Identification of Surfactin-producing Bacillus Velezensis C5A-1 and Evaluation of the Plant Growth-promoting Effects of Its Surfactin

doi:10.13560/j.cnki.biotech.bull.1985.2025-0364

Abstract

Abstract:

Objective This research is aimed to screen the high surfactin-producing strains, understand the genetic background and surfactin biosynthesis of the strains from the whole genome, and explore the functions of surfactin produced by the strains in plant growth-promoting. Method Surfactin-producing strains were isolated from plant rhizosphere soil by selective separation technique. The quantification and structural characterization of surfactin were performed using high-performance liquid chromatography and MALDI-TOF MS, respectively. The whole-genome was sequenced using Illumina + PacBio third-generation sequencing platform; and species identification, functional gene annotation, secondary metabolic gene clusters and surfactin biosynthesis were analyzed by bioinformatics method. Pot experiments were performed to detect the plant growth-promoting effect by surfactin. Result Strain C5A-1 with strong oil-displacing and antifungal activity was screened from poplar rhizosphere soil collected from Datong, Shanxi province. Surfactin homologues from C5A-1 were identified as C14- and C15-surfactin A, with the yield of 1 208.16 mg/L. The genomic average nucleotide identity (ANI) and digital DNA-DNA hybridization values between C5A-1 and B. velezensis FZB42 were 98.35% and 85.4%, respectively. C5A-1 genome size was 3 929 585 bp with 46.5% guanine-cytosine content, encoding 3 747 genes and harboring 12 secondary metabolite biosynthetic gene clusters. The surfactin biosynthetic gene cluster includes the core genes srfAA, srfAB, and srfAC. Surfactin produced by C5A-1 significantly enhanced the plant height, stem diameter, and aboveground and underground dry biomass of Populus tomentosa, Zea mays and Glycine max. Conclusion C5A-1 with high-producing surfactin was a novel strain belonging to B. velezensis, and the C14- and C15-surfactin A isoforms produced by C5A-1 demonstrated significant plant growth-promoting activity, which provides an optimized chassis cell factory for improving the surfactin yield by metabolic engineering strategies, and established a theoretical foundation for the practical application of surfactin in agricultural and forestry systems.

Key words: Bacillus velezensis, surfactin, MALDI-TOF MS, whole-genome sequencing, secondary metabolite biosynthetic gene clusters

LYU Zhen, GAN Tian, HUO Si-yu, ZHAO Chen-di, ZHAO Meng-yao, LI Ya-tao, MA Yu-chao, GENG Yu-qing. Identification of Surfactin-producing Bacillus Velezensis C5A-1 and Evaluation of the Plant Growth-promoting Effects of Its Surfactin[J]. Biotechnology Bulletin, 2025, 41(9): 265-276.

Figures/Tables 10

References 32

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样品名称 Sample name	采样地 Sampling location	植物种类 Plant species	经纬度 Longitude and latitude
A5A	湖南省岳阳市	番茄	112°90′E 28°68′N
A5C	云南省丽江市	苹果	100°25′E 26°86′N
C5A	山西省大同市	杨树	113°20′E 40°3′N
A6A	北京市昌平区	黄豆	116°25′E 40°7′N
A6B	湖北省仙桃市	白萝卜	112°20′E 30°18′N
A6D	湖南省常德市	蒜苗	108°93’E 34°23’N

样品名称 Sample name	采样地 Sampling location	植物种类 Plant species	经纬度 Longitude and latitude
A5A	湖南省岳阳市	番茄	112°90′E 28°68′N
A5C	云南省丽江市	苹果	100°25′E 26°86′N
C5A	山西省大同市	杨树	113°20′E 40°3′N
A6A	北京市昌平区	黄豆	116°25′E 40°7′N
A6B	湖北省仙桃市	白萝卜	112°20′E 30°18′N
A6D	湖南省常德市	蒜苗	108°93’E 34°23’N

Cluster	Type	Start	End	Similar cluster	Similarity （%）
1	NRPS	323409	387387	Surfactin	82
2	PKS-like	923849	926504	Butirosin A/B	7
3	Terpene	1049972	1067381	-	-
4	Lanthipeptide-class-ii	1188370	1217259	-	-
5	TransAT-PKS	1383378	1471714	Macrolactin H	100
6	TransAT-PKS	1691242	1791808	Bacillaene	100
7	NRPS	1865549	1999860	Fengycin	100
8	Terpene	2028497	2025381	-	-
9	T3PKS	2113698	2154799	-	-
10	TransAT-PKS	2282174	2375967	Difficidin	100
11	NRPS	300670	3052462	Bacillibactin	100
12	Other	3588771	3630190	Bacilysin	100

Cluster	Type	Start	End	Similar cluster	Similarity （%）
1	NRPS	323409	387387	Surfactin	82
2	PKS-like	923849	926504	Butirosin A/B	7
3	Terpene	1049972	1067381	-	-
4	Lanthipeptide-class-ii	1188370	1217259	-	-
5	TransAT-PKS	1383378	1471714	Macrolactin H	100
6	TransAT-PKS	1691242	1791808	Bacillaene	100
7	NRPS	1865549	1999860	Fengycin	100
8	Terpene	2028497	2025381	-	-
9	T3PKS	2113698	2154799	-	-
10	TransAT-PKS	2282174	2375967	Difficidin	100
11	NRPS	300670	3052462	Bacillibactin	100
12	Other	3588771	3630190	Bacilysin	100