Development of a Wettable Powder Formulation of Bacillus subtilis C08 and Its Biocontrol Efficacy Against Tomato Early Blight

doi:10.7525/j.issn.1673-5102.2026.02.009

Abstract

Abstract:

Tomato early blight， caused by Alternaria solani， represents a significant disease that severely impacts both the yield and quality of tomatoes， primarily due to its short latent period and rapid reinfection cycle. In contrast to conventional chemical control methods， biological control offers considerable potential due to its environmentally friendly and sustainable attributes. In this study， a strain of Bacillussubtilis， designated as C08， demonstrating strong antagonistic activity against A.solani， was isolated from the rhizosphere soil of healthy tomato plants. The fermentation conditions were optimized， and a wettable powder formulation was subsequently developed using the fermentation broth as the active ingredient. The results indicated that the formulation achieved high efficacy in controlling tomato early blight， significantly promoted plant growth， enhanced the activities of antioxidant enzymes such as superoxide dismutase（SOD） and peroxidase（POD）， and improved soil enzyme activities such as sucrase and urease. These findings suggest that strain C08 holds substantial promise as a biocontrol agent for the sustainable management of tomato early blight.

Key words: Bacillus subtilis, tomato early blight, wettable powder, biological control

CLC Number:

S476.1

Yunxin XU, Yuyan CONG, Yao ZHANG, Aoxue WANG. Development of a Wettable Powder Formulation of Bacillus subtilis C08 and Its Biocontrol Efficacy Against Tomato Early Blight[J]. Bulletin of Botanical Research, 2026, 46(2): 294-309.

Figures/Tables 22

Table 1

Fig.1

Fig.2

Table 2

Fig.3

Fig.4

Fig.5

Table 3

Table 4

Table 5

Table 6

Table 7

Table 8

Table 9

Fig.6

Table 10

Fig.7

Table 11

Fig.8

Table 12

Fig.9

Fig.10

References 32

[1]	李靓.基于产业链视角的蔬菜价格形成研究［D］.北京：中国农业大学，2018.
	LI J.Study on the formation of vegetable price based on industrial chain［D］.Beijing：China Agricultural University，2018.
[2]	李明远.番茄早疫病及其易混淆的一些病害［J］.蔬菜，2022（7）：82-84.
	LI M Y.Early blight of tomato and some easily confused diseases［J］.Vegetables，2022（7）：82-84.
[3]	李军安.保护地番茄早疫病防治新技术［J］.河南农业，2022（4）：27.
	LI J A.New techniques for the control of tomato early blight in protected cultivation［J］.Henan Agriculture，2022（4）：27.
[4]	宋根苗，蒋家珍，邱立红.噻霉酮和苯醚甲环唑混配对4种不同病原菌的增效作用［J］.植物保护，2012，38（4）：171-174.
	SONG G M， JIANG J Z， QIU L H.Synergistic activity of benziothiazolinone/difenoconazole complex against four different fungi［J］.Plant Protection，2012，38（4）：171-174.
[5]	SHATTUCK A， WERNER M， MEMPEL F，et al.Global pesticide use and trade database（GloPUT）：new estimates show pesticide use trends in low-income countries substantially underestimated［J］.Global Environmental Change，2023，81：102693.
[6]	POVEDA J， ROESCHLIN R A， MARANO M R，et al.Microorganisms as biocontrol agents against bacterial citrus diseases［J］.Biological Control，2021，158：104602.
[7]	VANITTANAKOM N， LOEFFLER W， KOCH U，et al.Fengycin-a novel antifungal lipopeptide antibiotic produced by Bacillus subtilis F-29-3［J］.The Journal of Antibiotics，1986，39（7）：888-901.
[8]	ZOU Y K， ZHANG J N， YANG D L，et al.Effects of different land use patterns on nifH genetic diversity of soil nitrogen-fixing microbial communities in Leymus Chinensis steppe［J］.Acta Ecologica Sinica，2011，31（3）：150-156.
[9]	CWALINA-AMBROZIAK B， NOWAK M K.The effects of biological and chemical controls on fungal communities colonising tomato （Lycopersicon esculentum Mill.） plants and soil［J］.Folia Horticulturae，2012，24（1）：13-20.
[10]	杨文娟，余君，杨春雷，等.烟草化感自毒物质降解复合菌剂的优化及应用效果评价［J］.微生物学报，2024，64（4）：1044-1063.
	YANG W J， YU J， YANG C L，et al.Optimization and evaluation of a compound bacterial agent degrading autotoxins of tobacco［J］.Acta Microbiologica Sinica，2024，64（4）：1044-1063.
[11]	LAKOWITZ A， GODARD T， BIEDENDIECK R，et al.Mini review：recombinant production of tailored bio-pharmaceuticals in different Bacillus strains and future perspectives［J］.European Journal of Pharmaceutics and Biopharmaceutics，2018，126：27-39.
[12]	秦旭，冯艳娟，羊国根，等.小麦全蚀病菌拮抗细菌的筛选及生防效果［J］.植物保护学报，2023，50（3）：716-724.
	QIN X， FENG Y J， YANG G G，et al.Screening and evaluation of antagonistic bacteria as biocontrol agents against fungal pathogen Gaeumannomyces tritici ［J］.Journal of Plant Protection，2023，50（3）：716-724.
[13]	马朝旭，蒋晓，李林辉，等.桑椹菌核病的绿色防控研究进展［J］.西华师范大学学报（自然科学版），2025，46（2）：127-133.
	MA Z X， JIANG X， LI L H，et al.Research progress on green prevention and control of mulberry sclerotial disease［J］.Journal of China West Normal University （Natural Sciences），2025，46（2）：127-133.
[14]	尹杨燕，彭昊，李小宁，等.枯草芽孢杆菌体外抑菌试验及其抑菌蛋白的鉴定［J］.畜牧与兽医，2022，54（12）：57-63.
	YIN Y Y， PENG H， LI X N，et al.Antibacterial test in vitro of Bacillus subtilis and identification of its antibacterial proteins［J］.Animal Husbandry and Veterinary Medicine，2022，54（12）：57-63.
[15]	SRIVASTAVA R K， SINGH R K， PRASAD R D.Relative antogonistic effect of different isolates of Trichoderma viridi and Trichoderma harzanium against Rhizoctonia solani ［J］.National Academy Science Letters，2012，35（1）：49-52.
[16]	PANDIN C， LE COQ D， DESCHAMPS J，et al.Complete genome sequence of Bacillus velezensis QST713：a biocontrol agent that protects Agaricus bisporus crops against the green mould disease［J］.Journal of Biotechnology，2018，278：10-19.
[17]	MILIJAŠEVIĆ-MARČIĆ S， STEPANOVIĆ M， TODOROVIĆ B，et al.Biological control of green mould on Agaricus bisporus by a native Bacillus subtilis strain from mushroom compost［J］.European Journal of Plant Pathology，2017，148（3）：509-519.
[18]	MWANGI R W， KARIUKI S T， WAGARA I N.Biocontrol of green mould disease of oyster mushroom （Pleurotus ostreatus） using Bacillus amyloliquefaciens ［J］.Journal of Biology，Agriculture and Healthcare，2017，7（10）：25-30.
[19]	GENG P L， HU Y M， ZHOU G P，et al.Characterization of three autolysins with activity against cereulide-producing Bacillus isolates in food matrices［J］.International Journal of Food Microbiology，2017，241：291-297.
[20]	贺海滨，苏峻冬，滕凯，等.解淀粉芽孢杆菌研究进展［J］.现代农业科技，2021（2）：101-104.
	HE H B， SU J D， TENG K，et al.Research progress on Bacillus amyloliquefaciens ［J］.Modern Agricultural Science and Technology，2021（2）：101-104.
[21]	刘露，李丽，闫洪雪，等.巨大芽孢杆菌的应用研究进展［J］.北方农业学报，2016，44（4）：117-120.
	LIU L， LI L， YAN H X，et al.Research progress of application of Bacillus megaterium ［J］.Journal of Northern Agriculture，2016，44（4）：117-120.
[22]	陈志谊.芽孢杆菌类生物杀菌剂的研发与应用［J］.中国生物防治学报，2015，31（5）：723-732.
	CHEN Z Y.Research and application of bio-fungicide with Bacillus spp.［J］.Chinese Journal of Biological Control，2015，31（5）：723-732.
[23]	王丽丽，朱诗君，狄蕊，等.微生物菌肥菌剂对番茄生长发育和产量品质的影响［J］.土壤与作物，2022，11（1）：88-95.
	WANG L L， ZHU S J， DI R，et al.Effects of bio-organic fertilizer and microbial agent on the growth，yield and quality of tomato［J］.Soil and Crops，2022，11（1）：88-95.
[24]	ZHANG X X， ZHANG R J， GAO J S，et al.Thirty-one years of rice-rice-green manure rotations shape the rhizosphere microbial community and enrich beneficial bacteria［J］.Soil Biology and Biochemistry，2017，104：208-217.
[25]	KHAN A R， MUSTAFA A， HYDER S，et al. Bacillus spp.as bioagents：uses and application for sustainable agriculture［J］.Biology，2022，11（12）：1763.
[26]	XIE Z Y， LI M M， WANG D K，et al.Biocontrol efficacy of Bacillus siamensis LZ88 against brown spot disease of tobacco caused by Alternaria alternata ［J］.Biological Control，2021，154：104508.
[27]	杜丹超，朱莉，蒲占湑，等.淡紫紫孢菌ZJPL08可湿性粉剂研制及对柑橘木虱的毒力分析［J］.浙江大学学报（农业与生命科学版），2024，50（5）：747-757.
	DU D C， ZHU L， PU Z X，et al.Development of Purpureocillium lilacinum ZJPL08 wettable powder and its toxicity to Diaphorina citri Kuwayama［J］.Journal of Zhejiang University （Agriculture and Life Sciences），2024，50（5）：747-757.
[28]	赵春燕，安良聪，赵秋玲，等.贝莱斯芽孢杆菌JK-1可湿性粉剂研制及对辣椒褐腐病的防治效果［J］.天津农业科学，2023，29（2）：52-56.
	ZHAO C Y， AN L C， ZHAO Q L，et al.Preparation of wettable powder for Bacillus velezensis JK-1 and control effect on brown rot of cayenne pepper［J］.Tianjin Agricultural Sciences，2023，29（2）：52-56.
[29]	敖明远，张晓云，郭庆港，等.防治蔬菜灰霉病枯草芽胞杆菌BAB-1水分散片剂的研制［J］.中国生物防治学报，2022，38（2）：404-413.
	AO M Y， ZHANG X Y， GUO Q G，et al.Development of Bacillus subtilis BAB-1 water dispersible tablet to control vegetable gray mold［J］.Chinese Journal of Biological Control，2022，38（2）：404-413.
[30]	赵新贝，王娟，上官妮妮，等.番茄灰霉病生防细菌TD-7的鉴定、发酵条件优化及其防治效果［J］.中国生物防治学报，2019，35（2）：226-239.
	ZHAO X B， WANG J， SHANGGUAN N N，et al.Identification，fermentation condition optimization and control efficiency of biocontrol bacterium TD-7 against the tomato grey mould［J］.Chinese Journal of Biological Control，2019，35（2）：226-239.
[31]	姚蒋庞，唐婧红，肖洋，等.响应面法制备黑曲霉浸膏水分散片剂及抗病促生作用［J］.生物学杂志，2022，39（3）：88-93.
	YAO J P， TANG J H， XIAO Y，et al.Preparation of Aspergillus niger extract water dispersible tablets by response surface methodology and study on its anti-disease and growth-promoting effects［J］.Journal of Biology，2022，39（3）：88-93.
[32]	陈雨薇，王喜刚，郭成瑾，等.贝莱斯芽胞杆菌GZ8-6对马铃薯根际土壤酶活性的影响及促生作用［J］.植物保护，2024，50（1）：48-55.
	CHEN Y W， WANG X G， GUO C J，et al.Effects of Bacillus velezensis GZ8-6 on the enzyme activities in potato rhizosphere soil and potato growth［J］.Plant Protection，2024，50（1）：48-55.

发病等级 Disease severity	分级标准 Rating criteria
0	叶片无病斑
1	病斑占叶片1/4以下
2	病斑占叶片1/4至1/2
3	病斑占叶片1/2至3/4且近一半叶片干枯
4	病斑占叶片3/4以上且超一半叶片干枯

载体 Carrier	悬浮率 Suspension rate/%	润湿时间 Wetting time/s	活菌数 Viable cell count/（10⁸ CFU·g^-1）
高岭土 Kaolin	48.76±0.72^a	68.66±0.48^c	2.13±0.05^a
硅藻土 Diatomite	36.12±0.25^b	82.61±1.48^a	1.84±0.13^b
白炭黑 White carbon black	50.23±0.64^a	65.11±0.64^c	1.82±0.22^b
膨润土 Bentonite	29.60±0.42^c	74.74±0.71^b	1.41±0.04^c

润湿剂 Wetting agent	悬浮率 Suspension rate/%	润湿时间 Wetting time/s	活菌数 Viable cell count/（10⁸ CFU·g^-1）
达润DCM-82 Darun DCM-82	45.33±2.07^b	43.77±2.17^b	1.91±0.51^b
十二烷基苯磺酸钠（DBS） Sodium dodecyl benzene sulfonate	53.39±1.28^a	38.76±1.83^c	2.17±0.08^a
吐温-80 Tween-80	51.52±1.40^a	64.30±1.13^a	1.82±0.22^b
达润D317 Darun D317	41.39±1.70^b	42.33±1.84^b	1.41±0.04^c

分散剂 Dispersant	悬浮率 Suspension rate/%	润湿时间 Wetting time/s	活菌数 Viable cell count/（10⁸ CFU·g^-1）
十二烷基硫酸钠（SDS） Sodium dodecyl sulfate	65.33±2.07^b	51.77±1.17^a	1.91±0.41^b
聚乙二醇8000 PEG8000	53.39±1.28^c	42.76±1.73^b	1.88±0.08^b
木质素磺酸钙 Calcium lignosulfonate	75.52±1.23^a	44.30±1.13^b	2.18±0.03^a
羧甲基纤维素钠 Sodium carboxymethyl cellulose	61.39±1.70^b	52.33±1.84^a	1.42±0.04^c

比例 Ratio	悬浮率 Suspension rate/%	润湿时间 Wetting time/s	活菌数 Viable cell count/（10⁸ CFU·g^-1）
1∶9	71.33±2.37^b	66.77±1.17^a	1.91±0.41^b
2∶8	78.52±0.53^a	56.21±1.52^b	2.23±0.18^b
3∶7	70.12±1.33^b	53.30±1.13^c	2.26±0.03^a
4∶6	71.39±1.41^b	53.33±1.84^c	1.61±0.04^c
5∶5	65.22±1.48^c	51.21±1.04^c	1.63±0.84^c
6∶4	66.39±1.51^c	50.79±0.54^c	1.57±0.54^d
7∶3	71.28±0.21^b	47.75±0.22^d	1.55±0.34^d
8∶2	63.39±1.55^c	44.98±0.29^d	1.53±0.07^d
9∶1	58.39±0.41^d	39.37±0.54^e	1.52±0.54^d