Systemic Nematicides for the Control of Tricbodorid Nematodes and of Potato Spraing Disease caused by Tobacco Rattle Virus

Field-grown tubers of potato were examined for infection by Tobacco rattle virus (TRV) and consequent production of corky ringspot or spraing symptoms. A microarray study identified genes that are differentially expressed in tuber tissue in response to TRV infection and to spraing production, suggesting that hypersensitive response (HR) pathways are activated in spraing-symptomatic tubers. This was confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) of a selected group of HR-related genes and by histochemical staining of excised tuber tissue with spraing symptoms. qRT-PCR of TRV in different regions of the same tuber slice showed that nonsymptomatic areas contained higher levels of virus relative to spraing-symptomatic areas. This suggests that spraing formation is associated with an active plant defense that reduces the level of virus in the infected tuber. Expression of two of the same plant defense genes was similarly upregulated in tubers that were infected with Potato mop-top virus, a virus that also induces spraing formation.

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VipariNama: RNA viral vectors to rapidly elucidate the relationship between gene expression and phenotype

Plant Physiology ◽

10.1093/plphys/kiab197 ◽

2021 ◽

Author(s):

Arjun Khakhar ◽

Cecily Wang ◽

Ryan Swanson ◽

Sydney Stokke ◽

Furva Rizvi ◽

...

Keyword(s):

Gene Expression ◽

Transcription Factors ◽

Viral Vectors ◽

Viral Vector ◽

Tobacco Rattle Virus ◽

Plant Size ◽

Great Promise ◽

Attractive Alternative ◽

Gibberellin Biosynthesis ◽

In Planta

Abstract Synthetic transcription factors have great promise as tools to help elucidate relationships between gene expression and phenotype by allowing tunable alterations of gene expression without genomic alterations of the loci being studied. However, the years-long timescales, high cost, and technical skill associated with plant transformation have limited their use. In this work we developed a technology called VipariNama (ViN) in which vectors based on the Tobacco Rattle Virus (TRV) are used to rapidly deploy Cas9-based synthetic transcription factors and reprogram gene expression in planta. We demonstrate that ViN vectors can implement activation or repression of multiple genes systemically and persistently over several weeks in Nicotiana benthamiana, Arabidopsis (Arabidopsis thaliana), and tomato (Solanum lycopersicum). By exploring strategies including RNA scaffolding, viral vector ensembles, and viral engineering, we describe how the flexibility and efficacy of regulation can be improved. We also show how this transcriptional reprogramming can create predictable changes to metabolic phenotypes, such as gibberellin biosynthesis in N. benthamiana and anthocyanin accumulation in Arabidopsis, as well as developmental phenotypes, such as plant size in N. benthamiana, Arabidopsis, and tomato. These results demonstrate how ViN vector-based reprogramming of different aspects of gibberellin signaling can be used to engineer plant size in a range of plant species in a matter of weeks. In summary, VipariNama accelerates the timeline for generating phenotypes from over a year to just a few weeks, providing an attractive alternative to transgenesis for synthetic transcription factor-enabled hypothesis testing and crop engineering.

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Optimisation of tobacco rattle virus-induced gene silencing in Arabidopsis

Functional Plant Biology ◽

10.1071/fp05096 ◽

2006 ◽

Vol 33 (4) ◽

pp. 347 ◽

Cited By ~ 46

Author(s):

Changchun Wang ◽

Xinzhong Cai ◽

Xuemin Wang ◽

Zhong Zheng

Keyword(s):

Gene Silencing ◽

High Throughput ◽

Function Analysis ◽

Tobacco Rattle Virus ◽

Phytoene Desaturase ◽

Growth Stages ◽

Virus Induced Gene Silencing ◽

Gene Functions ◽

Genes Encoding ◽

Gene Function Analysis

Arabidopsis thaliana (L.) Heynh. is a model plant species in which to study plant gene functions. Recently developed virus-induced gene silencing (VIGS) offers a rapid and high-throughput technique platform for gene function analysis. In this paper we report optimisation of tobacco rattle virus (TRV)-induced gene silencing in Arabidopsis. The parameters potentially affecting the efficiency of VIGS in Arabidopsis were investigated. These included the concentration and pre-incubation of Agrobacterium inocula (agro-inocula), the concentration of acetosyringone included in agro-inocula, the Agrobacterium inoculation (agro-inoculation) method, the ecotypes and the growth stages of Arabidopsis plants for agro-inoculation, and the growth temperature of agro-inoculated plants. The optimised VIGS procedure involves preparing the agro-inocula with OD600 of 2.0, pre-incubating for 2 h in infiltration buffer containing 200 μm acetosyringone, agro-inoculating by vacuum infiltration, and growth of agro-inoculated plants at 22 −24°C. Following this procedure consistent and highly efficient VIGS was achieved for the genes encoding phytoene desaturase (PDS) and actin in Arabidopsis. The silencing phenotype lasts for at least 6 weeks, and is applicable in at least seven ecotypes, including Col-0, Cvi-0, Sd, Nd-1, Ws-0, Bay-0 and Ler. TRV-induced VIGS was expressed not only in leaves, but also in stems, inflorescences and siliques. However, VIGS was not transmissible through seed to the subsequent generation. The optimised procedure of the TRV-induced gene silencing should facilitate high-throughput functional analysis of genes in Arabidopsis.

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Biological activity of transcripts synthesized in Vitro from full-length and mutated DNA copies of tobacco rattle virus RNA 2

Virology ◽

10.1016/0042-6822(89)90222-5 ◽

1989 ◽

Vol 173 (1) ◽

pp. 68-76 ◽

Cited By ~ 19

Author(s):

Gerco C. Angenent ◽

Eelke Posthumus ◽

John F. Bol

Keyword(s):

Biological Activity ◽

Tobacco Rattle Virus ◽

Full Length ◽

Virus Rna

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The genomic RNA1 and RNA2 sequences of the tobacco rattle virus isolates found in Polish potato fields

Virus Research ◽

10.1016/j.virusres.2014.02.019 ◽

2014 ◽

Vol 185 ◽

pp. 110-113 ◽

Cited By ~ 7

Author(s):

Zhimin Yin ◽

Magdalena Pawełkowicz ◽

Krystyna Michalak ◽

Mirosława Chrzanowska ◽

Ewa Zimnoch-Guzowska

Keyword(s):

Tobacco Rattle Virus ◽

Virus Isolates

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Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

International Journal of Molecular Sciences ◽

10.3390/ijms21228741 ◽

2020 ◽

Vol 21 (22) ◽

pp. 8741

Author(s):

Güven Edgü ◽

Lena Julie Freund ◽

Stefanie Hartje ◽

Eckhard Tacke ◽

Hans-Reinhard Hofferbert ◽

...

Keyword(s):

Disease Management ◽

Management Practices ◽

Tobacco Rattle Virus ◽

Simultaneous Detection ◽

Isothermal Amplification ◽

Diagnostic Methods ◽

Tuber Quality ◽

Loop Mediated Isothermal Amplification ◽

Tuber Necrosis ◽

Corky Ringspot

Potato is an important staple food crop in both developed and developing countries. However, potato plants are susceptible to several economically important viruses that reduce yields by up to 50% and affect tuber quality. One of the major threats is corky ringspot, which is a tuber necrosis caused by tobacco rattle virus (TRV). The appearance of corky ringspot symptoms on tubers prior to commercialization results in ≈ 45% of the tubers being downgraded in quality and value, while ≈ 55% are declared unsaleable. To improve current disease management practices, we have developed simple diagnostic methods for the reliable detection of TRV without RNA purification, involving minimalized sample handling (mini), subsequent improved colorimetric loop-mediated isothermal amplification (LAMP), and final verification by lateral-flow dipstick (LFD) analysis. Having optimized the mini-LAMP-LFD approach for the sensitive and specific detection of TRV, we confirmed the reliability and robustness of this approach by the simultaneous detection of TRV and other harmful viruses in duplex LAMP reactions. Therefore, our new approach offers breeders, producers, and farmers an inexpensive and efficient new platform for disease management in potato breeding and cultivation.

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