Crop Biotechnology Research Group, College of Science, Health, Engineering and Education, WA State Agricultural Biotechnology Centre, Murdoch University, Perth, Australia.
Sci Rep. 2021 May 27;11(1):11156. doi: 10.1038/s41598-021-90363-8.
Dicers and dicer-like enzymes play an essential role in small RNA processing in eukaryotes. Nematodes are thought to encode one dicer, DCR-1; only that for Caenorhabditis spp. is well-characterised. Using genomic sequences of eight root-knot nematodes (Meloidogyne spp.), we identified putative coding sequences typical of eukaryotic DICERS. We noted that the primary and secondary structures of DICERS they encode were different for different Meloidogyne species and even for isolates of the same species, suggesting paralogy for the gene. One of the genes for M. incognita (Midcr-1.1) expressed in eggs, juvenile stage 2 and adults, with the highest expression in the adult females. All the Meloidogyne DICERS had seven major domains typical of those for Caenorhabditis spp. and humans with very similar protein folding. RNAi of Midcr-1.1 in J2s using seven dsRNAs, each based on sequences encoding the domains, induced mild paralysis but measurable knockdown was detected in J2s treated with five of the dsRNAs. For four of the dsRNAs, the RNAi effect lasted and reduced the nematode's infectivity. Also, host plant delivery of dsRNAs complementary to coding sequences of the Dicer Dimerisation domain impaired development, reducing nematode infection by 71%. These results confirm the importance of the gene to nematode health.
Dicers 和 dicer-like 酶在真核生物的小 RNA 加工中发挥着重要作用。线虫被认为编码一个 dicer,DCR-1;只有 Caenorhabditis spp. 的 well-characterised。使用 8 种根结线虫(Meloidogyne spp.)的基因组序列,我们鉴定出了具有典型真核 DICERS 的编码序列。我们注意到,它们编码的 DICERS 的一级和二级结构因不同的 Meloidogyne 物种甚至同一物种的分离株而不同,这表明该基因存在基因平行现象。其中一个在卵、2 龄幼虫和成虫中表达的 M. incognita(Midcr-1.1)基因,在成虫雌性中表达量最高。所有的 Meloidogyne DICERS 都有 7 个主要结构域,与 Caenorhabditis spp. 和人类的结构域非常相似,具有非常相似的蛋白质折叠。使用 7 条 dsRNA 在 J2s 中对 Midcr-1.1 进行 RNAi,每条 dsRNA 都基于编码结构域的序列,诱导轻微的麻痹,但在用 5 条 dsRNA 处理的 J2s 中检测到可测量的敲低。对于 4 条 dsRNA,RNAi 效果持续并降低了线虫的感染力。此外,dsRNA 互补物对 Dicer Dimerisation 结构域编码序列的宿主植物传递损害了发育,使线虫感染减少了 71%。这些结果证实了该基因对线虫健康的重要性。