Satellite Spitzenkörper in growing hyphal tips

Summary Growing hyphal tips of higher fungi contain an organized assemblage of secretory vesicles and other cell components collectively known as the Spitzenkörper. Until now, the Spitzenkörper has been portrayed as a single spheroid complex located near the apical cell wall. This study demonstrates the occurrence of multiple Spitzenkörper in growing hyphal apices imaged by video-enhanced phase-contrast microscopy. In addition to the main Spitzenkörper, smaller satellite Spitzenkörper arise a few micrometers behind the apical pole. Four developmental stages were identified: (a) the satellites first appeared as faint phase-dark plaques next to the plasma membrane, (b) gradually increased in size and assumed an ovoid profile, (c) they migrated to the hyphal apex, and (d) finally they merged with the main Spitzenkörper. After the merger, the main Spitzenkörper temporarily increased in size. Satellites were observed in 14 fungi, most of which had relatively large (5–10 m diam.), fast-growing hyphae (2–33 m/min elongation rate). The average frequency of in-focus satellites was 7+/min forFusarium culmorum and 11+/min forTrichoderma viride. As with the main Spitzenkörper, satellites were present only in growing cells. They were transient and remained visible for 3–8 s before merging with the main Spitzenkörper. Within the hyphae, satellites travelled up to six times faster than the average cell elongation rate. Multiple satellites sometimes occurred simultaneously; up to three were seen within a hyphal apex at the same time. Localized cell enlargement occurred next to stationary satellites, suggesting that satellite Spitzenkörper are functional as sources of new cell surface before they reach the main Spitzenkörper; therefore, they account for some variations in the profiles of the growing hyphae. By electron microscopy, satellites consisted of small clusters of apical vesicles surrounding a group of microvesicles located next to the plasma membrane. The identification and behavior of the satellites represent clear evidence of directional mass transport of vesicles toward the hyphal apex. Our observations indicate that satellites are a common phenomenon in growing hyphal apices of septate fungi and that they contribute to growth of the hyphal apex.Abbreviations VSC vesicle supply center     

Vitamin C stabilization as a consequence of the plasma membrane redox system

Summary Ascorbate is stabilized in the presence of HL-60 cells. Our results showed that cAMP derivatives and agents that increase cAMP stimulate the ability of HL-60 cells to stabilize ascorbate. On the other hand, tunicamycin, a glycosilation-interfering agent, inhibited this ability. The ascorbate stabilization in the presence of HL-60 cells has been questioned as a simple chemical effect. Further properties and controls about the enzymatic nature of this stabilization are described and discussed. This data, together with hormonal regulation, support the hypothesis that an enzymatic redox system located at the plasma membrane is responsible of the extracellular ascorbate stabilization by HL-60 cells.Abbreviations AFR ascorbate free radicals - FCS fetal calf serum - Sp-cAMPS Sp-cyclic adenosine monophosphothionate - Rp-cAMPS Rp-cyclic adenosine monophosphothionate     

Factors affecting Citrus tree isozyme-gene expression

Ten enzymatic systems of Citrus species and cultivars have been evaluated for identification purposes and for genetic variability studies. The following factors that could affect their expression were studied: season of sampling, location, rootstock, position of the branch, infection, and age of the tree. Differences involving the presence-absence of the Cu/Zn SOD within the same tree were found. This difference is mainly related to the position of the leaf relative to the sunlight. No change was observed at any of the ten enzymatic systems assayed regarding the location, the rootstock, the growing condition, the season, or the infection with most virus and virus-like pathogens. Viroids induced noticeable changes on 6PG and PRXa zymograms in C. medica. A new peroxidase (not present in healthy plants) was detected that could be related to appearance of symptoms. This may induce errors when trees without sanitary control are characterized by this enzymatic system. On the other hand, it provides a new possibility for studying the plant response to the presence of viroids. An effect of age, from 3 months up to 12 years, was observed on citrange Troyer and mandarin Cleopatra PRX, MDH and 6PG patterns. An important change occurs around the first year, most likely related to the end of the seedling stage. This is followed by a long transition phase, the end of which (around 9 years later) coincides with a change in the PRX pattern. These age-related changes seem to involve post-translational modifications of pre-existing isozymes.     

Chromosome localization and characterization of a family of long interspersed repetitive DNA elements from the genus Zea

This paper describes the characterization and chromosomal distribution of new long repetitive sequences present in all species of the genus Zea. These sequences constitute a family of moderately repetitive elements ranging approximately from 1350 to 1700 copies per haploid genome in modern maize (Zea mays ssp. mays) and teosinte (Zea diploperennis), respectively. The elements are long, probably larger than 9 kb, and they show a highly conserved internal organization among Zea subspecies and species. The elements are present in all maize chromosomes in an interspersed pattern of distribution, are absent from centromeric and pericentric heterochromatin, and with some clustering in the distal regions of chromosome arms.     

The vegetable rennet of Cynara cardunculus L. contains two proteinases with chymosin and pepsin-like specificities

The flowers of cardoon (genus Cynara) are traditionally used in Portugal for cheese making. In this work the vegetable rennet of the species Cynara cardunculus L. was characterized in terms of enzymic composition and proteolytic specificity of its proteinases (cardosin A and cardosin B). Cardosin A was found to cleave insulin B chain at the bonds Leu15-Tyr16, Leu17-Val18 and Phe25-Tyr26. In addition to the bonds mentioned cardosin B cleaves also Glu13-Ala14, Ala14-Leu15 and Phe24-Phe25 indicating that it has a broader specificity. The kinetic parameters for the hydrolysis of the synthetic peptide Leu-Ser-Phe(NO₂)-Nle-Ala-Leu-oMe were also determined and compared to those of chymosin and pepsin. The results obtained indicate that in terms of specificity and kinetic parameters cardosin A is similar to chymosin whereas cardosin B is similar to pepsin. It appears therefore that the enzyme composition of cardoon rennet closely resembles that of calf rennet.     

The binding motifs for Ac transposase are absolutely required for excision of Ds1 in maize

A reverse genetic system for studying excision of the transposable elementDs1 in maize plants has been established previously. In this system, theDs1 element, as part of the genome of maize streak virus (MSV), is introduced into maize plants via agroinfection. In the presence of theAc element, excision ofDs1 from the MSV genome results in the appearance of viral symptoms on the maize plants. Here, we used this system to study DNA sequences requiredin cis for excision ofDs1. TheDs1 element contains theAc transposase binding motif AAACGG in only one of its subterminal regions (defined here as the 5′ subterminal region). We showed that mutation of these motifs abolished completely the excision capacity ofDs1. This is the first direct demonstration that the transposase binding motifs are essential for excision. Mutagenesis with oligonucleotide insertions in the other (3′) subterminal region resulted in elements with either a reduced or an increased excision efficiency, indicating that this subterminal region also has an important function.     

A new mite-plant association: mites living amidst the adhesive traps of a carnivorous plant

Non-antagonistic interactions between arthropods and leaves of insectivorous plants with adhesive traps so far have never been reported. The mites are common prey of such plants, but we have found a new subspecies of the mite Oribatula tibialis living on the leaves of Pinguicula longifolia. Because of its small size and the low glandular density of the host, the mite moves without being trapped by the mucilaginous droplets of the leaf surface. P. longifolia provides shelter and food for the mite, while the plant may also benefit because of its fungivorous and scavenging activities. This new interaction is another dramatic example of widespread miteplant associations.     

Role of endogenous gibberellins in germination of melon (Cucumis melo) seeds

The effect of the plant growth retardants ancymidol. mefluidide and uniconazole on germination of two melon accessions differing in their ability to germinate at 14°C was examined. The accessions were the cold sensitive Noy Yizre'el and the cold tolerant Persia 202. The three growth retardants were able to delay the germination of intact Noy Yizre'el seeds, but did not affect that of intact Persia 202 seeds. On the other hand germination of decoated seeds of both accessions was unaffected by these inhibitors at normal oxygen concentration, but was inhibited at 5% oxygen. When gibberellin-like activity was measured by a dwarf rice biological assay following HPLC fractionation, it was found that seeds of Persia 202 contained much more gibberellin-like activity than Noy Yizre'el seeds. Among the extracted compounds several endogenous gibberellins were identified by combined gas chromatography-mass spectrometry (GC-MS). They included GA₄, GA₂₀, GA₁ and GA₃ in Noy Yizre'el and GA₃₄, GA₂₀, GA₁ and GA₈ in Persia 202. It is suggested that the better germination of intact Persia 202 seeds, compared to Noy Yizre'el seeds at low temperature and low oxygen concentration, is due to a higher endogenous level of GA and a better seed coat permeability to oxygen.     

Study of interaction between calcium and zinc ond-galactose intestinal transport

Zinc is an essential trace element necessary to life. This metal may exert some of its physiological effects by acting directly on cellular membranes, either by altering permeability or by modulating the activity of membrane-bound enzymes. On the other hand, calcium is an essential element in a wide variety of cellular activities. The aim of the present work was to study a possible interaction between zinc and calcium on intestinal transport ofd-galactose in jejunum of rabbit in vitro. In media with Ca²⁺, when ZnCl₂ was present at 0.5 or 1 mM, zinc was found to reduce thed-galactose absorption significantly. In Ca²⁺-free media, where CaCl₂ was omitted and replaced isotonically with choline chloride, the sugar transport was not modified by zinc. Verapamil at 10⁻⁶ M (blocking mainly Ca²⁺ transport) did not modify the inhibitory effect of zinc ond-galactose transport. When 10⁻⁶ M of A 23187 (Ca²⁺-specific ionophore) was added with/without Ca²⁺ to the media, ZnCl₂ produced no change in sugar transport. These results could suggest a possible interaction of calcium and zinc for the same chemical groups of membrane, which could affect the intestinal absorption of sugars.     

An unusual Group 2 LEA gene family in citrus responsive to low temperature

Six cDNAs representing unique cold-induced sequences have been cloned from the hardy citrus relative Poncirus trifoliata. Among these, pBCORc115 and pBCORc119 were found to belong to the same gene family. Sequencing data indicated that pBCORc115 and pBCORc119 each contained an open reading frame, coding for a 19.8 kDa protein (COR19) and a smaller 11.4 kDa protein (COR11) respectively. Inspection of the deduced amino acid sequences revealed three large repeats in COR19, but only one was present in the COR11. Two elements: a Q-clustered tract and a K-rich motif were identified in each repeat. The K-rich motifs were similar to those of cotton D-11 and Group 2 LEA proteins. A Serine-cluster, a common feature in many Group 2 LEA-like proteins, was also found in these proteins, but it was in an unusual position at the carboxy-terminus. A bipartite motif of basic residues, similar to known nuclear targeting sequences, was also present in COR19 and COR11, suggesting that members of this protein family may have a nuclear targeting function. The expression of COR19 mRNA in response to cold acclimation, drought, flooding, and salinization was examined. COR19 expression in leaf tissue was induced in response to cold acclimation, but repressed during drought and flooding stress.