A virus‐induced gene‐silencing system for functional genetics in a betalainic species, <i>Amaranthus tricolor</i> (Amaranthaceae)

Adhikary, Dinesh; Khatri-Chhetri, Upama; Tymm, Fiona J. M.; Murch, Susan J.; Deyholos, Michael K.

doi:10.1002/aps3.1221

Cited by 17 publications

(13 citation statements)

References 46 publications

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“…However, the molecular basis underlying betalain biosynthesis in A. tricolor remains largely unknown. Recently, virus-induced gene silencing was applied to elucidate the function of CYP76AD1 in producing betalain pigments in A. tricolor 29 . In addition, a comparative analysis of a transcriptome database constructed from different leaf samples of A. tricolor cv.…”

Section: Introductionmentioning

confidence: 99%

Elucidation of the core betalain biosynthesis pathway in Amaranthus tricolor

Chang

Chiu

Tsao

et al. 2021

Sci Rep

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Amaranthus tricolor L., a vegetable Amaranthus species, is an economically important crop containing large amounts of betalains. Betalains are natural antioxidants and can be classified into betacyanins and betaxanthins, with red and yellow colors, respectively. A. tricolor cultivars with varying betalain contents, leading to striking red to green coloration, have been commercially produced. However, the molecular differences underlying betalain biosynthesis in various cultivars of A. tricolor remain largely unknown. In this study, A. tricolor cultivars with different colors were chosen for comparative transcriptome analysis. The elevated expression of AmCYP76AD1 in a red-leaf cultivar of A. tricolor was proposed to play a key role in producing red betalain pigments. The functions of AmCYP76AD1, AmDODAα1, AmDODAα2, and AmcDOPA5GT were also characterized through the heterologous engineering of betalain pigments in Nicotiana benthamiana. Moreover, high and low L-DOPA 4,5-dioxygenase activities of AmDODAα1 and AmDODAα2, respectively, were confirmed through in vitro enzymatic assays. Thus, comparative transcriptome analysis combined with functional and enzymatic studies allowed the construction of a core betalain biosynthesis pathway of A. tricolor. These results not only provide novel insights into betalain biosynthesis and evolution in A. tricolor but also provide a basal framework for examining genes related to betalain biosynthesis among different species of Amaranthaceae.

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Section: Introductionmentioning

confidence: 99%

Elucidation of the core betalain biosynthesis pathway in Amaranthus tricolor

Chang

Chiu

Tsao

et al. 2021

Sci Rep

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“…Therefore, in the past decade, tremendous improvements in VIGS have been reported [ 4 – 8 , 12 ]. Efforts has been made to select the efficient reporter gene [ 10 , 26 – 28 ]. As far as we know, VIGS vectors with traceable and visualizable indicator for the prediction of positive gene silencing plants for reverse genetics are still lacking.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous silencing of two different Arabidopsis genes with a novel virus-induced gene silencing vector

Zhang

et al. 2021

Plant Methods

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Background Virus-induced gene silencing (VIGS) is a useful tool for functional characterizations of plant genes. However, the penetrance of VIGS varies depending on the genes to be silenced, and has to be evaluated by examining the transcript levels of target genes. Results In this report, we report the development of a novel VIGS vector that permits a preliminary assessment of the silencing penetrance. This new vector is based on an attenuated variant of Turnip crinkle virus (TCV) known as CPB that can be readily used in Arabidopsis thaliana to interrogate genes of this model plant. A CPB derivative, designated CPB1B, was produced by inserting a 46 nucleotide section of the Arabidopsis PHYTOENE DESATURASE (PDS) gene into CPB, in antisense orientation. CPB1B induced robust PDS silencing, causing easily visible photobleaching in systemically infected Arabidopsis leaves. More importantly, CPB1B can accommodate additional inserts, derived from other Arabidopsis genes, causing the silencing of two or more genes simultaneously. With photobleaching as a visual marker, we adopted the CPB1B vector to validate the involvement of DICER-LIKE 4 (DCL4) in antiviral defense against TCV. We further revealed the involvement of ARGONAUTE 2 (AGO2) in PDS silencing and antiviral defense against TCV in dcl2drb4 double mutant plants. These results demonstrated that DOUBLE-STRANDED RNA-BINDING PROTEIN 4 (DRB4), whose protein product (DRB4) commonly partners with DCL4 in the antiviral silencing pathway, was dispensable for PDS silencing induced by CPB1B derivative in dcl2drb4 double mutant plants. Conclusions The CPB1B-based vector developed in this work is a valuable tool with visualizable indicator of the silencing penetrance for interrogating Arabidopsis genes, especially those involved in the RNA silencing pathways.

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“…Therefore, in the past decade, tremendous improvements in VIGS have been reported [4-8, 12, 26]. Efforts has been made to select the e cient reporter gene [10,19,[27][28][29]. As far as we know, VIGS vectors with traceable and visualizable indicator for the prediction of positive gene silencing plants for reverse genetics are still lacking.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Silencing of Two Different Arabidopsis Genes With A Novel Virus-Induced Gene Silencing Vector

Zhang

et al. 2020

Preprint

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Background: Virus-induced gene silencing (VIGS) is a useful tool for functional characterizations of plant genes. However, the penetrance of VIGS varies depending on the genes to be silenced, and has to be evaluated by examining the transcript levels of target genes. Results: In this report, we report the development of a novel VIGS vector that permits a preliminary assessment of the silencing penetrance. This new vector is based on an attenuated variant of Turnip crinkle virus (TCV) known as CPB that can be readily used in Arabidopsis thaliana to interrogate genes of this model plant. A CPB derivative, designated CPB1B, was produced by inserting a 46 nucleotide section of the Arabidopsis PHYTOENE DESATURASE (PDS) gene into CPB, in antisense orientation. CPB1B induced robust PDS silencing, causing easily visible photobleaching in systemically infected Arabidopsis leaves. More importantly, CPB1B can accommodate additional inserts, derived from other Arabidopsis genes, causing the silencing of two or more genes simultaneously. With photobleaching as a visual marker, we adopted the CPB1B vector to evaluate the relative importance of several known RNA silencing pathway genes in PDS VIGS. This approach allowed us to validate the involvement of DICER-LIKE 4 (DCL4) and ARGONAUTE 2 (AGO2) in PDS silencing. Notably, double-stranded RNA-binding protein 4 (DRB4), whose protein product (DRB4) commonly partners with DCL4 in the antiviral silencing pathway, was dispensable for PDS silencing induced by CPB1B. Conclusions: The CPB1B-based vector developed in this work is a valuable tool with tracable and visualizable indicator of the silencing penetrance for interrogating Arabidopsis genes, especially those involved in the RNA silencing pathways.

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A virus‐induced gene‐silencing system for functional genetics in a betalainic species, Amaranthus tricolor (Amaranthaceae)

Cited by 17 publications

References 46 publications

Elucidation of the core betalain biosynthesis pathway in Amaranthus tricolor

Elucidation of the core betalain biosynthesis pathway in Amaranthus tricolor

Simultaneous silencing of two different Arabidopsis genes with a novel virus-induced gene silencing vector

Simultaneous Silencing of Two Different Arabidopsis Genes With A Novel Virus-Induced Gene Silencing Vector

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