一株红棕象甲寄生真菌的分离鉴定

为明确一株红棕象甲寄生真菌的归属,采用形态学及分子生物学方法,从产孢表型、分生孢子特征和ITS序列测定等方面对其进行了分析鉴定。结果表明:该菌分生孢子梗粗短,顶端具2～5个瓶状小梗,分生孢子呈长椭圆形或圆柱形,大小为4.8～9.3μm×2.2～3.1μm;ITS通用引物扩增该菌得到了预期的558bp条带,其与金龟子绿僵菌小孢变种的支持率高达100%。因此,将该红棕象甲寄生真菌菌株鉴定为金龟子绿僵菌小孢变种Metarhiziumanisopliaevar.anisopliae。     

Characterization and pathogenicity of Rhizoctonia isolates associated with cauliflower in Belgium

Sixty-two isolates of Rhizoctonia spp. were collected from Belgian cauliflower fields during 2005 and 2006. The majority of the isolates (60 out of 62) had multinucleate cells and were identified as Rhizoctonia solani . Characterization of anastomosis groups (AGs) was performed using pectic zymograms, PCR-RFLP and sequencing of the rDNA-ITS region. The most prevalent AG was AG 2-1 (55% of isolates), followed by AG 2-1 subset Nt (11%), AG 1-1C (8%), AG 5 (8%), AG 4 HGII (6%), AG 3 (5%) and AG 1-1B (3%). Pathogenic potential towards different vegetable crops and towards maize was determined. Damage to cauliflower and endive was caused by different AGs, with the isolates aggressive towards cauliflower belonging to AG 2-1, AG 2-1 subset Nt, AG 4 HGII, AG 1-1C, AG 1-1B and AG 2-2, and those aggressive towards endive belonging to AG 1-1B, AG 1-1C, AG 2-1 subset Nt, AG 2-2, AG 4 HGII and AG 5. The most aggressive isolates towards bean belonged to AG 2-1 subset Nt and AG 2-2, for lettuce to AG 1-1B and AG 2-1, on carrot to AG 4 HGII and towards maize to AG 2-2. Within the isolates of AG 2-1, variability was observed in PCR-RFLP pattern and in aggressiveness towards several crops, indicating this subgroup to be heterogeneous. This is the first study concerning the occurrence of R. solani AGs causing wirestem in Belgian cauliflower fields and the first report of aggressive isolates of AG 1-1C, AG 2-1 subset Nt and AG 4 HGII associated with cauliflower.     

辽宁地区辣椒疫病菌鉴定及同源性分析

对采集到的辣椒疫病病原菌通过形态学观察和rDNA-ITS序列分析进行鉴定及同源性研究。利用真菌通用引物ITS4、ITS5对病菌基因组进行PCR扩增,将测序结果与已知序列进行比对,并利用Clustalx1.83及MEGA4.0软件进行聚类分析。试验结果表明:病菌菌落为白色,菌丝呈鹅毛绒状,无隔膜,孢子囊卵圆形或梨形,乳突单生;测得病菌ITS序列为805～877bp,rDNA-ITS序列同源性高,致病菌病原均为辣椒疫霉菌(Phytophthora capsici Leonian)。     

山东省小麦禾谷孢囊线虫rDNA-ITS序列特征和基于ISSR的遗传多样性分析

小麦孢囊线虫病(CCN,Heterodera avenae)已成为山东省小麦生产上的一种重要病害,研究CCN群体间和群体内的遗传多样性可以为其综合防治提供理论依据。本实验研究了山东省17地市34个群体的r DNA-ITS区,并采用ISSR分子标记技术对其中10个地市的27个种群做了遗传多样性分析。结果表明,在r DNA-ITS系统发育树中,山东省34个小麦孢囊线虫群体与H.pratensis、H.australis及中国H.avenae群体亲缘关系较近。3个ISSR引物共扩增出31条条带,多态性条带百分率(PPB)为100%。菏泽、潍坊、烟台群体遗传多样性较高,枣庄、威海、淄博、滨州群体遗传多样性相对较低。M antel检测和聚类结果表明,群体间的遗传分化与地理距离并无显著的相关性,AM OVA分析结果显示,在总的遗传变异中17.7%的变异发生在群体间,82.3%的变异发生在群体内。研究结果显示山东省H.avenae具有较高的遗传多样性,且群体间已发生了一定程度的遗传分化。     

晚熟桃采后锈斑处病原菌的分离与鉴定

以晚熟桃"映霜红"为试材,通过对桃果面、果蒂处带菌量分析,并进一步对病原真菌分离纯化,明确晚熟桃采后0℃贮藏过程中锈斑处的主要致病菌及其致病性。结果表明,晚熟桃锈斑病一般在采前出现浅褐色锈斑;新采收的晚熟桃果蒂锈斑处微生物总量为(4.50±0.05)lg(CFU·g~(-1)),显著高于果面处(3.51±0.03)lg(CFU·g~(-1))(P0.05);在果面和果蒂处共分离出8种真菌,其中病原真菌A-1的分离率最高,达43.8%,该菌有伤接种致病率达100%。结合病原菌的生长特性、显微镜下的形态特征和病原菌rDNA-ITS检测结果,病原菌A-1鉴定为半知菌纲(Deuteromycetes)链孢霉目(Moniliales)黑霉科(Dematiaceae)链格孢属(Alternaria)链格孢(Alternaria alternata),该病原菌加剧了贮藏后期晚熟桃果蒂处褐腐病的发生。     

天祝县燕麦叶斑病的发生情况及其病原鉴定

叶斑病（Drechslera avenacea）是燕麦（Avena sativa）的重要病害之一,严重影响燕麦的品质和产量。天祝藏族自治县是甘肃省燕麦的主要种植区,为明确这一地区叶斑病的发生情况及其病原菌种类,本研究对天祝县5个村镇的燕麦田进行了叶斑病发生情况调查和病样采集,对病原菌进行了致病性测定、形态特征观察和rDNA-ITS序列分析。结果表明,5个调查区域中松山镇黑马圈河村和东大滩乡上泉村发病最严重,病叶率高达92%以上,病情指数60以上;安远镇柳树村、抓喜秀龙乡南泥沟村和东大滩乡酸茨沟村发病也较重,发病率在64%左右,病情指数23左右。经分离鉴定,发现引起天祝县燕麦叶斑病的病原菌共有4属5种,其中德氏霉属（Drechslera）2种,链格孢属（Alternaria）、附球菌属（Epicoccum）和枝孢菌属（Cladosporium）各1种。天祝县燕麦叶斑病发病较严重,共分离筛选出的5种病原菌,除德氏霉属的燕麦内脐蠕孢（Drechslera avenae）外,其它病原菌在国内燕麦上未见报道。     

Molecular analysis of the major Phytophthora species on cocoa

The internally transcribed spacer (ITS) regions of the ribosomal RNA (rRNA) gene cluster of 161 isolates of Phytophthora species involved in pod rot, stem canker and leaf blight of cocoa were analysed to determine inter- and intraspecific variation in this disease complex. The species P. palmivora , P. megakarya , P. capsici , P. citrophthora and P. nicotianae could all be clearly distinguished by PCR amplification of the ITS region followed by restriction analysis with Hae III, Hinf I, Pvu II and Alu I. This method provided a relatively rapid identification procedure for these species, and was able to distinguish isolates that had previously been misidentified by morphological methods. Sequence analysis showed that the four main cocoa-associated species formed two distinct groups, one comprising P. capsici and P. citrophthora , and the other P. palmivora and P. megakarya . Detailed sequence analysis and comparison with published literature suggested that P. capsici isolates from cocoa may be closely related to P. tropicalis , a species recently described from Cyclamen and Dianthus .     

Molecular analysis and virus transmission tests place Olpidium virulentus, a vector of Mirafiori lettuce big-vein virus and tobacco stunt virus, as a distinct species rather than a strain of Olpidium brassicae

The rDNA-ITS sequences of ten single-sporangium isolates of Olpidium virulentus (a noncrucifer strain of Olpidium brassicae), which transmits Mirafiori lettuce big-vein virus (MLBVV) and tobacco stunt virus (TStV), were compared with those of six single-sporangium isolates of O. brassicae. The sequence similarity within isolates of O. virulentus or O. brassicae was almost identical (98.5%–100.0%), but was low between the two species (79.7%–81.8%). In a phylogenetic analysis of the rDNA-ITS region, O. virulentus and O. brassicae fell into two distinct clusters, indicating that O. virulentus, a vector of MLBVV and TStV, is a distinct species rather than a strain of O. brassicae.     

玉米根围土壤中腐霉菌的分离鉴定及核糖体DNA-ITS序列分析

从兰州地区的红古、七里河等地采集玉米根围土壤标样,诱饵法与组织分离法相结合分离纯化到23株腐霉菌株,通过形态特征、培养特性、生长速度和对温度适应范围的研究,结合rDNA-ITS序列分析,将分离到的腐霉菌株鉴定为硬腐霉、终极腐霉和一个腐霉待定种。这是首次报道在我国分离到硬腐霉,为我国新记录种;终极腐霉也系首次在甘肃从玉米根围土壤中分离到。统计表明,3种腐霉菌种在兰州地区玉米根围土壤中的分离频率不同,终极腐霉出现次数最多,为优势种群,分离频率为60.9%;其次为腐霉待定种,分离频率为26.1%;而硬腐霉分离频率最低,为13.0%。     

引起乙烯褪绿蜜橘果实腐烂的主要致病真菌分离及鉴定

为明确贮藏过程中乙烯褪绿蜜橘果实腐烂的主要致病真菌,以重庆市北碚产区无核蜜橘为研究材料,利用乙烯利对其果实进行褪绿处理,观察贮藏期间褪绿蜜橘果实的发病症状,确定病害种类,统计发病率;并从发病果实中分离主要致病真菌,采用形态学特征观察和ITS序列分子生物学分析对其进行鉴定。结果表明,乙烯褪绿加剧贮藏蜜橘果实病害的发生。从发病果实中共分离出5株主要致病真菌菌株,分别编号为A、B、C、D、E;结合形态学特征和5株致病真菌的ITS序列分析结果,A、B菌株均被鉴定为胶孢炭疽菌Colletotrichum gloeosporioides,但这2株菌株具有不同的菌落形态及致病特性;C菌株被鉴定为焦腐病菌Lasiodiplodia theobromae;D菌株被鉴定为交链格孢菌Alternaria alternata;E菌株被鉴定为棘孢曲霉Aspergillus aculeatus。A、B菌株引起的果实腐烂占乙烯褪绿蜜橘总腐烂果实的81.96%,是引起贮藏过程中乙烯褪绿蜜橘果实腐烂的主要致病真菌,其次为C菌株,D、E菌株为次要致病真菌。