Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus

Zhong, X.P.; Xue, Yuan; Wu, Ze; Khan, Muhammad Ali; Chen, Jin; Li, Xiaoxin; Bi, Gong; Zhao, Yang; Wu, Jian; Wu, Chenyu; Yi, Mingfang

doi:10.1007/s00299-013-1530-2

Cited by 35 publications

(23 citation statements)

References 43 publications

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“…In VIGS systems, silencing phenotypes often appear 3-4 weeks after infection, but can sometimes appear at >6 weeks after inoculation (Di Stilio et al 2010;Tian et al 2014;Zhong et al 2014). In this study, N. benthamiana plants exhibited the photo-bleaching phenotype at approximately 17 dai (Figure 2).…”

Section: Discussionmentioning

confidence: 82%

“…Plants can be inoculated with PSV and CMV by rubbing the leaves with virus solution. Agrobacterium-infiltration and biolistic delivery of viral vectors (Pflieger et al 2013) are not necessary for PSV inoculation and other special methods such as vacuuminfiltration (Di Stilio et al 2010;Yan et al 2012;Zhong et al 2014) are dispensable to this VIGS system. Thus, the ease of inoculation would be advantageous for the large scale screening of endogenous gene functions.…”

Section: Discussionmentioning

confidence: 99%

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Peanut stunt virus-induced gene silencing in white lupin (Lupinus albus)

Yamagishi

Masuta

Suzuki

et al. 2015

Plant Biotechnology

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White lupin (Lupinus albus L.) plants develop cluster roots and have strong resistance to phosphorus starvation. Although many expressed sequences have been identified to explain the mechanisms used by white lupin to acquire phosphorus, the lack of a stable transformation technique has made it challenging to evaluate the functions of these expressed sequences. Virus-induced gene silencing (VIGS) is an attractive method for assaying gene function in species that are difficult to stably transform. Here, we show that the Peanut stunt virus (PSV) vector effectively induces silencing of endogenous genes in white lupin. It is unknown whether PSV is useful for VIGS; therefore, we first inoculated Nicotiana benthamiana plants with PSV harbouring fragments of the N. benthamiana phytoene desaturase gene (NbPDS). Two out of four distinct sequence fragments of NbPDS induced photo-bleaching in N. benthamiana, indicating that PSV can be used to knockdown endogenous gene sequences in a sequence-dependent manner. White lupin plants inoculated with PSV harbouring fragments of the L. albus PDS gene (LaPDS) developed photo-bleaching that was associated with a significant reduction in LaPDS mRNA accumulation. PSV spread systemically in leaves, roots, and cluster roots, and small interfering RNA of LaPDS was detected in these organs. This is the first study to demonstrate the use of VIGS by PSV, suggesting that this vector can be applied to supress endogenous gene expression in shoots and roots of white lupin and to clarify the mechanisms of phosphorus starvation resistance.

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Section: Discussionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Peanut stunt virus-induced gene silencing in white lupin (Lupinus albus)

Yamagishi

Masuta

Suzuki

et al. 2015

Plant Biotechnology

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“…This plant defense mechanism has been used for the development of a process for silencing of the endogenous plant genes (24). To silence a specifi c plant gene, a fragment of the target gene is cloned into a modifi ed virus vector and delivered to the plant (25)(26)(27). The virus replication mechanism multiplies the target gene fragment introduced in the virus vector in the plant cell, and the transcripts spread throughout the plant systemically (28).…”

Section: Introductionmentioning

confidence: 99%

Sugarcane Mosaic Virus-Based Gene Silencing in Nicotiana benthamiana

Ali

Nasir

Rafiq

et al. 2017

Iran. J. Biotechnol.

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Background: Potyvirus-based virus-induced gene silencing (VIGS) is used for knocking down the expression of a target gene in numerous plant species. Sugarcane mosaic virus (SCMV) is a monopartite, positive single strand RNA virus. Objectives: pBINTRA6 vector was modifi ed by inserting a gene segment of SCMV in place of Tobacco rattle virus (TRV) genome part 1 (TRV1 or RNA1) and the two nonstructural proteins of TRV2(RNA2). Materials and Methods: SCMV construct was inoculated into 3-4 weeks Nicotiana benthamiana plant leaves either by using a needleless syringe or applying pricking with a toothpick. Results: The construct (SCMV-RNA2) successfully induced post-transcriptional gene silencing (PTGS) of the target genes GFP and ChlI through agroinoculation proving that SCMV is a substitute of the RNA1, which plays a pivotal role in the systemic gene silencing. 2-3-weeks of post inoculation, target genes' silencing was observed in the newly developed noninoculated leaves. Conclusions:The newly developed construct expresses the knocked down of the endogenous as well as exogenous genes and only four weeks are required for the transient expression of the gene silencing based on SCMV-VIGS system.

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“…In eudicots, for example, VIGS has been applied to Aquilegia (Kramer et al 2007), Eschscholzia californica (Wege et al 2007), Thalictrum (Di Stilio et al 2010), and Catharanthus roseus (Liscombe and O'Connor 2011), using Tobacco rattle virus (TRV) as the vector, and to Gentiana spp., using Apple latent spherical virus (Nakatsuka et al 2015) and Broad bean wilt virus 2 (Tasaki et al 2016). In monocots, VIGS has been applied to both Gladiolus and Phalaenopsis, using TRV (Zhong et al 2014) and Cymbidium mosaic virus (Hsieh et al 2013), respectively. To use VIGS technology, the choice of viral vector is important, since VIGS is only possible when the viral vector can spread systemically in infected plants (Pflieger et al 2013).…”

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confidence: 99%

Virus-induced gene silencing (VIGS) in Lilium leichtlinii using the Cucumber mosaic virus vector

Tasaki

Terada

Masuta

et al. 2016

Plant Biotechnology

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Lilies (Lilium) are among the most important floriculture crops, and to accelerate research regarding lily genetics, the development of reverse-genetics tools is necessary. However, Agrobacterium-mediated transformation in Lilium is timeconsuming, since the plants require several years to progress from acclimation to flowering. Thus, virus-induced gene silencing (VIGS) is an attractive method for assaying gene function. In the present study, we modified a lily-derived strain of Cucumber mosaic virus (CMV-HL) as a VIGS vector and evaluated its effectiveness for inducing gene silencing in Lilium leichtlinii by introducing L. leichtlinii phytoene desaturase (LlPDS) gene fragments into an intercistronic region between the 3a and 3b genes of the CMV-HL RNA3 genome. At 30 days after inoculation (dpi) with LlPDS-containing CMV-HL, photobleaching was observed in the upper leaves of L. leichtlinii, and at 57 dpi, we observed that the natural orange color in flower tepals had faded. Reduced LlPDS expression and the detection of small interfering LlPDS RNA indicated that the color changes were the result of LlPDS gene silencing. In addition, the leaves also exhibited a mild photo-bleaching phenotype in the following year. Therefore, our results indicate that CMV-HL spreads systemically in the leaves and flowers of Lilium during the first year of infection, as well as in new shoots during the following year, and that the vector system can be successfully applied to induce short-term endogenous gene silencing in lilies.

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Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus

Cited by 35 publications

References 43 publications

<i>Peanut stunt virus</i>-induced gene silencing in white lupin (<i>Lupinus albus</i>)

<i>Peanut stunt virus</i>-induced gene silencing in white lupin (<i>Lupinus albus</i>)

Sugarcane Mosaic Virus-Based Gene Silencing in Nicotiana benthamiana

Virus-induced gene silencing (VIGS) in <i>Lilium leichtlinii</i> using the <i>Cucumber mosaic virus</i> vector

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