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| High-Level Accumulation of Exogenous Small RNAs Not Affecting Endogenous Small RNA Biogenesis and Function in Plants |
| SHEN Wan-xia, Neil A Smith, ZHOU Chang-yong, WANG Ming-bo |
1、College of Plant Protection, Southwest University, Chongqing 400715, P.R.China
2、CSIRO Plant Industry, Canberra, ACT 2601, Australia
3、National Citrus Engineering Research Center, Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing 400712, P.R.China |
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摘要 RNA silencing is a fundamental plant defence and gene control mechanism in plants that are directed by 20-24 nucleotide (nt) small interfering RNA (siRNA) and microRNA (miRNA). Infection of plants with viral pathogens or transformation of plants with RNA interference (RNAi) constructs is usually associated with high levels of exogenous siRNAs, but it is unclear if these siRNAs interfere with endogenous small RNA pathways and hence affect plant development. Here we provide evidence that viral satellite RNA (satRNA) infection does not affect siRNA and miRNA biogenesis or plant growth despite the extremely high level of satRNA-derived siRNAs. We generated transgenic Nicotiana benthamiana plants that no longer develop the specific yellowing symptoms generally associated with infection by Cucumber mosaic virus (CMV) Y-satellite RNA (Y-Sat). We then used these plants to show that CMV Y-Sat infection did not cause any visible phenotypic changes in comparison to uninfected plants, despite the presence of high-level Y-Sat siRNAs. Furthermore, we showed that the accumulation of hairpin RNA (hpRNA)-derived siRNAs or miRNAs, and the level of siRNA-directed transgene silencing, are not significantly affected by CMV Y-Sat infection. Taken together, our results suggest that the high levels of exogenous siRNAs associated with viral infection or RNAi-inducing transgenes do not saturate the endogenous RNA silencing machineries and have no significant impact on normal plant development.
Abstract RNA silencing is a fundamental plant defence and gene control mechanism in plants that are directed by 20-24 nucleotide (nt) small interfering RNA (siRNA) and microRNA (miRNA). Infection of plants with viral pathogens or transformation of plants with RNA interference (RNAi) constructs is usually associated with high levels of exogenous siRNAs, but it is unclear if these siRNAs interfere with endogenous small RNA pathways and hence affect plant development. Here we provide evidence that viral satellite RNA (satRNA) infection does not affect siRNA and miRNA biogenesis or plant growth despite the extremely high level of satRNA-derived siRNAs. We generated transgenic Nicotiana benthamiana plants that no longer develop the specific yellowing symptoms generally associated with infection by Cucumber mosaic virus (CMV) Y-satellite RNA (Y-Sat). We then used these plants to show that CMV Y-Sat infection did not cause any visible phenotypic changes in comparison to uninfected plants, despite the presence of high-level Y-Sat siRNAs. Furthermore, we showed that the accumulation of hairpin RNA (hpRNA)-derived siRNAs or miRNAs, and the level of siRNA-directed transgene silencing, are not significantly affected by CMV Y-Sat infection. Taken together, our results suggest that the high levels of exogenous siRNAs associated with viral infection or RNAi-inducing transgenes do not saturate the endogenous RNA silencing machineries and have no significant impact on normal plant development.
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Received: 19 March 2013
Accepted:
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| Fund: MOA’s Public Benefit Research Foundation of China (201203076) and the grants from the National Key Technology R&D Program (2007BAD47B03), and the project from Prof. Wang Mingbo was supported by an Australian Research Council Future Fellowship (FT0991956). |
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Corresponding Authors:
WANG Ming-bo, Tel: +61-2-62465197, E-mail:ming-bo.wang@csiro.au; ZHOU Chang-yong, Tel: +86-23-68349701, E-mail: cyzhou@swu.edu.cn; Neil A Smith, E-mail: Neil.Smith@csiro.au
E-mail: ming-bo.wang@csiro.au;
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| About author: SHEN Wan-xia, Tel: +86-23-68349002, Mobile: 18680791095, E-mail: wxshen136@yahoo.com |
Cite this article:
SHEN Wan-xia, Neil A Smith, ZHOU Chang-yong, WANG Ming-bo.
2014.
High-Level Accumulation of Exogenous Small RNAs Not Affecting Endogenous Small RNA Biogenesis and Function in Plants. Journal of Integrative Agriculture, 13(5): 1017-1023.
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