Establishment of virus-induced gene silencing system and functional analysis of ScbHLH17 in Senecio cruentus

Li, Yajun; Liu, Yuting; Qi, Fangting; Deng, Chengyan; Lu, Chenfei; Huang, Huaguo; Dai, Silan

doi:10.1016/j.plaphy.2019.12.024

Cited by 15 publications

(14 citation statements)

References 32 publications

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“…5b ). In the previous study, ScbHLH17 regulated anthocyanin accumulation in cineraria by influencing the expression of ABP genes [ 34 ]. Since two bHLH TFs, NtAN1a and NtAN1b, were proved to enhance anthocyanin accumulation in tobacco [ 36 ], we measured the expression of NtAN1a and NtAN1b in OE- ScAG tobacco lines to explore if ScAG and ScAGL11 regulate anthocyanin accumulation through bHLH TFs.…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, no MYB gene from this study displayed a differential expression pattern in the coloured and colourless regions. The transcript level of ScbHLH17 , which correlates with anthocyanin accumulation [ 34 ], showed no significant change between ScAG / ScAGL11 -silenced leaves and the CK lines. Recent studies established that MADS-box genes directly regulate the expression of the ABP structural gene instead of going through the MBW complex.…”

Section: Discussionmentioning

confidence: 99%

“…However, ScCHS2 (gene encoding chalcone synthase), ScDFR3 , and ScANS (gene encoding anthocyanidin synthase) were highly expressed in coloured flowers [ 1 ]. Considering the TFs regulating anthocyanin biosynthesis, ScbHLH17 (subfamily V bHLH TF) was isolated [ 34 ]. Indeed, silencing ScbHLH17 caused loss of anthocyanin accumulation in cineraria leaves, verifying that ScbHLH17 activated anthocyanin biosynthesis [ 34 ].…”

Section: Introductionmentioning

confidence: 99%

“…Considering the TFs regulating anthocyanin biosynthesis, ScbHLH17 (subfamily V bHLH TF) was isolated [ 34 ]. Indeed, silencing ScbHLH17 caused loss of anthocyanin accumulation in cineraria leaves, verifying that ScbHLH17 activated anthocyanin biosynthesis [ 34 ]. However, no other TFs were reported, and the molecular mechanism of bicolour formation in cineraria ray florets remained unknown.…”

Section: Introductionmentioning

confidence: 99%

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Functional analysis of the ScAG and ScAGL11 MADS-box transcription factors for anthocyanin biosynthesis and bicolour pattern formation inSenecio cruentusray florets

Liu

Luo

et al. 2022

Horticulture Research

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Cineraria (Senecio cruentus) is an ornamental plant with pure colour and bicolour cultivars, widely used for landscaping. Anthocyanin biosynthesis influences colouration patterns in cineraria. However, how anthocyanins accumulate and distribute in cineraria is poorly understood. This study investigated the molecular mechanisms underlying anthocyanin biosynthesis and bicolour formation in cineraria using pure colour and bicolour cultivars. Transcriptome and gene expression analysis showed five genes, including ScCHS2, ScF3H1, ScDFR3, ScANS, and ScbHLH17, were inhibited in the white cultivar and colourless regions of bicolour cultivars. In contrast, two MADS-box genes, ScAG and ScAGL11, showed significantly higher expression, in the colourless regions of bicolour cultivars. ScAG and ScAGL11 were localized in the nucleus and co-expressed with the bicolour trait. Further functional analysis verified that ScAG inhibits anthocyanin accumulation in tobacco (Nicotiana tabacum). However, virus-induced gene silencing (VIGS) experiments showed that silencing of ScAG and ScAGL11 increases anthocyanin content in cineraria leaves. Similar results were observed when silenced ScAG and ScAGL11 in the cineraria capitulum, accompanied by the smaller size of the colourless region, specifically in the ScAG/ScAGL11-silenced plants. The expression of ScCHS2, ScDFR3, and ScF3H1 increased in silenced cineraria leaves and capitulum. Furthermore, Y2H and BiFC experiments demonstrated that ScAG interacts with ScAGL11. Moreover, ScAG directly inhibited the transcription of ScF3H1 while ScAGL11 inhibited ScDFR3 expression by binding to their promoter separately. The findings reported herein indicate that ScAG and ScAGL11 negatively regulate anthocyanin biosynthesis in cineraria ray florets, and their differential expression in ray florets influenced the bicolour pattern appearance.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functional analysis of the ScAG and ScAGL11 MADS-box transcription factors for anthocyanin biosynthesis and bicolour pattern formation inSenecio cruentusray florets

Liu

Luo

et al. 2022

Horticulture Research

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“…Although several key physiological processes involved in PCNA have been ignored, more attention to this aspect is needed in the future. VIGS is a powerful and reliable tool to investigate gene functions and disease research in numerous crop plants (Li et al 2020). Meanwhile, in other studies on gene functions, VIGS was not considered the best method because it can be used in certain transgenic plants but not all stages of the life cycle.…”

Section: Discussionmentioning

confidence: 99%

TaPCNA plays a role in programmed cell death after UV-B exposure in wheat (

Zhang

Chen

et al. 2021

Funct. Plant Biol.

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Ultraviolet (UV)-B is a component of sunlight and shows a significant effect on DNA damage, which can be regulated by proliferating cell nuclear antigen (PCNA). The role of TaPCNA in wheat (Triticum aestivum L.) programmed cell death (PCD) under UV-B has not been investigated previously. Here, we explored the function of TaPCNA in wheat exposed to UV-B utilising Barley Stripe Mosaic Virus-virus-induced gene silencing (VIGS). The results showed that the expression of TaPCNA was downregulated, and curly wheat leaves with several spots were determined by VIGS. The growth rate and mesophyll cell length were significantly inhibited after TaPCNA was silenced. The activity of superoxide dismutase and the contents of soluble sugar and soluble protein decreased, whereas the activities of peroxidase and catalase and malondialdehyde content increased in TaPCNA-silenced and UV-B treatment groups. DNA laddering and propidium iodide staining results showed that DNA fragments and micronucleus accumulated after TaPCNA silencing with or without UV-B. Thus, TaPCNA participates in plant growth and DNA damage and PCD under UV-B. This study suggests an idea for the exploration of the function of certain genes in such complex wheat genomes and offers a theoretical basis to improve wheat agronomic traits.

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Development and application of the TRV-induced gene-silencing system in different Rhododendron species

Xu,

Cui,

Chen

et al. 2024

Plant Cell Tiss Organ Cult

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Rhododendron is one of the top ten traditional owers in China, with high ornamental and medicinal values. However, molecular research on this species has been impeded by the lack of e cient molecular genetic techniques. Virus-induced gene silencing (VIGS) technology is an effective tool for analyzing gene functions, which has been successfully applied to many plant species. But there have been no reports of VIGS system for Rhododendron. In this study, tobacco rattle virus (TRV) was chosen to infect Rhododendron micranthum using phytoene desaturase (PDS) as the reporter gene. After the inoculation of pTRV2-RmPDS by leaf injection, photobleaching phenomena appeared in the newly developed leaves and the qRT-PCR assay demonstrated that RmPDS was successfully silenced. Then three parameters including the developmental stage, the Agrobacterium concentration and the inoculation temperature were examined to optimize the system. The silencing e ciency was increased from 2.4% to 11.4% and the optimized conditions were as follows: the developmental stage of the two true-leaf stage, the adjustment of the inoculation solution to a nal OD 600 =1.5 and the inoculation temperature of 18 ℃. To further validate the system, the most optimal combination was used to infect other six rhododendron genotypes. R. mucronulatum, R. ovatum, R. × pulchrum, R. simsii and R. yedoense displayed the silenced phenotype of PDS as expected. We successfully established TRV-mediated VIGS technology in Rhododendron which could evaluate and characterize the function of plant genes without the need for cumbersome tissue culture. Key MessageIt was proved for the rst time that virus-induced gene silencing system could be applied in different Rhododendron species, which provided a new approach to analyze gene functions of Rhododendron.

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Establishment of virus-induced gene silencing system and functional analysis of ScbHLH17 in Senecio cruentus

Cited by 15 publications

References 32 publications

Functional analysis of the ScAG and ScAGL11 MADS-box transcription factors for anthocyanin biosynthesis and bicolour pattern formation inSenecio cruentusray florets

Functional analysis of the ScAG and ScAGL11 MADS-box transcription factors for anthocyanin biosynthesis and bicolour pattern formation inSenecio cruentusray florets

TaPCNA plays a role in programmed cell death after UV-B exposure in wheat (

Development and application of the TRV-induced gene-silencing system in different Rhododendron species

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