枯草芽孢杆菌Czk1与化学杀菌剂复配对橡胶树炭疽病菌的抑菌活性

谢立 贺春萍 梁艳琼 李锐 巩佳莉 翟纯鑫 吴伟怀 易克贤

摘要：【目的】探究枯草芽孢杆菌与化学杀菌剂协同防治橡胶炭疽病的可行性，构建由生防菌与化学杀菌剂组成的菌药复配剂，为橡胶炭疽病的可持续防治提供科学依据。【方法】以6种化学杀菌剂（咪鲜胺、百菌清、根康、嘧霉胺、丙环唑和嘧菌酯）及2株菌株（枯草芽孢杆菌Czk1和橡胶胶孢炭疽菌RC178）为试验材料，采用菌丝生长速率法测定6种化学杀菌剂对RC178的室内毒力，稀释平板涂布法测定6种杀菌剂与Czk1的相容性，抑菌圈法测定Czk1对RC178的室内毒力，Horsfall法明确菌药的复配方案。【结果】室内毒力测定研究结果显示，6种杀菌剂中咪鲜胺对RC178的抑菌效果最好，其有效抑制中浓度（EC50）仅为0.0778 μg/mL，其次为百菌清和根康，EC50分别为0.4694和0.4733 μg/mL。化学杀菌剂与Czk1的相容性测定结果表明，嘧霉胺、嘧菌酯、丙环唑、根康和咪鲜胺与Czk1的相容性均较好，其中根康在中高浓度下还能促进Czk1菌落数量的增长。将浓度为4.75×107 CFU/mL的Czk1分别与浓度为10.4733 μg/mL的根康和浓度为0.0778 μg/mL的咪鲜胺混配，Czk1∶根康=7∶3（v/v）时对RC178抑制的增效作用最强，增效比率（IR）为1.36;Czk1∶咪鲜胺=8∶2（v/v）时对RC178抑制的增效作用最强，IR为1.65。【结论】枯草芽孢杆菌Czk1与咪鲜胺和根康具有较好的相容性，Czk1发酵液分别与咪鲜胺和根康复配对橡胶炭疽病菌RC178的抑菌活性具有明顯增效作用，具有开发成菌药制剂的潜力。

关键词： 枯草芽孢杆菌;橡胶炭疽病;化学杀菌剂;协同防治

中图分类号： S476;S435.76                        文献标志码： A 文章编号：2095-1191（2020）10-2480-08

Antimicrobial activity of Bacillus subtilis Czk1 compounded with chemical fungicides against Colletotrichum gloeosporioides

XIE Li1，2， HE Chun-ping1*， LIANG Yan-qiong1， LI Rui1，  GONG Jia-li3，

ZHAI Chun-xin3， WU Wei-huai1，YI Ke-xian1

（1Environment and Plant Protection Institute， Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Integrated Pest Management on Tropical Grops， Ministry of Agriculture and Rural Affairs/Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests， Haikou  571101， China; 2Institute of Forestry， Hainan University，Haikou  570228， China; 3China College of Plant Protection， Nanjing Agricultural University， Nanjing  210095， China）

Abstract：【Objective】To explore the feasibility of synergistic control of rubber anthracnose by Bacillus subtilis and chemical fungicides， and construct a bacteria-fungicide compound composed of bio-control bacteria and chemical fungicides to provide a basis for the sustainable control of rubber anthracnose. 【Method】Six chemical fungicides， prochloraz， chlorothalonil， Genkang， pyrimethanil， propiconazole and azoxystrobin， and two strains of B. subtilis Czk1 and Colletotrichum gloeosporioides RC178 were used as test materials. The hypha growth rate method was used to determine the indoor toxicity of six chemical fungicides to RC178， the dilution plate coating method was used to determine the compatibility of six fungicides with Czk1， the inhibition zone method was used to determine the indoor toxicity of Czk1 to RC178， Horsfall method to clarify the compound plan of bacteria and medicine. 【Result】The results of laboratory toxicity testing showed that prochloraz had the best antibacterial effect on RC178 among the six fungicides， with an median effect concentration（EC50） of only 0.0778 μg/mL， followed by chlorothalonil and Genkang with EC50 of 0.4694 and 0.4733 μg/mL.Compatibility test results of chemical fungicides and Czk1 showed that pyrimethanil， azoxystrobin， propiconazole， Gen-kang and prochloraz had good compatibility with Czk1. Genkang at medium to high concentrations promoted the growth of the number of Czk1 colonies. Mixing Czk1 at  concentration of 4.75×107 CFU/mL with Genkang at concentration of 10.4733 and mixing 4.75×107 CFU/mL ber of Czk1 and prochloraz at 0.0778 μg/mL. When Czk1∶Genkang=7∶3（v/v）， the synergistic effect on the inhibition of RC178 was the strongest， the synergistic ratio（IR） was 1.36; when Czk1∶prochlorazide=8∶2（v/v）， the synergistic effect on the inhibition of RC178 was the highest， and IR was 1.65. 【Conclusion】B. subtilis Czk1 has good compatibility with prochloraz and Genkang. Czk1 fermentation liquor with prochloraz and with Genkang have obvious synergistic effect on the antibacterial activity of C. gloeosporioides RC178， which have the potential to develop a bacterial medicine preparation potential.

Key words： Bacillus subtilis; rubber anthracnose; chemical fungicide; synergistic control

Foundation item： National Key Research and Development Program of China（2018YFD0201100）; National Natural Rubber Industry Technical System Construction Special Project（CARS-33-BC1）

0 引言

【研究意义】橡胶树炭疽病主要由胶孢炭疽菌（Colletortrichum gloeosporioides Penz. Sace）和尖孢炭疽菌（C. acutatum Simmons）引起，是橡胶树上的一种主要叶部病害（李继锋，2010）。……