INVESTIGADORES
ZAVALA Jorge Alberto
artículos
Título:
Molecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuata. VII. Changes in the plants proteome.
Autor/es:
GIRI, A.P.; WÜNSCHE, H.; MITRA, S.; ZAVALA, J.A.; MUCK, A.; SVATO, A.; BALDWIN I.T.
Revista:
PLANT PHYSIOLOGY.
Referencias:
Año: 2006 vol. 142 p. 1621 - 1641
ISSN:
0032-0889
Resumen:
When Manduca sexta attacks Nicotiana attenuata, fatty acid-amino acid conjugates (FACs) in the larvaes oral secretions (OS) are
introduced into feeding wounds. These FACs trigger a transcriptional response that is similar to the response induced by insect
damage. Using two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization-time of flight, and liquid
chromatography-tandem mass spectrometry, we characterized the proteins in phenolic extracts and in a nuclear fraction of
leaves elicited by larval attack, and/or in leaves wounded and treated with OS, FAC-free OS, and synthetic FACs. Phenolic
extracts yielded approximately 600 protein spots, many of which were altered by elicitation, whereas nuclear protein fractions
yielded approximately 100 spots, most of which were unchanged by elicitation. Reproducible elicitor-induced changes in 90
spots were characterized. In general, proteins that increased were involved in primary metabolism, defense, and transcriptional
and translational regulation; those that decreased were involved in photosynthesis. Like the transcriptional defense responses,
proteomic changes were strongly elicited by the FACs in OS. A semiquantitative reverse transcription-PCR approach
based on peptide sequences was used to compare transcript and protein accumulation patterns for 17 candidate proteins. In six
cases the patterns of elicited transcript accumulation were consistent with those of elicited protein accumulation. Functional
analysis of one of the identified proteins involved in photosynthesis, RuBPCase activase, was accomplished by virus-induced
gene silencing. Plants with decreased levels of RuBPCase activase protein had reduced photosynthetic rates and RuBPCase
activity, and less biomass, responses consistent with those of herbivore-attacked plants. We conclude that the response of the
plants proteome to herbivore elicitation is complex, and integrated transcriptome-proteome-metabolome analysis is required
to fully understand this ubiquitous ecological interaction.Manduca sexta attacks Nicotiana attenuata, fatty acid-amino acid conjugates (FACs) in the larvaes oral secretions (OS) are
introduced into feeding wounds. These FACs trigger a transcriptional response that is similar to the response induced by insect
damage. Using two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization-time of flight, and liquid
chromatography-tandem mass spectrometry, we characterized the proteins in phenolic extracts and in a nuclear fraction of
leaves elicited by larval attack, and/or in leaves wounded and treated with OS, FAC-free OS, and synthetic FACs. Phenolic
extracts yielded approximately 600 protein spots, many of which were altered by elicitation, whereas nuclear protein fractions
yielded approximately 100 spots, most of which were unchanged by elicitation. Reproducible elicitor-induced changes in 90
spots were characterized. In general, proteins that increased were involved in primary metabolism, defense, and transcriptional
and translational regulation; those that decreased were involved in photosynthesis. Like the transcriptional defense responses,
proteomic changes were strongly elicited by the FACs in OS. A semiquantitative reverse transcription-PCR approach
based on peptide sequences was used to compare transcript and protein accumulation patterns for 17 candidate proteins. In six
cases the patterns of elicited transcript accumulation were consistent with those of elicited protein accumulation. Functional
analysis of one of the identified proteins involved in photosynthesis, RuBPCase activase, was accomplished by virus-induced
gene silencing. Plants with decreased levels of RuBPCase activase protein had reduced photosynthetic rates and RuBPCase
activity, and less biomass, responses consistent with those of herbivore-attacked plants. We conclude that the response of the
plants proteome to herbivore elicitation is complex, and integrated transcriptome-proteome-metabolome analysis is required
to fully understand this ubiquitous ecological interaction.