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Studies on callose and cutin during the expression of competence and determination for organogenic nodule formation from internodes of Humulus lupulus var. Nugget
Authors:Fortes Ana M  Testillano Pilar S  Del Carmen Risueño Maria  Pais Maria S
Institution:Centro de Biotecnologia Vegetal, Faculdade de Ciências, University Lisboa, Bloco C2, Piso 1, Campo Grande, 1749-016 Lisboa, Portugal; Plant Development and Nuclear Organization group, Centro de Investigaciones Biologicas, CSIC, Velazquèz 144, 28006 Madrid, Spain
Abstract:Callose and cutin deposition were followed by staining with Aniline Blue and Nile Red and by immunolocalization using antibodies raised against callose. Along with morphogenesis induction from internodes of Humulus lupulus var. Nugget, a temporal and spatial differential deposition of callose and cutin was observed. A cutin layer showing bright yellow autofluorescence appears, surrounding cells or groups of cells committed to express morphogenic competence. This cutin layer that evolves to a randomly organized network appeared underneath a callose layer and may create a specific cellular environment with altered permeability and altered receptors providing conditions for entering the cell cycle. The incipient callose accumulation in control explants cultured on basal medium suggests the involvement of callose in the initiation of the morphogenic programme leading to nodule formation. A scanning electron microscopic study during the organogenic process showed that before shoot bud regeneration, the cutin layer increases in thickness and acquires a smooth texture. This cutin layer is specific to nodular organogenic regions and disappeared with plantlet regeneration. This layer may control permeability to water and solute transfer throughout plantlet regeneration.
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