两株贝莱斯芽胞杆菌鉴定及其对薄壳山核桃黑斑病病原菌的拮抗效果  

Identification of Two Strains of Bacillus velezensis Isolated from Carya illinoensis Leaf and Their Antagonistic Effects on Pecan Black Spot Pathogen

作　　者：王迪[1,2] 高岩 吴小双 张威 舒金平[2] 张亚波[2] 翟凤艳[1] WANG Di;GAO Yan;WU Xiaoshuang;ZHANG Wei;SHU Jinping;ZHANG Yabo;ZHAI Fengyan(He’nan Institute of Science and Technology,Xinxiang 453003,China;Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou 311400,China;Jiaxing Forestry Technology Extension Station,Jiaxing 314001,China)

机构地区：[1]河南科技学院,新乡453003 [2]中国林业科学研究院亚热带林业研究所,杭州311400 [3]嘉兴市林业技术推广站,嘉兴314001

出　　处：《中国生物防治学报》2022年第6期1572-1581,共10页Chinese Journal of Biological Control

基　　金：浙江省科技计划项目(2019C02024);河南省高等学校重点科研项目(23B210002);浙江省木本油料树种高危病虫害生防菌剂的研发与应用(2022SY03)。

摘　　要：黑斑病为薄壳山核桃重要真菌性病害,为了实现薄壳山核桃黑斑病绿色防控,本研究旨在筛选优良的内生拮抗菌株,为黑斑病的生物防控提供生防资源。通过形态鉴定、生理生化特征测定和16S r DNA与Gyr A基因分析,对薄壳山核桃内生拮抗菌进行鉴定;采用平板对峙法测定内生拮抗菌对黑斑病病原真菌(5属21种26株)菌丝生长的抑制能力,分离筛选到2株高效拮抗菌ZJJDZY和ZJJDYB,鉴定为贝莱斯芽胞杆菌Bacillus velezensis。平板对峙法测定结果表明,2株拮抗菌均对12种12株炭疽菌Colletotrichum spp.、1种4株葡萄座腔菌Botryosphaeriadothidea、2种2株链格孢菌Alternariaspp.、4种7株新拟盘多毛孢Neopestalotiopsisspp.和1种1株拟茎点霉菌Phomopsissp.均具有拮抗效果。其中,对C.henanense PCZJJD18、C.sojae PCAHFY5、N.mesopotamica PCSCLZ2、A.alternata PCAHFY6、Phomopsis PCZJJD14sp.的抑制率为最高,抑制率在57.1%~60.6%,且两株拮抗菌抑菌能力相似;不同浓度的拮抗菌对不同属病原真菌的抑制效果不同,浓度为108CFU/mL时,拮抗效果最好,随着浓度降低,拮抗效果显著减弱。该两个菌株对多种薄壳山核桃黑斑病病原真菌具有拮抗作用,具有较佳的生防潜力。Black spot is an important pathogenic fungi disease infected pecan(Carya illinoensis).To develop green and efficient biological control method for pecan black spot disease,effective antagonistic endophytic strains were screened and identified as biocontrol resources against black spot disease.Based on morphological identification,physiological and biochemical characteristics,and phylogenetic analysis of 16S rDNA and GyrA gene,entophytic antagonistic strains isolated from pecan leaf were identified and their inhibition effects on mycelium growth of 26isolates fungi caused black spot disease was detected by plate confrontation method.Two antagonistic entophytic strains screened from pecan leaf,named ZJJDZY and ZJJDYB,were identified as Bacillus velezensis.Both ZJJDZY and ZJJDYB showed significant antagonistic effects on 12 Colletotrichum species,1 Botryosphaeria species,2 Alternaria species,4 Neopestalotiopsis species and 1 Phomopsis species.ZJJDZY and ZJJDYB demonstrated higher inhibition activity against PCZJJD18 C.henanense,PCAHFY5 C.sojae,PCSCLZ2 N.saprophytica,PCAHFY6 A.alternate and PCZJJD14 Phomopsis sp.,with the inhibitory rates ranging from 57.1% to 60.6%,and the antibacterial ability of ZJJDZY and ZJJDYB was similar.Inhibition effects of ZJJDZY and ZJJDYB against different pathogenic fungi varied significantly at different concentrations,and the best antagonistic effect was shown at the concentration of 10~8 CFU/mL.While,with the decrease of concentration,the antagonistic effect weakened significantly.Two entophytic antagonistic bacteria,ZJJDZY and ZJJDYB showed significant antagonistic effects against various pathogenic fungi causing black spot disease,which has the potential as biocontrol resource.

关 键 词：薄壳山核桃 黑斑病 鉴定 贝莱斯芽胞杆菌 生物防治 

分 类 号：S476[农业科学—农业昆虫与害虫防治]