Exploration of a multifunctional biocontrol agent Streptomyces sp. JCK-8055 for the management of apple fire blight.

Apple fire blight, caused by the bacterium Erwinia amylovora, is a devastating disease of apple and pear trees. Biological control methods have attracted much attention from researchers to manage plant diseases as they are eco-friendly and viable alternatives to synthetic pesticides. Herein, we isolated Streptomyces sp. JCK-8055 from the root of pepper and investigated its mechanisms of action against E. amylovora. Streptomyces sp. JCK-8055 produced aureothricin and thiolutin, which antagonistically affect E. amylovora. JCK-8055 and its two active metabolites have a broad-spectrum in vitro activity against various phytopathogenic bacteria and fungi. They also effectively suppressed tomato bacterial wilt and apple fire blight in in vivo experiments. Interestingly, JCK-8055 colonizes roots as a tomato seed coating and induces apple leaf shedding at the abscission zone, ultimately halting the growth of pathogenic bacteria. Additionally, JCK-8055 can produce the plant growth regulation hormone indole-3-acetic acid (IAA) and hydrolytic enzymes, including protease, gelatinase, and cellulase. JCK-8055 treatment also triggered the expression of salicylate (SA) and jasmonate (JA) signaling pathway marker genes, such as PR1, PR2, and PR3. Overall, our findings demonstrate that Streptomyces sp. JCK-8055 can control a wide range of plant diseases, particularly apple fire blight, through a combination of mechanisms such as antibiosis and induced resistance, highlighting its excellent potential as a biocontrol agent. KEY POINTS: • JCK-8055 produces the systemic antimicrobial metabolites, aureothricin, and thiolutin. • JCK-8055 treatment upregulates PR gene expression in apple plants against E. amylovora. • JCK-8055 controls plant diseases with antibiotics and induced resistance.

Response of particle-attached and free-living bacterial communities to Microcystis blooms.

The massive proliferation of Microcystis threatens freshwater ecosystems and degrades water quality globally. Understanding the mechanisms that contribute to Microcystis growth is crucial for managing Microcystis blooms. The lifestyles of bacteria can be classified generally into two groups: particle-attached (PA; > 3 µm) and free-living (FL; 0.2-3.0 µm). However, little is known about the response of PA and FL bacteria to Microcystis blooms. Using 16S rRNA gene high-throughput sequencing, we investigated the stability, assembly process, and co-occurrence patterns of PA and FL bacterial communities during distinct bloom stages. PA bacteria were phylogenetically different from their FL counterparts. Microcystis blooms substantially influenced bacterial communities. The time decay relationship model revealed that Microcystis blooms might increase the stability of both PA and FL bacterial communities. A contrasting community assembly mechanism was observed between the PA and FL bacterial communities. Throughout Microcystis blooms, homogeneous selection was the major assembly process that impacted the PA bacterial community, whereas drift explained much of the turnover of the FL bacterial community. Both PA and FL bacterial communities could be separated into modules related to different phases of Microcystis blooms. Microcystis blooms altered the assembly process of PA and FL bacterial communities. PA bacterial community appeared to be more responsive to Microcystis blooms than FL bacteria. Decomposition of Microcystis blooms may enhance cooperation among bacteria. Our findings highlight the importance of studying bacterial lifestyles to understand their functions in regulating Microcystis blooms. KEY POINTS: • Microcystis blooms alter the assembly process of PA and FL bacterial communities • Microcystis blooms increase the stability of both PA and FL bacterial communities • PA bacteria seem to be more responsive to Microcystis blooms than FL bacteria.

Novosphingobium cyanobacteriorum sp. nov., isolated from a eutrophic reservoir during the Microcystis bloom period.

A novel Gram-stain-negative, aerobic and rod-shaped bacterial strain, HBC54T, was isolated from periphyton during a Microcystis bloom. Based on the results of the 16S rRNA gene sequence analysis, strain HBC54T was closely related to Novosphingobium aerophilum 4Y4T (98.36 %), Novosphingobium aromaticivorans DSM 12444T (98.08 %), Novosphingobium huizhouense c7T (97.94 %), Novosphingobium percolationis c1T (97.65 %), Novosphingobium subterraneum DSM 12447T (97.58 %), Novosphingobium olei TW-4T (97.58 %) and Novosphingobium flavum UCT-28T (97.37 %). The average nucleotide identity and digital DNA-DNA hybridization values between HBC54T and its related type stains were below 78.97 and 23.7 %, which are lower than the threshold values for species delineation. The major fatty acids (>10.0 %) were identified as C14 : 0 2-OH, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and the respiratory quinone was ubiquinone Q-10. The main polar lipids detected in the strain were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, diphosphatidylglycerol and three unidentified phospholipids. The genomic DNA G+C content was 64.8 mol%. Strain HBC54T is considered to represent a novel species within the genus Novosphingobium, for which the name Novosphingobium cyanobacteriorum sp. nov. is proposed. The type strain is HBC54T (=KCTC 92033T=LMG 32427T).

Hymenobacter cyanobacteriorum sp. nov., isolated from a freshwater reservoir during the cyanobacterial bloom period.

A Gram-negative, red-colored, and rod-shaped bacterial strain, DH14T, was isolated from a eutrophic reservoir. The 16S rRNA gene sequence analysis showed that strain DH14T was most closely related to Hymenobacter terrigena (98.3% similarity) and Hymenobacter terrae (98.1%). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain DH14T and its related type strains were below 82.9% and 27.2%, respectively. Strain DH14T contained iso-C15:0 (32.6%), anteiso-C15:0 (14.0%), and summed feature 3 (C16:1 ω6c and/or C16:1 ω7c) (25.8%) as major cellular fatty acids. The main polar lipids were phosphatidylethanolamine, two unidentified aminophospholipids, and one unidentified lipid. The respiratory quinone was menaquinone 7 (MK-7). The genomic DNA G + C content was 62.1%. These evidences support the classification of strain DH14T as a novel species in the genus Hymenobacter, for which the name Hymenobacter cyanobacteriorum sp. nov. is proposed. The type strain is DH14T (= KCTC 92040T = LMG 32425T).

Panacibacter microcysteis sp. nov., isolated from a eutrophic reservoir during the Microcystis bloom period.

A Gram-stain-negative, rod-shaped bacterial strain DH6T was isolated from fresh water of the Daechung Reservoir during the Microcystis bloom period. The strain grew at pH 6.0-8.5, at temperature 17-40 °C, and at 0-1% (w/v) NaCl concentration. Comparison of 16S rRNA gene sequence indicated that strain DH6T exhibits the highest similarity with Panacibacter ginsenosidivorans Gsoil 1550T (96.6%). The average nucleotide identity (ANI), digital DNA-DNA hybridization (dDDH), and average amino acid identity (AAI) values of strain DH6T compared to its related type strains were below 74.2%, 22.3%, and 74.8%, respectively. The predominant fatty acids (> 5.0%) were identified as iso-C17:0 3-OH, iso-C13:0, iso-C15:0, C17:0 2-OH, iso-C11:0, anteiso-C13:0, and iso-C15:1 G. The polar lipid profile contained phosphatidylethanolamine, four unidentified aminolipids, and three unidentified lipids. The respiratory quinone was menaquinone 7 (MK-7). The genomic DNA G + C content was 42.6%. Collectively, strain DH6T should be classified as a novel species within the genus Panacibacter, for which the name Panacibacter microcysteis sp. nov. is proposed. The type strain is DH6T (= KCTC 82471T = LMG 32426T).

Flavobacterium inviolabile sp. nov. isolated from stream water.

A Gram-stain-negative, rod-shaped, aerobic and non-motile bacterium, designated P2-65T, was isolated from Moonsan stream water in the Republic of Korea. The temperature, NaCl concentration and pH ranges for growth of strain P2-65T were 10-37 °C, 0.0-3.0% (w/v) and 6.5-8.5 with optimum growth at 25-30 °C, 0.0-1.0% and 7.0-7.5, respectively. Comparison of 16S rRNA gene sequence showed that strain P2-65T was closely related to Flavobacterium cauense (95.4%) and Flavobacterium cheniae (95.3%). The major fatty acids were iso-C15:0, iso C17:0 3-OH, summed feature 3 (C16:1ω7c and/or C16:1ω6c), summed feature 9 (iso-C17:1 ω9c and/or 10-methyl C16:0) and iso-C15:0 3-OH. The predominant respiratory quinone was menaquinone-6 (MK-6). The major polar lipids detected in the strain were phosphatidylethanolamine, one aminophospholipid, one unidentified aminolipid and one unidentified polar lipid. The G + C content of the genomic DNA was 39.7%. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values for strain P2-65T with closely related Flavobacterium species were below 74.8% and 20%, respectively. Based on polyphasic features, strain P2-65T is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium inviolabile sp. nov. is proposed. The type strain is P2-65T (= KCTC 62055T = NBRC 112953T).

Sphingobacterium praediipecoris sp. nov. isolated from effluent of a dairy manure treatment plant.

A novel Gram-reaction-negative, rod-shaped, non-motile bacterium, designated as strain G2-10T was isolated from effluent of a dairy manure treatment plant. Growth occurred at 20-40 °C (optimum at 25-30 °C), pH 7.0-8.0 (optimum at pH 7.0). The range of NaCl concentration for growth was between 0% and 3% (w/v) (optimum 0-1%, w/v). Comparison of 16S rRNA gene sequence indicated that strain G2-10T was moderately related to the type strains of Sphingobacterium nematocida M-SX103T and Sphingobacterium suaedae T47T with a pair-wise sequence similarity of 94.3% and 94.0%, respectively. The major fatty acid constituents of strain G2-10T were identified as iso-C15:0 (37.6%), summed feature 3 (consisting of C16:1ω7c and/or C16:1ω6c, 29.6%) and iso-C17:0 3-OH (15.2%). Phosphatidylethanolamine was the major polar lipids of strain G2-10T. Sphingophospholipids were present. The isoprenoid quinone was composed of only MK-7. The DNA G + C content of strain G2-10T was found to be 42.5 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain G2-10T represents a novel species within the genus Sphingobacterium, for which the name Sphingobacterium praediipecoris is proposed. The type strain is G2-10T (= KCTC 52880T = NBRC 112848T).

Optimization of mycelial growth and cultivation of wild Ganoderma sinense.

Ganoderma sinense, a well-known medicinal macrofungus of Basidiomycetes, is widely used in traditional medicine for promoting health and longevity in East Asia. The fruiting bodies of G. sinense contain polysaccharides, ergosterol, and coumarin, which have antitumor, antioxidant, and anticytopenia activities. Mushroom cultivation requires suitable conditions for the formation of fruiting bodies and yield. However, little is known about the optimal culture conditions for mycelial growth and cultivation of G. sinense. In this study, the successful cultivation of a G. sinense strain collected from the wild was reported. The optimal culture conditions were identified by examining one factor at a time. The results of this study revealed that the nutritional requirements for the optimal mycelial growth of G. sinense were fructose (15 g/l) as the carbon source and yeast extract (1 g/l) as the nitrogen source. The optimal pH and temperature for G. sinense were 7 and 25-30°C, respectively. The mycelia grew fastest in treatment II (69% rice grains + 30% sawdust + 1% calcium carbonate). G. sinense produced fruiting bodies under all tested conditions and showed the highest biological efficiency (2.95%) in treatment B (96% sawdust, 1% wheat bran, 1% lime). In summary, under optimal culture conditions, G. sinense strain GA21 showed satisfactory yield and a high potential for commercial cultivation.