Preliminary Study on the Synergistic Effects of Bacillus subtilis KXZ-33 and Chemical Pesticides against Cotton Fusarium wilt

doi:10.6048/j.issn.1001-4330.2022.05.021

Abstract

Abstract:

【Objective】 Through the combination of biocontrol bacteria and chemical fungicides, the formula combination that can effectively control cotton Fusarium wilt was screened out, and the seed treatment was carried out with a view to providing technical support for the green prevention and control of cotton disease. 【Methods】 Fusarium oxysporum, the main pathogen of Fusarium wilt, was selected by indoor plate confrontation test. The virulence of chemical fungicides was tested by the method of toxic media, and the chemical fungicides used in coordination with biocontrol bacteria were screened by compatibility test. The control effect was evaluated by pot experiment. 【Results】 All the six biocontrol bacteria had inhibitory effects on Fusarium oxysporum, with the inhibition rates ranging from 56.69% to 79.53%, and Bacillus subtilis KXZ-33 had the best inhibition rate of 79.53%. The EC₅₀ of Thiobacillam-methyl, Carbendazim and Formetrix were 15.840,1, 18.018,0 and 12.794,0 mg/L, respectively. The maximum compatibility of KXZ-33 with 20 mg/L Thiobacillam-methyl was 90.46%, and the viable bacteria count was 2.18×10⁷ cfu/mL.The control effect of KXZ-33 and 10 mg/L Carbendazim on 7 d was 90.5%, and decreased to 27.0% on 28 d. KXZ-33 combined with 10 mg/L Thiobacillam-methyl was 87.7% for 7 days and 43.7% for 28 days. 【Conclusion】 Bacillus subtilis KXZ-33 seed treatment and spraying 50% methyl Thiobacillam-methyl 10 mg/L to control cotton Fusarium wilt disease can improve the control effect, make up for biocontrol bacteria in agricultural production of unstable control effect and at the same time reduce the use of chemical pesticides, thus further developing service production.

Key words: cotton; Fusarium wilt; biological control; chemical control; collaborative prevention and control

摘要：

【目的】分析生防细菌与化学杀菌剂复配效果,筛选出有效防治棉花枯萎病的配方组合处理种子,为棉花病害绿色防控提供技术支撑。【方法】以棉花枯萎病主要致病菌尖孢镰刀菌为研究对象,采用室内平板对峙试验筛选拮抗细菌,含毒介质法对化学杀菌剂进行毒力测定,相容性测定筛选出与生防细菌协同使用的化学杀菌剂;评价盆栽试验防病效果。【结果】6种生防细菌对尖孢镰刀菌均有抑制作用,抑制率在56.69%~79.53%,其中枯草芽孢杆菌KXZ-33对其抑制率最好,为79.53%。化学药剂甲基硫菌灵、多菌灵、福美双抑菌效果较好,EC₅₀分别为15.840 1、18.018 0、12.794 0 mg/L。KXZ-33与20 mg/L的甲基硫菌灵的相容性最高为90.46%,活菌数为2.18×10⁷ cfu/mL。KXZ-33与10 mg/L多菌灵在7 d防治效果最高为90.5%,28 d下降为27.0%;KXZ-33与10 mg/L甲基硫菌灵协同防治棉花枯萎病7 d防效为87.7%,28 d防病效果为43.7%。【结论】枯草芽孢杆菌KXZ-33种子处理与喷施50%甲基硫菌灵10 mg/L防治棉花枯萎病可以起到提高防效,弥补生防细菌在农业生产中的防效不稳定的同时可以减少化学农药的使用量。

关键词: 棉花, 枯萎病, 生物防治, 化学防治, 协同防治

CLC Number:

S513
S435.62

ZHAO Yingying, ZHANG Jungao, LI Jin, LIANG Jing, GAO Xiangyu, GU Aixing, LIE Bin. Preliminary Study on the Synergistic Effects of Bacillus subtilis KXZ-33 and Chemical Pesticides against Cotton Fusarium wilt[J]. Xinjiang Agricultural Sciences, 2022, 59(5): 1216-1222.

赵莹莹, 张军高, 李进, 梁晶, 高翔宇, 顾爱星, 雷斌. 枯草芽孢杆菌KXZ-33与化学药剂协同防控棉花枯萎病的效果评价[J]. 新疆农业科学, 2022, 59(5): 1216-1222.

Figures/Tables 4

Table 1 The inhibition rate of bacteria from cotton roots against F.oxysporum

处理 Treatment	抑菌率 Inhibition rate(%)
KXZ-33	79.53±10.20^a
KX-4	58.66±8.66^b
A57	68.50±7.3 $9 a b$
UB17-1	75.51±9.23^a
Twt34	71.61±6.1 $7 a b$
Hwt34	56.69±3.02^c

Table 1 The inhibition rate of bacteria from cotton roots against F.oxysporum

处理 Treatment	抑菌率 Inhibition rate(%)
KXZ-33	79.53±10.20^a
KX-4	58.66±8.66^b
A57	68.50±7.3 $9 a b$
UB17-1	75.51±9.23^a
Twt34	71.61±6.1 $7 a b$
Hwt34	56.69±3.02^c

Table 2 Laboratory toxicity test of chemical pesticides to F.oxysporum

化学杀菌剂 Fungicide	回归方程 Regression equation	EC₅₀ (mg/L)	R² Correlation coefficient
50%甲基硫菌灵可湿性粉剂 Thiophanate-methyl	y=0.609 8x+3.315 4	15.840 1	0.997 9
40%多菌灵悬浮剂 Carbendazim	y=1.221 6x+1.467 9	18.018 0	0.995 1
43%氟菌肟菌酯乳油 Fluomycetin	y=0.177 3x+4.070 8	188.827 7	0.951 2
80%代森锰锌可湿性粉剂 Mancozeb	y=0.258 2x+3.040 2	1 987.788 6	0.972 8
80%乙蒜素乳油 Ethylicin	y=0.668 6x+1.767 8	125.748 0	0.969 2
30%噁霉灵可湿性粉剂 Hymexazo	y=1.047 4x+0.533 0	89.221 3	0.997 3
50%福美双可湿性粉剂 Formetrix	y=0.550 3+3.597 3	12.794 0	0.978 6

Fig.1 Compatibility analysis of KXZ-33 with chemical agents

Table 4 Synergistic control effect of different chemical fungicides and KXZ-33 on Fusarium wilt of cotton

处理 Treatment	7 d		14 d		21 d		28 d
处理 Treatment	病情指数 Condition index	防病效果 Control effect(%)	病情指数 Condition index	防病效果 Control effect(%)	病情指数 Condition index	防病效果 Control effect(%)	病情指数 Condition index	防病效果 Control effect (%)
KXZ-33 10⁷cfu/mL	0.11±0.08^c	72.4±7.3^ab	0.36±0.09^bc	45.2±4.2^ab	0.46±0.04^bcd	37.0±4.3^ab	0.62±0.04^b	20.2±5.5^bc
50%甲基硫菌灵 20 mg/L	0.31±0.07^ab	21.0±0.1^b	0.35±0.09^bc	47.1±8.0^ab	0.38±0.03^cd	48.9±4.7^a	0.50±0.02^c	35.6±0.3^ab
KXZ-33 10⁷cfu/mL+20 mg/L 甲基硫菌灵	0.05±0.04^c	87.7±0.1^a	0.28±0.09^b	57.7±3.7^a	0.32±0.08^d	56.3±5.8^a	0.44±0.06^c	43.7±8.0^a
20 mg/L多菌灵	0.20±0.06^bc	50.0±0.1^ab	0.40±0.03^bc	38.4±5.6^ab	0.58±0.03^ab	20.5±3.4^b	0.64±0.04^b	17.8±6.4^cd
KXZ-33 10⁷ cfu/mL+10 mg/L 多菌灵	0.04±0.03^c	90.5±0.1^a	0.42±0.09^ab	35.2±2.0^ab	0.56±0.14^abc	24.3±3.1^b	0.57±0.123^a	27.0±4.6^bc
CK	0.40±0.04^a	-	0.66±0.06^a	-	0.73±0.08^a	-	0.78±0.06^a	-

References 20

[1]	Shaapulatov U M, Buriev Z T, Ulloa M. Characterization of small RNAs and their targets from Fusarium oxysporum infected and noninfected cotton root tissues[J]. Plant Molecular Biology Reporter, 2016, 34(3): 698-706. DOI URL
[2]	蔡梦杭. 新疆主要棉区棉花黄萎病菌培养特性及致病力分化研究[D]. 石河子: 石河子大学, 2019.
	CAI Menghang. Cultural Characteristics and Pathogenicity Differentiation among Strains of Verticillium dahliae from Cotton in Xinjiang[D]. Shihezi: Shihezi University, 2019.
[3]	朱琦, 张春竹, 顾爱星. 3株棉花黄萎病菌拮抗菌株生防促生效果研究[J]. 新疆农业大学学报, 2019, 42(6): 418-423.
	ZHU Qi, ZHANG Chunzhu, GU Aixing. Study on Biocontrol and Growth Promotion Effects of 3 Antibacterial Strains against Verticillium dahliae in Cotton[J]. Journal of Xinjiang Agricultural University, 2019, 42(6): 418-423.
[4]	曾艳玲, 刘韩, 黎肇家. 生防芽孢杆菌与杀菌剂的筛选复配及对栾树枝枯病防控效果[J]. 四川农业大学学报, 2021, 39(1): 55-62.
	ZENG Yanling, LIU Han, LI Zhaojia. Screening and Compounding of Biocontrol Bacillus and Fungicide and Their Control Effect on Branch Withered Disease on Koelreuteria paniculata[J]. Journal of Sichuan Agricultural University, 2021, 39(1): 55-62.
[5]	陈丽华, 何鹏飞, 袁德超. 一种防治棉花黄萎病的生物复合种衣剂的研制[J]. 棉花学报, 2018, 30(3): 282-291.
	CHEN Lihua, HE Pengfei, YUAN Dechao. Development of a Biological Complex Seed-Coating Agent to Control Verticillium Wilt[J]. Cotton Science, 2018, 30(3): 282-291.
[6]	Glare Travis R, Gwynm Roma L, Morandiez Maria E. Development of Biopesticides and Future Opportunities[J]. Methods in Molecular Biology, 2016, 1477: 211. DOI PMID
[7]	王蕊, 王腾, 李二峰. 生防芽孢杆菌在植物病害领域的研究进展[J]. 天津农学院学报, 2021, 28(4): 71-77.
	WANG Rui, WANG Teng, LI Erfeng. Research advances of biocontrol Bacillus in the field of plant diseases[J]. Journal of Tianjin Agricultural University, 2021, 28(4): 71-77.
[8]	黄慧婧, 罗坤. 芽孢杆菌与杀菌剂复配防治植物病害的研究进展[J]. 微生物学通报, 2021, 48(3): 938-947.
	HUANG Huijing, LUO Kun. Research progress on the control of plant diseases by the combination of Bacillus and fungicides[J]. Microbiology China, 2021, 48(3): 938-947.
[9]	杨秀荣, 孙淑琴, 田涛. 生防细菌B579与多菌灵协同防治立枯病的效果初探[J]. 山东农业科学, 2011,(11): 83-85.
	YANG Xiurong, SUN Shuqin, TIAN Tao. Preliminary Study on Synergy Control Effect of Biocontrol Bacterium B579 and Carbendazim on Rhizoctonia solani[J]. Shandong Agricultural Sciences, 2011,(11): 83-85.
[10]	蔡高磊, 张凡, 欧阳友香. 贝莱斯芽孢杆菌研究进展[J]. 北方园艺, 2018,(12): 162-167.
	CAI Gaolei, ZHANG Fan, OUYANG Youxiang. Research Advances of the Rare and Endemic Plant-Kmeria septentrionalis Dandy[J]. Northern Horticulture, 2018,(12): 162-167.
[11]	谷春艳, 王学峰, 苏贤岩. 解淀粉芽孢杆菌WH1G与氟啶胺协同他防治草莓灰霉病[J]. 农药, 2017, 56(12): 932-936.
	GU Chunyan, WANG Xuefeng, SU Xianyan. Synergistic Effect of Antagonistic Bacteria WH1G and Fluazinam against Strawberry Gray Mould[J]. Agrochemicals, 2017, 56(12): 932-936.
[12]	贺惠文, 纪兆林, 徐敬友. 地衣芽孢杆菌W10与化学药剂复配对桃褐腐病菌抑制活性的增效作用[J]. 亚热带植物科学, 2017, 46(4): 391-393.
	HE Huiwen, JI Zhaolin, XU Jingyou. Synergistic Effects of the Combination of Bacillus licheniformis W10 and Chemical Fungicides on Monilinia fructicola[J]. Subtropical Plant Science, 2017, 46(4): 391-393.
[13]	Soum Sanogo, Jinfa Zhang. Resistance screening techniques and disease management for Fusarium wilt in cotton[J]. Euphytica, 2016, 207(2): 255-271. DOI URL
[14]	周茂超, 黄艳娜, 段赛菲. 微生物种衣剂的研制及其对玉米苗期生长的影响. 中国农业科技导报. 2021, 23(4): 110-118.
	ZHOU Maochao, HUANG Yanna, DUAN Saifei. Development of Microbial Seed Coating Agents and Their Effects on the Growth of Maize Seedlings[J]. Journal of Agricultural Science and Technology, 2021, 23(4): 110-118.
[15]	Cheng Xingkaii, Ji Xiaoxue, Ge Yanahen, et al. Characterization of Antagonistic Bacillus methylotrophicus Isolated From Rhizosphere and Its Biocontrol Effects on Maize Stalk Rot[J]. Phytopathology, 2019, 109(4): 571-581. DOI PMID
[16]	李海薇, 侯仙, 王梦瑶. 海岛棉枯萎病病原菌分离及室内药剂筛选[J]. 中国农学通报, 2019, 35(9): 68-74.
	LI Haiwei, HUI Can, WANG Mengyao. Pathogen isolation of fusarium wilt of island cotton and indoor fungicides screening[J]. Chinese Agricultural Science Bulletin, 2019, 35(9): 68-74.
[17]	林司曦, 吴小芹, 丁晓磊. 短小芽孢杆菌Bacillus pumilus HR10增殖扩繁培养基组分优化[J]. 微生物学杂志, 2014, 34(6): 22-28.
	LIN Sixi, WU Xiaoqin, DING Xiaolei. Optimization of sporulation medium and fermentation conditions for Bacillus pumilus HR10[J]. Journal of Microbiology, 2014, 34(6): 22-28.
[18]	柳自清, 张博然, 刘叶. 短小芽孢杆菌KX-33做种衣剂对棉花枯萎病的的盆栽防效[J]. 新疆农业大学学报, 2020, 43(5): 323-329.
	LIU Ziqing, ZHANG Boran, LIU Ye. Effect of Bacillus pumilus KX-33 seed coating agent on verticillium wilt of cotton[J]. Journal of Xinjiang Agricultural University, 2020, 43(5): 323-329.
[19]	刘邮洲, 陈志谊, 刘永峰. 枯草芽孢杆菌sf628和米鲜胺锰盐协同作用防止番茄枯萎病[J]. 江苏农业学报, 2011, 27(6): 249-253.
	LIU Youzhou, CHEN Zhiyi, LIU Yongfeng. Synergistic efficacy of Bacillus subtilis sf 628 and prochloraz-manganese on tomato fusarium wilt[J]. Jiangsu Journal of Agricultural Sciences, 2011, 27(6)249-253.
[20]	崔凯. 甲基硫菌灵及其代谢物多菌灵防控黄瓜枯萎病发生的根际微生物效应[D]. 北京: 中国农业科学院, 2021.
	CUI Kai. The Effects of Thiophanate-methyl and Its Metabolite Carbendazim on the Cucumber Rhizosphere Microbiota in the Control of Cucumber Fusarium Wilt[D]. Beijing: Chinese Aacademy of Agricultural Sciences, 2021.