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PLoS One. 2015 Jun 15;10(6):e0130139. doi: 10.1371/journal.pone.0130139. eCollection 2015.

Development of a Mild Viral Expression System for Gain-Of-Function Study of Phytoplasma Effector In Planta.

Author information

1
Institute of Biotechnology, National Taiwan University, Taipei, Taiwan.
2
Departement of Plant Pathology and Microbiology, National Taiwan University, Taipei, Taiwan.
3
Department of Agronomy, National Taiwan University, Taipei, Taiwan.
4
Department of Horticulture and Landscape Architecture, National Taiwan University, Taipei, Taiwan.
5
Joint Center for Instruments and Researches, College of Bioresources and Agriculture, National Taiwan University, Taipei, Taiwan.
6
Institute of Biotechnology, National Taiwan University, Taipei, Taiwan; Genome and Systems Biology Degree Program, National Taiwan University, Taipei, Taiwan; Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.

Abstract

PHYL1 and SAP54 are orthologs of pathogenic effectors of Aster yellow witches'-broom (AYWB) phytoplasma and Peanut witches'-broom (PnWB) phytoplasma, respectively. These effectors cause virescence and phyllody symptoms (hereafter leafy flower) in phytoplasma-infected plants. T0 lines of transgenic Arabidopsis expressing the PHYL1 or SAP54 genes (PHYL1 or SAP54 plants) show a leafy flower phenotype and result in seedless, suggesting that PHYL1 and SAP54 interfere with reproduction stage that restrict gain-of-function studies in the next generation of transgenic plants. Turnip mosaic virus (TuMV) mild strain (TuGK) has an Arg182Lys mutation in the helper-component proteinase (HC-ProR182K) that blocks suppression of the miRNA pathway and prevents symptom development in TuGK-infected plants. We exploited TuGK as a viral vector for gain-of-function studies of PHYL1 and SAP54 in Arabidopsis plants. TuGK-PHYL1- and TuGK-SAP54-infected Arabidopsis plants produced identical leafy flower phenotypes and similar gene expression profiles as PHYL1 and SAP54 plants. In addition, the leafy flower formation rate was enhanced in TuGK-PHYL1- or TuGK-SAP54-infected Arabidopsis plants that compared with the T0 lines of PHYL1 plants. These results provide more evidence and novel directions for further studying the mechanism of PHYL1/SAP54-mediated leafy flower development. In addition, the TuGK vector is a good alternative in transgenic plant approaches for rapid gene expression in gain-of-function studies.

PMID:
26076458
PMCID:
PMC4468105
DOI:
10.1371/journal.pone.0130139
[Indexed for MEDLINE]
Free PMC Article

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