The Prospect of Hydrolytic Enzymes from Bacillus Species in the Biological Control of Pests and Diseases in Forest and Fruit Tree Production.

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, ß-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, ß-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.

Entomopathogenic Potential of Bacillus velezensis CE 100 for the Biological Control of Termite Damage in Wooden Architectural Buildings of Korean Cultural Heritage.

Biocontrol strategies are gaining tremendous attention in insect pest management, such as controlling termite damage, due to the growing awareness of the irreparable harm caused by the continuous use of synthetic pesticides. This study examines the proteolytic and chitinolytic activities of Bacillus velezensis CE 100 and its termiticidal effect through cuticle degradation. The proteolytic and chitinolytic activities of B. velezensis CE 100 systematically increased with cell growth to the respective peaks of 68.3 and 128.3 units/mL after seven days of inoculation, corresponding with the highest cell growth of 16 × 107 colony-forming units (CFU)/mL. The in vitro termiticidal assay showed that B. velezensis CE 100 caused a rapid and high rate of termite mortality, with a median lethal time (LT50) of >1 h and the highest mortality rates of 91.1% and 92.2% recorded at 11 h and 12 h in the bacterial broth culture and crude enzyme fraction, respectively. In addition to broken setae and deformed sockets, termites treated with the bacterial broth culture exhibited degraded epicuticles, while the crude enzyme fraction caused severe disintegration of both the epicuticle and endocuticle. These results indicate the tremendously higher potential of B. velezensis CE 100 in the biological control of subterranean termites compared to the previously used entomopathogenic bacteria.

Utilization of bamboo as biorefinery feedstock: Co-production of xylo-oligosaccharide with succinic acid and lactic acid.

Herein, we investigated the possibility of co-producing xylo-oligosaccharides (XOSs) from bamboo, as value-added products, along with succinic and lactic acids, as platform chemicals. Xylan was extracted from bamboo using the alkali method under mild conditions. From xylan, XOSs were produced by partial enzymatic hydrolysis at a conversion rate of 83.9%, and all reaction conditions resulted in similar degrees of polymerization. Hydrogen peroxide-acetic acid (HPAC) pretreatment effectively removed lignin from NaOH-treated bamboo, and the enzymatic hydrolytic yield of NaOH and HPAC-treated bamboo was 84.3% of the theoretical yield. The production of succinic and lactic acids from the hydrolysate resulted in conversion rates of approximately 63.2% and 91.3% of the theoretical yield using Corynebacterium glutamicum Δldh and Actinobacillus succinogenes, respectively, under facultative anaerobic conditions. This study demonstrates that bamboo has a high potential to produce value-added products using a biorefinery process and is an alternative resource for compounds typically derived from petroleum.

Control of Fungal Diseases and Fruit Yield Improvement of Strawberry Using Bacillus velezensis CE 100.

Due to the increasing health and environmental risks associated with the use of fungicides in agriculture, alternatives-such as using plant growth-promoting bacteria (PGPB) to suppress phytopathogens-that simultaneously improve plant yield, are important. This study evaluated the biocontrol efficiency of Bacillus velezensis CE100 against Macrophomina phaseolina and Fusarium oxysporum f. sp. fragariae, the respective causal agents for charcoal rot and fusarium wilt diseases in strawberry, and its potential to enhance strawberry growth and fruit production. B. velezensis CE 100 produced fungal cell wall-degrading enzymes, chitinases, and ß-1,3-glucanases; and inhibited the mycelial growth of M. phaseolina and F. oxysporum f. sp. fragariae by 64.7% and 55.2%, respectively. The mycelia of both phytopathogenic fungi showed severe swelling and rupturing of the hyphae compared to the smooth, normal growth in the control group. Moreover, B. velezensis CE100 produced up to 2.8 units/mL of indole-3-acetic acid (IAA) during incubation and enhanced root biomass in strawberries. Consequently, B. velezensis CE 100 not only increased the fruit yield of strawberries by controlling the fungal diseases but also through enhancing plant growth. The findings of this study indicate that B. velezensis CE100 could be a safe, ecofriendly biocontrol alternative to chemical fungicides in strawberry production.

Antifungal Activity of Bacillus velezensis CE 100 against Anthracnose Disease (Colletotrichum gloeosporioides) and Growth Promotion of Walnut (Juglans regia L.) Trees.

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 ß-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.

Biological Control of Leaf Blight Disease Caused by Pestalotiopsis maculans and Growth Promotion of Quercus acutissima Carruth Container Seedlings Using Bacillus velezensis CE 100.

Leaf blight disease caused by Pestalotiopsismaculans lead to deleterious losses in the quality of forest container seedlings. The use of plant growth-promoting bacteria provides a promising strategy to simultaneously control diseases and enhance forest seedling production. This study investigated the biocontrol of leaf blight disease and growth promotion potential of Bacillus velezensis CE 100 in Quercus acutissima Carruth seedlings. B. velezensis CE 100 produced cell wall degrading enzymes, such as chitinase, ß-l,3-glucanase, and protease, which caused cell wall lysis and hyphae deformation of P. maculans, leading to mycelial growth inhibition by 54.94%. Inoculation of B. velezensis CE 100 suppressed P. maculans infection and increased seedling survival rate by 1.6-fold and 1.3-fold compared to chemical fertilizer and control, respectively. In addition, B. velezensis CE 100 produced indole-3-acetic acid, which improved root development and nutrient uptake compared to chemical fertilizer and control. Especially, inoculation with B. velezensis CE 100 increased the total nitrogen content of Q. acutissima seedlings, improved the chlorophyll index in the leaves, and increased seedling biomass by 1.3-fold and 2.2-fold compared to chemical fertilizer and control, respectively. Thus, B. velezensis CE 100 could be applied in the eco-friendly production of high-quality forest seedlings.

The Role of Lysobacter antibioticus HS124 on the Control of Fall Webworm (Hyphantria cunea Drury) and Growth Promotion of Canadian Poplar (Populus canadensis Moench) at Saemangeum Reclaimed Land in Korea.

Populus canadensis Moench forests established in Saemangeum-reclaimed land have been invaded by Hyphantria cunea Drury, causing defoliation and stunted growth. This study investigated the biocontrol potential of cuticle degrading chitinase and protease secreted by Lysobacter antibioticus HS124 against H. cunea larvae. In addition, L. antibioticus HS124 was examined for indole-3-acetic acid phytohormone production for plant growth promotion. To determine the larvicidal activity in the laboratory experiments, crude enzymes, bacteria culture, CY medium, and water (control) were sprayed on the larvae reared on natural diet in insect rearing dishes. Treatment with crude enzymes and bacteria culture caused 76.7% and 66.7% larvae mortality, respectively. The larvae cuticle, mainly composed of chitin and proteins, was degraded by cuticle-degrading enzymes, chitinase, and protease in both the bacteria culture and crude enzyme treatments, causing swelling and disintegration of the cuticle. Field application of the bacteria culture was achieved by vehicle-mounted sprayer. Bacterial treatment caused morphological damage on the larvae cuticles and subsequent mortality. Foliar application of the bacteria culture reduced tree defoliation by H. cunea and enhanced growth compared to the control. Especially, L. antibioticus HS124 produced auxins, and increased growth of poplar trees.

Bacillus velezensis CE 100 Inhibits Root Rot Diseases (Phytophthora spp.) and Promotes Growth of Japanese Cypress (Chamaecyparis obtusa Endlicher) Seedlings.

Root rot diseases, caused by phytopathogenic oomycetes, Phytophthora spp. cause devastating losses involving forest seedlings, such as Japanese cypress (Chamaecyparis obtusa Endlicher) in Korea. Plant growth-promoting rhizobacteria (PGPR) are a promising strategy to control root rot diseases and promote growth in seedlings. In this study, the potential of Bacillus velezensis CE 100 in controlling Phytophthora root rot diseases and promoting the growth of C. obtusa seedlings was investigated. B. velezensis CE 100 produced ß-1,3-glucanase and protease enzymes, which degrade the ß-glucan and protein components of phytopathogenic oomycetes cell-wall, causing mycelial growth inhibition of P. boehmeriae, P. cinnamomi, P. drechsleri and P. erythoroseptica by 54.6%, 62.6%, 74.3%, and 73.7%, respectively. The inhibited phytopathogens showed abnormal growth characterized by swelling and deformation of hyphae. B. velezensis CE 100 increased the survival rate of C. obtusa seedlings 2.0-fold and 1.7-fold compared to control, and fertilizer treatment, respectively. Moreover, B. velezensis CE 100 produced indole-3-acetic acid (IAA) up to 183.7 mg/L, resulting in a significant increase in the growth of C. obtusa seedlings compared to control, or chemical fertilizer treatment, respectively. Therefore, this study demonstrates that B. velezensis CE 100 could simultaneously control Phytophthora root rot diseases and enhance growth of C. obtusa seedlings.

Antifungal Activity of Cyclic Tetrapeptide from Bacillus velezensis CE 100 against Plant Pathogen Colletotrichum 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.

The Effect of Bacillus licheniformis MH48 on Control of Foliar Fungal Diseases and Growth Promotion of Camellia oleifera Seedlings in the Coastal Reclaimed Land of Korea.

This study investigated the control of foliar fungal diseases and growth promotion of Camellia oleifera seedlings in coastal reclaimed land through the use of Bacillus licheniformis MH48. B. licheniformis MH48 can produce lytic enzymes chitinase and ß-1,3-glucanase that can inhibit foliar pathogens by 37.4 to 50.5%. Nevertheless, foliar diseases appeared in the seedlings with bacterial inoculation, and their survival rate decreased because they were unable to withstand salt stress. However, B. licheniformis MH48 significantly increased the total nitrogen and phosphorus contents in the soils through fixing atmospheric nitrogen and solubilizing phosphorus. The growth of seedlings with bacterial inoculation increased, particularly in root dry weight, by 7.42 g plant-1, which was 1.7-fold greater than that of the control. B. licheniformis MH48 produces the phytohormone auxin, which potentially stimulates seedling root growth. C. oleifera seedlings significantly increased in total nitrogen content to 317.57 mg plant-1 and total phosphorus content to 46.86 mg plant-1. Our results revealed the effectiveness of B. licheniformis MH48 not only in the control of foliar fungal diseases but also in the growth promotion of C. oleifera seedlings in coastal lands.

Isolation and characterization of metabolites from Bacillus licheniformis MH48 with antifungal activity against plant pathogens.

In this study, we isolated Bacillus licheniformis MH48 from rhizosphere soil and demonstrated that this strain shows significant antifungal activity against Rhizoctonia solani, Colletotrichum gloeosporioides, and Phytophthora capsici. Our results showed that a 50% concentration of bacterial cell-free culture filtrate of B. licheniformis MH48 shows strong activity against fungal pathogens. Benzoic acid produced by B. licheniformis MH48 was purified by various chromatographic techniques and identified by nuclear magnetic resonance and gas chromatography-mass spectrometry analysis. Benzoic acid displayed antifungal activity against R. solani and C. gloeosporides with minimum inhibitory concentration of 128 µg/mL against mycelial growth. Microscopic examination revealed that benzoic acid (50 µg/mL and 100 µg/mL) transformed C. gloeosporioides conidial morphology and inhibited conidial germination. In addition, benzoic acid (100 µg/mL and 200 µg/mL) degraded R. solani mycelia. Therefore, our results demonstrate that B. licheniformis MH48 strain shows potential for utility as a biological agent for the control of various fungal pathogens of plants.

Distribution of Mercury Concentrations in Tree Rings and Surface Soils Adjacent to a Phosphate Fertilizer Plant in Southern Korea.

This study aimed to determine mercury concentrations in tree rings and surface soils at distances of 4, 26 and 40 km from a fertilizer plant located in Yeosu City, Korea. Mercury concentrations in all tree rings were low prior to the establishment of the plant in 1977 and became elevated thereafter. The highest average mercury concentration in the tree rings was 11.96 ng g-1 at the Yeosu site located nearest to the plant, with the lowest average mercury concentration of 4.45 ng g-1 at the Suncheon site furthest away from the plant. In addition, the highest mercury content in the surface soil was 108.51 ng cm-3 at the Yeosu site, whereas the lowest mercury content in the surface soil was 31.47 ng cm-3 at the Suncheon site. The mercury levels decreased gradually with increasing distance from the plant.

Hydrology, suspended sediment dynamics and nutrient loading in Lake Takkobu, a degrading lake ecosystem in Kushiro Mire, northern Japan.

Suspended sediment and nutrient loadings from agricultural watersheds have lead to habitat degradation in Lake Takkobu. To examine their relationships with land-use activities, we monitored sediment, nutrient and water discharges into the lake for a 1-year sampling period. The Takkobu River contributed the largest portion of the annual water discharge into the lake, compared with the other tributaries. During dry conditions, lake water flowed into the Kushiro River, and conversely during flooding, Kushiro River water flowed into the lake. Inflows from the Kushiro River had a high proportion of inorganic matter, with high concentrations of total nitrogen and total phosphorus, attributed to agricultural land-use development and stream channelization practiced since the 1960s in the Kushiro Mire. Nutrient loadings from these two rivers were significantly higher during flooding than in dry conditions. However, there was no clear correlation between river discharge and nutrient concentrations. Since land-use activities in the Kushiro River and Takkobu River watersheds were concentrated near rivers, nutrients easily entered the drainage system under low flow conditions. In contrast, water discharged from small, forest-dominated watersheds contained a low proportion of inorganic matter, and low nutrient concentrations. The suspended sediment delivered to the lake during the sample period was estimated as approximately 607 tons, while the total nitrogen and total phosphorus inflows were about 10,466 and 1,433 kg, respectively. Suspended sediment input into the lake was 65%, and total nitrogen and total phosphorus were 40% and 48%, respectively, being delivered by the Kushiro River.