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Cherry Adapted Plum Pox Virus (PPV) CR Isolate Alters the Abundance of Proteins Involved in Photosynthesis and Stress-response of the Experimental Herbaceous Host Nicotiana benthamiana More Aggressively than the Mild PPV-C Isolate

ABSTRACT: Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide each year. Among the PPV strains identified so far, only PPV-C and PPV-CR are able to infect cherries under natural conditions. Herein, we evaluated the impact of strains differing by the pathogenic potential on herbaceous host Nicotiana benthamiana. At first glance, a faster accumulation of PPV capsid protein was noticed by semi-quantitative DAS-ELISA on tobacco leaves infected by PPV-CR (the RU-30sc isolate) in contrast to PPV-C (BY-101 isolate). This result was correlated perfectly with observed phenotypic symptoms. To assess the host response to infection more deeply, a comprehensive proteomic profiling was performed using reverse phase UHPLC followed by label-free mass spectrometry quantification. Thirty-one unique plant proteins were identified as significantly altered due to the infection. Precise evaluation of particular amount shifts of identified proteins in relation to infection has revealed that the aggressive PPV-CR isolate has a stronger influence on the abundance of photosynthesis-related proteins, mainly from Calvin cycle, as the mild PPV-C. This observation was accompanied by a significant reduction of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of remaining identified proteins, indicates activation of repair mechanism, stimulation of photosynthetic capacity, and modifications in amino acid and carbohydrate metabolism to affect plant growth and initiate energy formation via gluconeogenesis. Furthermore, we speculate that accumulation of H2O2, in PPV-CR infected leaves may activate glutathione synthesis, which plays a crucial role in further plant defense and development.

INSTRUMENT(S): Q-Tof Premier

ORGANISM(S): Plum Pox Virus Nicotiana Benthamiana

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Maksym Danchenko

LAB HEAD: Slavomira Novakova

PROVIDER: PXD010018 | Pride | 2018-08-13

REPOSITORIES: Pride

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			Action	DRS
	PPV-pride.DerivedFromSearch596756ba-5d98-41d3-ad8d-51fe29c626de.mgf	Mgf
	PPV-pride.DerivedFromSearch596756ba-5d98-41d3-ad8d-51fe29c626de.pride.mgf.gz	Mgf
	PPV-pride.mzid.gz	Mzid
	PPV-pride.pride.mztab.gz	Mztab
	PPV.zip	Other

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Project description:Objectives: Our work focuses on the responses of Solanaceous plants to viruses that cause economically important diseases in tree fruits. Using mock inoculated leaf tissue as a reference, we plan to compare the gene expression profiles of Nicotiana Benthamiana plants infected with one of three viruses; Plum Pox Potyvirus (PPV), Tomato Ringspot Nepovirus (ToRSV), and Prunus Nectrotic Ringspot Nepovirus (PNRSV). Our goals are as follows: (1) Identify genes that are induced/repressed in response to individual viruses. (2) Identify genes that are induced/repressed in response to all 3 viruses. (3) Compare results to existing potato array data to look for similarities in responses to other pathogens. Experimental Design: Nicotiana benthamiana plants were inoculated with one of three viruses: PPV, ToRSV, or PNRSV. 3 week old plants were inoculated by rubbing virus infected plant sap onto leaves dusted with carborundum. Control plants were mock inoculated using sap from healthy plants. All plants were maintained in a growth chamber at 22C for 18 days. 8 plants were inoculated with each virus or mock inoculated. This experiment was repeated twice. 4 biological replicates derived from 2 virus infected plants from each replica experiment (4 plants) are to be used for hybridizations. RNA from all mock inoculated plants was similarly pooled to create 4 biological replicates. Each replicate control will serve as a universal reference sample that is to be hybridized pair wise with each of the three virus infected samples. RNA extraction: After 18 days, un-inoculated leaves displaying clear symptoms were harvested and immediately frozen in liquid N2. Total RNA was purified using Trizol according to TIGRs listed protocol. RNA was subsequently treated with Turbo DNA-free RNase (Ambion cat#1907). Finally, total RNA was further purified on RNeasy columns (Qiagen) according to manufacturer’s instructions and quantified using a Nanodrop spectrophotometer. Keywords: Reference design 23 hybs total

Dataset Information

Cherry Adapted Plum Pox Virus (PPV) CR Isolate Alters the Abundance of Proteins Involved in Photosynthesis and Stress-response of the Experimental Herbaceous Host Nicotiana benthamiana More Aggressively than the Mild PPV-C Isolate

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