[논문]Pathological Properties of Cryptococcus pseudolongus on the Mycelia and Fruit Body of Lentinula edodes

Kwon, Hyuk Woo; Kim, Seong Hwan

doi:10.1080/12298093.2021.1873486

Pathological Properties of Cryptococcus pseudolongus on the Mycelia and Fruit Body of Lentinula edodes 원문보기

Mycobiology, v.49 no.2, 2021년, pp.173 - 182

Kwon, Hyuk Woo (Forest Mushroom Research Center, National Forestry Cooperative Federation) , Kim, Seong Hwan (Department of Microbiology and Institute of Biodiversity, Dankook University)

Abstract ▼ AI-Helper

Recently, Cryptococcus pseudolongus has been reported as a new pathogen of shiitake (Lentinula edodes). However, its pathological properties are not much known. To further understand its impact on the mushroom, we investigated the pathogen's interactions with the mycelium of shiitake, histopathological properties, host range, and sensitivity to diverse antifungal agents. The strain C. pseudolongus DUCC 4014 inhibited the mycelial growth of L. edodes strain (cultivar Sanjo 701ho) and caused browning in the mycelia confronted with the yeast on PDA. Spray inoculation of the yeast caused an abnormal browning symptom on the cap and/or gills of three shiitake cultivars grown on sawdust media in vinyl bags. Scanning electron microscopic images of the abnormally browned parts of shiitake fruit body illustrated that mushroom tissues were loosed and dispersed in the middle and edge of the cap and the arrangement of basidiospores borne on basidia in the gills was disturbed compared to those of normal shiitake fruit body. Spray inoculation also led to developing abnormal browning on the harvested fruit body, indicating C. pseudolongus could be a problem during mushroom storage. But the yeast was not able to induce abnormal browning on mushrooms of Pleurotus ferulae, Pleurotus fostreatus, and Agaricus bisporus. But it induced browning only on button mushroom (A. bisporus) when they were inoculated after wounding. Tests with 16 kinds of fungicides revealed that the cell growth of C. pseudolongus could be inhibited by benzalkonium chloride at MIC 7 ㎍/ml and benomyl at MIC 3 ㎍/ml.

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표/그림 (8)

그림 Figure 1. Inhibition and browning of the mycelia of shiitake (Sanjo 701ho) by Cryptococcus pseudolongus DUCC 4014 on PDA. (A): Top side of the media, (B): Bottom side of the media, (C): Confrontation area between the two fungi. Scale: 100 um.
그림 Figure 2. Inoculation test of Cryptococcus pseudolongus DUCC 4014 to three shiitake cultivars using a spray method. Sterile distilled water was used for the inoculum of the control test. The cell concentration of 1×10⁶ cells/ml was used for the inoculation. Scales: 10mm.
그림 Figure 3. Inoculation test of Cryptococcus pseudolongus DUCC4014 to three different mushrooms (Agaricus bisporus, Pleurotus ferulae, Pleurotus ostreatus) using spray and wound & drop methods. The cell concentration of 1×10⁶ cells/ml was used as inoculum. White arrows indicate the wounded position on the cap of the mushroom fruit bodies. Red circles indicate browning of the cut-inoculated area of the mushroom cap. Scales: 10mm.
그림 Figure 4. Stereo microscopic images of the Cryptococcus pseudolongus DUCC 4014 inoculated parts on the cap of three different shiitake cultivars. (A) Image of a part of the dissected mushroom cap with a wounded area; (B) Enlarged image in the cut-wounded area with a blade. Arrows in the A column indicate the wounded position on the cap of the mushroom fruit bodies. Arrows in the B column indicate inside of the cut-wounded area with a blade. Inoculation was performed after wounding with a blade cut. A drop of cells (1×10⁶ cells/ml) was inoculated onto the wounded area. Scales: 1mm.
그림 Figure 5. Scanning electron microscopic images of shiitake fruit body (Chamaram cultivar) inoculated by sterile water (a1-a3) and Cryptococcus pseudolongus DUCC 4014 (b1-b3) using a spray method. (A) Mimetic diagram of sampling position on the fruit body for SEM image analysis. Specimens of red boxed area in the control and inoculated fruit bodies were used for SEM observation, respectively; (B) SEM images of mushroom fruit bodies inoculated with sterile water (control, top pictures) and C. pseudolongus (bottom pictures). Scales: 100 μm (a-1, a-2, b-1, b-2) and 10 μm (a-3, b-3).
그림 Figure 6. Inoculation test using different Cryptococcus spp. to shiitake fruit bodies (Chamaram cultivar). Wound and spray methods were used for inoculation using the inoculum concentration of 1×10⁶ cells/ml. White arrows indicate the wounded position on the cap of the mushroom fruit bodies. Red arrows indicate inside of the cut-wounded area with a blade. Scales: 10 mm for fruit body and 1 mm for the stereo microscopic images.
그림 Figure 7. The effect of sixteen kinds of fungicides at three different concentrations on the cell growth of Cryptococcus pseudolongus DUCC 4014 on YMA. The tested C. pseudolongus cell concentration was adjusted from 1×10³ cells/ml to 1×10⁷ cells/ ml, respectively.
그림 Figure 8. Minimum inhibition concentration of benzalkonium chloride and benomyl to Cryptococcus pseudolongus DUCC 4014 on YMA. The concentration of the fungicides was made at 3 μg/ml to 10 μg/ml (benzalkonium chloride) and 0.5 μg/ml to 10 μg/ml (benomyl). C. pseudolongus cell concentration was adjusted from 1×10³ cells/ml to 1×10⁷ cells/ml.

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