Identification of a Biocontrol Strain Isolated from Tobacco Rhizosphere and Its Biocontrol Potential Analysis

doi:10.13560/j.cnki.biotech.bull.1985.2025-1076

Abstract

Abstract:

Objective The purpose of this study is to determine the taxonomic status of biocontrol strain YC25, evaluate its antibacterial ability, deeply analyze its genomic characteristics and potential of secondary metabolites, and elucidate its mechanism of induced disease resistance to tobacco (Nicotiana tabacum L.) plants, so as to provide theoretical basis and technical support for the development of efficient and environmentally friendly biocontrol agents for tobacco soil-borne diseases. Method The strain YC25 was identified by morphological, physiological, biochemical and molecular biological methods. The antibacterial ability was evaluated by confrontation test, and whole genome sequencing and secondary metabolite prediction were performed. The effect of YC25 treatment on gene expression in tobacco plants was analyzed by transcriptomics. Result The strain YC25 was identified as Bacillus spizizenii and showed broad-spectrum antagonistic activity against 9 plant pathogenic fungi. Field experiments showed that the combination of strain YC25 and metalaxyl effectively controlled tobacco root rot and reduced the amount of metalaxyl. The whole genome sequencing showed that the total length of the genome was 4 120 588 bp, the average GC content was 43.85%, and the coding genes were 4 045. A total of 13 secondary metabolite synthesis gene clusters were predicted, encoding antimicrobial active substances such as Surfactin, Bacillaene, Fengycin, Bacillibactin, Subtilosin and Bacilysin. Transcriptome analysis showed that a total of 205 differentially expressed genes (DEGs) were induced in tobacco after YC25 application, of which 119 were up-regulated and 86 were down-regulated. Several genes involved in plant stress response and immune response were up-regulated, including FLS2, CaM, CYP84A, ERF1, ERF038, bHLH041, bHLH92 and PR-1. Conclusion YC25 demonstrates the comprehensive ability to inhibit pathogens and induce plant resistance to disease, and thus has the potential to be further developed as a biological pesticide.

Key words: Bacillus spizizenii, antimicrobial activity, genome analysis, transcriptome analysis

WEI Xiao, LI Yan-yan, YUAN Qin-feng, YAO Jing-wu, CAO Chun-xia, HUANG Da-ye. Identification of a Biocontrol Strain Isolated from Tobacco Rhizosphere and Its Biocontrol Potential Analysis[J]. Biotechnology Bulletin, 2026, 42(7): 279-291.

Figures/Tables 12

References 46

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基因名称 Gene name	引物名称 Primer name	序列 Sequence（5'-3'）
GAPDH	N. tabacumGAPDH F	ATGAGAGAGTGCATATCGAT
GAPDH	N. tabacumGAPDH R	TTCACTGAAGAAGGTGTTGAA
PR-1	N. tabacum PR-1 F	ATTGGAGAGGACAACGTCCC
PR-1	N. tabacum PR-1 R	ACACACCTCCATAATACCACC
PR-3	N. tabacum PR-3 F	ACAGATTCTGCCGGACACAC
PR-3	N. tabacum PR-3 R	AAGGAAGGTCCCAAAAGGGT
NPR-1	N. tabacumNPR1 F	GGATGCGGATGACTTGTCTGA
NPR-1	N. tabacumNPR1 R	GGAACAAGATGACGAGAGGTG
PAL	N. tabacumPAL F	AAGCATCACCCCGGACAAAT
PAL	N. tabacumPAL R	GGTGATGTTCTGAGGGCGTA
GST	N. tabacumGST F	CCTCACCCCCTCCCTCATAA
GST	N. tabacumGST R	TCGCCTTGATCTGTTTGTAGACT
SOD	N. tabacumSOD F	TAGCACCACACGCACAGAAG
SOD	N. tabacumSOD R	TGAAGGCTTCTCTTCTGCCT
CAT	N. tabacumCAT F	TGAAGGCTTCTCTTCTGCCT
CAT	N. tabacumCAT R	TGGAGGCCTTGTCTGATCCT

基因名称 Gene name	引物名称 Primer name	序列 Sequence（5'-3'）
GAPDH	N. tabacumGAPDH F	ATGAGAGAGTGCATATCGAT
GAPDH	N. tabacumGAPDH R	TTCACTGAAGAAGGTGTTGAA
PR-1	N. tabacum PR-1 F	ATTGGAGAGGACAACGTCCC
PR-1	N. tabacum PR-1 R	ACACACCTCCATAATACCACC
PR-3	N. tabacum PR-3 F	ACAGATTCTGCCGGACACAC
PR-3	N. tabacum PR-3 R	AAGGAAGGTCCCAAAAGGGT
NPR-1	N. tabacumNPR1 F	GGATGCGGATGACTTGTCTGA
NPR-1	N. tabacumNPR1 R	GGAACAAGATGACGAGAGGTG
PAL	N. tabacumPAL F	AAGCATCACCCCGGACAAAT
PAL	N. tabacumPAL R	GGTGATGTTCTGAGGGCGTA
GST	N. tabacumGST F	CCTCACCCCCTCCCTCATAA
GST	N. tabacumGST R	TCGCCTTGATCTGTTTGTAGACT
SOD	N. tabacumSOD F	TAGCACCACACGCACAGAAG
SOD	N. tabacumSOD R	TGAAGGCTTCTCTTCTGCCT
CAT	N. tabacumCAT F	TGAAGGCTTCTCTTCTGCCT
CAT	N. tabacumCAT R	TGGAGGCCTTGTCTGATCCT

生理生化指标 Physiological and biochemical index	结果 Result	生理生化指标 Physiological and biochemical index	结果 Result
淀粉水解 Starch hydrolysis	+	L-苹果酸	+
接触酶	+	好氧生长	+
氧化酶	+	D-乳糖	-
蔗糖Sucrose	+	D-岩藻糖	-
D-甘露醇	+	L-鼠李糖	-
柠檬酸	+	pH 5.0	+
D-山梨糖醇	+	8% NaCl	+

生理生化指标 Physiological and biochemical index	结果 Result	生理生化指标 Physiological and biochemical index	结果 Result
淀粉水解 Starch hydrolysis	+	L-苹果酸	+
接触酶	+	好氧生长	+
氧化酶	+	D-乳糖	-
蔗糖Sucrose	+	D-岩藻糖	-
D-甘露醇	+	L-鼠李糖	-
柠檬酸	+	pH 5.0	+
D-山梨糖醇	+	8% NaCl	+

编号 Number	病原菌名称 Name of pathogens	抑制率 Inhibition rate （%）
A	尖孢镰孢菌 Fusarium oxysporum	67.45±1.47d
B	核盘菌 Sclerotinia sclerotiorum	72.55±1.47bcd
C	灰葡萄孢菌 Botrytis cinerea	83.92±2.93a
D	烟草炭疽菌 Colletotrichum micotianae	69.02±2.42cd
E	茶拟盘多毛孢菌 Pestalotiopsis theae	74.90±2.00bc
F	烟草疫霉菌 Phytophthora nicotianae	70.20±0.55bcd
G	互隔链格孢菌 Alternaria alternata	76.08±1.47b
H	立枯丝核菌 Rhizoctonia solani	74.90±1.47bc
I	多主棒孢菌 Corynespora cassiicola	72.94±0.96bcd