Double-stranded RNA-mediated interference of dumpy genes in Bursaphelenchus xylophilus by feeding on filamentous fungal transformants

doi:10.1016/j.ijpara.2016.01.008

	Double-stranded RNA-mediated interference of dumpy genes in Bursaphelenchus xylophilus by feeding on filamentous fungal transformants
	Wang, Meng 1; Wang, Diandong 2,4; Zhang, Xi 1; Wang, Xu 1; Liu, Wencui 1; Hou, Xiaomeng 1; Huang, Xiaoyin 1; Xie, Bingyan 2; Cheng, Xinyue 1,3
刊名	INTERNATIONAL JOURNAL FOR PARASITOLOGY
	2016
卷号	46 期号:5-6 页码:351-360
关键词	Pinewood nematode Bursaphelenchus xylophilus Silencing vector pDH-RH RNA interference (RNAi) feeding Dumpy gene Collagen Filamentous fungal transformant Agrobacterium tumefaciens-mediated transformation (ATMT)
ISSN号	0020-7519
DOI	10.1016/j.ijpara.2016.01.008
通讯作者	Wang, Meng
英文摘要	RNA interference (RNAi) is a valuable tool for studying gene function in vivo and provides a functional genomics platform in a wide variety of organisms. The pinewood nematode, Bursaphelenchus xylophilus, is a prominent invasive plant-parasitic nematode and has become a serious worldwide threat to forest ecosystems. Presently, the complete genome sequence of B. xylophilus has been published, and research involving genome-wide functional analyses is likely to increase. In this study, we describe the construction of an effective silencing vector, pDH-RH, which contains a transcriptional unit for a hairpin loop structure. Utilising this vector, double-stranded (ds)RNAs with sequences homologous to the target genes can be expressed in a transformed filamentous fungus via Agrobacterium tumefaciens-mediated transformation technology, and can subsequently induce the knockdown of target gene mRNA expression in B. xylophilus by allowing the nematode to feed on the fungal transformants. Four dumpy genes (Bx-dpy-2, 4, 10 and 11) were used as targets to detect RNAi efficiency. By allowing the nematode to feed on target gene-transformed Fusarium oxysporum strains, target transcripts were knocked down 34-87% compared with those feeding on the wild-type strain as determined by real-time quantitative PCR (RT-qPCR). Morphological RNAi phenotypes were observed, displaying obviously reduced body length; weak dumpy or small (short and thin) body size; or general abnormalities. Moreover, compensatory regulation and non-specific silencing of dpy genes were found in B. xylophilus. Our results indicate that RNAi delivery by feeding in B. xylophilus is a successful technique. This platform may provide a new opportunity for undertaking RNAi-based, genome-wide gene functional studies in vitro in B. xylophilus. Moreover, as B. xylophilus feeds on endophytic fungi when a host has died, RNAi feeding technology will offer the prospect for developing a novel control strategy for the nematode. Furthermore, this platform may also be applicable to other parasitic nematodes that have a facultative, fungivorous habit. (C) 2016 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
学科主题	Parasitology
语种	英语
出版者	ELSEVIER SCI LTD
WOS记录号	WOS:000375811600006
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/100822]
专题	蔬菜花卉研究所_植物保护研究室
作者单位	1.Beijing Normal Univ, Coll Life Sci, Beijing 100875, Peoples R China 2.Chinese Acad Agr Sci, Inst Vegetables & Flowers, Beijing 100081, Peoples R China 3.Minist Educ, Key Lab Biodivers Sci & Ecol Engn, Beijing 100875, Peoples R China 4.Yangtze Normal Univ, Chongqing 408100, Peoples R China
推荐引用方式 GB/T 7714	Wang, Meng,Wang, Diandong,Zhang, Xi,et al. Double-stranded RNA-mediated interference of dumpy genes in Bursaphelenchus xylophilus by feeding on filamentous fungal transformants[J]. INTERNATIONAL JOURNAL FOR PARASITOLOGY,2016,46(5-6):351-360.
APA	Wang, Meng.,Wang, Diandong.,Zhang, Xi.,Wang, Xu.,Liu, Wencui.,...&Cheng, Xinyue.(2016).Double-stranded RNA-mediated interference of dumpy genes in Bursaphelenchus xylophilus by feeding on filamentous fungal transformants.INTERNATIONAL JOURNAL FOR PARASITOLOGY,46(5-6),351-360.
MLA	Wang, Meng,et al."Double-stranded RNA-mediated interference of dumpy genes in Bursaphelenchus xylophilus by feeding on filamentous fungal transformants".INTERNATIONAL JOURNAL FOR PARASITOLOGY 46.5-6(2016):351-360.