External dsRNA Downregulates Anthocyanin Biosynthesis-Related Genes and Affects Anthocyanin Accumulation in Arabidopsis thaliana

Kiselev, Konstantin V.; Suprun, Andrey R.; Aleynova, Olga A.; Ogneva, Zlata V.; Kalachev, Alexander V.; Dubrovina, Alexandra S.

doi:10.3390/ijms22136749

Cited by 24 publications

(40 citation statements)

References 64 publications

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“…In our studies, the structural genes that regulated the phenylpropanoid biosynthesis pathways (43 genes), flavonoid biosynthesis pathways (22 genes), and anthocyanidin biosynthetic pathways (10 genes) were significantly expressed when the color of the leaves was changed (Figure 5). Among them, six PAL genes, nine 4CL genes, and six CHS genes were substantially expressed in red-colored leaves of A. triflorum, suggesting that these early biosynthetic genes lead to the production of flavonoids, as observed in other plant species with red leaves or fruits [39][40][41][42]. In addition, some downregulated PAL, 4CL, and CHS genes were also found in red leaves, suggesting that the color change in A. triflorum leaves was controlled by multiple single genes.…”

Section: Key Genes Related To Anthocyanin Synthesis In a Triflorum Le...mentioning

confidence: 77%

Metabolome and Transcriptome Analyses Unravels Molecular Mechanisms of Leaf Color Variation by Anthocyanidin Biosynthesis in Acer triflorum

et al. 2022

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Acer triflorum Komarov is an important ornamental tree, and its seasonal change in leaf color is the most striking feature. However, the quantifications of anthocyanin and the mechanisms of leaf color change in this species remain unknown. Here, the combined analysis of metabolome and transcriptome was performed on green, orange, and red leaves. In total, 27 anthocyanin metabolites were detected and cyanidin 3-O-arabinoside, pelargonidin 3-O-glucoside, and peonidin 3-O-gluside were significantly correlated with the color development. Several structural genes in the anthocyanin biosynthesis process, such as chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), and dihydroflavonol 4-reductase (DFR), were highly expressed in red leaves compared to green leaves. Most regulators (MYB, bHLH, and other classes of transcription factors) were also upregulated in red and orange leaves. In addition, 14 AtrMYBs including AtrMYB68, AtrMYB74, and AtrMYB35 showed strong interactions with the genes involved in anthocyanin biosynthesis, and, thus, could be further considered the hub regulators. The findings will facilitate genetic modification or selection for further improvement in ornamental qualities of A. triflorum.

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Section: Key Genes Related To Anthocyanin Synthesis In a Triflorum Le...mentioning

confidence: 77%

Metabolome and Transcriptome Analyses Unravels Molecular Mechanisms of Leaf Color Variation by Anthocyanidin Biosynthesis in Acer triflorum

et al. 2022

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“…Several studies show that it is possible to down-regulate plant transgenes, including GFP , GUS , and YFP [ 24 , 25 , 26 , 27 , 28 ] or plant endogenous genes, including R2R3-MYB transcription factors, sugar transporter gene, a downy mildew susceptibility gene, and a chalcone synthase gene [ 19 , 20 , 21 , 22 , 23 ]. However, the number of reports on the effect of exogenous RNAs on the expression of specific plant gene targets is small and there are scarce data on the specificity and effectiveness of this approach.…”

Section: Discussionmentioning

confidence: 99%

“…This strategy is based on the exogenously induced RNAi (exoRNAi), which is induced by the application of the synthetic exogenous RNAs to the plant surfaces and is being developed as a promising alternative to transgenic plants and VIGS for plant pathogen protection. At the same time, there are few reports on the successful application of this strategy to downregulate expression of plant endogenous genes [ 19 , 20 , 21 , 22 , 23 ] or plant transgenes [ 24 , 25 , 26 , 27 , 28 ]. Several studies have reported downregulation of transgenes encoding green fluorescent protein (GFP), β-glucuronidase (GUS), yellow fluorescent protein (YFP), or neomycin phosphotransferase II ( NPTII ) during external application of dsRNA, siRNA, or hpRNA solutions of the leaf surface of tobacco or Arabidopsis [ 24 , 25 , 26 , 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

The Specificity of Transgene Suppression in Plants by Exogenous dsRNA

Kiselev

Suprun

Aleynova

et al. 2022

Plants

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The phenomenon of RNA interference (RNAi) is widely used to develop new approaches for crop improvement and plant protection. Recent investigations show that it is possible to downregulate plant transgenes, as more prone sequences to silencing than endogenous genes, by exogenous application of double-stranded RNAs (dsRNAs) and small interfering RNAs (siRNAs). However, there are scarce data on the specificity of exogenous RNAs. In this study, we explored whether plant transgene suppression is sequence-specific to exogenous dsRNAs and whether similar effects can be caused by exogenous DNAs that are known to be perceived by plants and induce certain epigenetic and biochemical changes. We treated transgenic plants of Arabidopsis thaliana bearing the neomycin phosphotransferase II (NPTII) transgene with specific synthetic NPTII-dsRNAs and non-specific dsRNAs, encoding enhanced green fluorescent protein (EGFP), as well as with DNA molecules mimicking the applied RNAs. None of the EGFP-dsRNA doses resulted in a significant decrease in NPTII transgene expression in the NPTII-transgenic plants, while the specific NPTII-dsRNA significantly reduced NPTII expression in a dose-dependent manner. Long DNAs mimicking dsRNAs and short DNA oligonucleotides mimicking siRNAs did not exhibit a significant effect on NPTII transgene expression. Thus, exogenous NPTII-dsRNAs induced a sequence-specific and RNA-specific transgene-suppressing effect, supporting external application of dsRNAs as a promising strategy for plant gene regulation.

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“…Anthocyanin and stilbene content analysis was performed by the method HPLC-MS as described [ 45 , 47 , 48 ]. Briefly, for anthocyanins 100 mg fresh cells tissue were subsequently homogenized using a mortar and a pestle in 1 mL of 1% ( v/v ) hydrochloric acid in methanol.…”

Section: Methodsmentioning

confidence: 99%

Effect of VaMyb40 and VaMyb60 Overexpression on Stilbene Biosynthesis in Cell Cultures of Grapevine Vitis amurensis Rupr.

Ananev

Suprun

Aleynova

et al. 2022

Plants

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Stilbenes are plant defense compounds known to rapidly accumulate in grapevine and some other plant species in response to microbial infection and several abiotic stresses. Stilbenes have attracted considerable attention due to valuable biological effects with multi-spectrum therapeutic application. However, there is a lack of information on natural signaling pathways and transcription factors regulating stilbene biosynthesis. It has been previously shown that MYB R2R3 transcription factor genes VaMyb40 and VaMyb60 were up-regulated in cell cultures of wild-growing grapevine Vitis amurensis Rupr. in response to UV irradiation. In this study, the effects of VaMyb40 or VaMyb60 overexpression in cell cultures of V. amurensis on their capability to produce stilbenes were investigated. Overexpression of the VaMyb60 gene led to a considerable increase in the content of stilbenes in three independently transformed transgenic lines in 5.9–13.9 times, while overexpression of the VaMyb40 gene also increased the content of stilbenes, although to a lesser extent (in 3.4–4.0 times) in comparison with stilbene levels in the empty vector-transformed calli. Stilbene content and stilbene production in the VaMyb60-transgenic calli reached 18.8 mg/g of dry weight (DW) and 150.8 mg/L, respectively. Using HPLC analysis, we detected eight individual stilbenes: t-resveratrol diglucoside, t-piceid, t-resveratrol, ε-viniferin, δ-viniferin, cis-resveratrol, cis-piceid, t-piceatannol. T-resveratrol prevailed over other stilbenoid compounds (53.1–89.5% of all stilbenes) in the VaMyb-overexpressing cell cultures. Moreover, the VaMyb40- and VaMyb60-transformed calli were capable of producing anthocyanins up to 0.035 mg/g DW, while the control calli did not produce anthocyanins. These findings show that the VaMyb40 and VaMyb60 genes positively regulate the stilbene biosynthesis as strong positive transcription regulators and can be used in biotechnological applications for stilbene production or high-quality viticulture and winemaking.

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External dsRNA Downregulates Anthocyanin Biosynthesis-Related Genes and Affects Anthocyanin Accumulation in Arabidopsis thaliana

Cited by 24 publications

References 64 publications

Metabolome and Transcriptome Analyses Unravels Molecular Mechanisms of Leaf Color Variation by Anthocyanidin Biosynthesis in Acer triflorum

Metabolome and Transcriptome Analyses Unravels Molecular Mechanisms of Leaf Color Variation by Anthocyanidin Biosynthesis in Acer triflorum

The Specificity of Transgene Suppression in Plants by Exogenous dsRNA

Effect of VaMyb40 and VaMyb60 Overexpression on Stilbene Biosynthesis in Cell Cultures of Grapevine Vitis amurensis Rupr.

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