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山东牛心柿炭疽病菌的分离鉴定及致病性 总被引：1，自引：0，他引：1
【目的】对侵染牛心柿引起炭疽病的病原菌进行分离鉴定,分析其致病性,初步构建其侵染谱,为柿树病害的病原诊断和防治提供理论依据和技术支持。【方法】通过组织分离法和单孢分离法从感病牛心柿的果实、叶片和嫩梢中分离病原菌,根据菌落和孢子形态特征及其 rDNA-ITS 序列分析进行种类鉴定,采用以菌饼进行伤口和非伤口接种、孢子悬浮液进行喷雾接种的离体接种试验分析其致病性。【结果】从感病牛心柿果实、叶片和嫩梢中分离获得9个病原菌分离物,菌落及孢子形态特征显示均为炭疽菌。以真菌转录间隔区通用引物 ITS6和 ITS4为引物,以菌丝总 DNA 为模板,扩增获得病原菌 rDNA-ITS基因片段,经 rDNA-ITS序列分析和系统进化树的构建,将该病原物鉴定为柿树炭疽菌,将其 rDNA-ITS 基因序列提交到 GenBank 数据库(基因登录号 KF010811)。牛心柿炭疽病菌和次郎炭疽病菌对不同品种柿树的致病性相同,均无差异性,对果实的致病性比叶片强;接种7天后,叶片无病斑或病斑较小,菌饼无伤口接种几乎不发病,在叶片上不产生病斑,果实上病斑较小;不同的接种方式在果实和叶片上产生的病斑大小为：菌饼伤口接种＞孢子悬浮液接种＞菌饼无伤口接种;牛心柿炭疽病菌和次郎炭疽菌对桃、苹果、梨等果树的叶片和果实均不致病。【结论】柿炭疽病菌具有一定的专性寄生性,对除柿树外的其他果树不致病,且在遗传上有一定的保守性,因地理距离产生的变异很小,这些特性对于柿炭疽病的防治具有重要意义,在一定程度上可以避免或削弱因病菌的遗传变异而产生的抗药性,从而有利于炭疽病的防治。在柿树有伤口时最易侵染致病,雨天或高湿度的情况下经孢子侵染致病的可能性居其次。在田间管理时注意避免造成柿树伤口感染,或者避免在下雨天气进行修剪,可最大程度地减轻或消除柿树炭疽病菌的侵染以及柿炭疽病的发生和流行,从而在一定程度上解决该病流行而造成的减产问题。 相似文献
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湖南省油茶炭疽病病原鉴定 总被引：3，自引：0，他引：3
《林业科学》2017,(8)
【目的】研究湖南省油茶炭疽病病原,为该病害的防控提供依据。【方法】采集湖南省长沙、株洲、浏阳、永州、怀化、常德、常宁等油茶主产区9个样地的典型油茶炭疽病叶,分离纯化叶片病健交界处的真菌菌株,观察菌株在(PDA)培养基上的菌落、分生孢子及附着孢的形态特征,确定为炭疽属真菌。进一步对炭疽属菌株进行柯赫氏法则验证,确定油茶炭疽病病原菌菌株。对获得病原菌的r DNA-ITS、CAL和GAPDH 3个基因进行PCR扩增和测序,得到的基因序列与GenBank中已有模式炭疽属菌株序列按照3个基因一致的顺序拼接后采用PAUP和MrBayes软件构建系统发育树。【结果】从湖南省9个采样地共分离获得62株炭疽属菌株,致病性测试表明这62株菌均能对油茶嫩叶和果致病,但不同菌株的发病时间具有一定差异;根据形态特征及多基因系统发育分析,最终明确湖南省油茶炭疽病的病原种类。【结论】湖南省油茶炭疽病的病原菌包含果生炭疽菌、暹罗炭疽菌、胶孢炭疽菌、山茶炭疽菌和哈锐炭疽菌5种炭疽属真菌,其中果生炭疽菌分布范围最广,分离率最高,达到64.5%。 相似文献
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《经济林研究》2014,(1)
为了给生产中柿树次郎甜柿炭疽病的诊断及防治提供理论指导,通过组织分离法和单孢分离法从感病次郞甜柿的果实和嫩梢中获得了2个分离物,菌落及孢子形态特征显示该分离物为炭疽菌。以通用引物ITS6和ITS4,通过PCR扩增,获得了rDNA-ITS基因片段。测序结果显示,从柿的果实和嫩梢分离获得的染病次郎甜柿分离物ITS序列完全一致,片段大小为598 bp,该片段与感病浙江无核柿(AY787483)和新西兰分离物(GQ329690)的同源性分别为100%和99.8%。系统进化树分析结果显示,染病次郎甜柿分离物与浙江无核柿(AY787483,AY791890)和新西兰分离物(GQ329687,GQ329688和GQ329690)处在同一分支,据此,将该病原物鉴定为柿树炭疽菌Colletotrichum horii,并将其ITS基因序列提交到GenBank数据库(基因登录号:JQ957543)。 相似文献
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银杏是中国特有的珍贵树种也是重要的城市绿化树种。近年来,叶斑病已成为沈阳市银杏树的常发病害,严重影响了银杏的观赏价值,甚至引起银杏死亡。为了明确沈阳市内银杏叶斑病的病原菌种类,采集了市内银杏发病叶片,采用组织分离法对病原物进行了分离和纯化,并进行了致病性测定,通过形态学和rDNA ITS序列分析,将沈阳市银杏叶斑病的病原菌鉴定为链格孢Alternaria alternate。 相似文献
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《林业科学研究》2021,34(5)
[目的 ]鉴定引起河南省洛阳市嵩县天池山国家森林公园臭椿炭疽病病原菌,为该病害的防控提供依据。[方法 ]采用常规组织分离法进行病原分离,利用形态学和多基因系统发育分析相结合的方法对分离菌株鉴定并进行致病性测定。[结果 ]从病组织中共分离获得11株菌株,所有菌株在PDA培养基上菌落特征表现一致,正面菌落先白色后灰色,背面黑色,分生孢子为圆柱状,两端钝圆,单孢,无色,初步鉴定分离菌株为炭疽菌Colletotrichum spp.。选取代表菌株CH-1和CH-3接种臭椿叶片,有伤条件下接种均能引起臭椿叶片发病,发病叶片症状与田间一致,柯赫氏法则验证成立;经多位点基因(ITS、ACT、TUB2、CHS-1和GAPDH)系统发育分析,发现分离菌株与果生炭疽菌C. fructicola聚集在同一个分支上,支持率达到99%。[结论 ]本研究首次采用形态学结合多基因系统发育研究,明确发生在河南省洛阳市嵩县天池山国家森林公园的臭椿炭疽病为果生炭疽菌C. fructicola所致。 相似文献
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油松枯梢病病原菌的分离鉴定 总被引：1，自引：0，他引：1
《中国森林病虫》2016,(3)
为明确近年来华北地区出现的大量油松枯梢病病原菌及其分类地位,对发病植物材料采用组织分离法分离病原菌;根据柯赫法则用针刺法接种田间健康植物进行致病性测定;通过形态学观察和ITS序列分析鉴定病原菌。结果表明,分离得到的菌株是油松枯梢病的致病菌,鉴定为半知菌亚门腔孢纲球壳孢目球壳孢属真菌Sphaeropsis sapinea。 相似文献
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【目的】分离和鉴定非刚竹属5种竹子丛枝病病原菌,并分析其与麦角菌科其他真菌间的系统发育关系,为竹子丛枝病病原菌的鉴定、病害的防控和麦角菌科物种间亲缘关系的确定提供科学依据。【方法】采用组织分离法从非刚竹属5种竹子丛枝病样品上分离病原菌,利用显微镜观察病原菌的形态结构并测量分生孢子大小,结合形态学与分子生物学方法鉴定病原菌物种,基于ITS rDNA和LSU rDNA序列构建麦角菌科的系统发育树。【结果】中华业平竹、巴山木竹和薄箨茶竿竹丛枝病组织分离的病原菌形态结构与竹针孢座囊菌一致,分生孢子在PDA平板上形成表面凹凸不平的褶皱、略有光泽、边缘不整齐的浅黄色菌落,分生孢子针形,大小为(22.4～52.0)μm×(2.2～6.0)μm。黄麻竹和牡竹丛枝病组织分离的病原菌菌丝在PDA平板上形成表面平整、边缘整齐的圆形菌落,病原菌培养72天后产生短棒状分生孢子,大小为(5.4～10.5)μm×(1.2～2.8)μm。ITS rDNA和LSU rDNA序列分析和系统发育分析表明,分离自中华业平竹、巴山木竹和薄箨茶竿竹丛枝病样品的病原菌与针孢座囊菌聚为一支,支持率为99%;分离自黄麻竹和牡竹丛枝病样品的病原菌与竹异香柱菌聚为一支,支持率为100%。【结论】中华业平竹、巴山木竹和薄箨茶竿竹丛枝病的病原菌为竹针孢座囊菌,该菌与麦角菌属的亲缘关系近。黄麻竹和牡竹丛枝病的病原菌为箣竹异香柱菌,该菌与肉瘤座菌属的亲缘关系近,与香柱菌属的亲缘关系远。 相似文献
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Syamak Hanifeh Doustmorad Zafari Mohammad Javad Soleimani Abbasali Ravanlou 《Forest Pathology》2019,49(1)
During spring and summer of 2016, dieback symptoms including blights of leaves, twigs, and acorns were observed on current year shoots of Quercus infectoria in the Zagros oak forests of Iran. The fungus isolated from damaged tissues was identified as Discula quercina based on both morphological characteristics and ITS sequencing. To fulfil the Koch's postulates, a representative isolate was inoculated onto shoots of Q. infectoria and Quercus libani in both laboratory and forest conditions. Responses of the two oak species to inoculation with D. quercina were examined under laboratory and forest conditions in a completely randomized experiment. Discula quercina was clearly involved in oak dieback, and Q. infectoria was more susceptible than Q. libani to damage by the pathogen. This is the first record of the occurrence and pathogenicity of the fungus Discula quercina on Quercus infectoria. The fungus is considered as an emerging pathogen on oak trees in Zagros forests in Iran. Furthermore, the pathogenicity of the Discula quercina on Q. libani under laboratory and forest conditions increases the potential importance of this pathogen in Zagros forests. 相似文献
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To examine the effects of salicylic acid (SA), the ethylene precursor 1‐aminocyclopropan‐1‐carboxylic acid (ACC) and methyl jasmonate (MJ), which induce defence responses against pathogen in plants, on foliar endophytic fungi, we performed exogenous treatments of Quercus serrata leaves with these chemicals and subsequently isolated endophytic fungi. SA and ACC significantly reduced the isolation frequency of the most dominant fungus, a Xylaria species as identified by sequence analysis of the internal transcribed spacer (ITS) region of the ribosomal DNA. In contrast, the frequency of the second most frequent fungus increased by SA and ACC. A few other fungi were also affected positively or negatively. Shannon diversity indices decreased by all treatments because of the uneven isolation frequencies of the endophytes. Our results imply that host defence responses induced by the chemical treatments reduced the number of infections of some endophytes while the frequency of some other endophytes increased benefiting from the reduction of competitors. 相似文献
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First detection of Neoscytalidium dimidiatum associated with canker disease in Egyptian Ficus trees 
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下载免费PDF全文 In September 2013, a canker disease of Ficus nitida and F. benjamina was reported for the first time in Assiut governorate, Egypt. Infected samples were collected from various locations. Pure cultures of a fungus were isolated on potato dextrose agar at 25°C from diseased plants. Morphological investigation and DNA sequencing showed that the causal agent was Neoscytalidium dimidiatum. A pathogenicity test conducted using 2‐year‐old plants of each host gave 70%–80% infection, and the pathogen was reisolated from the inoculated plants. A pure culture of N. dimidiatum was deposited in the culture collection of the Assiut University Mycological Centre (AUMC) under the code AUMC 9293 and the ITS sequence was placed in NCBI under accession number KX985929 . 相似文献
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张妍 《内蒙古林业调查设计》2020,43(2):96-99,75
文章对采集到的一株野生大型真菌子实体进行了菌种分离,并对获得的菌丝进行了rDNA ITS序列分析。结合形态鉴定,对分离株r DNA ITS片段进行PCR扩增,扩增产物长度为649 bp,登录NCBI与GenBank中已知的菌种进行Blastn比对,序列相似性在99%~91%之间,下载相似性高的菌种的ITS序列,通过ClustalW进行序列比对,MEGA5.2构建系统发育树,通过树图分析,鉴定出分离的菌株为紫丁香蘑纯菌种。然后,对紫丁香蘑液体菌种培养进行了初步探索,其中配方3生长速度最快,适宜紫丁香蘑液体菌剂培养。 相似文献
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真菌ITS区序列结构及其应用 总被引：15，自引：0，他引：15
分析了真核生物以及原核生物核糖体基因内部转录间隔区序列结构.核rDNA的内转录间隔区ITS序列包含被5.8S rDNA所分隔的ITS1和ITS2两个片段.列举了根据真菌ITS区序列设计的扩增通用引物与特异引物.在此基础上,综述了真菌ITS序列结构在种间亲缘关系研究、植物病原真菌的检测、未知病原真菌的鉴定、近似种或疑难种的确定等方面的应用. 相似文献
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Mohamed Maizatul‐Suriza Jaabi Suhanah Ahmad Zairun Madihah Abu Seman Idris Hasmah Mohidin 《Forest Pathology》2021,51(1):e12660
Marasmius palmivorus is a marasmioid fungal species that exhibits parasitic behaviour, although most marasmioids are rarely parasitic. The fungus has been reported to cause fruit bunch rot disease of oil palm and coconut, but only a few studies on its pathogenic behaviour are available, particularly on oil palm. Hence, there is a need to assess the ability of the fungus to act as a pathogen and to study its molecular evolution and taxonomy. Nine isolates of M. palmivorus were successfully isolated from basidiocarps and diseased fruitlets of oil palm collected from oil palm plantations and were morphologically characterized on potato dextrose agar (PDA) followed by molecular identification based on nucleotide sequence alignments of internal transcribed spacer (ITS) regions of ribosomal RNA (rRNA) gene clusters with sequences from GenBank. Koch's Postulates confirmed that M. palmivorus could infect oil palm fruitlets with symptoms similar to those observed for bunch rot disease. Molecular phylogenetic studies using nucleotide sequences of ITS and the nuclear ribosomal large subunit (nLSU) showed that isolates sharing the same phenotypic characteristics of Marasmius palmivorus and Marasmiellus palmivorus are monophyletic and share a common ancestor. The fungus has also been shown to be more closely related to the genus Marasmius than Marasmiellus; therefore, we support retention of the taxon name of the pathogen causing bunch rot disease of oil palm as Marasmius palmivorus. 相似文献
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红树林内生真菌1403的形态与分子鉴定(英文) 总被引：3，自引：1，他引：2
CHENG Zhong-shan TANG Wen-cheng SU Zhi-jian CAI Yi SUN Shi-feng CHEN Qi-jin WANG Fang-hai LIN Yong-cheng She Zhi-gang LLP Vrijmoed 《林业研究》2008,19(3)
红树林内生真菌1403是从中国南海分离的一株镰刀菌,在液体发酵条件下能够产生灰黄霉素和葸醌类的抗生素。基于1403在形态学方面具有以下特征:产小型分生孢子的分生孢子梗较长,小型分生孢子呈椭圆形,大型分生孢子线形或略微弯曲,本文将它与有类似形态特征的镰刀菌进行了比较,发现1403与植物病原菌Fusarium verticillioides(有性态为Gibberella moniliforme)表面上很相似,气生菌丝均可产生大量假头状或链状的小型分生孢子,但深入的研究却发现1403与F.verticillioides具有不同的性状,如产生的多生分生孢子梗有时与单生分生孢子梗长度相当,产生环状菌丝,以及菌丝发生融合等现象,进一步以最大简约法和贝叶斯分析法将1403与其它相似镰刀菌进行系统发育比较,综合18S、28S、ITS三水平上的分析结果可以将1403归于镰刀菌亚组Gibberella fujikuroi,并最终在EF—1α水平上将其定为Fusarium proliferatum. 相似文献
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Hiroto Suhara Nitaro Maekawa Takashi Kubayashi Ryuichiro Kondo 《Journal of Wood Science》2005,51(1):83-88
In order to monitor the basidiomycetous fungus Phlebia brevispora isolated from butt rot of Chamaecyparis obtusa (Japanese cypress) in 1997 in Nagasaki Prefecture, a sensitive polymerase chain reaction (PCR)-based assay was developed to specifically detect the fungus on-site. A species-specific primer for P. brevispora was derived from the internal transcribed spacer (ITS) region (containing 5.8S ribosomal DNA, ITS1 and ITS2) sequences of the fungus. The PCR assay was able to detect down to 1fg DNA (per 1µl PCR reaction mixture) and down to 0.2mg mycelium of P. brevispora (per 1g of decayed wood). The samples for on-site monitoring were collected in 2002 from the decayed tree stump in which P. brevispora had first been isolated. From the decayed tree tissue, P. brevispora could be detected by PCR assay even when its mycelium could not isolated from the tree tissue by culturing. This indicates that the PCR amplification using the specific primer developed here is a useful method for monitoring P. brevispora on-site. 相似文献