Engineered Pine Endophytic Fungus Expressing dsRNA Targeting Lethal Genes to Control the Plant Parasitic Nematode Bursaphelenchus xylophilus

Wei Zhang; Ruijiong Wang; Yongxia Li; Dongzhen Li; Xuan Wang; Xiaojian Wen; Yuqian Feng; Zhenkai Liu; Shuai Ma; Xingyao Zhang

doi:10.1094/PHYTO-07-24-0203-R

Engineered Pine Endophytic Fungus Expressing dsRNA Targeting Lethal Genes to Control the Plant Parasitic Nematode Bursaphelenchus xylophilus

Phytopathology. 2024 Dec 24. doi: 10.1094/PHYTO-07-24-0203-R. Online ahead of print.

Authors

Wei Zhang^{1

2}, Ruijiong Wang³, Yongxia Li^{1

4

5}, Dongzhen Li⁶, Xuan Wang^{1

7}, Xiaojian Wen⁸, Yuqian Feng^{1

9}, Zhenkai Liu¹⁰, Shuai Ma¹¹, Xingyao Zhang¹²

Affiliations

¹ Chinese Academy of Forestry Ecology and Nature Conservation Institute, Beijing, China.
² Nanjing Forestry University, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing, China; [email protected].
³ Chinese Academy of Forestry Ecology and Nature Conservation Institute, Beijing, China; [email protected].
⁴ Nanjing Forestry University, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing, China.
⁵ Kunyushan Forest Ecosystem National Observation and Research Station, Yantai, China; [email protected].
⁶ Chinese Academy of Forestry Ecology and Nature Conservation Institute, Beijing, China; [email protected].
⁷ Nanjing Forestry University, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing, China; [email protected].
⁸ Chinese Academy of Forestry Ecology and Nature Conservation Institute, Beijing, China; [email protected].
⁹ Nanjing Forestry University, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing, China; [email protected].
¹⁰ Chinese Academy of Forestry Ecology and Nature Conservation Institute, Beijing, China; [email protected].
¹¹ Chinese Academy of Forestry, Beijing, China; [email protected].
¹² Chinese Academy of Forestry Ecology and Nature Conservation Institute, Beijing, China; [email protected].

PMID: 39718567
DOI: 10.1094/PHYTO-07-24-0203-R

Abstract

The pine wood nematode (PWN), Bursaphelenchus xylophilus, is one of the most serious invasive forest pests, responsible for pine wilt disease (PWD). Currently, there are no effective, environmentally friendly control methods available. RNA interference (RNAi) technology has been extensively utilized to screen functional genes in eukaryotes and to explore sustainable pest management approaches through genetic engineering. In this study, we identified 353 predicted lethal genes in PWN by comparing its genome with those of lethal genes from Caenorhabditis elegans. We selected five predicted lethal genes (Bxy1177, Bxy1239, Bxy1104, Bxy667, and BxyAK1) with identification values exceeding 60% to evaluate their nematicidal effects on PWN. We tested the double-stranded RNA (dsRNA) of these genes using two methods: firstly, soaking in a synthesized dsRNA solution in vitro, or secondly, feeding on a dsRNA-engineered endophytic fungus, Fusarium babinda. Following dsRNA ingestion, either through soaking or fungal feeding, the expression of genes Bxy1177, Bxy667, Bxy1104, and BxyAK1 was significantly suppressed. Notably, nematode populations that consumed fungi expressing dsL1177 and dsAK1 showed substantial declines over time. These findings provide novel insights and a practical foundation for employing endophytic fungi-expressed dsRNA in sustainable pest management strategies.

Keywords: Biological Control; Biotechnology; Disease Control and Pest Management.