Whole Genome Sequencing of Pseudomonas chlororaphis ZL3 and Antifungal Activity of atoB Gene Recombinant Strain Against Botrytis cinerea

doi:10.16420/j.issn.0513-353x.2024-0879

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (11): 3044-3056.doi: 10.16420/j.issn.0513-353x.2024-0879

• Plant Protection • Previous Articles Next Articles

Whole Genome Sequencing of Pseudomonas chlororaphis ZL3 and Antifungal Activity of atoB Gene Recombinant Strain Against Botrytis cinerea

YUAN Shengnan, YUAN Jiaqi, WANG Mengxi, WANG Yan, WANG Meiqin, and WANG Chunwei^*()

College of Plant Protection， Shanxi Agricultural University，Taigu， Shanxi 030801， China

Received:2025-04-03 Revised:2025-08-06 Online:2025-11-27 Published:2025-11-27
Contact: and WANG Chunwei

Abstract

Abstract:

In order to clarify the antibacterial mechanism of Pseudomonas chlororaphis ZL3 against Botrytis cinerea，the gene atoB related to metabolic pathways of terpenoids and polyketones was screened by whole genome sequencing analysis. The protein structure of gene atoB was predicted. The recombinant strain Escherichia coli BL21（DE3）/pET32-atoB was constructed. The inhibitory effects of E. coli BL21 （DE3）/pET32-atoB on seven pathogenic fungi were determined by using two-sealed-base-plate method. The control effects of E. coli BL21（DE3）/pET32-atoB against cherry gray mold was determined. The effects of E. coli BL21（DE3）/pET32-atoB on cherry defense enzyme activities were detected by using ultraviolet spectrophotometry. The results showed that the total length of strain ZL3 was 6 715 612 bp，with one chromosome and no plasmid，and the GC content was 63.04%. The genome contained 5 956 coding genes. A total of 2 873，5 032 and 3 446 genes were annotated in GO，COG and KEGG databases，respectively. The recombinant strain E. coli BL21（DE3）/pET32-atoB had good antifungal effects on seven pathogenic fungi，and presented the highest inhibition effects on B. cinerea with an inhibition rate of 83.21%. After treatment with recombinant strain，the catalase（CAT），chitinase（CHI）and phenylalanine aminolyase（PAL）activities in cherry. The incidence of cherry gray mold decreased by 43.75%，while the diameter of the disease spots decreased by 39.01%.

Key words: cherry, Pseudomonas chlororaphis, whole genome sequencing, atoB gene, Botrytis cinerea, biological control, antifungal activity

YUAN Shengnan, YUAN Jiaqi, WANG Mengxi, WANG Yan, WANG Meiqin, and WANG Chunwei. Whole Genome Sequencing of Pseudomonas chlororaphis ZL3 and Antifungal Activity of atoB Gene Recombinant Strain Against Botrytis cinerea[J]. Acta Horticulturae Sinica, 2025, 52(11): 3044-3056.

Figures/Tables 10

Fig. 1 Genome circle map of Pseudomonas chlororaphis ZL3

Fig. 2 GO functional annotation analysis of Pseudomonas chlororaphis ZL3 genome

Fig. 3 COG functional annotation analysis of Pseudomonas chlororaphis ZL3 genome

Fig. 4 KEGG functional annotation analysis of Pseudomonas chlororaphis ZL3 genome

Fig. 5 The predicted secondary structure of atoB encoded proteins Blue：α helix；Green：β corner；Purple：Irregular curl；Red：Extension chain

Fig. 6 PCR validation of the atoB gene（A）and the recombinant strain E. coli BL21（DE3）/pET32-atoB（B）

Fig. 7 Conditions for expression of the recombinant protein atoB

Fig. 8 Inhibition effects of E. coli BL21（DE3）/pET32-atoB on different pathogens

Fig. 9 Effects of E. coli BL21（DE3）/pET32-atoB on incidence and spot diameter of cherry Different lowercase letters indicate significant differences among different treatments at 0.05 level

Fig. 10 Effects of E. coli BL21（DE3）/pET32-atoB on enzyme activity of cherry * P < 0.05，** P < 0.01

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Whole Genome Sequencing of Pseudomonas chlororaphis ZL3 and Antifungal Activity of atoB Gene Recombinant Strain Against Botrytis cinerea

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