Plant diseases and insect pest damage cause tremendous losses in forestry and fruit tree production. Even though chemical pesticides have been effective in the control of plant diseases and insect pests for several decades, they are increasingly becoming undesirable due to their toxic residues that affect human life, animals, and the environment, as well as the growing challenge of pesticide resistance. In this study, we review the potential of hydrolytic enzymes from Bacillus species such as chitinases, β-1,3-glucanases, proteases, lipases, amylases, and cellulases in the biological control of phytopathogens and insect pests, which could be a more sustainable alternative to chemical pesticides. This study highlights the application potential of the hydrolytic enzymes from different Bacillus sp. as effective biocontrol alternatives against phytopathogens/insect pests through the degradation of cell wall/insect cuticles, which are mainly composed of structural polysaccharides like chitins, β-glucans, glycoproteins, and lipids. This study demonstrates the prospects for applying hydrolytic enzymes from Bacillus sp. as effective biopesticides in forest and fruit tree production, their mode of biocidal activity and dual antimicrobial/insecticidal potential, which indicates a great prospect for the simultaneous biocontrol of pests/diseases. Further research should focus on optimizing the production of hydrolytic enzymes, and the antimicrobial/insecticidal synergism of different Bacillus sp. which could facilitate the simultaneous biocontrol of pests and diseases in forest and fruit tree production.

Walnut anthracnose caused by Colletotrichum gloeosporioides is a deleterious disease that severely affects the production of walnut (Juglans regia L.). The aim of this study was to assess the antifungal and growth promotion activities of Bacillus velezensis CE 100 as an alternative to chemical use in walnut production. The crude enzyme from B. velezensis CE 100 exhibited chitinase, protease, and β-l,3-glucanase activity and degraded the cell wall of C. gloeosporioides, causing the inhibition of spore germination and mycelial growth by 99.3% and 33.6% at 100 µL/mL, respectively. The field application of B. velezensis CE 100 culture broth resulted in a 1.3-fold and 6.9-fold decrease in anthracnose disease severity compared to the conventional and control groups, respectively. Moreover, B. velezensis CE 100 produced indole-3-acetic acid (up to 1.4 µg/mL) and exhibited the potential for ammonium production and phosphate solubilization to enhance the availability of essential nutrients. Thus, field inoculation of B. velezensis CE 100 improved walnut root development, increased nutrient uptake, enhanced chlorophyll content, and consequently improved total biomass by 1.5-fold and 2.0-fold compared to the conventional and control groups, respectively. These results demonstrate that B. velezensis CE 100 is an effective biocontrol agent against anthracnose disease and a potential plant growth-promoting bacteria in walnut tree production.

Colletotrichum gloeosporioides is the most prevalent phytopathogen, causing anthracnose disease that severely affects the production of various fruit trees, including walnut and jujube. In this study, the volatile organic compounds (VOCs) from Bacillus velezensis CE 100 disrupted the cell membrane integrity of C. gloeosporioides and reduced the spore germination by 36.4% and mycelial growth by 20.0% at a bacterial broth concentration of 10%, while the control group showed no antifungal effect. Based on the headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS) analysis, seven VOCs were identified from the headspace of B. velezensis CE 100. Out of the seven VOCs, 5-nonylamine and 3-methylbutanoic acid were only detected in the headspace of B. velezensis CE 100 but not in the control group. Both 5-nonylamine and 3-methylbutanoic acid showed significant antifungal activity against the spore germination and mycelial growth of C. gloeosporioides. Treatment with 100 µL/mL of 5-nonylamine and 3-methylbutanoic acid suppressed the spore germination of C. gloeosporioides by 10.9% and 30.4% and reduced mycelial growth by 14.0% and 22.6%, respectively. Therefore, 5-nonylamine and 3-methylbutanoic acid are the potential antifungal VOCs emitted by B. velezensis CE 100, and this is the first report about the antifungal activity of 5-nonylamine against C. gloeosporioides.

The aim of this study was to investigate the antifungal activity of a cyclic tetrapeptide from Bacillus velezensis CE 100 against anthracnose-causing fungal pathogen Colletotrichum gloeosporioides. Antifungal compound produced by B. velezensis CE 100 was isolated and purified from ethyl acetate extract of B. velezensis CE 100 culture broth using octadecylsilane column chromatography. The purified compound was identified as cyclo-(prolyl-valyl-alanyl-isoleucyl) based on mass spectrometer and nuclear magnetic resonance analyses. This is the first report of the isolation of a cyclic tetrapeptide from B. velezensis CE 100 culture filtrate. Cyclic tetrapeptide displayed strong antifungal activity at concentration of 1000 µg/mL against C. gloeosporioides mycelial growth and spore germination. Our results demonstrate that the antifungal cyclic tetrapeptide from B. velezensis CE 100 has potential in bioprotection against anthracnose disease of plants caused by C. gloeosporioides.

This study investigated the control of Fusarium root rot and development of coastal pine (Pinus thunbergii) seedlings in a container nursery by using Bacillus licheniformis MH48. High-quality seedlings without infectious diseases cause vigorous growth. Fusarium root rot caused by Fusarium oxysporum is responsible for serious damage to coastal pine seedlings in nurseries. B. licheniformis MH48 produced enzymes that degraded the fungal cell walls, such as chitinase and β-1,3-glucanase. These lytic enzymes exhibited destructive activity toward F. oxysporum hyphae, which were found to play key roles in the suppression of root rot. In addition, B. licheniformis MH48 increased the nitrogen and phosphorus in soils via fixed atmospheric nitrogen and solubilized inorganic phosphate. B. licheniformis MH48 produced the phytohormone auxin, which stimulated seedling root development, resulting in increased nutrient uptake in seedlings. Both the bacterial inoculation and the chemical fertilizer treatments significantly increased seedling growth and biomass, and the bacterial inoculation had a greater effect on seedling development. Based on the results from this study, B. licheniformis MH48 showed potential as a biological agent against Fusarium root rot and as a promoter of growth and development of Pinus thunbergii seedlings.