灰葡萄孢菌引起的向日葵叶斑和盘腐及热胁迫对病菌菌核形成的影响  

Leaf blotch and head rot caused by Botrytis cinerea on sunflower and effect of heat stress on sclerotium formation of the pathogen

作　　者：白滨[1,2] 文朝慧 何苏琴 柳利龙[2,6] 张爱琴 王青 BAI Bin;WEN Zhaohui;HE Suqin;LIU Lilong;ZHANG Aiqin;WANG Qing(Gansu Academy of Agricultural Sciences,Lanzhou 730070,Gansu,China;Laboratory of Quality&Safety Risk Assessment for Agro-products(Lanzhou),Ministry of Agriculture and Rural Affairs,Lanzhou 730070,Gansu,China;Technical Centre of Lanzhou Customs District,Lanzhou 730010,Gansu,China;Institute of Plant Protection,Gansu Academy of Agricultural Sciences,Lanzhou 730070,Gansu,China;Scientific Observing and Experimental Station of Crop Pests in Tianshui,Ministry of Agriculture and Rural Affairs,Tianshui 741200,Gansu,China;Institute of Agricultural Quality Standards and Testing Technology,Gansu Academy of Agricultural Sciences,Lanzhou 730070,Gansu,China)

机构地区：[1]甘肃省农业科学院,甘肃兰州730070 [2]农业农村部农产品质量安全风险评估实验室(兰州),甘肃兰州730070 [3]兰州海关技术中心,甘肃兰州730010 [4]甘肃省农业科学院植物保护研究所,甘肃兰州730070 [5]农业农村部天水作物有害生物科学观测实验站,甘肃天水741200 [6]甘肃省农业科学院农业质量标准与检测技术研究所,甘肃兰州730070

出　　处：《微生物学通报》2024年第5期1405-1424,共20页Microbiology China

基　　金：国家农产品质量安全风险评估计划(GJFP20220107)。

摘　　要：【背景】2022年8-9月,从甘肃省兰州市种植的食用向日葵田中采集到零星发病的叶斑和盘腐病害标样。【目的】明确叶斑病和盘腐病的病原及其生物学特性。【方法】采用单孢分离法进行病原菌分离;通过Koch’s法则明确分出病菌的致病性;采用形态学和分子生物学方法对病原菌进行种类鉴定;通过平板法测定试验菌株的适宜生长温度;观察经30℃(生长抑制温度)培养后试验菌株产菌核特性的变化。【结果】分离得到12株菌落形态不同的葡萄孢属菌株。试验菌株SKY-A-SKY-D接种食用向日葵离体叶片和花盘可引起与自然发病相似的症状,接种发病的叶片和花盘病组织上原接种菌的分出率达100%。试验菌株在PDA平板上15℃和20℃培养10-40 d,大型分生孢子无色至淡褐色,单胞,卵圆形、长椭圆形、球形、近球形、棒状或不规则形,[6.0-14.2(-20.1)]μm×[6.0-10.4(-14.9)]μm;菌核表生,黑色,球形、近球形、椭圆形或不规则形,(0.5-11.1)mm×(0.5-5.0)mm;Ⅰ型微菌核红褐色至黑褐色,表生或埋生,(26.9-492.5)μm×(14.9-149.3)μm,微菌核的组成细胞大小为(12.7-35.0)μm×(11.9-25.3)μm;Ⅱ型微菌核近黑色,埋生,(35.8-373.1)μm×(23.9-229.9)μm,微菌核的组成细胞大小为(8.2-16.4)μm×(8.2-14.9)μm;假微菌核由特化的附着胞构成,暗褐色至近黑色,埋生,(32.0-447.8)μm×(19.4-358.2)μm。试验菌株的菌丝适宜生长温度为20-25℃;菌核产生温度为5-25℃。30℃培养7 d后转至20℃培养14 d,可诱导2个不产菌核的菌株SKY-B和SKY-C产生菌核,突变株在继代培养时保持其产菌核能力。BLASTn分析结果显示,试验菌株SKY-A–SKY-D的rDNA-ITS序列(国家微生物科学数据中心登录号为NMDCN00038NP–NMDCN00038NS)与已知灰葡萄孢菌(Botrytiscinerea)序列的相似性达99.8%–100.0%。g3pdh、hsp60和rpb2多基因系统发育分析结果显示,4个试验菌株被聚在B.cinerea的不同亚群里。【结论】引起向日�[Background]In August and September 2022,the plant samples of leaf blotch and head rot were collected from sporadic diseased fields of confectionery sunflower(Helianthus annuus)in Lanzhou,Gansu,China.[Objective]To identify and characterize the pathogens of leaf blotch and head rot.[Methods]We isolated the pathogens by the single spore isolation method,measured the pathogenicity of the isolates based on Koch’s postulates,and identified the isolates by morphological observation and molecular biological methods.Furthermore,we determined the optimum growth temperatures of the isolates by the plate culture method,and the changes of sclerotium formation characteristics of the tested isolates were observed after cultured at 30℃(growth inhibition temperature).[Results]Twelve Botrytis isolates with different colony morphology were isolated from the diseased plant samples.The inoculation of four isolates,SKY-A to SKY-D,on detached sunflower leaves and heads induced the symptoms similar to those of natural diseases in the field,and the re-isolation rates of inoculated isolates from the infected leaves and heads were 100%.When the four isolates were cultured on PDA plates at 15℃and 20℃for 10–40 d,macroconidia were pale to light brown,unicellular,oval,oblong,spherical,subspherical,clavate or irregular shaped,(6.0–14.2(–20.1))μm×(6.0–10.4(–14.9))μm.The sclerotia were superficial,black,spherical,subspherical,ellipse or irregularly shaped,(0.5–11.1)mm×(0.5–5.0)mm.Type-I microsclerotia were reddish brown to dark brown,superficial or submerged,(26.9–492.5)μm×(14.9–149.3)μm,with the cell sizes of(12.7–35.0)μm×(11.9–25.3)μm.Type-II microsclerotia were nearly black,submerged,(35.8–373.1)μm×(23.9–229.9)μm,with the cell sizes of(8.2–16.4)μm×(8.2–14.9)μm.Pseudo-microsclerotia consisting of specialized appressoria were dark brown to nearly black,submerged,(32.0–447.8)μm×(19.4–358.2)μm.The optimal temperature range for mycelial growth and the temperature range for sclerotium for

关 键 词：向日葵 叶斑 盘腐 灰葡萄孢菌 菌核 微菌核 假微菌核 热胁迫 

分 类 号：S435.655[农业科学—农业昆虫与害虫防治]