Initial decomposition ofNymphoides peltata (Gmel.) O. Kuntze (Menyanthaceae), as studied by the leaf-marking method

Summary In this paper the leaf-marking method as used for the study of the development and initial decomposition of floating leaves is described and the reliability of the various measurements is tested and/or discussed. Some general results obtained withNymphoides peltata (Gmel.) O. Kuntze in tanks and in the field are presented and crltically discussed. Autolysis followed by microbial decay was in all cases the most important factor by which leaves disintegrated. In the field plots animals were responsible for the disappearance of 22% of the total leaf area produced during a growth season. This is, however, the combined effect of consumption and damage succeeded by microbial decay. Real grazing can be estimated to be no more than 10% of the production of floating leaves. Fungi can have an important role in initial decomposition, especially after the flowering period, as is demonstrated forSeptoria villarsiae Desm. All damage types show temporal and, in the case of animals, also spatial distribution patterns.     

Transport of amino acids and ammonium in mycelium of Agaricus bisporus.

Mycelium of Agaricus bisporus took up methylamine (MA), glutamate, glutamine and arginine by high-affinity transport systems following Michaelis-Menten kinetics. The activities of these systems were influenced by the nitrogen source used for mycelial growth. Moreover, MA, glutamate and glutamine uptakes were derepressed by nitrogen starvation, whereas arginine uptake was repressed. The two ammonium-specific transport systems with different affinities and capacities were inhibited by NH(+)(4), with a K(i) of 3.7 microM for the high-velocity system. The K(m) values for glutamate, glutamine and arginine transport were 124, 151 and 32 microM, respectively. Inhibition of arginine uptake by lysine and histidine showed that they are competitive inhibitors. MA, glutamate and glutamine uptake was inversely proportional to the intracellular NH(+)(4) concentration. Moreover, increase of the intracellular NH(+)(4) level caused by PPT (DL-phosphinotricin) resulted in an immediate cessation of MA, glutamine and glutamate uptake. It seems that the intracellular NH(+)(4) concentration regulates its own influx by feedback-inhibition of the uptake system and probably also its efflux which becomes apparent when mycelium is grown on protein. Addition of extracellular NH(+)(4) did not inhibit glutamine uptake, suggesting that NH(+)(4) and glutamine are equally preferred nitrogen sources. The physiological importance of these uptake systems for the utilization of nitrogen compounds by A. bisporus is discussed.     

Determination of deuterium-labeled tryptophan in proteins by means of high-performance liquid chromatography and thermospray mass spectrometry

In order to develop direct methods for determining the extent of metabolic incorporation of isotopically labeled amino acids into a protein, the determination of deuterated tryptophan in [2H5]tryptophan-bacteriorhodopsin was investigated. The isotopically modified protein was subjected to alkaline hydrolysis. After phenyl isothiocyanate derivatization of the hydrolysate, the mixture was separated by reversed-phase liquid chromatography. Field desorption mass spectrometry and thermospray mass spectrometry were investigated for their ability to determine the ratio between [2H5]tryptophan and total tryptophan in the collected fractions. In order to check the procedure a set of known tryptophan/[2H5]tryptophan mixtures were passed through the same derivatization, HPLC separation, and lyophilization procedure as used for the biological samples.     

Evolution of sprouting versus seeding inAspalathus linearis

We have tested the hypothesis that reseeding is a plesiomorphic character state and that sprouting is a derived state inAspalathus linearis, and that the latter is an adaptation to ensure fire-survival in a fireprone environment. Samples of five seeder and four sprouter populations of A.linearis were examined by horizontal starch gel electrophoresis to assess the amount of genetic differentiation within and between sprouting and seeding populations, and to determine the extent of gene flow between the populations. Leaf extracts were surveyed for ten enzymes and gene products revealed genetic variation at 13 (76%) of 17 protein coding loci. Allele frequency differences were found between sprouting and seeding populations and genetic distance values show that the sprouters are grouped separate from the seeders, thus providing support for the morphological data on which the above mentioned hypothesis is based. It is evident that evolution operates at the population level inA. linearis.     

Retrograde Traffic Out of the Yeast Vacuole to the TGN Occurs via the Prevacuolar/Endosomal Compartment

A large number of trafficking steps occur between the last compartment of the Golgi apparatus (TGN) and the vacuole of the yeast Saccharomyces cerevisiae. To date, two intracellular routes from the TGN to the vacuole have been identified. Carboxypeptidase Y (CPY) travels through a prevacuolar/endosomal compartment (PVC), and subsequently on to the vacuole, while alkaline phosphatase (ALP) bypasses this compartment to reach the same organelle. Proteins resident to the TGN achieve their localization despite a continuous flux of traffic by continually being retrieved from the distal PVC by virtue of an aromatic amino acid–containing sorting motif. In this study we report that a hybrid protein based on ALP and containing this retrieval motif reaches the PVC not by following the CPY sorting pathway, but instead by signal-dependent retrograde transport from the vacuole, an organelle previously thought of as a terminal compartment. In addition, we show that a mutation in VAC7, a gene previously identified as being required for vacuolar inheritance, blocks this trafficking step. Finally we show that Vti1p, a v-SNARE required for the delivery of both CPY and ALP to the vacuole, uses retrograde transport out of the vacuole as part of its normal cellular itinerary.