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Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition

Colaborador(es): Nahirñak, Vanesa | Almasia, Natalia Inés | Fernández, Paula Virginia | Hopp, Horacio Esteban | Carrari, Fernando | Vazquez Rovere, C.
ISSN: 0032-0889.Tipo de material: Artículos y capítulos. Recurso electrónico.Tema(s): GREEN FLUORESCENT PROTEIN | SN1 PROTEIN, SOLANUM TUBEROSUM | VEGETABLE PROTEIN | CELL DIVISION | CELL MEMBRANE | CELL WALL | CHEMISTRY | CYTOLOGY | GENE EXPRESSION REGULATION | GENE SILENCING | GENETICS | INFRARED SPECTROSCOPY | MASS FRAGMENTOGRAPHY | METABOLISM | MOLECULAR GENETICS | NUCLEOTIDE SEQUENCE | PHYSIOLOGY | PLANT EPIDERMIS | PLANT LEAF | POTATO | SOLANACEAE | TRANSGENIC PLANT | GAS CHROMATOGRAPHY-MASS SPECTROMETRY | MOLECULAR SEQUENCE DATA | PLANT LEAVES | PLANT PROTEINS | PLANTS, GENETICALLY MODIFIED | SOLANUM TUBEROSUM | SPECTROSCOPY, FOURIER TRANSFORM INFRARED | ARABIDOPSIS | NICOTIANA BENTHAMIANA | Recursos en línea: Haga clic para acceso en línea | LINK AL EDITOR. En: Plant Physiology vol.158, no.1 (2012), p.252-263Resumen: Snakin-1 [SN1] is an antimicrobial cysteine-rich peptide isolated from potato [Solanum tuberosum] that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70 percent to 90 percent increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense.
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Snakin-1 [SN1] is an antimicrobial cysteine-rich peptide isolated from potato [Solanum tuberosum] that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70 percent to 90 percent increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense.

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