title: Protein profile of Beta vulgaris leaf apoplastic fluid and changes induced by Fe deficiency and Fe resupply.
abstract: The fluid collected by direct leaf centrifugation has been used to study the proteome of the sugar beet apoplastic fluid as well as the changes induced by Fe deficiency and Fe resupply to Fe-deficient plants in the protein profile. Plants were grown in Fe-sufficient and Fe-deficient conditions, and Fe resupply was carried out with 45 μM Fe(III)-EDTA for 24 h. Protein extracts of leaf apoplastic fluid were analyzed by two-dimensional isoelectric focusing-SDS-PAGE electrophoresis. Gel image analysis revealed 203 consistent spots, and proteins in 81% of them (164) were identified by nLC-MS/MS using a custom made reference repository of beet protein sequences. When redundant UniProt entries were deleted, a non-redundant leaf apoplastic proteome consisting of 109 proteins was obtained. TargetP and SecretomeP algorithms predicted that 63% of them were secretory proteins. Functional classification of the non-redundant proteins indicated that stress and defense, protein metabolism, cell wall and C metabolism accounted for approximately 75% of the identified proteome. The effects of Fe-deficiency on the leaf apoplast proteome were limited, with only five spots (2.5%) changing in relative abundance, thus suggesting that protein homeostasis in the leaf apoplast fluid is well-maintained upon Fe shortage. The identification of three chitinase isoforms among proteins increasing in relative abundance with Fe-deficiency suggests that one of the few effects of Fe deficiency in the leaf apoplast proteome includes cell wall modifications. Iron resupply to Fe deficient plants changed the relative abundance of 16 spots when compared to either Fe-sufficient or Fe-deficient samples. Proteins identified in these spots can be broadly classified as those responding to Fe-resupply, which included defense and cell wall related proteins, and non-responsive, which are mainly protein metabolism related proteins and whose changes in relative abundance followed the same trend as with Fe-deficiency.
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Laura
authors: L
authors: Ceballos-Laita
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Elain
authors: E
authors: Gutierrez-Carbonell
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Giuseppe
authors: G
authors: Lattanzio
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Saul
authors: S
authors: Vázquez
authors: Laboratory of Computational and Structural Biology, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain ; Fundación ARAID Zaragoza, Spain.
authors: Bruno
authors: B
authors: Contreras-Moreira
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Anunciación
authors: A
authors: Abadía
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Javier
authors: J
authors: Abadía
authors: Plant Stress Physiology Group, Department of Plant Nutrition, Aula Dei Experimental Station, Consejo Superior de Investigaciones Científicas Zaragoza, Spain.
authors: Ana-Flor
authors: AF
authors: López-Millán
keywords: iron deficiency; leaf apoplast; proteome; sugar beet; two-dimensional electrophoresis