广西山药炭疽病病原菌的鉴定与ITS序列分析

 本文对山药炭疽病在广西的危害、症状特点以及病原菌的鉴定进行了研究。2005年从广西5个病区采集的25个标样均分离到类似的分离物, 根据病原菌的形态特征和致病性, 并结合其rDNA-ITS区域的序列分析, 将广西山药炭疽病的病原菌鉴定为胶孢炭疽菌(Colletotrichum gloeosporioides)。     

杨树炭疽病菌对多菌灵及3种DMIs杀菌剂的敏感性

采用菌丝生长速率法,测定了可引起杨树炭疽病的59株胶孢炭疽菌和4株炭疽菌对多菌灵、苯醚甲环唑、戊唑醇和三唑酮4种杀菌剂的敏感性。结果表明:59株胶孢炭疽菌对多菌灵、苯醚甲环唑、戊唑醇和三唑酮的EC50值范围分别在0.037 1~0.130 1、0.102 5~1.680、0.069 1~1.917及3.053~38.59 μg/mL之间,平均EC50值分别为（0.066 4±0.013 1）、（0.374 1±0.254 8）、（0.681 2±0.442 1）和（19.82±6.200）μg/mL。胶孢炭疽菌对多菌灵的敏感性频率分布呈连续单峰曲线,表明尚未出现敏感性下降的群体;而对苯醚甲环唑和戊唑醇的敏感性频率分布呈现双峰,表明已出现敏感性下降的群体。4株炭疽菌对多菌灵、苯醚甲环唑、戊唑醇和三唑酮的最小EC50值和最大EC50值分别相差1.33、13.19、13.66和2.14倍,其中菌株Ca-4对苯醚甲环唑和戊唑醇的EC50值均大于4 μg/mL。不同寄主来源的胶孢炭疽菌对同种杀菌剂的敏感性之间无显著差异（P>0.05）。Spearman’s秩相关分析表明,胶孢炭疽菌对苯醚甲环唑和戊唑醇的敏感性之间呈显著正相关性（ρ=0.665 5,Pρ=0.489 6,PP>0.05）。研究结果对合理使用杀菌剂防治杨树炭疽病具有借鉴意义和参考价值。     

越南进境人参果炭疽病病原菌的分离鉴定

2008年4月从广州出入境检验检疫局送检的越南进境人参果样品(报检号:P02297)中发现果实表面密布近圆形褐色病斑,病斑直径约为3mm,中央黄褐色,边缘深红褐色,多个病斑可愈合成片.室温下放置1周,病果即完全腐烂,全果果皮及果肉均转成褐色.采用组织分离培养法获得分离物,经对分离物培养性状与形态特征观察、致病性测定,并辅助于分子生物学检测,将病害鉴定为人参果炭疽病,病原菌为胶孢炭疽菌(Colletotrichum gloeosporioides).Blast分析表明从病原菌基因组中扩增到的1.8 kbp 18S rDNA 片段与胶孢炭疽菌18S rRNA基因同源率为99%.迄今为止人参果炭疽病在国内外均未见报道,这是我国首次从进境人参果中截获并检验鉴定该病害.     

猫豆炭疽病病原分离与鉴定

猫豆炭疽病主要危害猫豆的叶片、茎蔓和豆荚,导致猫豆品质和产量降低.本研究对猫豆炭疽病的病原进行分离和致病性测定,并通过形态学特征和ITS rDNA序列分析将该病原鉴定为胶孢炭疽菌[Colletotrichum gloeosporioides (Penz.)],该病原菌危害猫豆在国内为首次报道.     

广西油茶炭疽病病原菌鉴定及生物学特性

为明确广西油茶炭疽病病原菌种类及其生物学特性,通过形态学方法对分离获得的病原菌进行初步鉴定,以核糖体内转录间隔区、微管蛋白基因、几丁质合成酶基因、肌动蛋白基因、3-磷酸甘油醛脱氢酶基因进行多基因系统分析鉴定;并用十字交叉法、菌丝干重法和血球计数法对其生物学特性进行研究。结果表明,经形态学和多基因联合系统结合分析鉴定,广西油茶炭疽病病原菌为胶孢炭疽菌Colletotrichum gloeosporioides Penz.。该病原菌生长温度范围为10~35℃,最适生长温度为28℃;产孢温度范围为15~32℃,最适产孢温度为32℃;生长p H范围为3~11,最适生长p H为4;可促进菌丝生长并适合产孢的碳源为菊糖,阿拉伯树胶粉抑制菌丝生长但可促进产孢;对菌丝生长及产孢均有促进作用的氮源为酵母粉和牛肉膏,而硫酸铵和硝酸铵则表现抑制作用。     

无花果炭疽病菌的生物学特性及8种杀菌剂对其抑制作用

 通过十字交叉法和血球计数板计数法测定无花果炭疽病菌的生物学特性及其对8种杀菌剂的敏感性,结果表明,菌丝体生长、产孢的最适温度都为25～30℃,最适生长pH值为5~6。同时明确了不同碳、氮源对无花果炭疽病菌菌落生长及其产孢的影响。氟硅唑和戊唑醇抑制毒力最高,EC₅₀分别为0.025 7 μg·mL^-1和0.183 μg·mL^-1,分别为醚菌酯的1 395.55和195.99倍;其次为苯醚甲环唑和异菌脲,EC₅₀分别为0.187 9 μg·mL^-1和0.434 9 μg·mL^-1;醚菌酯毒力较小,不适合用于防治无花果炭疽病。     

云南葡萄产区葡萄炭疽病病原鉴定及致病力分析

为了明确引起云南葡萄产区炭疽病的病原种类,利用形态鉴定和特异性引物分子检测相结合的方法对从云南省主要葡萄产区采集的60株炭疽病菌菌株进行了鉴定。葡萄炭疽病菌菌株的菌落形态和生长速率与对照菌株尖孢炭疽菌Colletotrichum acutatum差异不明显,但其分生孢子大小显著小于尖孢炭疽菌,附着胞深褐色,球形或不规则形。胶孢炭疽菌Colletotrichum gloeosporioides特异性引物CgInt/ITS4从供试葡萄炭疽病菌菌株基因组DNA中扩增出1条约500 bp的特异性条带,而尖孢炭疽菌特异性引物CaInt2/ITS4对葡萄炭疽病菌无扩增条带。研究表明,引起云南葡萄主产区炭疽病的病原为胶孢炭疽菌;供试胶孢炭疽菌对红提葡萄均有致病力,但菌株致病力差异较大,对番茄和草莓存在交叉侵染的能力,且对多菌灵的敏感性较尖孢炭疽菌高。     

柿树炭疽病菌的生物学特性及几种杀菌剂对其的抑制作用

通过十字交叉法和血球计数板法测定了柿树炭疽病菌Collelotrichum gloeosporioides的生物学特性;采用菌丝生长速率法测定了7种常用杀菌剂对该病菌的抑制作用;选择5种市售杀菌剂进行了田间药效试验。结果表明:该病菌菌丝生长的最适温度为20~30 ℃,最适产孢温度为25~30 ℃,最适生长的pH值为5~6;在7种常用杀菌剂中,多菌灵、苯醚甲环唑、戊唑醇和氟硅唑对柿树炭疽病菌具有明显的抑制作用,EC50值分别为0.118 8、0.138 0、0.159 9和1.297 2 μg/mL。田间试验结果显示,25%咪鲜胺乳油600倍液、10%苯醚甲环唑水分散粒剂1 000倍液和25%戊唑醇水乳剂1 500倍液的防治效果分别为89.60%、88.56%、87.82%。     

胶孢炭疽菌生防放线菌gz-8的鉴定及生物活性初步评价

从山东省临沂地区土壤中分离到一株放线菌gz-8,发现其对胶孢炭疽菌Colletotrichum gloeosporioides具有较强的抑制作用,皿内抑菌活性达72.5%。通过形态特征和生理生化测定,结合16S rDNA序列分析,鉴定该菌株为栗褐链霉菌Streptomyces badius。抗菌谱测定结果表明,菌株gz-8对6种植物病原真菌均具有较强的抑菌活性,抗菌谱较广。进一步对菌株gz-8的粗提物进行了生物活性评价,发现其发酵液粗提物可以明显抑制胶孢炭疽菌的菌丝生长（EC50=3.18 μg/mL）和分生孢子萌发（IC50=1.92 μg/mL）,且活性明显高于菌体粗提物;同时发酵液粗提物具有良好的热稳定性及pH稳定性;1 000 mg/L的发酵液粗提物对感染胶孢炭疽病芒果果实的防效达到35.94%,与同浓度下百菌清的防效（34.11%）无显著差异。     

福建省大豆炭疽病病原菌的分离与鉴定

为明确引起福建省大豆炭疽病的病原菌种类,2018—2019年从福建省福州、三明、莆田、泉州和南平市采集具有大豆炭疽病症状的大豆豆荚,采用组织分离法分离病原菌,结合形态学特征和多基因系统发育分析对病原菌进行鉴定。结果表明,从采集的豆荚样本中共分离获得152株炭疽菌菌株,经形态学初步鉴定分属于3种类型,从3种类型的病原菌中分别选取3株代表菌株进行多基因系统发育分析,这9株代表菌株分别与Colletotrichum plurivorum、平头炭疽菌C.truncatum和胶孢炭疽菌C.gloeosporioides聚在一起。形态学和多基因系统发育分析结果表明,引起福建省大豆炭疽病的病原菌有3种,分别为C.plurivorum、平头炭疽菌和胶孢炭疽菌,其占比分别为10.53%、50.00%、39.47%。致病性测定结果表明,C.plurivorum、平头炭疽菌和胶孢炭疽菌对大豆均有致病作用,但不同的菌株致病力存在差异。表明平头炭疽菌是引起福建省大豆炭疽病的主要病原菌,而C.plurivorum为引起我国大豆炭疽病的新纪录种。     

辽宁省西瓜炭疽病病原菌鉴定及生物学特性研究

针对西瓜炭疽病病原菌进行了鉴定及其生物学特性研究。通过病原菌形态学鉴定,柯赫氏法则证病,rDNA ITS测序,鉴定该病致病菌为瓜类炭疽菌（Colletotrichumorbiculare）。生物学特性测定结果表明：病菌菌丝生长最适培养基为PSA; 最适碳、氮源分别为蔗糖和NH4Cl; 最适温度为25℃; 最适pH为6; 黑暗条件利于病菌生长。病菌产孢最适培养基为西瓜茎叶煎汁培养基; 最适碳、氮源分别为蔗糖和NH42SO4; 最适温度为25℃; 最适pH为6; 黑暗条件利于孢子产生。菌丝致死温度为55℃,10min; 分生孢子致死温度为52℃,10min。     

苦瓜炭疽病病原菌鉴定

苦瓜炭疽病是江西农业大学周边菜地重要病害之一，目前，国内外对该病害研究较少。本文按照柯赫氏法则对该病害病原进行了分离培养鉴定，鉴定结果为瓜类炭疽菌Colletotrichum orbiculared（Berk．＆ Mont）Arx。文中对病害的发生情况、病菌培养和形态特征等性状作了记述。     

油茶炭疽病病原鉴定及绿色防治药剂筛选

油茶炭疽病是我国油茶重要病害,为筛选出有效的防治药剂,本研究从我国油茶主产区采集具有典型症状的炭疽病病叶分离病原菌,结合病原菌形态特征和多基因系统发育分析对其进行了系统鉴定,并采用菌丝生长速率法测定病原菌对化学杀菌剂敏感性。结果表明,本研究分离出的17株油茶炭疽病菌均属于刺盘孢属胶孢复合群(Gloeosporioides complex)真菌,包括山茶刺盘孢Colletotrichum camelllae、果生刺盘孢Colletotrichum fructicola、暹罗刺盘孢Colletotrichum siamense、胶孢刺盘孢Colletotrichum gloeosporioides、卡瓦刺盘孢Colletotrichum kahawae、哈锐刺盘孢Colletotrichum horri等。测试的13种杀菌剂中,油茶炭疽病菌对咪鲜胺药剂最敏感,平均EC₅₀值为0.035 μg/mL;其次为氟啶胺、多菌灵、咯菌腈、吡唑醚菌酯、苯醚甲环唑、氟环唑、苯并烯氟菌唑和戊唑醇,平均EC₅₀依次为0.076、0.169、0.202、0.243、0.342、0.490、0.534和1.401 μg/mL;此外,测试的油茶炭疽病菌对嘧菌酯、啶酰菌胺、氟醚菌酰胺和氟吡菌酰胺四种药剂均不敏感,平均EC₅₀值大于100 μg/mL。     

Molecular identification of Colletotrichum gloeosporioides causing yam anthracnose in Nigeria

Four forms of Colletotrichum representing three distinct virulence phenotypes were found associated with foliar anthracnose of yam in Nigeria: the highly virulent (= severity of disease) slow-growing grey (SGG); the moderately virulent fast-growing salmon (FGS); the weakly virulent fast-growing grey (FGG); and the moderately virulent fast-growing olive (FGO) morphotype. Isolates of the four forms were identified as C. gloeosporioides , based on morphology. The reaction of monoconidial cultures on casein hydrolysis medium (CHM), PCR-RFLP and sequence analysis of the internal transcribed spacer region of the ribosomal DNA (ITS1-5·8S-ITS2) were used to establish the identity of the yam anthracnose pathogen(s). All yam isolates were distinguished from C. acutatum by the absence of protease activity on CHM. On ITS PCR and enzymatic digestion of PCR products, all FGS, FGO and SGG isolates produced RFLP patterns identical to those of C. gloeosporioides reference isolates, while FGG isolates revealed unique ITS RFLP banding patterns. Sequence analysis of the ITS1 region and of the entire ITS region revealed that SGG, FGS and FGO isolates were highly similar (98–99% nucleotide identity) and showed 97–100% identity to C. gloeosporioides . Less than 93% similarity of these fungal isolates to reference C. acutatum and C. lindemuthianum isolates was observed. The molecular study confirmed that foliar anthracnose of yam is caused by C. gloeosporioides . While a high similarity was found among most C. gloeosporioides fungi from yam, isolates of the FGG form did not cluster with any previously described Colletotrichum species, and probably represent a distinct species.     

四川省山药黑斑病病原菌的鉴定

为鉴定引起山药茎蔓黑斑病的病原菌,本研究利用柯赫氏法则验证其致病性,根据菌株的形态特征和基于rpb2基因序列分析进行分类鉴定,并初步测定其寄主范围。结果显示:分离的2株病原菌分生孢子椭圆形,单胞,(3.9～5.1)μm×(1.8～2.7)μm,产孢细胞瓶梗形,分生孢子器近圆形或梨形,无刚毛,具孔口,有时孔口延长形成乳突,NaOH颜色反应呈阳性,参考Phoma鉴定手册,可将其划分到section Peyronellaea。将供试菌的rpb2序列进行BLAST比对分析,其与GenBank中的Epicoccum latusicollum具有最高的同源性,支持率为97%,结合形态学特征可将其鉴定为E.latusicollum,该菌经人工接种后也能侵染番茄、辣椒和花菜幼苗。     

河南省一例西瓜果实腐烂病病原鉴定

西瓜营养丰富、甘味多汁、清爽解渴,堪称“夏季水果之王”.我国西瓜生产面积和产量均居世界第一,西瓜已成为我国重要的经济作物之一.近年来,由于连作重茬,导致西瓜病害日益加重,严重制约西瓜产业的发展[1].瓜类炭疽菌[Colletotrichum orbiculare( Berk.&Mont.)Arx]引起的西瓜腐烂病几乎在世界各瓜区均有发生,一旦发病就会造成严重的损失[2].以往对炭疽病病原的分类、鉴定,主要是基于形态学特性、致病性测定的传统方法.     

Epidemiology,histopathology and aetiology of olive anthracnose caused by Colletotrichum acutatum and C. gloeosporioides in Portugal

Anthracnose is an important disease affecting mature olive fruits, causing significant yield losses, and poor fruit and oil quality. In Portugal, high anthracnose incidence was recorded during 2003–2007 with 41% of 908 orchards surveyed displaying disease symptoms. In another 14% of the orchards, the pathogen was recorded in symptomless plants. Disease severity was on average 36%, frequently reaching 100%. In Portugal, anthracnose is endemic to neglected orchards of susceptible cultivars, but under favourable conditions it can also severely affect less susceptible cultivars. Pathogens were genetically heterogeneous, with Colletotrichum acutatum genetic group A2 as the most frequent (80%), followed by group A4 (12%) and group A5 along with C. gloeosporioides (3–4%), while groups A3 and A6 of C. acutatum were sporadic. Important geographic variations were observed in the frequencies of these populations, accompanied by year‐to‐year populational shifts. Epidemiology and histopathology studies showed the presence of the pathogens on vegetative organs year‐round, particularly on olive leaves and branches, and on weeds. These represent inoculum reservoirs where secondary conidiation occurs, and conidia are then dispersed by spring rains reaching flowers and young fruits or by autumn rains reaching pre‐mature fruits. Unripe fruits were colonized without showing symptoms up to penetration of the cuticle, but further colonization and symptom production was completed only as fruits matured. These findings challenge current control practices, particularly the timing of fungicide treatment, and contribute to improved disease management.     

4种杀菌剂防治淮山炭疽病药效试验

选用4种杀菌剂防治淮山炭疽病田间药效筛选试验。结果表明:10%世高WG和80%代森锰锌WP(大生M-45)校正防治效果在药后7 d为86.7%、72.9%;药后10 d为78.0%、63.83%。其中10%世高WG具有内吸性能好、渗透性强,双向传导杀菌活性,持药期长和耐雨水性好等特点,在生产上可作为防治淮山炭疽病首选药剂。