areas within the 16S-23S rDNA spacer region
were located in five closely related Pseudomonas fluorescens and P. putida strains. DNA banding profiles of 16 different pseudomonads were generated using PCR primers flanking this region, followed
by DGGE of the PCR products. Distinct banding profiles were observed for each strain, and specificity could be increased by
designing additional primers within the spacer region. A specific primer (513-1) was used to selectively amplify and detect
a plant-disease suppressive bacterium, P. fluorescens strain 513, in soil. Six field soils from different locations were used with the 513-1 primer to test the specificity of this
technique. Five soils did not yield any gel bands, but one soil led to a faint 250-bp band, similar in size to that of P. fluorescens 513. Resolution of strain 513 from the indigenous strain in this soil was achieved by DGGE of the amplified DNA fragments.
The results therefore demonstrate the utility of PCR-DGGE analysis for strain-specific identification of pseudomonads in environmental
samples.
Received 17 January 2000/ Accepted in revised form 10 May 2000 相似文献
18.
在湖南省常德市西洞庭管区种植的朝鲜蓟（Cynara scolumus L.）上发现了一种新的病害,其症状表现为地上部分从外层叶片开始逐步枯萎,随后根部和主茎杆的髓部腐烂变褐,最后整株枯萎。从田间感病朝鲜蓟茎杆的病健交界处用NA培养基分离,获得10个菌株,分别指定为HNXDT001~010,并进行了致病性测定、形态观察和细菌学特征分析,同时对HNXDT002菌株进行分子生物学鉴定。结果表明,该系列菌株在NA培养基上均形成灰白色圆形菌落,稍突起,有光泽,半透明。在显微镜下菌体呈短杆状,两端钝圆,具有2～8根周生鞭毛,革兰氏阴性。10个菌株通过针刺法接种均可导致朝鲜蓟茎杆、胡萝卜、辣椒、白菜、土豆、番茄和莴苣茎杆软腐,经科赫法则验证为致病病原菌。该菌株的16S rDNA序列和果胶酶基因片段测序(分别用16S rDNA通用引物16SF/16SR和果胶酶基因引物Y1/Y2扩增)与系统发育学分析表明,其16S rDNA序列(GenBank Accession No. JF721958)与胡萝卜软腐果胶杆菌胡萝卜亚种（Pectobacterium carotovorum subsp. carotovorum）菌株ATCC15713 (GenBank Accession No. U80197)序列同源性高达99％;果胶酶基因序列(GenBank Accession No. JF721960)与胡萝卜软腐果胶杆菌胡萝卜亚种（Pectobacterium carotovorum subsp. carotovorum）PC1菌株(GenBank Accession No. CP001657)序列同源性为93%。结果表明：朝鲜蓟细菌性根茎腐烂病病原为胡萝卜软腐果胶杆菌胡萝卜亚种。 相似文献
19.
Edward M. Onkendi Lucy N. Moleleki 《European journal of plant pathology / European Foundation for Plant Pathology》2014,139(3):557-566
Using a DNA-based typing method, 48 bacterial strains isolated from infected potato (Solanum tuberosum) tubers originating from Kenya were characterized. The pel gene specific primers showed that all the 48 bacterial strains were pectolytic. Subspecies-specific primers EXPCCF/EXPCCR and Br1f/L1r identified 66 % of the strains as Pectobacterium carotovorum subsp. carotovorum while 34 % were identified as Pectobacterium carotovorum subsp. brasiliense based on their characteristic band sizes of 550 and 322 bp, respectively. Amplification of the 16S-23S rDNA (ITS) region did not yield observable differences in banding patterns between the Kenyan strains. However, PCR-RFLP analysis together with partial nucleotide sequences of the housekeeping mdh and gapA genes confirmed the results obtained by the specific primers. Phylogenetic analysis of the concatenated partial gene sequences grouped Pectobacterium carotovorum subsp. carotovorum and Pectobacterium carotovorum subsp. brasiliense Kenyan strains together with those identified in other parts of the world with 90 % and 99 % bootstrap support values, respectively. Pathogenicity assays using representative Kenyan strains demonstrated varied levels of tuber maceration ability. The Pectobacterium carotovorum subsp. carotovorum and Pectobacterium carotovorum subsp. brasiliense Kenyan strains were shown to be less aggressive in causing soft rot when compared to type strains. This study describes for the first time the genetic diversity of pectolytic bacteria causing soft rot disease of potatoes in Kenya. 相似文献
20.
John Adriko Ernest Rashid Mbega Carmen Nieves Mortensen Ednar Gadelha Wulff Wilberforce Kateera Tushemereirwe Jerome Kubiriba Ole Søgaard Lund 《European journal of plant pathology / European Foundation for Plant Pathology》2014,138(2):293-306
A PCR-based system was developed to reliably and robustly identify group I and II members of the genus Xanthomonas. Primer sets developed from three gene targets namely fyuA, ITS and gumD were evaluated in the study. Primer sets were evaluated using DNA extracted from 45 Xanthomonas strains representing 25 species broadly covering the genus. Fifteen non-Xanthomonas strains of plant-associated bacteria including phylogenetically closely related species Stenotrophomonas maltophilia and Xylella fastidiosa were also tested. The primers targeting fyuA amplified DNA from all xanthomonads except X. theicola, while the ITS primers amplified a DNA fragment of 254 bp in all 45 Xanthomonas strains; whereas no amplification was observed for non-xanthomonads. The gumD primers allowed efficient amplification of DNA in 38 out of 39 isolates from Group II, whereas no or very weak amplification occurred with DNA from Group I members. Internal controls of primers targeting bacterial 16S rDNA or plant 26S mitochondrial rDNA were successfully applied in multiplex PCRs for testing bacterial cultures or plant tissue, respectively. The findings give us a PCR based approach that can reliably and effectively differentiate xanthomonads from non-xanthomonads as well as separating the strains belonging to the two described groups of the genus Xanthomonas. The study thus offers valuable tools for disease surveillance and management. It can effectively be applied in rapid assessment of new disease occurrences, for which no specific detection tools could be in place. 相似文献