The virological model plant, Nicotiana benthamiana expresses a single functional RDR6 homeolog

<p>Pepper yellow leaf curl disease caused by Pepper yellow leaf curl Indonesia virus (PepYLCIV) has become a challenge to chili pepper cultivation. Development of resistant variety by utilizing recessive resistance gene is expected to control the disease in the field. This study aimed to validate three plant genes, namely deltaCOP, hsc70, and BAM1, in PepYLCIV infection by applying Virus-induced Gene Silencing (VIGS) in a model plant, wild type Nicotiana benthamiana. PepYLCIV and construct of Tobacco rattle virus (TRV) which induced silencing of each gene were co-inoculated into N. benthamiana plants through agroinfiltration. Gene expression and the relative amount of viral DNA were determined by quantitative reverse transcription PCR (qRT-PCR) and quantitative PCR (qPCR), respectively, at 15 days post inoculation. The results showed a decreased level of deltaCOP, hsc70, and BAM1 expressions to 66.4%, 53.0%, and 47.0%, respectively, compared to that in the control (100%). Silencing of the three genes decreased the accumulation of PepYLCIV to 0.1%, 18.4%, and 63.0%, respectively, compared to that in the control. deltaCOP and hsc70 genes were indicated to be involved in the viral infection and could be good candidate genes for obtaining chili pepper varieties resistant to PepYLCIV. This result affirmed that the reverse genetics technique could be an alternative approach for identifying plant genes involved in viral infection, including PepYLCIV. The use of an infectious clone in this study allows the virus inoculations could be carried out without rearing and maintaining its natural vector, hence reduces the risk of virus transmission to healthy plants.</p>

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Screening Isolates of Soybean mosaic virus for Infectivity in a Model Plant, Nicotiana benthamiana

Plant Disease ◽

10.1094/pdis-04-14-0405-re ◽

2015 ◽

Vol 99 (4) ◽

pp. 442-446 ◽

Cited By ~ 5

Author(s):

L. Gao ◽

R. Zhai ◽

Y. K. Zhong ◽

A. Karthikeyan ◽

R. Ren ◽

...

Keyword(s):

Genetic Transformation ◽

Mosaic Virus ◽

Nicotiana Benthamiana ◽

Soybean Mosaic Virus ◽

Narrow Host Range ◽

Model Plant ◽

Resistance Strategies ◽

The Family ◽

New Hosts ◽

Soybean Leaves

Soybean mosaic virus (SMV), belonging to the genus Potyvirus of the family Potyviridae, has a relatively narrow host range almost exclusively confined to leguminous hosts. While disease management through genetic transformation can be an effective approach, soybean remains recalcitrant to routine genetic transformation. In this context, it is important to identify new hosts for SMV that can be used to develop effective transgenic resistance strategies. Transformation in Nicotiana benthamiana is simple and highly efficient; hence, here we demonstrate the infectivity of SMV strain SC7 in N. benthamiana plants. To identify an SMV strain infectious in N. benthamiana, we mechanically inoculated N. benthamiana plants with 37 isolates from 21 (SC1 to SC21) SMV strains. Plants inoculated with isolates of strain SC7 produced mosaic symptoms on leaves. However, N. benthamiana plants inoculated with the 20 other SMV strains showed no visible symptoms. Furthermore, soybean cv. Nannong 1138-2 inoculated with sap prepared from symptomatic N. benthamiana leaves showed typical SMV mosaic symptoms 2 weeks after inoculation. In addition, SMV was detected in symptomatic N. benthamiana and soybean leaves by RT-PCR, DAS-ELISA, and further identified by sequencing. Together, the results indicate that N. benthamiana plants could support multiplication of SMV strain SC7. The findings of this study would be useful for the investigation of SMV resistance using the model plant N. benthamiana.

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Agroinoculation of a full-length cDNA clone of cotton leafroll dwarf virus (CLRDV) results in systemic infection in cotton and the model plant Nicotiana benthamiana

Virus Research ◽

10.1016/j.virusres.2013.04.007 ◽

2013 ◽

Vol 175 (1) ◽

pp. 64-70 ◽

Cited By ~ 11

Author(s):

Verónica C. Delfosse ◽

María F. Casse ◽

Yamila C. Agrofoglio ◽

Iván Bonacic Kresic ◽

Horacio E. Hopp ◽

...

Keyword(s):

Cdna Clone ◽

Nicotiana Benthamiana ◽

Systemic Infection ◽

Full Length ◽

Dwarf Virus ◽

Full Length Cdna ◽

Model Plant ◽

Cotton Leafroll Dwarf Virus

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SseF, a type III effector protein from the mammalian pathogen Salmonella enterica, requires resistance-gene-mediated signalling to activate cell death in the model plant Nicotiana benthamiana

New Phytologist ◽

10.1111/j.1469-8137.2012.04124.x ◽

2012 ◽

Vol 194 (4) ◽

pp. 1046-1060 ◽

Cited By ~ 25

Author(s):

Şuayib Üstün ◽

Petra Müller ◽

Ralf Palmisano ◽

Michael Hensel ◽

Frederik Börnke

Keyword(s):

Cell Death ◽

Resistance Gene ◽

Nicotiana Benthamiana ◽

Salmonella Enterica ◽

Effector Protein ◽

Type Iii Effector ◽

Model Plant ◽

Type Iii ◽

Activate Cell

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Nicotiana benthamiana as model plant for Meloidogyne graminicola infection

Nematology ◽

10.1163/15685411-00003154 ◽

2018 ◽

Vol 20 (5) ◽

pp. 491-499 ◽

Cited By ~ 1

Author(s):

Diana Naalden ◽

Ruben Verbeek ◽

Godelieve Gheysen

Keyword(s):

Life Cycle ◽

Host Range ◽

Nicotiana Benthamiana ◽

Cell Biology ◽

Broad Host Range ◽

Meloidogyne Graminicola ◽

Model Plant ◽

Host Pathogen Interaction ◽

Host Pathogen ◽

Insight Into

Summary Nicotiana benthamiana is widely used as a model plant to analyse cell biology and to obtain insight into the molecular host-pathogen interaction because it is susceptible to many pathogens. Since N. benthamiana can be transformed easily, it is also used to study pathogens for which it is not a known host. Meloidogyne graminicola has a fairly broad host range of mainly monocots and some dicots but no data were available on the ability of M. graminicola to infect N. benthamiana. In this study, we show that M. graminicola is able to infect and complete its life cycle in N. benthamiana, although our experiments demonstrate a lower susceptibility compared to rice. In addition, M. graminicola was also able to develop in N. tabacum but the reproduction was very low. Therefore, we conclude that N. benthamiana can be considered as a host, while this is not the case for N. tabacum.

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Simple and efficient heterologous expression of necrosis-inducing effectors using the model plant Nicotiana benthamiana

10.1101/2021.05.02.442377 ◽

2021 ◽

Author(s):

Bayantes Dagvadorj ◽

Peter Solomon

Keyword(s):

Heterologous Expression ◽

High Throughput ◽

Posttranslational Modifications ◽

Nicotiana Benthamiana ◽

Fungal Pathogens ◽

Host Plants ◽

Plant Pathogens ◽

Expression Systems ◽

In Planta ◽

Model Plant

Plant fungal pathogens cause devastating diseases on cereal plants and threaten global food security. During infection, these pathogens secrete proteinaceous effectors that promote disease. Some of these effectors from necrotrophic plant pathogens induce a cell death response (necrosis), which facilitates pathogen growth in planta. Characterisation of these effectors typically requires heterologous expression and microbial expression systems such as bacteria and yeast are the predominantly used. However, microbial expression systems often require optimization for any given effector and are, in general, not suitable for effectors involving cysteine bridges and posttranslational modifications for activity. Here, we describe a simple and efficient method for expressing such effectors in the model plant Nicotiana benthamiana. Briefly, an effector protein is transiently expressed and secreted into the apoplast of N. benthamiana by Agrobacterium-mediated infiltration. Two-to-three days subsequent to agroinfiltration, the apoplast from the infiltrated leaves is extracted and can be directly used for phenotyping on host plants. The efficacy of this approach was demonstrated by expressing the ToxA, Tox3 and Tox1 necrosis-inducing effectors from Parastagonospora nodorum. All three effectors produced in N. benthamiana were capable of inducing necrosis in wheat lines, and two of three showed visible bands on Coomassie-stained gel. These data suggest that N. benthamiana-agroinfiltration system is a feasible tool to obtain fungal effectors, especially those that require disulfide bonds and posttranslational modifications. Furthermore, due to the low number of proteins typically observed in the apoplast (compared to intracellular), this simple and high-throughput approach circumvents the requirement to lyse cells and further purify the target proteins that is required in other heterologous systems. Because of its simplicity and potential for high-throughput, this method is highly amenable to the phenotyping of candidate protein effectors on host plants.

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Construction of an Arabidopsis/Nicotiana inter-order graft for studies of scion-rootstock interaction

10.21203/rs.2.10041/v1 ◽

2019 ◽

Author(s):

Zhuying Deng ◽

Mengting Jiang ◽

Mi Wang ◽

Dacheng Liang

Keyword(s):

Nicotiana Benthamiana ◽

Union Formation ◽

Order Model ◽

Vascular Connection ◽

Critical Step ◽

Model Plant ◽

Stressed Group ◽

Plant Bioreactor ◽

Graft Incompatibility

Abstract Background Scion–rootstock union formation is a critical step towards functional assemblage of heterogeneous plants. However, scion-rootstock interaction often results in graft incompatibility during the process of assemblage. So far, the lack of model heterografts involving both clear genetic backgrounds and taxonomically distant species greatly impedes insights into the mechanisms underlying scion-rootstock interaction. Results In this report, we established an Arabidopsis (At)/Nicotiana benthamiana (Nb) heterografting system in which the model plant At and the model plant Nb for plant bioreactor was used as scion and rootstock respectively, to explore the interaction between the two model plants. Regarding to the At scion phenotypes, the At-Nb connection can be characterized into three groups: the mild-stressed, the albino and the dormant grafts. Examination of symplastic and apoplastic flow indicated that a functional inter-order grafting was established in the mild-stressed group, but not in the dormant group. What’s more, the free GFP movement in both At/At homograft and the At/Nb graft implicated that macromolecules moved across the heterograft union of the mild-stressed graft, but congealed at the union of dormant graft. These results accentuated the role of vascular connection in the establishment of compatible heterografts. Conclusions The present study established an inter-order model graft involving Arabidopsis and Nicotiana. The interactions from these two species resulted in three distinct grafting groups, which offer us a novel vista to explore many important issues such as grafting compatibility and biomolecule movement.

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