广东省江门市‘新会柑’叶斑病病原菌鉴定

近年来广东省江门市‘新会柑’上出现一种叶斑病,症状表现为叶尖和叶缘出现不规则的、带有黄色晕圈的棕褐色斑点。为明确引起该病的病原菌,作者利用组织分离法对病样进行分离纯化,采用ITS序列和LSU基因进行系统发育学分析,同时结合形态特征观察对纯化后的菌株进行鉴定,结合柯赫氏法则对其进行验证。结果确定病原菌为番茄匍柄霉Stemphylium lycopersici。     

莱芜生姜茎腐病的病原鉴定

利用组织分离技术和菌丝尖端纯化技术对莱芜地区生姜茎腐病的病原菌进行了分离和纯化.并按照柯赫法则,利用真菌形态学鉴定方法对病原菌进行了鉴定.结果表明,引起山东莱芜生姜产区的生姜茎腐病的病原菌为群结腐霉(Pythium myriotylum Drechsler).     

贵州省余庆县茶褐枯病病原菌的鉴定

为鉴定引起茶褐枯病的病原菌,本研究对病原菌进行分离、纯化和培养,通过柯赫氏法则验证菌株的致病性,并观察病原菌的形态特征,依据病原菌rDNA-ITS、ACT、CAL和TUB2基因进行多基因系统发育分析。结果显示:病原菌分生孢子呈淡蓝色,表面光滑,无隔膜,圆柱状,两端钝或向底部变窄,水滴状斑点,大小为(11.7~29.5)μm×(3.9~7.7)μm,平均为(19.4±4.4)μm×(5.4±0.8)μm,分生孢子梗形成于气生菌丝上,透明,具有隔膜;附着胞为棒状等不规则形状,颜色呈棕色到深棕色,单生。基于病原菌形态学鉴定和多基因系统发育分析结果,将病原菌确定为山茶刺盘孢Colletotrichum camelliae。     

引起我国甜樱桃叶斑病的间座壳菌种类鉴定

为明确引起我国甜樱桃叶斑病的病原菌种类,于2019年在北京市、四川省、山东省和辽宁省对与已报道的其他叶斑病症状明显不同的一种甜樱桃叶斑病进行调查并采集病叶,通过组织分离法进行病原菌分离纯化,结合形态学特征和多基因(ITS、EF1-α、TUB2和CAL)系统发育分析对病原菌的种类进行鉴定,并采用离体叶片接种法进行致病性测定。结果显示,新发现的这种甜樱桃叶斑病通常从叶边缘产生圆形或不规则灰褐色病斑,病斑周围具黄色晕圈,有时形成穿孔。从病样中共分离纯化得到10株单孢系菌株,结合其菌落形态、分生孢子形态和大小等,以及多基因系统发育分析结果,最终将10株菌株鉴定为甜樱间座壳Diaporthe eres(5株)和大豆间座壳D. sojae(5株),致病性测定结果显示甜樱间座壳和大豆间座壳对甜樱桃叶片均具有致病性,完成柯赫氏法则验证。     

云南省蓝莓茎基腐病病原菌鉴定及其生物学特性分析

为明确云南省蓝莓茎基腐病的病原菌种类及其生物学特性,从玉溪市江川区蓝莓种苗种植基地采集具有茎基腐病典型症状的蓝莓病株进行病原菌分离、纯化和致病性测定,对病原菌形态学特征进行观察并基于Act、H3、EF-1α和β-tub基因序列分析进行分子生物学鉴定,同时对病原菌的生物学特性进行室内测定。结果表明,分离到的病原菌纯化菌落为深棕色,菌丝白色至红褐色;大型分生孢子梗无色透明,长150～400μm,分枝末端产生2～4个瓶梗;囊泡球形或近球形,大小为15～23μm×6～15μm;大型分生孢子无色透明,直圆柱形,一端稍窄,大小为45～80μm×4～8μm。基于Act、H3、EF-1α和β-tub基因构建系统发育树,菌株BC-1、BC-2与冬青丽赤壳Calonectria ilicicola均聚在同一分支,结合形态学特征与系统发育分析结果,将云南省蓝莓茎基腐病病原菌鉴定为冬青丽赤壳C. ilicicola。病原菌对可溶性淀粉的利用效果最好,菌丝生长的最适氮源为蛋白胨,最适生长温度为25℃,最适pH范围为4～7。     

蓝莓间座壳茎溃疡病病原鉴定及生物学特性研究

在蓝莓园进行病样采集时,发现一种蓝莓茎部病害,病斑呈红褐色,溃疡状,长椭圆形。为明确该病害的病原菌种类,对病枝进行组织分离,经单孢纯化后得到菌落形态一致的7株致病菌,对供试菌株的ITS与EF1-α基因联合构建系统发育树,供试菌株与Diaporthe phaseolorum处于同一分支,结合形态学特征,鉴定该病原菌为Diaporthe phaseolorum。对病原菌的生物学特性研究结果表明,该菌最适培养基为PDA,最适生长温度为25℃,光照对菌丝生长无明显影响。本文首次报道Diaporthe phaseolorum引起蓝莓茎溃疡病。     

引起葡萄叶斑病的链格孢种类的初步鉴定

为明确北京市大兴区、顺义区和山西省襄汾县葡萄树上叶斑病的病原菌,利用组织分离法对病样进行分离纯化,采用多基因系统发育学分析(ITS,Alt a 1和gpd)和形态学对纯化后菌株进行鉴定,并结合柯赫氏法则对其进行验证。结果表明,共获得37株链格孢属真菌菌株,分属链格孢Alternaria alternata、葡萄链格孢A. viniferae和乔木链格孢A. arborescens,柯赫氏法则检测结果显示接种菌株对所接叶片与果穗均具致病性,说明这3种链格孢种均为葡萄叶斑病的病原菌;葡萄链格孢为主要的病原菌,有28株,故将该类病害命名为葡萄链格孢叶斑病;此外,该类菌株在2～40℃之间均能生长,25℃为最适生长温度。     

无梗五加圆斑病病原菌的鉴定

首次报道了无梗五加圆斑病的发生危害、症状特点,并进行了病原菌的分离、纯化和鉴定。根据致病性测定、病原菌形态和培养性状观察,确认引起无梗五加圆斑病的病原菌为真核茎点霉（Phoma eupyrena Sacc.）。     

海南省温郁金炭疽病的病原鉴定

温郁金炭疽病是一种重要的真菌性病害,该病主要危害叶片,初期产生水渍状的淡黄色病斑,后逐渐扩大成圆形、椭圆形或不规则形的灰褐色病斑,上面有黑色小点,病斑外面环绕一层黄色晕圈,严重时多个病斑联合造成叶片卷缩和枯死,2016年我们从海南省的温郁金种植基地采集炭疽病样本,经病原菌的分离纯化、柯赫氏法则验证、病原菌的形态特征观察及多基因谱系分析方法,确定其病原菌为郁金炭疽菌Colletotrichum curcumae(Syd.)E.J.ButlerBisby。     

糜子细菌性条斑病病原菌鉴定及其对13种杀菌剂的敏感性

为有效防治糜子细菌性条斑病,采用平板划线分离法对2016年自黑龙江省齐齐哈尔市采集的罹细菌性条斑病的糜子植株进行病原菌分离与纯化,依据柯赫氏法则对其致病性进行测定;采用Biolog自动微生物鉴定系统、16S rDNA和16S-23S rDNA间隔区序列分析及病原菌的生理生化反应对病原菌进行鉴定;并测定该病原菌对13种常用杀菌剂的敏感性。结果显示,从糜子叶片和叶鞘病斑中共分离纯化获得10株培养性状一致的菌株,经柯赫氏法则验证均为细菌性条斑病病原菌。经Biolog鉴定,3株代表菌株与标准菌株燕麦噬酸菌Acidovorax avenae的相似值分别为0.716、0.705、0.723,序列分析显示,这3株代表菌株与燕麦噬酸菌燕麦亚种A. avenae subsp. avenae的一致性为99%以上;该病原菌的革兰氏染色反应及碳源利用等生理生化反应与对照菌株燕麦噬酸菌ICMP3183表现一致。因此,确定糜子细菌性条斑病病原菌为燕麦噬酸菌。糜子细菌性条斑病病原菌代表菌株MZ121101对80%乙蒜素乳油、30%乙蒜素乳油及0.3%四霉素水剂的敏感性较高,抑制中浓度EC50介于389.31～939.94μg/mL之间,这2种药剂可作为防治糜子细菌性条斑病的首选药剂。     

Anthracnose disease of Amomum villosum caused by Colletotrichum fructicola in China

In May 2019, leaf spots were found on Amomum villosum in Guangxi Province, China. Light yellow-green spots first appeared at the tips or edges of leaves. A fungus was isolated from the leaves and identified as Colletotrichum fructicola based on morphological characteristics and molecular identification using the DNA sequences from ITS, TUB2, ACT, and GAPDH sequences. Similar leaf spots developed after leaves were inoculated with the isolate. This is the first report that C. fructicola causes leaf spots on A. villosum.

     

加那利海枣叶斑病病原鉴定*

对加那利海枣叶斑病的症状进行了观察。结果表明,加那利海枣叶斑病主要发生在叶片和叶柄。初期症状是在叶片和叶柄出现水渍状褪绿色的褐色小点,病斑扩大成浅褐色椭圆形或不规则病斑,病斑周围有黄色的晕圈,后期变褐色至灰褐色,中央产生小黑点。按照柯赫氏法则对该病害的病原菌进行分离和致病性测定。通过形态学观察和rDNA ITS序列分析鉴定,将病原菌鉴定为异色拟盘多毛孢［Pestalotiopsis versicolor (Speg.) Steyaert］。     

Evidence for oxidative stress involved in physiological leaf spot formation in winter and spring barley

ABSTRACT A leaf spot disease with unknown etiology has become more pronounced in spring and winter barley in Germany in recent years. The symptoms are similar to net blotch and Ramularia leaf spots, but the causal agents of these diseases are not identified. The symptom expression varied much on cultivars. Cultivars most affected by the disease of both spring and winter barley showed a significantly higher level of superoxide (O(2) ) production and lipid peroxidation (malondialdehyde), but a lower level of antioxidant potential expressed as superoxide dismutase (SOD) activity, catalase activity, and integral water-soluble antioxidant capacity (ACW) than insensitive cultivars. A high positive correlation between O(2) production and leaf spot development between ear emergence and milk ripeness was established in the most sensitive winter barley cv. Anoa (r(2) = 0.9622) and spring barley cv. Barke (r(2) = 0.9434). Leaf H(2)O(2) levels increased with the severity of leaf spots. The histochemical localization of O(2) and H(2)O(2) in the tissues adjacent to leaf spots indicated that these two active oxygen species (AOS) are involved in the formation of leaf spots. Reduction of symptom severity by applying strobilurin and azole fungicides was always associated with elevated SOD activity and ACW content and suppressed O(2) production. However, peroxidase activities were significantly higher in sensitive cultivars and in more severely affected tissues and decreased by applying fungicides. Thus, it is assumed that a possible genetic mechanism based on the imbalanced AOS metabolism contributes to formation of physiological leaf spots.     

First report of leaf spot on lettuce caused by <Emphasis Type="Italic">Curvularia aeria</Emphasis>

In 2017, leaf spots were found on lettuce growing in fields in Songkhla Province, southern Thailand. The fungus isolated from the spot lesions on the leaves was identified as Curvularia aeria (Bat., J.A.Lima and C.T.Vasconc.) Tsuda based on morphological characteristics and DNA sequences of the ITS region of the rRNA gene. After a conidial suspension of the isolate was sprayed on lettuce seedlings, the leaf spots developed on lettuce seedlings, and the fungus was reisolated; leaves of plants inoculated with water did not develop spots. This is the first report of C. aeria causing leaf spot on lettuce.     

Bacterial leaf blight of sweet pepper (Capsicum annuum) caused by Pseudomonas cichorii in Japan

Sweet peppers (cv. Tosajishi beauty) with leaf blight symptoms were observed in Kochi Prefecture, 2003. Initially, small spots formed on leaves, and later enlarged to brown to dark brown spots, and eventually blighted leaves fell. Several bacteria that caused the same symptoms were isolated and subsequently reisolated. These bacteria were identified as Pseudomonas cichorii on the basis of bacterial characteristics and the 16S rRNA gene sequence. This is the first report of bacterial leaf blight in the sweet pepper in the world.     

Increased content of phenolic compounds in pear leaves after infection by the pear rust pathogen

European pear rust induced by the fungus Gymnosporangium sabinae (Dicks) G. causes yellow to bright orange leaf spots on leaves of pear trees. The aim of this study is to identify and quantify polyphenolic compounds found in pear leaf extracts. Identified were: ten hydroxycinnamates, eight flavonols, nine flavan-3-ols and three arbutin derivatives. Polymeric procyanidins were additionally determined by UPLC-FL. The total content of phenolics in the control healthy green leaf extract was 11,889.98 mg but in the infected leaves it reached 28,573.89 mg in the samples with yellow spots and 11,480.06 mg/100 g dry matter (dm) in the green part of leaves. The yellow spots in pear rust leaves were characterized by increased content of flavanol (catechins and procyanidins) and arbutin compounds compared with the green part of the infected leaves and control healthy leaves.     

Host range and phytotoxicity of <Emphasis Type="Italic">Stemphylium solani</Emphasis>, causing leaf blight of garlic (<Emphasis Type="Italic">Allium sativum</Emphasis>) in China

Since 2004, a new leaf blight disease on garlic of high severity has been observed in Dangyang County, Hubei province, China. Initial symptoms consisted of multiple, small, irregular to oval, white leaf spots, which enlarge to produce sunken purple lesions, sometimes surrounded by a bright yellow margin. As the disease progressed, lesions expanded and merged, resulting in withering of leaf tips. After isolation and pathogenicity testing, the causal agent of leaf blight of garlic was identified as Stemphylium solani from cultural and morphological characteristics, and subsequent analysis of the internal transcribed spacer region of ribosomal DNA. When fungal plugs of two S. solani isolates were inoculated onto 11 garlic cultivars and 20 other crop species, leaf spots appeared on all inoculated plants, but two garlic cultivars (Qingganruanye and Ruanruanye) and three crop species (Capsicum annuum, Brassica napus and Amaranthus mangostanus) showed the smallest leaf spots. In cross-inoculation experiments, no indications of host specificity were observed, but S. solani isolated from garlic was generally the most virulent on five plant species, while the isolate from leek (Allium odorum) was generally the least virulent. Toxicity testing of the crude culture filtrates indicated that garlic isolates produced toxin(s) that were not heat-labile and induced different levels of phytotoxicity toward various garlic cultivars and crops.     

Synergism of hydrogen fluoride and leaf necrosis on freesias

Continuous fumigation with HF at a concentration of 0.5 µg/m³ and intermittent fumigation with 0.3 µg/m³ aggravated leaf necrosis in freesia. In the cultivars Rose Marie and Royal Blue, symptoms of leaf necrosis develop earlier and increase in severety with these concentrations of HF. The agent of this disease has not yet been identified nor isolated. The normal symptoms are initially chlorosis and later necrosis but HF causes necrotic spots and stripes as the first foliar symptoms in the places where under uncontaminated conditions chlorotic spots would appear.     

Aetiology of leaf spot of garlic and onion caused by Stemphylium vesicarium in Spain

Surveys between 1989 and 1993 in the major garlic production areas of Spain identified a new leaf spot disease, characterized by white and purple lesions followed by extensive necrosis. Isolation and pathogenicity tests with fungal isolates taken from these spots indicated that Stemphylium vesicarium was the causal agent. Pseudothecia of the teleomorph stage, Pleospora sp., were found on leaf debris from affected plants. Inoculation of garlic and onion plants with residues carrying mature pseudothecia, or with ascospore suspensions obtained from the pseudothecia, resulted in the development of white and purple leaf spots. Wetness periods longer than 24 h were required for symptom development under controlled conditions. Isolates of S. vesicarium from garlic, onion and asparagus caused disease in all three hosts. In garlic, cv. Blanco de Vallelado was most susceptible, while lines B4P17 and B6P1, and cvs Iberose and Golourose were less susceptible to the disease.     

烟叶成熟衰老程度与对赤星病感病性的关系

 5个栽培品种的盆栽植株,于13片真叶展开时人工接种,比较各叶位叶片的感病性。结果表明:烟叶自下而上逐渐成熟衰老,随着叶色由绿转黄,叶绿素含量依次递减,其汁液可溶性糖及浸出液还原糖含量递增,感病性相应地提高。以叶色为指标,绿色叶片对病害免疫;轻微转黄的绿叶,高度抗病,只能形成褐色小点,而不能产生分生孢子,也不能再扩展;中度转黄的绿叶上形成0.2cm以下的小斑,也不产生孢子,多数不能再扩展;黄色叶片为高感型,能形成0.5cm以上的典型病斑,并产生大量孢子。以1%葡萄糖液为介质,孢子萌发率以接种烟株的病情指数高于1%蔗糖液。以黄叶液为孢子液介质,其发芽率及接种发病率均高于黄绿叶及绿叶汁液。这说明烟叶成熟衰老程度与感病性高度相关。