Overexpression and VIGS system for functional gene validation in oriental melon (Cucumis melo var. makuwa Makino)

Liao, Jing; Wang, Cheng Hui; Xing, Qiao Juan; Li, Yue Peng; Liu, Xian Feng; Hua, Qi

doi:10.1007/s11240-019-01568-9

Cited by 23 publications

(17 citation statements)

References 61 publications

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“…A new TRSV vector was recently reported [17]. More recently, the TRV-VIGS system has been used in cucumber and oriental melon [18,19]. Comparing ALSV, TRSV and TRV with CGMMV, the first three belong to multipartite virus families with a bipartite genome, while CGMMV is a monopartite virus [22], and therefore is easier to manipulate.…”

Section: Discussionmentioning

confidence: 99%

“…In the past decades, a large number of viral vectors had been developed as powerful tools for the functional verification of genes in plants [9][10][11][12][13][14][15][16]. To date, three different RNA viruses have been developed as vectors for VIGS in cucurbit species, including apple latent spherical virus (ALSV) [11], tobacco ringspot virus (TRSV) [17] and tobacco rattle virus (TRV) [18,19]. However, very few applications of these vectors have been reported, implying that they have not been widely adopted for cucurbit gene function analyses.…”

Section: Introductionmentioning

confidence: 99%

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A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants

et al. 2020

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Background: Cucurbits produce fruits or vegetables that have great dietary importance and economic significance worldwide. The published genomes of at least 11 cucurbit species are boosting gene mining and novel breeding strategies, however genetic transformation in cucurbits is impractical as a tool for gene function validation due to low transformation efficiency. Virus-induced gene silencing (VIGS) is a potential alternative tool. So far, very few ideal VIGS vectors are available for cucurbits. Results: Here, we describe a new VIGS vector derived from cucumber green mottle mosaic virus (CGMMV), a monopartite virus that infects cucurbits naturally. We show that the CGMMV vector is competent to induce efficient silencing of the phytoene desaturase (PDS) gene in the model plant Nicotiana benthamiana and in cucurbits, including watermelon, melon, cucumber and bottle gourd. Infection with the CGMMV vector harboring PDS sequences of 69-300 bp in length in the form of sense-oriented or hairpin cDNAs resulted in photobleaching phenotypes in N. benthamiana and cucurbits by PDS silencing. Additional results reflect that silencing of the PDS gene could persist for over two months and the silencing effect of CGMMV-based vectors could be passaged. Conclusions: These results demonstrate that CGMMV vector could serve as a powerful and easy-to-use tool for characterizing gene function, controlling viral pathogens or even performing resistance breeding in cucurbits. Moreover, this study will possess considerable important reference value for developing different viral vectors.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants

et al. 2020

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“…A new TRSV vector was recently reported (Zhao et al, 2016). More recently, the TRV-VIGS system has been used in cucumber and oriental melon (Bu et al, 2019; Liao et al, 2019). Comparing ALSV, TRSV and TRV with CGMMV, the first three belong to multipartite virus families with a bipartite genome, while CGMMV is a monopartite virus (Ugaki et al, 1991), and therefore is easier to manipulate.…”

Section: Discussionmentioning

confidence: 99%

“…TRSV cDNA clones are not infectious in watermelon or pumpkin (Zhao et al, 2016). Although TRV has been widely used as VIGS vector since it has a wide host range (Ratcliff et al, 2001), its application for VIGS on cucurbits except cucumber and oriental melon have not been reported (Bu et al, 2019; Liao et al, 2019). Both ALSV and CGMMV vectors can be successfully used on common cucurbit plants such as watermelon, melon, cucumber and bottle gourd (Igarashi et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

“…In the past decades, a large number of viral vectors had been developed as powerful tools for the functional verification of genes in plants (Ruiz et al, 1998; Liu et al, 2002; Ding et al, 2006; Igarashi et al, 2009; Zhang et al, 2010; Sempere et al, 2011; Liu et al, 2016; Wang et al, 2016). To date, three different RNA viruses have been developed as vectors for VIGS in cucurbit species, including apple latent spherical virus (ALSV) (Igarashi et al, 2009), tobacco ringspot virus (TRSV) (Zhao et al, 2016) and tobacco rattle virus (TRV) (Bu et al, 2019; Liao et al, 2019). However, very few applications of these vectors have been reported, implying that they have not been widely adopted for cucurbit gene function analyses.…”

Section: Introductionmentioning

confidence: 99%

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A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants

Liu

Liang

Aranda

et al. 2019

Preprint

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One sentence summary: A CGMMV-based vector enables gene function 21 studies in cucurbits, an extremely low efficiency species for genetic 22 42 hairpin cDNAs resulted in photobleaching phenotypes in N. benthamiana and 43 cucurbits by PDS silencing. Additional results reflect that silencing of the PDS gene 44 could persist for over two months and the silencing effect of CGMMV-based vectors 45 could be passaged. These results demonstrate that CGMMV vector could serve as a 46 powerful and easy-to-use tool for characterizing gene function in cucurbits. 47 48 Keywords: Cucumber green mottle mosaic virus; viral vector; virus-induced gene 49 3 silencing; cucurbit plants 50 68 (VIGS) is a good alternative to gene transformation because of its simplicity, high 69 efficiency, and high throughput. 70 Gene silencing comprises transcriptional gene silencing (TGS) and 71 post-transcriptional gene silencing (PTGS). VIGS, a type of PTGS, is a natural 72 defense reaction that exists in a broad range of organisms. It confers resistance to 73 foreign nucleic acid invasion through PTGS at the RNA level. Because it can silence a 74 specific gene, leading to the loss of function of this gene, the potential of VIGS as a 75 tool to analyze gene function has been quickly recognized (Baulcombe, 1999). 76 4 In the past decades, a large number of viral vectors had been developed as 77 powerful tools for the functional verification of genes in plants (Ruiz et al., 1998; Liu 78 et al., 2002; Ding et al., 2006; Igarashi et al., 2009; Zhang et al., 2010; Sempere et al., 79 2011; Liu et al., 2016; Wang et al., 2016). To date, three different RNA viruses have 80 (CP). Only the 129 KDa and 186 KDa of replication-related proteins are translated 100 directly from the genomic RNA, whereas the 29-KDa MP and the 17.4-KDa CP are 101 translated from two subgenomic RNAs. There is an overlap between the MP and CP 102 ORFs (Ugaki et al., 1991). Viral vectors based on CGMMV for expressing foreign 103 genes have been constructed. Multiple cloning sites (MCS) were inserted adjacent to 104 the CP ORF, and the CP stop codon was altered to express the hepatitis B surface 105 antigen and a Dengue virus Epitope so that 20 and 44 foreign amino acids, 106 respectively, were expressed (Ooi et al., 2006; Teoh et al., 2009). Tobacco mosaic 107 virus (TMV), another member of the genus Tobamovirus, has been widely studied as a 108 5 model in this genus. TMV has successfully been developed as a VIGS vector by 109 including an additional duplicated copy of the CP subgenomic promoter (SGP) in the 110 viral genome (Kumagai et al., 1995).The CGMMV genome is similar to that of TMV, 111 and thus it was thought that methods similar to those used for TMV could be used to 112 create vectors based on CGMMV; unfortunately, results in this regard varied largely 113 129 Agricultural Sciences (Xinjiang, China), Tianjin Academy of Agricultural Sciences 130 (Tianjin, China) and Ningbo Academy of Agricultural Sciences (Ningbo, China), 131 respectively. All cucurbit seeds were soaked in ste...

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Silencing GhCOI1 in Gladiolus hybridus increases susceptibility to Alternaria brassicicola and impairs inducible defenses

Seng

et al. 2019

Plant Cell Tiss Organ Cult

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Overexpression and VIGS system for functional gene validation in oriental melon (Cucumis melo var. makuwa Makino)

Cited by 23 publications

References 61 publications

A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants

A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants

A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants

Silencing GhCOI1 in Gladiolus hybridus increases susceptibility to Alternaria brassicicola and impairs inducible defenses

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