Purification and partial characterization of an extracellular melanoprotein from the fungus Venturia inaequalis

The fungus Venturia inaequalis clone No. 36 isolated from Malus domestica cv. Gloster excretes a melanoprotein of 36 kDa in relatively high amounts during growth in liquid culture. The protein was isolated from the culture medium and purified to homogeneity. It was shown to contain melanin. After raising an antiserum against the isolated protein, the protein could be shown to be located in the apoplast fluid of the V. inaequalis infected Malus domestica cv. Elstar. Partial sequencing of the protein revealed no significant sequence homologies to so far sequenced proteins. The melanoprotein binds ferrous and ferric iron. Moreover, it could be shown that the binding of ferric iron (but not of ferrous iron) leads to a change in the absorbance of the protein suggesting a modification of the protein by ferric, but not by ferrous, iron. In addition to iron, the protein also binds copper, but does not bind manganese or nickel. A possible function of this protein in the recruiting and transport of iron and copper and/or in the protection of the fungus by metal-ion mediated oxidative stress is discussed. © 2005 Verlag der Zeitschrift für Naturforschung

Up-regulation of pathogenesis-related proteins in the apoplast of Malus domestica after application of a non-pathogenic bacterium

The intercellular washing fluid (IWF) of Malus domestica cv. Holsteiner Cox before and after application of the non-pathogenic bacterium Pseudomonas fluorescens Bk3 to the leaves was investigated in a comparative manner. SDS-PAGE in combination with ESI Q-ToF mass spectrometry, and homology search in relevant data bases revealed the highly up-regulated expression of several pathogenesis-related plant proteins in the apoplast of the leaves treated with P. fluorescens. These proteins were β-1,3-glucanase, chitinase, thaumatin-like protein, ribonuclease-like protein, and a hevein-like protein. Moreover, a 9 kDa non-specific lipid transfer protein was significantly reduced after the application of P. fluorescens. The possible relevance of a pre-treatment of apple cultivars with the non-pathogenic bacterium P. fluorescens Bk3, as an alternative method to the treatment with fungicides, for increasing the resistance of susceptible apple cultivars against an infection with the fungus Venturia inaequalis is discussed

A genetic symbiotic algorithm applied to the one-dimensional cutting stock problem

This work presents a genetic symbiotic algorithm to minimize the number of objects and the setup in a one-dimensional cutting stock problem. The algorithm implemented can generate combinations of ordered lengths of stock (the cutting pattern) and, at the same time, the frequency of the cutting patterns, through a symbiotic process between two distinct populations, solutions and cutting patterns. Working with two objectives in the fitness function and with a symbiotic relationship between the two populations, we obtained positive results when compared with other methods described in the literature.<br>Neste trabalho desenvolvemos um algoritmo genético simbiótico com objetivo de minimizar o número de objetos processados e o setup num problema de corte unidimensional. Nosso algoritmo genético gera seus próprios padrões em conjunto com soluções para o problema, através de um processo simbiótico entre duas populações distintas, a de soluções e a de padrões. Trabalhando com os dois objetivos na função de aptidão e com a relação simbiótica entre as duas populações, obtivemos resultados competitivos em relação aos métodos descritos na literatura