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不同植物中来源于竹花叶病毒及其伴随卫星 RNA 的小干扰 RNA 的全局分析。

Global analyses of small interfering RNAs derived from Bamboo mosaic virus and its associated satellite RNAs in different plants.

机构信息

Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan.

出版信息

PLoS One. 2010 Aug 2;5(8):e11928. doi: 10.1371/journal.pone.0011928.

Abstract

BACKGROUND

Satellite RNAs (satRNAs), virus parasites, are exclusively associated with plant virus infection and have attracted much interest over the last 3 decades. Upon virus infection, virus-specific small interfering RNAs (vsiRNAs) are produced by dicer-like (DCL) endoribonucleases for anti-viral defense. The composition of vsiRNAs has been studied extensively; however, studies of satRNA-derived siRNAs (satsiRNAs) or siRNA profiles after satRNA co-infection are limited. Here, we report on the small RNA profiles associated with infection with Bamboo mosaic virus (BaMV) and its two satellite RNAs (satBaMVs) in Nicotiana benthamiana and Arabidopsis thaliana.

METHODOLOGY/PRINCIPAL FINDINGS: Leaves of N. benthamiana or A. thaliana inoculated with water, BaMV alone or co-inoculated with interfering or noninterfering satBaMV were collected for RNA extraction, then large-scale Solexa sequencing. Up to about 20% of total siRNAs as BaMV-specific siRNAs were accumulated in highly susceptible N. benthamiana leaves inoculated with BaMV alone or co-inoculated with noninterfering satBaMV; however, only about 0.1% of vsiRNAs were produced in plants co-infected with interfering satBaMV. The abundant region of siRNA distribution along BaMV and satBaMV genomes differed by host but not by co-infection with satBaMV. Most of the BaMV and satBaMV siRNAs were 21 or 22 nt, of both (+) and (-) polarities; however, a higher proportion of 22-nt BaMV and satBaMV siRNAs were generated in N. benthamiana than in A. thaliana. Furthermore, the proportion of non-viral 24-nt siRNAs was greatly increased in N. benthamiana after virus infection.

CONCLUSIONS/SIGNIFICANCE: The overall composition of vsiRNAs and satsiRNAs in the infected plants reflect the combined action of virus, satRNA and different DCLs in host plants. Our findings suggest that the structure and/or sequence demands of various DCLs in different hosts may result in differential susceptibility to the same virus. DCL2 producing 24-nt siRNAs under biotic stresses may play a vital role in the antiviral mechanism in N. benthamiana.

摘要

背景

卫星 RNA(satRNA)是病毒寄生虫,仅与植物病毒感染有关,在过去 30 年中引起了广泛关注。在病毒感染后,病毒特异性小干扰 RNA(vsiRNA)由 Dicer 样(DCL)内切核酸酶产生,用于抗病毒防御。vsiRNA 的组成已被广泛研究;然而,satRNA 衍生的 siRNA(satsiRNA)或 satRNA 共感染后的 siRNA 谱研究有限。在这里,我们报告了感染竹花叶病毒(BaMV)及其两个卫星 RNA(satBaMV)在 Nicotiana benthamiana 和 Arabidopsis thaliana 中与感染相关的小 RNA 谱。

方法/主要发现:用水、单独的 BaMV 或共接种干扰或非干扰 satBaMV 的 N. benthamiana 或 A. thaliana 叶片采集 RNA 提取,然后进行大规模 Solexa 测序。在单独接种 BaMV 或共接种非干扰 satBaMV 的高度敏感的 N. benthamiana 叶片中,积累了高达约 20%的总 siRNA 作为 BaMV 特异性 siRNA;然而,在共感染干扰 satBaMV 的植物中仅产生约 0.1%的 vsiRNA。siRNA 分布在 BaMV 和 satBaMV 基因组上的丰富区域因宿主而异,但不因共感染 satBaMV 而异。大多数 BaMV 和 satBaMV siRNA 为 21 或 22 nt,具有(+)和(-)极性;然而,在 N. benthamiana 中产生的 22-nt BaMV 和 satBaMV siRNA 比例高于 A. thaliana。此外,感染病毒后,N. benthamiana 中非病毒 24-nt siRNA 的比例大大增加。

结论/意义:受感染植物中 vsiRNA 和 satsiRNA 的总体组成反映了病毒、satRNA 和宿主植物中不同 DCL 的共同作用。我们的研究结果表明,不同宿主中各种 DCL 的结构和/或序列需求可能导致对同一病毒的不同敏感性。在生物胁迫下产生 24-nt siRNA 的 DCL2 可能在 N. benthamiana 的抗病毒机制中发挥重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab55/2914070/32837ff20627/pone.0011928.g001.jpg

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