Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode

Pine wilt disease (PWD), caused by the pine wood nematode (PWN, <i>Bursaphelenchus xylophilus</i>), is one of the most serious threats to pine forests worldwide. The fungus <i>Esteya vermicola</i>, with its lunate conidia capable of parasitizing the PWN, has shown promise as...

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Main Authors: Lanwen Zhang, Yongxia Li, Xiaojian Wen, Xuan Wang, Wei Zhang, Dongzhen Li, Yuqian Feng, Zhenkai Liu, Xingyao Zhang
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/2/434
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author Lanwen Zhang
Yongxia Li
Xiaojian Wen
Xuan Wang
Wei Zhang
Dongzhen Li
Yuqian Feng
Zhenkai Liu
Xingyao Zhang
author_facet Lanwen Zhang
Yongxia Li
Xiaojian Wen
Xuan Wang
Wei Zhang
Dongzhen Li
Yuqian Feng
Zhenkai Liu
Xingyao Zhang
author_sort Lanwen Zhang
collection DOAJ
description Pine wilt disease (PWD), caused by the pine wood nematode (PWN, <i>Bursaphelenchus xylophilus</i>), is one of the most serious threats to pine forests worldwide. The fungus <i>Esteya vermicola</i>, with its lunate conidia capable of parasitizing the PWN, has shown promise as an efficient biological control agent against PWD. Solid-state fermentation (SSF) is preferred for large-scale applications in the field, as it facilitates microbial agent transport and ensures a long shelf life. However, research on enhancing the yield of lunate conidia from <i>E. vermicola</i> through SSF is limited. In this study, we initially achieved a yield of 3.04 × 10<sup>8</sup> conidia/g using a basic SSF medium composed of wheat bran, corn flour, and soybean flour. To improve this yield, we employed an orthogonal experimental design (OED) to identify the optimal medium composition, which required a wheat bran-to-corn flour-to soybean flour ratio of 7:2:1 (<i>w</i>/<i>w</i>/<i>w</i>), a substrate-to-water ratio of 1:0.7 (<i>w</i>/<i>v</i>), and the addition of 1.33% (<i>w</i>/<i>w</i>) glucose, 1.33% (<i>w</i>/<i>w</i>) yeast extract fermentation, and 1.33% (<i>w</i>/<i>w</i>) MgSO<sub>4</sub>. Using the response surface methodology (RSM), we calculated the optimal fermentation conditions, which were 24.9 °C, 78.0% relative humidity (RH), an inoculation volume of 16.3% (<i>v</i>/<i>w</i>), and a fermentation time of 7.1 days. Under these conditions, the yield of lunate conidia reached a maximum of 16.58 × 10<sup>8</sup> conidia/g, a 4.45-fold increase after optimization. This study improved the yield of <i>E. vermicola</i> lunate conidia and provides insights for developing biopesticides based on this strain.
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spelling doaj-art-d13335971eee4e36a10f23945bcfe0fe2025-08-20T02:01:23ZengMDPI AGMicroorganisms2076-26072025-02-0113243410.3390/microorganisms13020434Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood NematodeLanwen Zhang0Yongxia Li1Xiaojian Wen2Xuan Wang3Wei Zhang4Dongzhen Li5Yuqian Feng6Zhenkai Liu7Xingyao Zhang8Key Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, ChinaPine wilt disease (PWD), caused by the pine wood nematode (PWN, <i>Bursaphelenchus xylophilus</i>), is one of the most serious threats to pine forests worldwide. The fungus <i>Esteya vermicola</i>, with its lunate conidia capable of parasitizing the PWN, has shown promise as an efficient biological control agent against PWD. Solid-state fermentation (SSF) is preferred for large-scale applications in the field, as it facilitates microbial agent transport and ensures a long shelf life. However, research on enhancing the yield of lunate conidia from <i>E. vermicola</i> through SSF is limited. In this study, we initially achieved a yield of 3.04 × 10<sup>8</sup> conidia/g using a basic SSF medium composed of wheat bran, corn flour, and soybean flour. To improve this yield, we employed an orthogonal experimental design (OED) to identify the optimal medium composition, which required a wheat bran-to-corn flour-to soybean flour ratio of 7:2:1 (<i>w</i>/<i>w</i>/<i>w</i>), a substrate-to-water ratio of 1:0.7 (<i>w</i>/<i>v</i>), and the addition of 1.33% (<i>w</i>/<i>w</i>) glucose, 1.33% (<i>w</i>/<i>w</i>) yeast extract fermentation, and 1.33% (<i>w</i>/<i>w</i>) MgSO<sub>4</sub>. Using the response surface methodology (RSM), we calculated the optimal fermentation conditions, which were 24.9 °C, 78.0% relative humidity (RH), an inoculation volume of 16.3% (<i>v</i>/<i>w</i>), and a fermentation time of 7.1 days. Under these conditions, the yield of lunate conidia reached a maximum of 16.58 × 10<sup>8</sup> conidia/g, a 4.45-fold increase after optimization. This study improved the yield of <i>E. vermicola</i> lunate conidia and provides insights for developing biopesticides based on this strain.https://www.mdpi.com/2076-2607/13/2/434<i>Esteya vermicola</i><i>Bursaphelenchus xylophilus</i>solid-state fermentationorthogonal experimental designresponse surface methodology
spellingShingle Lanwen Zhang
Yongxia Li
Xiaojian Wen
Xuan Wang
Wei Zhang
Dongzhen Li
Yuqian Feng
Zhenkai Liu
Xingyao Zhang
Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
Microorganisms
<i>Esteya vermicola</i>
<i>Bursaphelenchus xylophilus</i>
solid-state fermentation
orthogonal experimental design
response surface methodology
title Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
title_full Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
title_fullStr Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
title_full_unstemmed Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
title_short Screening and Optimization of Solid-State Fermentation for <i>Esteya vermicola</i>, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
title_sort screening and optimization of solid state fermentation for i esteya vermicola i an entomopathogenic fungus against the major forest pest pine wood nematode
topic <i>Esteya vermicola</i>
<i>Bursaphelenchus xylophilus</i>
solid-state fermentation
orthogonal experimental design
response surface methodology
url https://www.mdpi.com/2076-2607/13/2/434
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