An RNA interference (RNAi) target with potential to control Dothistroma needle blight

Q3 Agricultural and Biological Sciences New Zealand Plant Protection Pub Date : 2023-10-04 DOI:10.30843/nzpp.2023.76.11770
Ashleigh M. Mosen, Yanan Guo, Berit Hassing, Carl H. Mesarich, Rosie E. Bradshaw
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Abstract

Spray-induced gene silencing (SIGS) involves exogenous spray applications of double-stranded RNA (dsRNA) molecules targeting specific genes, such as those essential to pathogens. This technique has shown great potential for controlling fungal pathogens of horticultural and agricultural plants, but few studies have evaluated its application to forest pathogens. Here we report the first demonstration of exogenous application of dsRNA molecules targeting a gene required for virulence in the foliar pine pathogen Dothistroma septosporum, the causal agent of Dothistroma needle blight (DNB). Constructs expressing sense and antisense dsRNAs were generated targeting a 509 bp fragment of the dothistromin toxin regulatory gene (DsAflR) in D. septosporum. DsAflR is required for the production of dothistromin, a phytotoxin essential for full pathogen virulence. We found that exogenous dsRNA is taken up and the RNA silencing mechanism is functional in D. septosporum through a series of in vitro and in planta trials, such as monitoring the uptake of fluorescently labelled DsAflR-dsRNA into the cell by confocal microscopy and quantifying the extent of gene silencing by quantitative Polymerase Chain Reaction (qPCR). In vitro applications of DsAflR-dsRNA to D. septosporum reduced the accumulation of DsAflR mRNA compared to controls and, in some cases, were found to be statistically significant, despite a high level of variability in the results. Our study also demonstrated a reduction in DNB symptoms on infected pine needles sprayed with DsAflR-dsRNA, which was further supported by a significant reduction in fungal biomass. This preliminary study highlights the potential for SIGS as a future management scheme for controlling the forest pathogen D. septosporum, as well as other forest pathogens.
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一种具有控制白斑病潜力的RNA干扰(RNAi)靶点
喷雾诱导的基因沉默(SIGS)涉及外源喷雾应用双链RNA (dsRNA)分子靶向特定基因,例如病原体必需的基因。该技术在控制园艺和农业植物的真菌病原体方面显示出巨大的潜力,但很少有研究评价其在森林病原体中的应用。在这里,我们首次报道了外源应用dsRNA分子靶向针叶松病菌(Dothistroma septosporum)毒力所需基因的实验。Dothistroma septosporum是Dothistroma needle blight (DNB)的病原体。以septosporum中降硫霉素毒素调控基因(DsAflR)的509 bp片段为靶点,构建了表达正义和反义dsRNAs的构建体。DsAflR是产生去硫氰酸酯所必需的,去硫氰酸酯是一种对病原体完全毒力必不可少的植物毒素。我们通过一系列的体外和植物试验,如用共聚焦显微镜监测荧光标记的DsAflR-dsRNA进入细胞的情况,用定量聚合酶链反应(qPCR)定量基因沉默的程度,发现d.s septosporum吸收了外源dsRNA,并且RNA沉默机制发挥了作用。与对照组相比,在体外将DsAflR- dsrna应用于隔孢菌可以减少DsAflR mRNA的积累,并且在某些情况下,尽管结果存在高度可变性,但发现具有统计学意义。我们的研究还表明,在喷洒了DsAflR-dsRNA的感染松针上,DNB症状有所减轻,真菌生物量的显著减少进一步支持了这一点。这项初步研究强调了SIGS作为控制森林病原菌septosporum以及其他森林病原菌的未来管理方案的潜力。
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来源期刊
New Zealand Plant Protection
New Zealand Plant Protection Agricultural and Biological Sciences-Insect Science
CiteScore
1.10
自引率
0.00%
发文量
8
期刊介绍: New Zealand Plant Protection is the journal of the New Zealand Plant Protection Society. It publishes original research papers on all aspects of biology, ecology and control of weeds, vertebrate and invertebrate pests, and pathogens and beneficial micro-organisms in agriculture, horticulture, forestry and natural ecosystems of relevance to New Zealand.
期刊最新文献
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