Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered Zucchini Yellow Mosaic Virus

Kang, Moon-Jin; Seo, Jang Kyun; Choi, Hoseong; Choi, Hong Soo; Kim, Kook Hyung

doi:10.5423/ppj.nt.08.2015.0173

Cited by 14 publications

(7 citation statements)

References 13 publications

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“…Such techniques are already used in plant physiology and biotechnology studies, especially for evaluating phloem functions in different plant species ( Grignon et al, 1989 ; Pradel et al, 1999 ; Zhang et al, 2010 ; Knoblauch et al, 2015 ). In the case of cucurbits the problem with the application of liquid substances into mature leaves generally required some damage of leaf surface by using, e.g., fine sandpaper or carborundum as an abrasive ( Grignon et al, 1989 ; Pradel et al, 1999 ; Shalitin and Wolf, 2000 ; Arazi et al, 2001 ; Zhang et al, 2010 ; Dunham et al, 2014 ; Kang et al, 2016 ). In contrast, the infiltration of luffa leaves is a more delicate method as it uses the natural ability of leaves to absorb water.…”

Section: Discussionmentioning

confidence: 99%

Mature Luffa Leaves (Luffa cylindrica L.) as a Tool for Gene Expression Analysis by Agroinfiltration

et al. 2017

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We exploited the potential of cucurbits for ectopic gene expression. Agroinfiltration is a simple and commonly used method to obtain transient expression of foreign genes in plants. In contrast to in vitro transformation techniques, agroinfiltration can be used for genetic modification of mature plant tissues. Although the cucurbits are commonly used as model plants for molecular biology and biotechnology studies, to date there are no literature sources on the possibility of transient gene expression in mature cucurbit tissues. Our research has shown that mature leaves of Luffa cylindrica L. (luffa), in contrast to other cucurbit species, can be successfully transiently transformed with Agrobacterium tumefaciens. We efficiently transformed luffa leaves with a reporter gene encoding β-glucuronidase (GUS). The GUS activity in transiently transformed leaf tissues was detected within 24 h after the infiltration with bacteria. Additionally, we have shown that the activity of a transiently expressed the GUS gene can be monitored directly in the EDTA-exudates collected from the cut petioles of the agroinfiltrated leaves. The results suggest that luffa leaves can be useful as a plant expression system for studies of physiological and biochemical processes in cucurbits.

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Section: Discussionmentioning

confidence: 99%

Mature Luffa Leaves (Luffa cylindrica L.) as a Tool for Gene Expression Analysis by Agroinfiltration

et al. 2017

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“…The plasmid inoculation approach was performed as previously described ( Kang et al, 2016 ; Seo et al, 2009 ). Plasmid DNA of pSMV-SX was prepared using the Plasmid Maxi Kit (Qiagen, Shanghai, China).…”

Section: Methodsmentioning

confidence: 99%

Complete Genome Sequencing and Infectious cDNA Clone Construction of Soybean Mosaic Virus Isolated from Shanxi

et al. 2021

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Soybean mosaic virus (SMV) is the predominant viral pathogen that affects the yield and quality of soybean. The natural host range for SMV is very narrow, and generally limited to Leguminosae . However, we found that SMV can naturally infect Pinellia ternata and Atractylodes macrocephala . In order to clarify the molecular mechanisms underlying the cross-family infection of SMV, we used double-stranded RNA extraction, rapid amplification of cDNA ends polymerase chain reaction and Gibson assembly techniques to carry out SMV full-length genome amplification from susceptible soybeans and constructed an infectious cDNA clone for SMV. The genome of the SMV Shanxi isolate (SMV-SX) consists of 9,587 nt and encodes a polyprotein consisting of 3,067 aa. SMV-SX and SMV-XFQ008 had the highest nucleotide and amino acid sequence identities of 97.03% and 98.50%, respectively. A phylogenetic tree indicated that SMV-SX and SMV-XFQ018 were clustered together, sharing the closest relationship. We then constructed a pSMV-SX infectious cDNA clone by Gibson assembly technology and used this clone to inoculate soybean and Ailanthus altissima ; the symptoms of these hosts were similar to those caused by the virus isolated from natural infected plant tissue. This method of construction not only makes up for the time-consuming and laborious defect of traditional methods used to construct infectious cDNA clones, but also avoids the toxicity of the Potyvirus special sequence to Escherichia coli , thus providing a useful cloning strategy for the construction of infectious cDNA clones for other viruses and laying down a foundation for the further investigation of SMV cross-family infection mechanisms.

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“…Foreign genes have been added to the potyviral genome via a pentapeptide insertion at the N-terminus of P1 (Rajamäki et al, 2005) and two proteolytic sites between P1 and HC-Pro (Cui and Wang, 2016) or between the NIb and CP genes (Kelloniemi et al, 2006;Bedoya et al, 2012;Lee et al, 2011), and these additions have not affected viral infectivity. To date, the complex process of viral infection and plant-virus-vector interactions have been studied using more than 13 in vitro-or in vivo-transcribed, fluorescently tagged potyviruses including potato virus A (Rajamäki et al, 2005, Kelloniemi et al, 2006, tobacco etch virus (Majer et al, 2013), potato virus Y (Matevz et al, 2015), plum pox virus (Lansac et al, 2005;Cui and Wang 2016), tobacco vein banding mosaic virus (Gao et al, 2012), clover yellow vein virus (Masuta et al, 2000), pepper mottle virus (Lee et al, 2011), tobacco vein mottling virus (Dietrich and Maiss , 2003), lettuce mosaic virus (German-Retana et al, 2003;Bordat et al, 2015), turnip mosaic virus (Beauchemin et al, 2005), soybean mosaic virus (Seo et al, 2009), zucchini yellow mosaic virus (Kang et al, 2016) and papaya ringspot virus (Tuo et al, 2017). Recently, the Antirrhinum majus MYBtype Rosea1 transcription factor, a non-fluorescent marker that activates anthocyanin accumulation in infected tissues, has been used to tag several potyviruses (Cordero et al, 2017;Bedoya et al, 2012).…”

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confidence: 99%

Two agroinfection-compatible fluorescent protein-tagged infectious cDNA clones of papaya leaf distortion mosaic virus facilitate the tracking of virus infection

Tuo¹,

Pu²,

Zhao³

et al. 2018

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Papaya leaf distortion mosaic virus (PLDMV, the genus Potyvirus) is an emerging threat to papaya production. Here, agroinfection-compatible fluorescent protein-tagged PLDMV infectious cDNA clones driven by the Cauliflower mosaic virus 35S promoter were successfully constructed using one-step Gibson assembly. The clones were directly transformed into Agrobacterium tumefaciens to prevent potential problems such as plasmid instability during propagation in Escherichia coli. Ninety-five percent of papaya seedlings infected with PLDMV-GFP or PLDMV-mCherry developed systemic symptoms typical of those caused by wild-type PLDMV. Green and mCherry red fluorescence was observed in leaves, stems, and roots of infected papaya plants. The fluorescent protein-tagged agroinfectious PLDMV cDNA clones were stable in papaya for more than 90 days and during six serial passages at 30-day intervals. The availability of these infectious clones will contribute to research on PLDMV-host interactions and can be applied in the papaya breeding program for PLDMV resistance.

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Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered Zucchini Yellow Mosaic Virus

Cited by 14 publications

References 13 publications

Mature Luffa Leaves (Luffa cylindrica L.) as a Tool for Gene Expression Analysis by Agroinfiltration

Mature Luffa Leaves (Luffa cylindrica L.) as a Tool for Gene Expression Analysis by Agroinfiltration

Complete Genome Sequencing and Infectious cDNA Clone Construction of Soybean Mosaic Virus Isolated from Shanxi

Two agroinfection-compatible fluorescent protein-tagged infectious cDNA clones of papaya leaf distortion mosaic virus facilitate the tracking of virus infection

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