incarnatum previously reported by Akino and Kondo [34]. For molecular analysis, the DNA sequences of the translation elongation factor-1α (EF-1α), which amplified using primers EF1/EF2 (GenBank Accession No. KC478361), also had 100% sequence identity to F. cf.

incarnatum strains (GenBank Accession No. JF270205 and GQ339786) (data not shown), confirming it to be F. cf. incarnatum as shown by the above mycological characteristics. In the pathogenicity tests with different conidial inoculum concentrations of the Fusarium isolate obtained from diseased cactus, the initial rot symptoms appeared on the root discs inoculated with 106 conidia/mL after 2 d. After 6 d of incubation, rot symptoms developed on whole root discs at a high conidial concentration (106 conidia/mL) of the fungal isolate CT4-1 ( Fig. 2). The root discs inoculated with 104 conidia/mL of the fungal isolate rarely showed rot symptom development, only slight discoloration during 6 d of incubation, and no symptoms were find more observed in the non-inoculated control. In the pot experiment, severe root rot also developed in ginseng roots inoculated with the fungal isolate

C4-1 at inoculum concentrations of 1% and 5%; however, only mild and no rot symptoms were induced by the fungus with 0.2% inoculum concentration and the noninoculated control, respectively ( Fig. 2). In total, 392 microbial isolates obtained from various areas including rotten ginseng roots, Selleck PFI-2 crop fields, and mountain areas were screened for antifungal activity against F. cf. incarnatum C4-1, among which 10 bacterial isolates were selected as potential antagonists. These antagonistic bacteria and two additional bacterial isolates with no antifungal activity (for comparison) were screened again for antifungal activity against the fungal pathogen using a dual culture method. Among the tested bacterial isolates, B2-5 and B8 most inhibited the pathogen’s mycelial growth ( Fig. 3). The isolate B2-5 was selected and used for further biocontrol studies because B8 had a phytotoxic effect on the

ginseng root tissues (data not shown). ADAMTS5 The bacterial isolate B2-5 was Gram-positive, rod-shaped, and bacillus-like with peritrichous flagella (Fig. 4), showing the typical characteristics of Bacillus species as in a previous study [33]. Biological analysis showed that the isolate B2-5 utilized 24 carbon sources including sorbitol, but did not utilize 25 carbon sources including D-arabinose, revealing a 96.6% similarity to Bacillus subtilis and Bacillus amyloliquefaciens (data not shown). The 16S rRNA gene sequences of B2-5 (GenBank Accession No. KC478362) were found to have the highest similarity to B. amyloliquefaciens subsp. plantarum (NCBI Accession No. CP000560) of 99.80% (data not shown). Therefore, the bacterial isolate B2-5 was identified as B. amyloliquefaciens subsp. plantarum. The effects of the bacterial isolate on antifungal activity against the pathogen were tested at temperatures of 15°C, 18°C, 21°C, 25°C, and 28°C.