The participation of plant cell organelles in compatible and incompatible potato virus Y-tobacco and -potato plant interaction

Otulak, Katarzyna; Garbaczewska, Grażyna

doi:10.1007/s11738-013-1389-4

Cited by 18 publications

(16 citation statements)

References 45 publications

(52 reference statements)

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“…Because PVY replicates in the cytoplasm (Kopek et al ., ; Otulak and Garbaczewska, ), we constructed binary vectors harbouring a synthetic LshCas13a gene driven by the UBQ10 ( Arabidopsis ubiquitin‐10) promoter, and sgRNAs with oligo (A)‐rich tails (serving as signal for nuclear export) (Stewart, ) driven by the AtU6 promoter. We next introduced the LshCas13a/sgRNA constructs into potato plants by stable Agrobacterium ‐mediated transformation (Figure b).…”

Section: Resultsmentioning

confidence: 99%

Generation of virus‐resistant potato plants by RNA genome targeting

Zhan

Zhang

Zhong

et al. 2019

Plant Biotechnology Journal

138

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Summary CRISPR /Cas systems provide bacteria and archaea with molecular immunity against invading phages and foreign plasmids. The class 2 type VI CRISPR /Cas effector Cas13a is an RNA ‐targeting CRISPR effector that provides protection against RNA phages. Here we report the repurposing of CRISPR /Cas13a to protect potato plants from a eukaryotic virus, Potato virus Y ( PVY ). Transgenic potato lines expressing Cas13a/sg RNA (small guide RNA ) constructs showed suppressed PVY accumulation and disease symptoms. The levels of viral resistance correlated with the expression levels of the Cas13a/sg RNA construct in the plants. Our data further demonstrate that appropriately designed sg RNA s can specifically interfere with multiple PVY strains, while having no effect on unrelated viruses such as PVA or Potato virus S . Our findings provide a novel and highly efficient strategy for engineering crops with resistances to viral diseases.

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Section: Resultsmentioning

confidence: 99%

Generation of virus‐resistant potato plants by RNA genome targeting

Zhan

Zhang

Zhong

et al. 2019

Plant Biotechnology Journal

138

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“…Our previous studies determining the hypersensitive response in potato cv. Rywal described ultrastructural alterations with various organelles changes [ 29 , 30 ], but precise ultrastructural analyses of apoplast area rearrangements in PVY infection are still lacking. Our paper concentrated on PVY NTN impact on cell wall ultrastructure modification and localization of proteins associated with structure and/or modification of the cell wall.…”

Section: Discussionmentioning

confidence: 99%

“…Potato (infected and mock-inoculated) leaf samples (2 mm × 2 mm sections) were fixed in 2% ( v / v ) paraformaldehyde and 2% ( v / v ) glutharaldehyde in 0.1 M cacodylate buffer (pH 7.2) for 2 h and washed 4 times in cacodylate buffer [ 30 ]. Samples were post-fixed in 2% ( v / v ) OsO 4 for 2 h at 4 °C, dehydrated in an ethanol series (10–99%) and propylene oxide, embedded in epoxy resin (EPON, Fluka, Switzerland) and polymerized at 60 °C for 24 h. Ultra-thin (90 nm) sections were stained with uranyl acetate/lead citrate (Sigma-Aldrich, St. Louis, MO, USA) and examined by transmission electron microscopy [ 76 ].…”

Section: Methodsmentioning

confidence: 99%

“…Controls consisted of mock-inoculated tissue and pre-immune serum. After rinsing with PBS-Tween20 buffer slides were treated with secondary goat anti-rabbit IgG conjugated with FITC (fluorescein isothiocyanate) goat anti-rat IgG in PBS at RT in the dark for 2 h. An Olympus AX70 Provis (Olympus Poland, Warsaw, Poland) with a UM61002 filter set and equipped with an Olympus SC35 camera was used for fluorescence imaging [ 30 , 74 ].…”

Section: Methodsmentioning

confidence: 99%

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Plant Cell Wall Dynamics in Compatible and Incompatible Potato Response to Infection Caused by Potato Virus Y (PVYNTN)

Otulak

Kozieł

Lockhart

2018

IJMS

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The cell wall provides the structure of the plant, and also acts as a barier against biotic stress. The vein necrosis strain of Potato virus Y (PVYNTN) induces necrotic disease symptoms that affect both plant growth and yield. Virus infection triggers a number of inducible basal defense responses, including defense proteins, especially those involved in cell wall metabolism. This study investigates the comparison of cell wall host dynamics induced in a compatible (potato cv. Irys) and incompatible (potato cv. Sárpo Mira with hypersensitive reaction gene Ny-Smira) PVYNTN–host–plant interaction. Ultrastructural analyses revealed numerous cell wall changes induced by virus infection. Furthermore, the localization of essential defensive wall-associated proteins in susceptible and resistant potato host to PVYNTN infection were investigated. The data revealed a higher level of detection of pathogenesis-related protein 2 (PR-2) in a compatible compared to an incompatible (HR) interaction. Immunofluorescence analyses indicated that hydroxyproline-rich glycoproteins (HRGP) (extensin) synthesis was induced, whereas that of cellulose synthase catalytic subunits (CesA4) decreased as a result of PVYNTN infection. The highest level of extensin localization was found in HR potato plants. Proteins involved in cell wall metabolism play a crucial role in the interaction because they affect the spread of the virus. Analysis of CesA4, PR-2 and HRGP deposition within the apoplast and symplast confirmed the active trafficking of these proteins as a step-in potato cell wall remodeling in response to PVYNTN infection. Therefore, cell wall reorganization may be regarded as an element of “signWALLing”—involving apoplast and symplast activation as a specific response to viruses.

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“…Potato (infected and mock-inoculated) leaf samples (2 mm × 2 mm sections) from mock-inoculated plants and 14 and 30 dpi were fixed in 2% (v/v) paraformaldehyde and 2% (v/v) glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) for 2 h and washed four times in cacodylate buffer [27,43]. Samples were post-fixed in 2% (v/v) OsO 4 for 2 h at 4 • C, dehydrated in an ethanol series (10%-99%) and propylene oxide, embedded in epoxy resin (EPON, Fluka, Switzerland), and polymerized at 60 • C for 24 h. Then, leaf sections (50-70 nm thick) were mounted on Formvar-coated nickel grids that were treated exactly according to the whole procedure of immunogold localization, as presented in [27].…”

Section: Electron Microscopy Materials Preparation Immunogold Localizmentioning

confidence: 99%

The Expression of Potato Expansin A3 (StEXPA3) and Extensin4 (StEXT4) Genes with Distribution of StEXPAs and HRGPs-Extensin Changes as an Effect of Cell Wall Rebuilding in Two Types of PVYNTN–Solanum tuberosum Interactions

Otulak

Kozieł

Lockhart

et al. 2020

Viruses

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The plant cell wall acts not only as a physical barrier, but also as a complex and dynamic structure that actively changes under different biotic and abiotic stress conditions. The question is, how are the different cell wall compounds modified during different interactions with exogenous stimuli such as pathogens? Plants exposed to viral pathogens respond to unfavorable conditions on multiple levels. One challenge that plants face under viral stress is the number of processes required for differential cell wall remodeling. The key players in these conditions are the cell wall genes and proteins, which can be regulated in specific ways during the interactions and have direct influences on the rebuilding of the cell wall structure. The cell wall modifications occurring in plants during viral infection remain poorly described. Therefore, this study focuses on cell wall dynamics as an effect of incompatible interactions between the potato virus Y (PVYNTN) and resistant potatoes (hypersensitive plant), as well as compatible (susceptible plant) interactions. Our analysis describes, for the first time, the expression of the potato expansin A3 (StEXPA3) and potato extensin 4 (StEXT4) genes in PVYNTN-susceptible and -resistant potato plant interactions. The results indicated a statistically significant induction of the StEXPA3 gene during a susceptible response. By contrast, we demonstrated the predominantly gradual activation of the StEXT4 gene during the hypersensitive response to PVYNTN inoculation. Moreover, the in situ distributions of expansins (StEXPAs), which are essential cell wall-associated proteins, and the hydroxyproline-rich glycoprotein (HRGP) extensin were investigated in two types of interactions. Furthermore, cell wall loosening was accompanied by an increase in StEXPA deposition in a PVYNTN-susceptible potato, whereas the HRGP content dynamically increased during the hypersensitive response, when the cell wall was reinforced. Ultrastructural localization and quantification revealed that the HRGP extensin was preferably located in the apoplast, but deposition in the symplast was also observed in resistant plants. Interestingly, during the hypersensitive response, StEXPA proteins were mainly located in the symplast area, in contrast to the susceptible potato where StEXPA proteins were mainly observed in the cell wall. These findings revealed that changes in the intracellular distribution and abundance of StEXPAs and HRGPs can be differentially regulated, depending on different types of PVYNTN–potato plant interactions, and confirmed the involvement of apoplast and symplast activation as a defense response mechanism.

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The participation of plant cell organelles in compatible and incompatible potato virus Y-tobacco and -potato plant interaction

Cited by 18 publications

References 45 publications

Generation of virus‐resistant potato plants by RNA genome targeting

Generation of virus‐resistant potato plants by RNA genome targeting

Plant Cell Wall Dynamics in Compatible and Incompatible Potato Response to Infection Caused by Potato Virus Y (PVYNTN)

The Expression of Potato Expansin A3 (StEXPA3) and Extensin4 (StEXT4) Genes with Distribution of StEXPAs and HRGPs-Extensin Changes as an Effect of Cell Wall Rebuilding in Two Types of PVYNTN–Solanum tuberosum Interactions

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