A Genetic Network for Systemic RNA Silencing in Plants

Chen, Weiwei; Zhang, Xian; Fan, Yaya; Li, Bin; Ryabov, Eugene V.; Shi, Nongnong; Zhao, Mei; Yu, Zhiming; Cheng, Qiqun; Qin, Zheng; Zhang, Pengcheng; Wang, Huizhong; Jackson, Stephen D.; Cheng, Qi; Liu, Yule; Gallusci, Philippe; Hong, Yiguo

doi:10.1104/pp.17.01828

Cited by 57 publications

(56 citation statements)

References 54 publications

(46 reference statements)

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“…Briefly, a nontranslatable 200-to 400-bp fragment corresponding to the coding region of each gene was PCR amplified and cloned into MluI/SalI or ClaI/EagI sites of the PVX vector (Bruce et al, 2011) to produce PVX/SlMET1, PVX/SlCMT4, PVX/SlNCED, PVX/SlZ-ISO, PVX/SlABI3, PVX/SlPSY1, and PVX/SlDET1. To generate the SlMET1 RNAi construct pRNAi-SlMET1, a 250-bp SlMET1 fragment was PCR amplified using tomato cDNA as template and cloned in the sense and antisense orientations into the pRNAi-LIC vector (Chen et al, 2018b). Primers used for making these constructs are listed in Supplemental Table S1.…”

Section: Vigs and Rnai Constructsmentioning

confidence: 99%

METHYLTRANSFERASE1 and Ripening Modulate Vivipary during Tomato Fruit Development

et al. 2020

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Vivipary, wherein seeds germinate prior to dispersal while still associated with the maternal plant, is an adaptation to extreme environments. It is normally inhibited by the establishment of dormancy. The genetic framework of vivipary has been well studied; however, the role of epigenetics in vivipary remains unknown. Here, we report that silencing of METHYLTRANSFERASE1 (SlMET1) promoted precocious seed germination and seedling growth within the tomato (Solanum lycopersicum) epimutant Colorless non-ripening (Cnr) fruits. This was associated with decreases in abscisic acid concentration and levels of mRNA encoding 9-cis-epoxycarotenoid-dioxygenase (SlNCED), which is involved in abscisic acid biosynthesis. Differentially methylated regions were identified in promoters of differentially expressed genes, including SlNCED. SlNCED knockdown also induced viviparous seedling growth in Cnr fruits. Strikingly, Cnr ripening reversion suppressed vivipary. Moreover, neither SlMET1/SlNCED-virus-induced gene silencing nor transgenic SlMET1-RNA interference produced vivipary in wildtype tomatoes; the latter affected leaf architecture, arrested flowering, and repressed seed development. Thus, a dual pathway in ripening and SlMET1-mediated epigenetics coordinates the blockage of seed vivipary.

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Section: Vigs and Rnai Constructsmentioning

confidence: 99%

METHYLTRANSFERASE1 and Ripening Modulate Vivipary during Tomato Fruit Development

et al. 2020

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“…There are several pathways for the generation of 22 nt siRNAs or miRNAs in plant cells. DCL2 processes 22 nt siRNAs from long dsRNAs, and its activity is critical for both the production of secondary siRNAs and the transitive silencing of transgenes (Mlotshwa et al 2008;Parent et al 2014;Taochy et al 2017;Chen et al 2018). Alternatively, 22 nt miRNAs can be generated by processing by DCL1 of an asymmetric precursor miRNA to generate a 21/22 nt miRNA (Manavella et al 2012), by imprecise cutting by DCL1 (Li et al 2016), or by mono-uridylation by HEN1 Supressor1 (HESO1) of a 21 nt miRNA (Fei et al 2018).…”

Section: Introductionmentioning

confidence: 99%

Topically delivered 22 nt siRNAs enhance RNAi silencing of endogenous genes in two species

Hendrix

Zheng

Bauer

et al. 2020

Preprint

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Abstract22 nt miRNAs or siRNAs have been shown to specifically induce production of transitive (secondary) siRNAs for targeted mRNAs. An abrasion method to deliver dsRNAs into leaf cells of intact plants was used to investigate the activities of 21 and 22 nt siRNAs in silencing genes in Nicotiana benthamiana and Amaranthus cruentus. We confirmed that both 21 and 22 nt siRNAs were able to silence a green fluorescent protein (GFP) transgene in treated leaves of N. benthamiana, but systemic silencing of GFP occurred only when the guide strand contained 22 nt. Silencing in the treated leaves of N. benthamiana was demonstrated for 3 endogenous genes: magnesium cheletase subunit I (CHL-I), magnesium cheletase subunit H (CHL-H), and GUN4. However, systemic silencing of these endogenous genes was not observed. Very high levels of transitive siRNAs were produced for GFP in response to treatment with 22 nt siRNAs, but only low levels were produced in response to a 21 nt siRNA. The endogenous genes tested also had more transitive siRNAs produced in response to 22 nt siRNAs, but the response varied from weak (CHL-I) to strong (CHL-H). 22 nt siRNAs produced greater local silencing phenotypes than 21 nt siRNAs for GFP, CHL-H and GUN4 in N. benthamiana. The special activity of 22 nt siRNAs in producing a greater local phenotype and induction of elevated levels of transitive siRNAs was also shown in A. cruentus for the CHL-H gene. These experiments suggest a functional role for transitive siRNAs in amplifying the RNAi response.

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“…These dsRNAs are processed into 21- to 24-nucleotide long small RNAs (sRNA), known as small interfering RNAs (siRNAs), by the RNase III -type ribonucleases (DICER or DICER-LIKE). The RNA-induced silencing complex recruits these siRNAs to direct the degradation of complementary RNAs [16,17]. In plants, RNA-dependent RNA polymerases (RdR, previously, s ilencing de fective (SDE)1/ s uppressor of g ene s ilencing (SGS)2) have been shown to be essential for virus-induced post-transcriptional gene silencing [18,19,20].…”

Section: Introductionmentioning

confidence: 99%

Suppression of RNA-Dependent RNA Polymerase 6 Favors the Accumulation of Potato Spindle Tuber Viroid in Nicotiana Benthamiana

Adkar-Purushothama

Perreault

2019

Viruses

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To date, two plant genes encoding RNA-dependent RNA polymerases (RdRs) that play major roles in the defense against RNA viruses have been identified: (i) RdR1, which is responsible for the viral small RNAs (vsRNAs) found in virus-infected plants, and, (ii) RdR6, which acts as a surrogate in the absence of RdR1. In this study, the role of RdR6 in the defense against viroid infection was examined by knock-down of RdR6 followed by potato spindle tuber viroid (PSTVd) infection. The suppression of RdR6 expression increased the plant’s growth, as was illustrated by the plant’s increased height. PSTVd infection of RdR6 compromised plants resulted in an approximately three-fold increase in the accumulation of viroid RNA as compared to that seen in control plants. Additionally, RNA gel blot assay revealed an increase in the number of viroids derived small RNAs in RdR6 suppressed plants as compared to control plants. These data provide a direct correlation between RdR6 and viroid accumulation and indicate the role of RDR6 in the plant’s susceptibility to viroid infection.

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A Genetic Network for Systemic RNA Silencing in Plants

Cited by 57 publications

References 54 publications

METHYLTRANSFERASE1 and Ripening Modulate Vivipary during Tomato Fruit Development

METHYLTRANSFERASE1 and Ripening Modulate Vivipary during Tomato Fruit Development

Topically delivered 22 nt siRNAs enhance RNAi silencing of endogenous genes in two species

Suppression of RNA-Dependent RNA Polymerase 6 Favors the Accumulation of Potato Spindle Tuber Viroid in Nicotiana Benthamiana

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