Evidence for a phytoreovirus associated with tobacco exhibiting leaf curl symptoms in South Africa

ABSTRACT Three forms of tobacco leaf curl (termed classes I, II, and III, based on symptomatology) recently have been described in southern Africa. Numerous attempts to isolate virus particles responsible for a nongeminivirus-induced leaf curl disease (class I) of tobacco in South Africa have been unsuccessful. Recently, 12 dsRNA segments were isolated from tobacco exhibiting class I leaf curl symptoms, suggesting a possible reovirus genome. The objective of our study was to confirm whether the dsRNA segments are associated with a reovirus. Isolation of icosahedral particles with an outer core 60 to 65 nm in diameter and an inner core 40 to 45 nm in diameter was achieved. Twelve distinct nonpolyadenylated dsRNAs were isolated from purified virions, and the total molecular masses of the dsRNAs ranged from 17.86 to 18.40 x 10(6) Da in polyacrylamide and agarose gels, respectively. Using hybridization analysis, dsRNAs were identified as non-homologous distinct segments. Comparisons with other known reoviruses revealed a unique banding pattern that was most similar to the wound tumor virus (WTV), the type species of the genus Phytoreovirus. Hybridizations of WTV cloned DNA probes (segments S4 and S6 to S9) and dsRNAs from infected tobacco indicated no significant sequence similarity, whereas indirect enzyme-linked immunosorbent assay with a polyclonal antiserum to WTV showed strong positive cross-reactivity to tobacco virions. Our results indicate a virus with features consistent with those of phytoreoviruses. This is the first report of a plant reovirus in tobacco, the first record in Africa, and the second example of a field-isolated dicot phytoreovirus.     

烟草靶斑病菌菌丝融合群及ITS序列分析

 烟草靶斑病是2006年我国新报道发生的一种叶部病害^［1］,其病原的无性世代为立枯丝核菌(Rhizoctonia solani Kühn),有性世代为瓜亡革菌(Thanatephorus cucumeris (Frank)Donk)。该病菌主要危害叶部形成病斑,对烟草的产量和品质影响显著,目前该病害主要分布在辽宁省丹东和铁岭地区,并呈现出迅速蔓延趋势。烟草靶斑病最早由巴西报道,此后,哥斯达黎加、美国、南非和津巴布韦也相继发生^［2,3］。     

东北地区烟草靶斑病菌(Rhizoctonia solani)融合群、致病力分化及品种抗病性研究

2013-2014年吉林省、黑龙江省烟区首次大面积发生烟草靶斑病,损失严重。采集吉林省柳河县、黑龙江省林口县和宾县的烟草靶斑病病叶进行分离纯化,得到了12个菌株。将所获菌株分别与立枯丝核菌标准融合群菌株AG-2、AG-3、AG-4进行对峙培养,结果表明:此12个菌株均与立枯丝核菌AG-3标准菌株发生融合反应,不与其他融合群发生融合;随机选取3个县各1个代表性菌株,提取菌丝基因组DNA并对其ITS序列进行分析与比对,菌株HLJ-2、JL-1、HLJ-7的ITS序列与辽宁省烟草靶斑病菌菌株LF-2、LJT-8(AG-3融合群)的同源性达99%~100%,因此推断,吉林省、黑龙江省烟草靶斑病菌与辽宁省烟草靶斑病菌应属于同一个融合群,即立枯丝核菌AG-3融合群(Rhizoctonia solani AG-3)。根据各菌株在鉴别寄主上表现的抗性不同,可将吉林省、黑龙江省所有菌株分为3个致病类型,即致病类型Ⅰ、致病类型Ⅱ和致病类型Ⅲ。同一省份不同菌株间致病力有差异,黑龙江省菌株3种致病类型均有,以致病类型Ⅱ为主;吉林省菌株只有致病类型Ⅰ和致病类型Ⅱ。分别选取3个省的强致病力菌株,用离体叶片接种法鉴定了我国21个主栽烟草品种对靶斑病的室内抗病性,结果表明:没有免疫或抗病品种,‘龙江981’、‘NC297’对供试的3个强致病力菌株均表现中抗,‘中烟202’、‘云烟97’、‘NC55’、‘KRK26’和‘G28’分别对不同菌株表现中抗。     

Preliminary investigation of the causal agent(s) of a disease causing leaf curl of tobacco in South Africa

A high incidence (86%) of potyvirus infection was noted in tobacco plants exhibiting a form of leaf curl in South Africa. Despite leaf curl being reported in the literature to be of geminiviral aetiology, no geminiviruses were detected. Furthermore, no other virus particles were detected by virus purification, TEM and serology. Twelve species of dsRNA were consistently isolated from these tobacco plants, but were absent from other forms of leaf curl-affected and healthy tobacco. Aphid and mechanical inoculation demonstrated that the purified potyvirus(es) did not cause leaf curl symptoms, but rather mild mottle and mosaic symptoms in tobacco. Partial characterization of the potyvirus preparation showed a possible relationship to a South African strain of potato virus Y. Because potyvirus-inoculated plants did not manifest leaf curl symptoms, and because leaf curl symptoms were noted in some plants not infected with a potyvirus, it was concluded that the potyvirus is not involved in the leaf curl aetiology, but causes a latent infection, the symptoms of which are masked. The pattern of the dsRNA banding, induction of enations and lack of mechanical and seed transmission are common to plant reoviruses. The possibility of a phytoreovirus involvement in this form of leaf curl is currently being investigated. The results from this study suggest that tobacco leaf curl disease worldwide, with regard to geminiviruses, be re-evaluated.     

烟草叶斑病病原菌的鉴定及其生物学特性

为明确贵州省新发现的烟草叶斑病病原菌种类和生物学特性,2019年自田间采集具有典型发病症状的烟叶,采用组织分离法对其进行分离纯化,对分离菌株进行致病性测定,观察病原菌形态学特征,基于ITS、LSU、tub2和rpb2基因序列进行分子生物学鉴定,同时对病原菌的生物学特性进行测定。结果显示,烟草叶斑病病斑呈圆形或椭圆形,病斑中间呈浅褐色,边缘呈棕色,周围环绕黄色晕圈。经分离纯化共获得3株病原菌,形成的菌落为深褐色,气生菌丝呈白色;分子孢子器呈球形或扁球形,为深褐色,大小为79.3～91.7μm×148.4～167.3μm;分生孢子呈椭圆形,光滑,无隔膜,具0～3个油球,大小为3.5～5.6μm×1.7～2.9μm。根据形态学特征和分子生物学特征将其鉴定为Didymella segeticola。该病原菌菌丝生长适宜温度为20～25℃;适宜pH为6～10,最适碳源为乳糖,最适氮源为蛋白胨,通气和光暗交替环境有利于其生长。     

Occurrence of bacterial brown spot of dry beans in the Transvaal province of South Africa

Bacterial brown spot, caused by Pseudomonas syringae pv. syringae , caused epidemics on dry beans ( Phaseolus vulgaris ) on the Transvaal Highveld in South Africa during 1992. Crop losses of up to 55% were reported in plantings of the dry bean cultivar Bonus. The pathogen was detected in commercial seed stocks.     

烟草赤星病菌对苯醚甲环唑和氟环唑敏感性测定及田间防效试验

为明确山东省主要烟草产区烟草赤星病菌Alternaria alternata对苯醚甲环唑和氟环唑的敏感性,采用菌丝生长速率法检测苯醚甲环唑和氟环唑对86株烟草赤星病菌菌株的EC_(50),用Spearman’s秩相关系数法分析供试菌株对2种杀菌剂敏感性的相关性,并评价其对烟草赤星病的田间防效。结果表明,苯醚甲环唑对烟草赤星病菌菌丝生长EC_(50)值介于5.3883～8.0316 mg/L之间,平均值为6.9594 mg/L,敏感性频率分布呈连续性偏正态分布,田间烟草赤星病菌对苯醚甲环唑仍然较为敏感,可建立田间烟草赤星病菌对苯醚甲环唑的敏感基线。氟环唑对烟草赤星病菌菌丝生长的EC_(50)值介于0.1253～0.2517 mg/L,平均值为0.1773 mg/L,敏感性频率分布呈连续性偏正态分布,可建立田间烟草赤星病菌对氟环唑的敏感基线。相关性分析结果表明,烟草赤星病菌对苯醚甲环唑和氟环唑敏感性之间不存在明显的相关性。田间试验结果表明,30%苯醚甲环唑悬浮剂对烟草赤星病具有较好的防效,其有效成分150 g/hm~2处理的防效最高,为73.99%;12.5%氟环唑悬浮剂对烟草赤星病的最高防效为70.57%。表明2种三唑类杀菌剂对烟草赤星病防效较好。     

啶酰菌胺等8种杀菌剂对烟草叶斑病菌的生物活性

采用生物测定方法分析了烟草叶斑病菌Didymella segeticola在菌丝生长阶段对8种杀菌剂(啶酰菌胺、苯醚甲环唑、丙环唑、氟硅唑、多菌灵、咪鲜胺、菌核净和代森锰锌)的敏感性,同时通过离体叶片法测定了8种杀菌剂对烟草叶斑病的保护和治疗作用。结果表明：供试8种杀菌剂对D. segeticola菌丝生长表现出不同的抑制活性,同时对其引起的病害具有一定的保护和治疗作用。抑菌活性最强的是啶酰菌胺,其平均EC₅₀值为(0.047 0±0.012 0) mg/L;其次依次为苯醚甲环唑[(0.079 0±0.005 0) mg/L]、咪鲜胺[(0.29±0.08) mg/L]、丙环唑[(0.69±0.12) mg/L]、菌核净[(1.08±0.33) mg/L]、多菌灵[(1.22±0.29) mg/L]、氟硅唑[(1.38±0.07)mg/L];代森锰锌的抑菌活性最弱[(22.80±10.51) mg/L]。进一步研究表明,氟硅唑、苯醚甲环唑、丙环唑、啶酰菌胺、菌核净和多菌灵对烟草叶斑病保护作用较强,25 mg/L药剂质量浓度处理下防效均>82%;100 mg/...     

烟草角斑病发病因素及上部叶经济损失估计

2002～2008年烟草角斑病与气象因素关系分析结果表明,6月份降雨量和雨日数与采收末期烟草角斑病病情指数关系较为密切,可作为病情预测的主要因子。品种龙江911较品种NC89更易感病;玉米茬种植烟草较烟草前茬烟株发病率和病情指数极显著偏高;接种条件下,中部叶与上部叶病情指数差异不显著,但中部叶和上部叶病情指数均显著高于下部叶;上部叶经济损失与病情指数之间存在着极显著直线回归关系,病情指数每增加1,单株上部叶经济损失增加0.004 9元;按目前的防治成本,达到经济损害时的病情指数为5.2。     

贵州烟草棒孢霉叶斑病生防菌鉴定及防治药剂筛选

为有效防治烟草棒孢霉叶斑病,采用Biolog全自动微生物鉴定系统鉴定了本实验室前期工作中筛选得到的生防菌株YC2140,进一步对其进行了16S rRNA和gyr B序列分子鉴定,并选取生产上常用的5种杀菌剂和YC2140菌株进行了盆栽和田间防效试验。结果显示,经Biolog鉴定和分子鉴定,YC2140菌株为荧光假单胞菌Pseudomonas fluorescens。盆栽试验结果表明,在药剂推荐施用剂量(449.78 mL/hm²或449.78 g/hm²)下,对烟草棒孢霉叶斑病的防治效果由高到低依次为500 g/L氟啶胺悬浮剂(防治效果78.77%)、450 g/L咪鲜胺水乳剂(77.85%)、430 g/L戊唑醇悬浮剂(67.12%)、1×10⁸ cfu/mL YC2140发酵液(61.64%)和50%啶酰菌胺水分散粒剂(46.12%)。相同施药剂量下的田间试验结果表明,防治效果由高到低依次为450 g/L咪鲜胺水乳剂(43.33%)、500 g/L氟啶胺悬浮剂(40.97%)、430 g/L戊唑醇悬浮剂(27.94%...     

南天竹一种新的叶斑病病原菌的分离鉴定

<正>南天竹(Nandina domestica)为小檗科南天竹属,常绿灌木,形态优雅,果实球形鲜红且经冬不落,是赏果观叶的园林绿化植物。南天竹具有重要的药用价值,有“镇咳止喘、兴奋神经、强壮身体”之效[1]。在生长过程中,南天竹叶片易受炭疽病(Colletotrichum)危害[2],也可受叶斑病(Pestalotia microspore)、褐斑病(Cercospora nandinae)、     

广西烤烟棒孢霉叶斑病病原分子鉴定及其生物学特性补充

本文对广西烤烟棒孢霉叶斑病的病原进行鉴定,并对其生物学特性进行初步研究。根据该病原菌的形态特征、致病性及rDNA-ITS序列测定,将引起广西烤烟棒孢霉叶斑病的病原菌鉴定为山扁豆生棒孢(Corynespora cas-siicola)。生物学特性测定结果表明:27.5℃、培养初始pH为6、以麦芽糖作为C源、硝酸钾作为N源及完全光照等条件最适合其菌丝生长;30℃、以乳糖作为C源、硝酸钾作为N源及完全黑暗等条件最有利于该菌分生孢子形成;27.5℃、湿度为100%+水膜、12h光暗交替等条件最有利于分生孢子萌发;分生孢子的致死温度为52℃。这是该病在广西的首次报道。     

薏苡叶枯病和叶斑病调查与病原鉴定

对福建省龙岩市新罗区薏苡叶部病害进行了调查。用组织分离法,PSA平板培养经纯化后作病原鉴定,在其上发现了两种病害:薏苡叶斑病为一种新病害,病原鉴定为薏苡尾孢(Cercospora sp.);薏苡叶枯病为福建省首次记录,其病原为薏苡平脐蠕孢(Bipolaris coicis)。     

烟草赤星病防治研究进展

在烟草赤星病综合防治研究中 ,国内外对烟草进行了诱导寄主抗病性、生物防治、化学药剂防治、转基因抗病育种等方面的研究。本文综述了近年烟草赤星病综合防治的研究成果 ,提出利用生物技术获得抗病品种 ,及采用生物防治、诱导抗病性进行烟草赤星病综合治理     

A bacterial leaf spot of wisteria in Italy1

Bacterial leaf spot was observed on Wisteria sinensis in the south of Italy. Lesions on leaflets were small, round, dark brown, surrounded by an extensive yellow halo. Upon artificial inoculation, lesions similar to naturally occurring ones were reproduced. The causal organism was rod-shaped, Gram-negative, aerobic, motile by polar flagella, fluorescent. On the basis of morphological, biochemical and physiological tests, the bacterium was identified as Pseudomonas syringae pv. syringae.