核桃枝枯病病原菌鉴定及其生物学特性

In May 2018,a new shoot dieback disease was found in several walnut orchards in Rizhao,Shandong Province.The fungal pathogen was obtained on PDA plate from diseased walnut shoots using a tissue isolation method.The purified fungal isolates formed white to grey colonies on PDA plate and three single spore isolates SDWa1-SDWa3 were obtained.Based on morphological characteristics and analysis of rDNA internal transcribed spacer (ITS),calmodulin (CAL),translation elongation factor 1-α (TEF1) and β-tubulin (TUB) sequences,the fungal isolates were identified as Diaporthe nobilis,a new pathogen on walnut.The test of Koch's rule confirmed that D.nobilis was the pathogen of walnut shoot dieback.To determine optimal growing conditions for the pathogen,its biological characteristics test was conducted.The results showed that the optimal growth temperature and pH value were at 25℃ and pH 5-9,respectively.The optimum carbon source was glucose,and favourable nitrogen source was peptone.     

辣木枝枯病病原菌鉴定及其生物学特性

为明确海南省辣木枝枯病病原菌及其生物学特性,采用组织分离法获得菌株MO157,通过柯赫氏法则、形态学特征及分子生物学技术对该菌株进行鉴定,并测定了其菌丝生长的最佳条件。结果表明,菌株MO157回接10 d后,辣木茎干表面周围呈黄褐色,随后发病部位表现出水渍状病斑,与自然发病辣木的病状相符;菌落近圆形,菌丝初期呈白色绒毛状,后期呈浅黄色,分生孢子顶胞钩状,3~5个隔膜,厚垣孢子呈球形,在菌丝间串生;菌株MO157的ITS序列与Gen Bank中木贼镰刀菌Fusarium equiseti的相似性为100%,结合形态特征与分子鉴定最终将其确定为木贼镰刀菌(Gen Bank登陆号:KX197955)。该菌菌丝生长适宜温度25~30℃,最适温度28℃;pH 6~8时菌丝生长速率增快,pH 7时最适菌丝生长;PDA培养基和PSA培养基最适合该菌生长,以蔗糖为碳源时利用率最高;以牛肉浸膏为氮源时利用率最高;该菌致死温度为65℃,10 min;光照对菌丝生长影响较小,不同处理间菌落直径无显著差异。     

核桃枝枯病病原菌鉴定及其生物学特性

In May 2018,a new shoot dieback disease was found in several walnut orchards in Rizhao,Shandong Province.The fungal pathogen was obtained on PDA plate from diseased walnut shoots using a tissue isolation method.The purified fungal isolates formed white to grey colonies on PDA plate and three single spore isolates SDWa1-SDWa3 were obtained.Based on morphological characteristics and analysis of rDNA internal transcribed spacer (ITS),calmodulin (CAL),translation elongation factor 1-α (TEF1) and β-tubulin (TUB) sequences,the fungal isolates were identified as Diaporthe nobilis,a new pathogen on walnut.The test of Koch's rule confirmed that D.nobilis was the pathogen of walnut shoot dieback.To determine optimal growing conditions for the pathogen,its biological characteristics test was conducted.The results showed that the optimal growth temperature and pH value were at 25℃ and pH 5-9,respectively.The optimum carbon source was glucose,and favourable nitrogen source was peptone.     

蓝莓枝枯病病原菌鉴定

 蓝莓 (Vaccinium spp.),是一类属于杜鹃花科 (Ericaceae) 越橘属 (Vaccinium) 的多年生小浆果类果树,广泛地应用于医药、保健、化妆品和环境保护等各方面,联合国粮农组织将其果实列为人类世界五大健康食品之一,也是我国南方酸性土丘陵地区值得发展的经济作物。全国已超过 10 个省份开始大面积的蓝莓商业化栽培,预计未来 10 年将超过 10 万 hm^{2 [1]}。但是,随着生产面积的不断扩大,蓝莓病害也日趋严重,严重影响和制约蓝莓产业的发展。2010年至今,在云南省昆明市周边及丽江市蓝莓种植区发生一种病害,主要为害嫩枝、枝条和主干,发病初期,感病枝条上产生红褐色圆或椭圆形坏死斑,随后逐渐扩大,造成枝条干枯死亡,并可扩展造成主干枯萎。调查发现,该病发病率通常在 10％～20％,重者可达 25％ 以上,可引起蓝莓整株枯萎死亡。本文通过病原菌分离、致病性测定及传统、分子鉴定,明确了该病的病原,以期为病害防治提供理论依据。     

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。     

蓝莓叶斑病病原菌鉴定及其生物学特性

为明确山东省蓝莓叶斑病病原菌的种类及其生物学特性,采用组织分离法获得菌株B1,通过形态学特征并结合r DNA-ITS序列分析对该菌株B1进行鉴定,以离体和活体叶片接种法测定菌株B1的致病性,并对其生物学特性进行研究。结果表明,菌株B1菌落呈白色绒毛状,背面淡黄色,分生孢子纺锤形,5个细胞,中间3个细胞为褐色,具有2～4根顶端附属丝;菌株B1的ITS序列与GenBank中棒状拟盘多毛孢Pestalotiopsis clavispora的相似性达99%以上,结合形态特征与rDNAITS序列分析将病原菌鉴定为棒状拟盘多毛孢,GenBank登录号为MG009201。菌株B1接种蓝莓叶片后产生褐色病斑,后期密生黑色分生孢子盘,与蓝莓田间自然发病症状一致。该菌株在5～35℃、pH 5～11范围内均可生长,最适温度范围为25～30℃,最适pH范围为5～9;但仅在25～30℃时产生分生孢子,最适产孢温度为25℃;光照条件对菌丝生长无明显影响,但连续黑暗有利于产孢;以葡萄糖为碳源时利用率最高,对乳糖利用效果最差;最适氮源为硝酸钠、硫酸铵和蛋白胨,几乎不能利用尿素。     

龙船花赤枯病病原菌鉴定及其生物学特性测定

采用离体接种法对龙船花叶部病害的一种真菌进行了鉴定和生物学特性测定。结果表明,致病性测定与田间症状相似。通过对病原菌形态学、rDNA-ITS和β-微管蛋白序列的综合分析,确定该病原菌为龙船花拟盘多毛孢(Pestalotiopsis ixorae)。生物学特性测定表明,菌丝生长和分生孢子萌发最适温度为25℃,最适pH分别为6和7;光照更有利于菌丝生长;碳源以可溶性淀粉和D-甘露糖、氮源以硝酸钠最利于菌丝生长;分生孢子在液态水中萌发率最高,为85.3%;菌丝致死温度为50℃10 min,分生孢子致死温度为50℃20 min或55℃5 min。     

苹果枝枯病病原菌鉴定及生物学特性

In July 2017, a new shoot dieback disease, caused brown necrotic spots with black granules on the shoots in the late stage, was found in several apple orchards in Yantai, Shandong Province. The fungal pathogen was isolated on PDA plate from the infected shoots using the tissue isolation, which formed white to grey colonies bearing abundant fruiting bodies on Czapek solid media with lactose as carbon source. The purified fungal isolates La1-La3 were obtained from single spore isolation after spore tendrils exuding from pycnidia generated on the plates. This study was to identify and characterize the causal agent . Based on morphological characteristics and analysis of rDNA internal transcribed spacer (ITS), β-tubulin (TUB) and translation elongation factor 1-α (TEF1) nucleotide sequences, the fungal isolates were identified as Lasiodiplodia pseudotheobromae, a new pathogen on apple tree. The test of Koch's rule confirmed that L. pseudotheobromae was the pathogen of this disease. Effects of different temperature, pH value, carbon and nitrogen sources were tested against the growth of isolated fungus in vitro. The results showed that the optimal growth temperature and pH value were at 30℃ and pH 6-9, respectively, while the optimum carbon source was sucrose, and favourable nitrogen sources were sodium nitrate, ammonium nitrate and peptone.     

苹果枝枯病病原菌鉴定及生物学特性

<正>苹果(Malus pumila Mill.)是世界上栽培最为广泛的水果之一,由于其生态适应性强、营养价值高、耐贮性好及供应周期长,相当多的国家都将其列为主要消费果品[1],而我国是世界上苹果种植面积最大、产量最高的国家[2]。我国苹果枝干病害以苹果轮纹病和腐烂病为主,近年来随着气候环境的改变,一些新的病害也逐渐产生。2017年7月对烟台市几个苹果果园调查时,发现一种新病害。该病害主要危害苹果枝条,产生褐色坏死病斑,严重时导致枝条枯死,发     

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。     

蓝莓茎溃疡病病原菌鉴定及品种抗病性研究

在浙江及福建蓝莓产区发现一种蓝莓茎部病害,病斑呈棕黄色至棕黑色,严重时引起蓝莓整株枯死。对病样进行分离得到具有致病性的36个分离株。通过对菌株的形态学研究及系统发育分析,将致病菌鉴定为Diaporthe hongkongensis,本文系国内外首次报道D.hongkongensis危害蓝莓。对代表菌株的生物学特性进行研究表明:菌落生长最适温度为25℃;最适培养基为PDA;菌落生长对光照不敏感。采用菌饼刺伤接种活体枝条的方法对10个蓝莓主栽品种进行抗病性测定,结果显示:不同蓝莓品种对该病菌表现出敏感差异,其中‘爱国者’、‘瑞卡’、‘日出’、‘奥林匹亚’、‘都克’、‘托柔’、‘蓝丰’相对抗病;‘北陆’、‘埃利奥特’、‘达柔’相对感病。     

云南核桃叶枯病病原鉴定及其生物学特性

为明确引起云南核桃叶枯病的病原及其生物学特性,本文对分离自叶枯病病斑上的一株拟盘多毛孢进行了形态学和分子生物学鉴定,并通过柯赫氏法则验证其致病性;采用生长速率法、悬滴法和载玻片法研究其菌丝生长和孢子萌发的生物学特性。结果表明,该菌株为小孢拟盘多毛孢Pestalotiopsis microspora,是引起云南核桃叶枯病的新病原。该病原菌的最适生长温度为20～25℃,最适光周期L//D=12 h//12 h,最适生长pH为4～8。病原菌的分生孢子萌发受pH和湿度的影响较大,在中性及碱性环境下不萌发,最适萌发pH为4～5;在高于90%的相对湿度下才能萌发。分生孢子在20～25℃下萌发率最高,全光照最适于孢子萌发;核桃叶汁液对分生孢子萌发有显著的促进作用。     

云南省荞麦叶枯病病原菌鉴定及其生物学特性

为明确云南省荞麦叶枯病病原菌种类,采用常规组织分离法获得病原菌菌株LW2015.3,通过形态学特征及分子生物学技术对菌株LW2015.3进行鉴定,并研究其生物学特性。结果表明,荞麦叶枯病病原菌的分生孢子呈倒棍棒状或倒梨状,褐色,具3～8个横隔膜,0～4个纵斜隔膜,大小为16.5～45.0μm×5.0～13.5μm,厚垣孢子呈球形,直径为6.0～12.0μm;该菌株ITS序列系统发育进化分析结果表明,菌株LW2015.3与链格孢Alternaria alternata(登录号:MG195995.1)的同源性为100%,结合形态学特征与分子鉴定结果确定云南省荞麦叶枯病病原菌为链格孢A. alternata(登录号:KT362732.1)。该病原菌菌丝生长适宜温度为20～30℃,25℃为最适温度;当p H为6～9时菌丝生长速率加快,pH 7最适菌丝生长;PDA培养基和PSA培养基最适菌丝生长;该病原菌对以麦芽糖为碳源和以硝酸钠为氮源时的利用率最高;菌丝的致死温度为50℃,10 min;不同光照条件对菌丝生长的影响有显著差异,连续光照最有利于菌丝生长。     

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)、     

辽宁梨树间座壳菌枝枯病的病原鉴定

 在辽宁兴城“锦丰”梨栽植园出现一种新的枝干病害,该病害主要发生在主干及侧枝,主要症状为主干呈现黑斑、枝条枯萎。本研究对该病害进行了系统调查及病原物分离培养;经形态学观察、柯赫氏法则验证,ITS、EF1-α、CAL、TUB和HIS基因联合建树分析,确定引起“锦丰”梨枝枯病的病原菌为间座壳属真菌Diaporthe nobilis。     

南瓜叶枯病病原菌生物学特性研究

本文对南瓜叶枯病病原菌瓜链格孢（Alternaria cucumerina）进行了生物学特性研究,结果表明：温度、pH、光照对其生长和产孢都有一定的影响。该病原菌菌丝生长、产孢和萌发的适宜温度为28 ℃,pH为7时菌丝生长最快,pH为8时最易产孢。25 ℃下连续黑暗培养产孢最多。碳、氮源对菌丝生长和产孢量有一定的影响,碳源中甘露醇组菌丝生长最快,山梨醇处理产孢量最多。氮源中牛肉浸膏最适宜菌丝生长和产孢,硫酸铵最差。该菌致死温度为50 ℃。