Rescue of tomato spotted wilt virus entirely from complementary DNA clones

Feng, Mingfeng; Cheng, Ruixiang; Chen, Minglong; Guo, Rong; Li, Luyao; Feng, Zhike; Wu, Jianyan; Xie, Li; Hong, Jian; Zhang, Zhongkai; Kormelink, Richard; Tao, Xiaorong

doi:10.1073/pnas.1910787117

Cited by 61 publications

(83 citation statements)

References 57 publications

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“…Full‐length ALSV RNA1 (GenBank accession no. AB030940) and RNA2 (AB030941) sequences were synthesized by the GenScript (Nanjing, China) and cloned individually into the pCB301‐2 × 35S‐RZ‐NOS vector (Feng et al , 2020) to produce pALSV1 and pALSV2, respectively. A partial sequence (300 bp) of AtPME or AtGBL was RT‐PCR amplified from a total RNA sample using specific primers and cloned into the pALSV2 vector to produce pALSV2‐AtPME or pALSV2‐AtGBL.…”

Section: Methodsmentioning

confidence: 99%

Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection

Huang

Hong

et al. 2020

Molecular Plant Pathology

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Tomato spotted wilt virus (TSWV) is one of the most devastating plant viruses and often causes severe crop losses worldwide. Generally, mature plants become more resistant to pathogens, known as adult plant resistance. In this study, we demonstrated a new phenomenon involving developmentally regulated susceptibility of Arabidopsis thaliana to TSWV. We found that Arabidopsis plants become more susceptible to TSWV as plants mature. Most young 3-week-old Arabidopsis were not infected by TSWV. Infection of TSWV in 4-, 5-, and 6-week-old Arabidopsis increased from 9%, 21%, and 25%, respectively, to 100% in 7-to 8-week-old Arabidopsis plants. Different isolates of TSWV and different tospoviruses show a low rate of infection in young Arabidopsis but a high rate in mature plants. When Arabidopsis dcl2/3/4 or rdr1/2/6 mutant plants were inoculated with TSWV, similar results as observed for the wild-type Arabidopsis plants were obtained. A cell-to-cell movement assay showed that the intercellular movement efficiency of TSWV NSm:GFP fusion was significantly higher in 8-week-old Arabidopsis leaves compared with 4-week-old Arabidopsis leaves. Moreover, the expression levels of pectin methylesterase and β-1,3glucanase, which play critical roles in macromolecule cell-to-cell trafficking, were significantly up-regulated in 8-week-old Arabidopsis leaves compared with 4-week-old Arabidopsis leaves during TSWV infection. To date, this mature plant susceptibility to pathogen infections has rarely been investigated. Thus, the findings presented here should advance our knowledge on the developmentally regulated mature host susceptibility to plant virus infection. K E Y W O R D S Arabidopsis thaliana, developmentally regulated susceptibility, mature-dependent pathogen infection, Tomato spotted wilt virus

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Section: Methodsmentioning

confidence: 99%

Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection

Huang

Hong

et al. 2020

Molecular Plant Pathology

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“…These observations raise the intriguing question of whether these proteins can overcome the antisense problem to facilitate rescue of mammalian nonsegmented NSVs from genomic cDNA copies. For segmented NSVs of plants and some animal counterparts, the current reverse genetics system schemes involve transcription of viral agRNAs that serve as templates for both genome replication and translation of NC proteins [ 20 , 21 , 24 , 29 , 30 ]. In these cases, adaptation of the gRNA approach would require additional plasmids to provide the NC proteins, and these in trans expressed proteins may be less efficient in recruiting the RNA templates for NC assembly [ 52 ].…”

Section: Discussionmentioning

confidence: 99%

“…Similar strategies were applicable to a barley yellow striate mosaic cytorhabdovirus, although infections of its native monocot hosts require insect vector-mediated transfer of the recombinant virus recovered from N. benthamiana leaves [ 28 ]. Recently, refined positive-strand agRNA strategies were also adapted to rescue a tri-segmented tospovirus [ 29 ] and a seven-segmented emaravirus [ 30 ]. In these two cases, expression of viral core proteins (N and L) from in trans provided supporting plasmids is unnecessary since the agRNA transcripts permit direct ribosomal translation to produce both core proteins in cis [ 29 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

“…Recently, refined positive-strand agRNA strategies were also adapted to rescue a tri-segmented tospovirus [ 29 ] and a seven-segmented emaravirus [ 30 ]. In these two cases, expression of viral core proteins (N and L) from in trans provided supporting plasmids is unnecessary since the agRNA transcripts permit direct ribosomal translation to produce both core proteins in cis [ 29 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

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Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA

2020

Viruses

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Generation of recombinant negative-stranded RNA viruses (NSVs) from plasmids involves in vivo reconstitution of biologically active nucleocapsids and faces a unique antisense problem where the negative-sense viral genomic RNAs can hybridize to viral messenger RNAs. To overcome this problem, a positive-sense RNA approach has been devised through expression of viral antigenomic (ag)RNA and core proteins for assembly of antigenomic nucleocapsids. Although this detour strategy works for many NSVs, the process is still inefficient. Using Sonchus yellow net rhabdovirus (SYNV) as a model; here, we develop a negative-sense genomic RNA-based approach that increased rescue efficiency by two orders of magnitude compared to the conventional agRNA approach. The system relied on suppression of double-stranded RNA induced antiviral responses by co-expression of plant viruses-encoded RNA silencing suppressors or animal viruses-encoded double-stranded RNA antagonists. With the improved approach, we were able to recover a highly attenuated SYNV mutant with a deletion in the matrix protein gene which otherwise could not be rescued via the agRNA approach. Reverse genetics analyses of the generated mutant virus provided insights into SYNV virion assembly and morphogenesis. This approach may potentially be applicable to other NSVs of plants or animals.

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“…However, there is still no evidence that CCA1 influences TSWV infectivity. The first report of an infectious TSWV clone ( Feng et al, 2020 ) may help to elucidate whether CCA1 can interact with the native TSWV M RNA in planta .…”

Section: The Core Circadian Clock Component Cca1 Suppresses Photomorpmentioning

confidence: 99%

Emerging Molecular Links Between Plant Photomorphogenesis and Virus Resistance

et al. 2020

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Photomorphogenesis refers to photoreceptor-mediated morphological changes in plant development that are triggered by light. Multiple photoreceptors and transcription factors (TFs) are involved in the molecular regulation of photomorphogenesis. Likewise, light can also modulate the outcome of plant–virus interactions since both photosynthesis and many viral infection events occur in the chloroplast. Despite the apparent association between photosynthesis and virus infection, little is known about whether there are also interplays between photomorphogenesis and plant virus resistance. Recent research suggests that plant–virus interactions are potentially regulated by several photoreceptors and photomorphogenesis regulators, including phytochromes A and B (PHYA and PHYB), cryptochromes 2 (CRY2), phototropin 2 (PHOT2), the photomorphogenesis repressor constitutive photomorphogenesis 1 (COP1), the NAM, ATAF, and CUC (NAC)-family TF ATAF2, the Aux/IAA protein phytochrome-associated protein 1 (PAP1), the homeodomain-leucine zipper (HD-Zip) TF HAT1, and the core circadian clock component circadian clock associated 1 (CCA1). Particularly, the plant growth promoting brassinosteroid (BR) hormones play critical roles in integrating the regulatory pathways of plant photomorphogenesis and viral defense. Here, we summarize the current understanding of molecular mechanisms linking plant photomorphogenesis and defense against viruses, which represents an emerging interdisciplinary research topic in both molecular plant biology and virology.

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Rescue of tomato spotted wilt virus entirely from complementary DNA clones

Cited by 61 publications

References 57 publications

Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection

Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection

Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA

Emerging Molecular Links Between Plant Photomorphogenesis and Virus Resistance

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