Talano Melina Andrea, Agostini Elizabeth, Medina María I, Reinoso Herminda, Tordable María del Carmen, Tigier Horacio A, de Forchetti Silvia Milrad
Departamento de Biología Molecular, FCEFQyN, Universidad Nacional de Río Cuarto, Ruta No. 36, Km 601 (CP 5800), Río Cuarto, Córdoba, Argentina.
J Plant Physiol. 2006 May;163(7):740-9. doi: 10.1016/j.jplph.2005.06.009. Epub 2005 Sep 12.
A highly basic peroxidase isoenzyme was shown to be released to the culture medium of tomato (Lycopersicon esculentum) hairy roots grown in Murashige-Skoog (MS) liquid medium when it was supplemented with 100 mM NaCl. In this paper we demonstrate that this enzyme is ionically bound to cell walls and that the release was a consequence of the continuous agitation of the tissue in a high ionic strength medium with salt addition. In order to establish the physiological role of this isoenzyme we partially purified it, and we analysed its kinetic properties as coniferyl alcohol peroxidase. The peroxidase isoenzyme showed a high catalytic efficiency for this substrate, which suggests that it would be associated with the ligno-suberization process. To confirm the involvement of this isoenzyme in that process, we studied the pattern of ligno-suberization of the tissue under different conditions of growth. Our results suggest that this basic peroxidase would be indeed involved in ligno-suberization since its leakage from cell walls, induced by 100 mM NaCl in liquid MS, caused less ligno-suberization of exo and endodermis. On the contrary, more ligno-suberization was seen in cell walls when the hairy roots were grown in a salt-supplemented MS solid medium without contact with it, a condition in which the release of the isoenzyme would be avoided. Thus, through the changes produced by the release of the enzyme from its site of action, we could demonstrate the physiological role of this peroxidase in the processing of root cell walls, being part of control mechanisms of ion and water fluxes through the root.
当在Murashige-Skoog(MS)液体培养基中添加100 mM NaCl时,一种高度碱性的过氧化物酶同工酶会释放到番茄(Lycopersicon esculentum)毛状根的培养基中。在本文中,我们证明这种酶与细胞壁离子结合,并且这种释放是在添加盐的高离子强度培养基中组织持续搅拌的结果。为了确定这种同工酶的生理作用,我们对其进行了部分纯化,并分析了其作为松柏醇过氧化物酶的动力学特性。该过氧化物酶同工酶对该底物表现出高催化效率,这表明它与木质化-栓质化过程有关。为了证实这种同工酶参与该过程,我们研究了不同生长条件下组织的木质化-栓质化模式。我们的结果表明,这种碱性过氧化物酶确实参与了木质化-栓质化过程,因为在液体MS中由100 mM NaCl诱导的其从细胞壁的泄漏导致外皮层和内皮层的木质化-栓质化减少。相反,当毛状根在添加盐的MS固体培养基中生长且不与之接触时,在细胞壁中观察到更多的木质化-栓质化,在这种条件下可以避免同工酶的释放。因此,通过酶从其作用位点释放所产生的变化,我们可以证明这种过氧化物酶在根细胞壁加工中的生理作用,它是通过根的离子和水通量控制机制的一部分。