Susceptibility and possible resistance mechanisms in the palm species Phoenix dactylifera, Chamaerops humilis and Washingtonia filifera against Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae).

Rhynchophorus ferrugineus, known as the Red Palm Weevil (RPW), is reported as a pest of up to 40 palm species. However, the susceptibility degree and the defense mechanisms of these species against this weevil are still poorly known. In Europe, the RPW is a major pest of Phoenix canariensis while other palm species, including the congeneric Phoenix dactylifera, seem to be less suitable hosts for this insect. The aim of our study was to compare the defensive response of P. dactylifera, Chamaerops humilis and Washingtonia filifera against R. ferrugineus and try to define the mechanisms of resistance that characterize these species. Bioassays were carried out to evaluate the mortality induced on RPW larvae by extracts from the leaf rachis of the studied palm species. Tests at semi-field scale were also conducted, based either on forced palm infestation, with larvae of RPW, or on natural infestation, with adult females. Rachis extracts from C. humilis and W. filifera caused 100% larval mortality after 2 days of exposure, while extracts of P. dactylifera did not impair larval survival. Independently of the effect of the leaf extracts, the weevils were unable to naturally infest the three palm species, although larval survival was high after forced infestation of the plants. We concluded that the observed lack of infestation of P. dactylifera by RPW is due to factors other than antibiosis. In W. filifera and C. humilis, although the presence of antixenosis mechanisms cannot be excluded, resistance to R. ferrugineus seems to rely on the presence of antibiosis compounds.

Kaempferide triglycoside: a possible factor of resistance of carnation (Dianthus caryophyllus) to Fusarium oxysporum f. sp. dianthi.

A kaempferide triglycoside has been found as a constitutive component in an uninfected carnation (Dianthus caryophyllus) of the cultivar Novada. The chemical structure has been determined mainly by the use of spectroscopic methods, including 2D NMR experiments. It showed a strong activity in restricting fungal parasite development, which could contribute to the known ability of carnation cv. Novada to resist to Fusarium oxysporum f. sp. dianthi infection.

Direct measurement by laser flash photolysis of intramolecular electron transfer in the three-electron reduced form of ascorbate oxidase from zucchini.

Ascorbate oxidase, which has been fully reduced by its substrate, can rapidly transfer a single electron to the laser-generated triplet state of 5-deazariboflavin. Subsequent to this, intramolecular electron transfer occurs resulting in the oxidation of the blue type I copper center. This latter process proceeds via biphasic kinetics, with observed rate constants of 9500 s-1 and 1400 s-1, both of which are protein concentration independent. This indicates that the initial oxidation reaction involves the type II, III trinuclear center, probably occurring via parallel reactions of two of the three copper atoms. The rate constants for intramolecular electron transfer in the three-electron reduced enzyme are one to two orders of magnitude larger than previously observed for the one-electron reduced enzyme, indicating a dramatic effect of the redox state of the enzyme on the intramolecular communication between the copper centers.

Oxidative turnover increases the rate constant and extent of intramolecular electron transfer in the multicopper enzymes, ascorbate oxidase and laccase.

Using laser flash photolysis of lumiflavin/EDTA solutions containing ascorbate oxidase, we find that the rate constant for intramolecular electron transfer varies from one enzyme preparation to another and is generally a more sensitive measure of the state of the active site than are steady-state assays. Thus, type I copper is initially reduced in a second-order reaction followed by first-order reoxidation by the type II-III trinuclear copper center. The observed rate constant for this intramolecular process in presumably native enzyme is 160 s-1 at pH 7, whereas an enzyme preparation which had less than 20% activity had a rate constant of 2.6 s-1. Other samples of relatively active enzyme showed biphasic intramolecular kinetics intermediate between the above values. The inactive enzyme sample could be reactivated by dialysis against ascorbate or by treatment with ferricyanide, resulting in a corresponding increase in the intramolecular rate constant for type I copper reoxidation to a value comparable to that of native enzyme. Using this same methodology, we have determined that the type I copper in Japanese lacquer tree laccase is reoxidized by the type II-III trinuclear copper center in a first-order (intramolecular) process with rate constants of 1 s-1 at pH 7.0 and 4.9 s-1 at pH 6.0, values which are approximately two orders of magnitude smaller than for ascorbate oxidase. The intramolecular rate constant and enzyme activity for laccase also increased, but only by a factor of 2-6, when the enzyme was treated with ascorbate or ferricyanide, respectively. We further found that intramolecular electron transfer in laccase was completely inhibited by fluoride ion, in contrast to ascorbate oxidase which is unaffected by this ion. These differences in behavior for these two very similar enzymes are rather remarkable, when it is considered that the distance between copper atoms is constrained by the location of the protein-derived copper ligands in the three-dimensional structure, and that the redox potentials of the enzymes are similar. Our results may be interpreted in terms of an interconversion between active and inactive enzyme in which there is a rearrangement of the type II-III trinuclear copper center, resulting in a lowering of the redox potential and a block in electron transfer. Turnover restores the active enzyme conformation.(ABSTRACT TRUNCATED AT 400 WORDS)

Flavonoid Accumulation Is Correlated with Adventitious Roots Formation in Eucalyptus gunnii Hook Micropropagated through Axillary Bud Stimulation.

Eucalyptus gunnii Hook microcuttings, obtained in vitro through axillary bud stimulation, show different rooting responses on the same rooting medium depending on the physiological state induced by cytokinins used in the previous multiplication medium. 6-Furfurylamino purine and 6-(4-hydroxy-3-methylbut-2-enylamino)purine induced a physiological state characterized by high sensitivity of microcuttings to the rooting stimulus exerted by the auxin 3-indolebutyric acid, but N(6)-benzyladenine did not produce the same effect. The former physiological state was characterized by an increased accumulation of two endogenous flavonoids (identified as quercetin glycosides) which may be markers of a well defined physiological state. They could have some direct influence on the rooting processes of the explants cultivated in vitro.