贵州省食用玫瑰叶斑病病原菌鉴定及其防治

Pathogens Identification and Control of Edible Roses Leaf Spot Diseases in Guizhou

摘要: 为了明确贵州省植物园和黔南州惠水县的墨红月季上叶斑病的病原菌种类，采集月季叶斑病的病样，采用单孢分离法和致病性测定，结合依据形态学结合ITS、GADPH、ACT、CHS-1等多基因序列分析，确定其病原菌分类地位；在此基础上，通过平板稀释和平板对峙法，结合生理生化实验和16S rRNA和gyrA多基因序列分析，筛选和鉴定对月季炭疽病有拮抗作用的拮抗细菌；结合菌丝生长速率法筛月季叶斑病选防治效果较好的化学药剂；选用商品化微生物制剂绿康威、12%腈菌·三唑酮EC + 65%代森锌、64%杀毒矾及生防菌GUHS97进行田间防效实验。结果表明：1) 黑斑病病原菌为蔷薇盘二孢(Marssinina rosae)，炭疽病病原菌博宁炭疽菌(Colletotrichum boninense)；2) 筛选到1株叶际有益芽胞杆菌GUHS97，鉴定该菌为解淀粉芽胞杆菌(Bacillus amyloliquefaciens)；3) 通过室内药剂筛选，3%中生菌素WP、10% 苯醚甲环唑WDG、25%吡唑醚菌酯SC和10%精甲·咯菌腈SC抑制效果最好，EC₅₀值分别为0.8440 mg∙L⁻¹、1.3854 mg∙L⁻¹、1.8667 mg∙L⁻¹和3.5945 mg∙L⁻¹，其次为8%宁南霉素AS和25%溴菌腈EC，EC₅₀值分别为18.9624 mg∙L⁻¹和25.4813 mg∙L⁻¹；将25%吡唑醚菌酯SC和25%溴菌腈EC以1:7、1:8、和1:10比例复配，对月季炭疽病菌具有更强的抑制作用，表现出明显的增效作用；通过田间小区实验表明，GUHS97对黑斑病的相对防效为59.12%，防病效果良好且明显优于化学药剂。以上研究结果为食用月季叶部病害的识别和生物、化学防治提供科学依据。

Abstract: In order to identify the pathogenic species of edible roses leaf spot diseases in Guizhou Botanical Garden and Huishui County, Guizhou Province, China, the samples of leaf spot diseases were collected. By using single spore isolation method and pathogenicity determination, combined with morphological analysis combined with ITS, GADPH, ACT, CHS-1 gens, the taxonomic status of its pathogenic bacteria was determined. On this basis, through plate dilution and plate confrontation method, combined with physiological and biochemical experiments and 16S rRNA and gyrA gens, screening and identification of antagonistic bacteria on edible rose, then combined with the mycelial growth rate method to screen the chemical agents with better control effect on rose leaf spot, commercial microbial preparation Lvkangwei, Myclobutanil 12% EC + Dithane 65% WP, Oxadixyl mancozeb 64% WP and GUHS97 were used for field control effect experiments. The results showed that: 1) The pathogen causing leaf black spot was confirmed to be Marssinina rosae, and the pathogen causing anthracnose was confirmed to be Colletotrichum boninense; 2) In this study, a strain GUHS97 was screened from the healthy rose leaves in the heavily infected field and was identified as Bacillus amyloliquefaciens; 3) Indoor toxicity test showed that 9 fungicides had a certain inhibitory effect on C. boninense. The inhibition effects of zhongshengmycin 3% WP, difenoconazole 10% WDG, pyraclostrobin 25% SC and metalaxyl∙fludioxonil 62.5% SC were higher with the EC₅₀ values of 0.8440 mg∙L⁻¹、1.3854 mg∙L⁻¹、1.8667 mg∙L⁻¹ and 3.5945 mg∙L⁻¹, respectively, followed by ningnanmycin 8% AS and bromothalonil 25% EC, with the EC₅₀ values of 18.9624 mg∙L⁻¹ and 25.4813 mg∙L⁻¹, respectively. Pyraclostrobin 25% SC and bromothalonil 25% EC at the mixture ratio of 1:7, 1:8 and 1:10 showed obvious synergistic effect; 4) Furthermore, GUHS97 exhibited 59.12% control over leaf spot in field experiments and was more effective than other treatments including fungicide application. Overall, these results provide a scientific basis for the identification of edible rose leaf diseases and their control.