Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation
Pinosylvin stilbenes are natural phenolic compounds found in the Pinaceae family and act as phytoalexins. Some pinosylvin stilbenes have strong nematicidal activity against pine wood nematodes (PWNs: <i>Bursaphelenchus xylophilus</i>). Here, we established the efficient production of two...
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MDPI AG
2022-10-01
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| author | Yi Rae Kim Jung Yeon Han Yong Eui Choi |
| author_facet | Yi Rae Kim Jung Yeon Han Yong Eui Choi |
| author_sort | Yi Rae Kim |
| collection | DOAJ |
| description | Pinosylvin stilbenes are natural phenolic compounds found in the Pinaceae family and act as phytoalexins. Some pinosylvin stilbenes have strong nematicidal activity against pine wood nematodes (PWNs: <i>Bursaphelenchus xylophilus</i>). Here, we established the efficient production of two pinosylvin stilbenes, dihydropinosylvin monomethylether (DPME) and pinosylvin monomethylether (PME), by cell suspension culture of <i>Pinus koraiensis</i> after fungal elicitation. DPME and PME were found in small amounts (less than 40 µg/g DW) in the stem bark and leaves of <i>P. koraiensis</i> plants. Cell suspension cultures were established from the cultures of calli derived from mature zygotic embryos of <i>P. koraiensis</i> in 1/2 Litvay medium containing 2.2 μM 2,4-D and 2.2 μM BA. Two types of fungal elicitors, fungal cell extract (CE) and fungal medium filtrate (MF), were prepared from three species of fungi (<i>Penicillium chrysogenum, P. pinophilum</i>, and <i>P. roquefortii</i>). CE and MF treatments strongly stimulated the production of PME and DPME in cultured cells. The production of PME in suspension cells of <i>P. chrysogenum, P. pinophilum</i>, and <i>P. roquefortii</i> MF treatments after 3 days was 5734 µg/g DW, 4051 µg/g DW, and 6724 µg/g DW, respectively. Pinosylvin synthase (<i>PkSTS</i>) and pinosylvin <i>O</i>-methyltransferase (<i>PkPMT</i>) are key genes in DPME and PME biosynthesis. qPCR analysis revealed that the expression of the <i>PkSTS</i> and <i>PkPMT</i> in cultured cells was highly enhanced after fungal elicitor treatment. The cell extracts after MF treatment resulted in 92.5 ± 7.8% immobilization of the adult PWNs and 63.7 ± 3.5% immobilization of the juvenile PWNs within 24 h. However, control cell extracts without MF treatment showed 11.3 ± 1.4% nematicidal activity against adult PWNs. Our results suggest that pinosylvin stilbenes can be produced from the cell culture of <i>P. koraiensis</i> after fungal elicitor treatment and can be used as nematicidal compounds against PWNs. |
| first_indexed | 2024-03-09T18:44:08Z |
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| spelling | doaj.art-71641e1ae2e0455a8f6400bd93a591342023-11-24T06:25:28ZengMDPI AGPlants2223-77472022-10-011121293310.3390/plants11212933Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal ElicitationYi Rae Kim0Jung Yeon Han1Yong Eui Choi2Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon 200-701, KoreaDepartment of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon 200-701, KoreaDepartment of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon 200-701, KoreaPinosylvin stilbenes are natural phenolic compounds found in the Pinaceae family and act as phytoalexins. Some pinosylvin stilbenes have strong nematicidal activity against pine wood nematodes (PWNs: <i>Bursaphelenchus xylophilus</i>). Here, we established the efficient production of two pinosylvin stilbenes, dihydropinosylvin monomethylether (DPME) and pinosylvin monomethylether (PME), by cell suspension culture of <i>Pinus koraiensis</i> after fungal elicitation. DPME and PME were found in small amounts (less than 40 µg/g DW) in the stem bark and leaves of <i>P. koraiensis</i> plants. Cell suspension cultures were established from the cultures of calli derived from mature zygotic embryos of <i>P. koraiensis</i> in 1/2 Litvay medium containing 2.2 μM 2,4-D and 2.2 μM BA. Two types of fungal elicitors, fungal cell extract (CE) and fungal medium filtrate (MF), were prepared from three species of fungi (<i>Penicillium chrysogenum, P. pinophilum</i>, and <i>P. roquefortii</i>). CE and MF treatments strongly stimulated the production of PME and DPME in cultured cells. The production of PME in suspension cells of <i>P. chrysogenum, P. pinophilum</i>, and <i>P. roquefortii</i> MF treatments after 3 days was 5734 µg/g DW, 4051 µg/g DW, and 6724 µg/g DW, respectively. Pinosylvin synthase (<i>PkSTS</i>) and pinosylvin <i>O</i>-methyltransferase (<i>PkPMT</i>) are key genes in DPME and PME biosynthesis. qPCR analysis revealed that the expression of the <i>PkSTS</i> and <i>PkPMT</i> in cultured cells was highly enhanced after fungal elicitor treatment. The cell extracts after MF treatment resulted in 92.5 ± 7.8% immobilization of the adult PWNs and 63.7 ± 3.5% immobilization of the juvenile PWNs within 24 h. However, control cell extracts without MF treatment showed 11.3 ± 1.4% nematicidal activity against adult PWNs. Our results suggest that pinosylvin stilbenes can be produced from the cell culture of <i>P. koraiensis</i> after fungal elicitor treatment and can be used as nematicidal compounds against PWNs.https://www.mdpi.com/2223-7747/11/21/2933<i>Pinus koraiensis</i>pinosylvin stilbenespinosylvin monomethyletherdihydropinosylvin monomethylethercallus culturefungal elicitor treatment |
| spellingShingle | Yi Rae Kim Jung Yeon Han Yong Eui Choi Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation Plants <i>Pinus koraiensis</i> pinosylvin stilbenes pinosylvin monomethylether dihydropinosylvin monomethylether callus culture fungal elicitor treatment |
| title | Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation |
| title_full | Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation |
| title_fullStr | Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation |
| title_full_unstemmed | Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation |
| title_short | Production of Nematicidal Pinosylvin Stilbenes in Cell Suspension Cultures of <i>Pinus koraiensis</i> by Fungal Elicitation |
| title_sort | production of nematicidal pinosylvin stilbenes in cell suspension cultures of i pinus koraiensis i by fungal elicitation |
| topic | <i>Pinus koraiensis</i> pinosylvin stilbenes pinosylvin monomethylether dihydropinosylvin monomethylether callus culture fungal elicitor treatment |
| url | https://www.mdpi.com/2223-7747/11/21/2933 |
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