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沉默宿主因子 eIF(iso)4E 基因赋予李树抗李痘病毒。

Silencing of the host factor eIF(iso)4E gene confers plum pox virus resistance in plum.

机构信息

Department of Biology, University of Western Ontario, London, Ontario, Canada.

出版信息

PLoS One. 2013;8(1):e50627. doi: 10.1371/journal.pone.0050627. Epub 2013 Jan 28.

DOI:10.1371/journal.pone.0050627
PMID:23382802
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3557289/
Abstract

Plum pox virus (PPV) causes the most economically-devastating viral disease in Prunus species. Unfortunately, few natural resistance genes are available for the control of PPV. Recessive resistance to some potyviruses is associated with mutations of eukaryotic translation initiation factor 4E (eIF4E) or its isoform eIF(iso)4E. In this study, we used an RNA silencing approach to manipulate the expression of eIF4E and eIF(iso)4E towards the development of PPV resistance in Prunus species. The eIF4E and eIF(iso)4E genes were cloned from plum (Prunus domestica L.). The sequence identity between plum eIF4E and eIF(iso)4E coding sequences is 60.4% at the nucleotide level and 52.1% at the amino acid level. Quantitative real-time RT-PCR analysis showed that these two genes have a similar expression pattern in different tissues. Transgenes allowing the production of hairpin RNAs of plum eIF4E or eIF(iso)4E were introduced into plum via Agrobacterium-mediated transformation. Gene expression analysis confirmed specific reduced expression of eIF4E or eIF(iso)4E in the transgenic lines and this was associated with the accumulation of siRNAs. Transgenic plants were challenged with PPV-D strain and resistance was evaluated by measuring the concentration of viral RNA. Eighty-two percent of the eIF(iso)4E silenced transgenic plants were resistant to PPV, while eIF4E silenced transgenic plants did not show PPV resistance. Physical interaction between PPV-VPg and plum eIF(iso)4E was confirmed. In contrast, no PPV-VPg/eIF4E interaction was observed. These results indicate that eIF(iso)4E is involved in PPV infection in plum, and that silencing of eIF(iso)4E expression can lead to PPV resistance in Prunus species.

摘要

李痘病毒 (PPV) 是李属植物中造成经济损失最严重的病毒性病害。遗憾的是，目前可用于防治 PPV 的天然抗性基因较少。某些 Potyvirus 的隐性抗性与真核翻译起始因子 4E（eIF4E）或其同工型 eIF(iso)4E 的突变有关。在本研究中，我们利用 RNA 沉默技术来调控 eIF4E 和 eIF(iso)4E 的表达，以期在李属植物中培育出对 PPV 的抗性。我们从李树（Prunus domestica L.）中克隆了 eIF4E 和 eIF(iso)4E 基因。李树 eIF4E 和 eIF(iso)4E 编码序列的核苷酸水平同源性为 60.4%，氨基酸水平同源性为 52.1%。定量实时 RT-PCR 分析表明，这两个基因在不同组织中的表达模式相似。通过农杆菌介导的转化法将允许产生李树 eIF4E 或 eIF(iso)4E 发夹 RNA 的转基因导入李树。基因表达分析证实了转 eIF4E 或 eIF(iso)4E 基因的李树中特定的 eIF4E 或 eIF(iso)4E 表达降低，并且与 siRNA 的积累相关。将这些转基因植株用 PPV-D 株系进行攻毒，通过测量病毒 RNA 的浓度来评估抗性。82%的 eIF(iso)4E 沉默的转基因植株对 PPV 具有抗性，而沉默 eIF4E 的转基因植株则未表现出对 PPV 的抗性。还证实了 PPV-VPg 与李树 eIF(iso)4E 之间存在物理相互作用。相比之下，没有观察到 PPV-VPg/eIF4E 相互作用。这些结果表明，eIF(iso)4E 参与了李树中 PPV 的感染，沉默 eIF(iso)4E 的表达可以导致李属植物对 PPV 的抗性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/0a1c0b3b63b5/pone.0050627.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/8cf4923573a3/pone.0050627.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/83724c2e8fdd/pone.0050627.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/1e968cd080c6/pone.0050627.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/8c9a6fb28974/pone.0050627.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/1fb9f1a25b26/pone.0050627.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/0a1c0b3b63b5/pone.0050627.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/8cf4923573a3/pone.0050627.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/83724c2e8fdd/pone.0050627.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/1e968cd080c6/pone.0050627.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/8c9a6fb28974/pone.0050627.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/1fb9f1a25b26/pone.0050627.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d0/3557289/0a1c0b3b63b5/pone.0050627.g006.jpg

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