Trypsin Inhibitors during Germination of Vigna sinensis Seeds

The trypsin inhibiting activity of cotyledonary extracts of serido beans (Vigna sinensis) falls continuously during germination to a value which represents 5 % of the value for the dry seed. The specific activity is constant over the first three days and then decreases, reaching a level at the seventh day of germination which represents 20 % of the initial value. It is also shown that the axial parts control the disappearance of the trypsin inhibiting activity of the cotyledons. The results presented here seem to indicate a role for these substances in the process of germination.     

Immunodetection of Botrytis-specific Invertase in Infected Grapes

Invertase (β-fructofuranosidase, EC 3.2.1.26) activity was shown to be stimulated in grape berries after infection with Botrytis cinerea . By using organism-specific extraction methods, evidence was found proving that both partners contribute to the increase in activity. Qualitative analysis of the extracts by SDS–PAGE showed a new invertase species in the botrytised material, with a molecular weight similar to that of Botrytis invertase (BIT). A method allowing the preparative isolation of homogeneous invertase from liquid cultures of B. cinerea in only three steps (ethanol precipitation, DEAE anion exchange and hydroxyapatite chromatography) with 50% recovery was developed. BIT appeared to be strongly glycosylated; interestingly, the amount of glycan seemed to vary and had a remarkable influence on the chromatographic behaviour of the enzyme. After chemical deglycosylation with TFMS, the BIT peptide was used for the production of polyclonal antibodies in chickens (anti-BIT-IgY). The antibodies recognised glycosylated as well as deglycosylated BIT, a partial denaturation of the protein being necessary for reaction. This indicates that the deglycosylation procedure had been successful and that the antibodies were in fact directed to the peptide moiety. However, after a short incubation at 70°C, native BIT was detectable by anti-BIT-IgY, while still active. Western blotting with extracts of diseased berries confirmed the fungal origin of the new invertase form. The anti-BIT-IgY proved highly specific, although some cross-reaction with a protein in Monilia laxa extracts occurred. The importance of careful immunogen preparation in the production of specific antibodies, and the potential of BIT as a target molecule for the immunological detection of B. cinerea are discussed.     

Pre‐wintering conditions and post‐winter performance in a solitary bee: does diapause impose an energetic cost on reproductive success?

1. Diapause is a dynamic process of low metabolic activity that allows insects to survive periods of harsh conditions. Notwithstanding the lowered metabolism, and because diapausing insects have no access to food, diapause has an energetic cost that may affect post‐diapause performance. 2. Previous studies on the solitary bee Osmia lignaria have shown that prolonged pre‐wintering periods (the time during which individuals already in diapause remain at warm temperatures) are associated with elevated lipid consumption, fat body depletion, and body weight loss. The present study investigated whether prolonged pre‐wintering also affects reproduction, i.e. whether the costs associated with diapause could have an effect on post‐diapause performance in this species. 3. Females were exposed to a range of pre‐wintering conditions, and ovary development and individual post‐wintering performance were monitored throughout their adult life span. 4. No evidence of an effect of pre‐wintering duration on post‐diapause reproductive success was found. Expected differences in the timing of establishment were not observed because ovary maturation was, surprisingly, not arrested during pre‐wintering. Prolonged pre‐wintering duration did not result in decreased life span, probably because emerging females could rapidly replenish their metabolic reserves through feeding. However, there was a very strong effect of the duration of the pre‐emergence period on the likelihood of nest establishment. 5. Longevity, the main factor determining fecundity in Osmia, is subjected to high levels of intrinsic variability, even among females of similar size exposed to identical conditions during development and nesting.     

Phylogenetic relationships between the families Capsalidae and Dionchidae (Platyhelminthes, Monogenea, Monopisthocotylea) indicated by the comparative ultrastructural study of spermiogenesis

Comparative ultrastructural observations were carried out on the spermiogenesis of the capsalid Caballerocotyla manteri Price and the dionchid Dionchus remorae MacCallum. At the beginning of spermiogenesis the zones of differentiation (ZD) jut out in all directions. A large mitochondrion shaped like a perforated bead, and through which the elongated nucleus passes, is found facing each ZD. Later the ZD become parallel and are embedded within the common cytoplasmic mass. Cortical longitudinal microtubules are present in the ZD at the outset of spermiogenesis, but they later disappear. The spermatozoon is long and filiform. It shows two parallel axonemes of the 9 +"1" flatworm pattern, the nucleus and mitochondrion, and no cortical microtubule. The ultrastructure of spermiogenesis and spermatozoon is remarkably similar in the two species studied, as in other capsalids previously described. Two characteristics, the perforated bead shape of the spermatid mitochondrion and the progressive disappearance of the microtubules of the ZD, may be considered as synapomorphies which indicate close phylogenetic relationships between the families Dionchidae and Capsalidae. This interpretation coincides with Llewellyn's (1971) scheme of the evolution of the monogeneans.     

The Sooty Falcon Falco concolor on the Red Sea coast of Saudi Arabia: distribution, numbers and conservation

Ground and aerial surveys of breeding Sooty Falcons Falco concolor were conducted along the western coast of Saudi Arabia and its offshore islands from August to October 1991 and 1992. An estimated population of 260–381 pairs was found. The known world breeding population is still below 1000 pairs. Most rocky islands were occupied except the largest islands of the Farasan archipelago. Among the many low islets surrounded with mangrove, only one small group near Al Lith harboured breeding Sooty Falcons. The distribution pattern, density and breeding success of the falcons seemed to be strongly influenced by food availability (autumn migration of small and medium-size birds) and the deterring presence of terrestrial predators. Human disturbance and trapping are still of local importance.     

CHANGES IN CHEMICAL, SENSORY AND RHEOLOGICAL CHARACTERISTICS OF MANCHEGO CHEESES DURING RIPENING

Manchego cheese is a high-fat pressed ewe's-milk cheese made in Castilla-La Mancha (Spain) and produced by enzymatic coagulation. The minimum ripening time before marketing required by the Regulatory Board of the Manchego Cheese Appellation of Origin is 60 days.
This paper describes the physicochemical, proteolysis, sensory and texture characteristics of Manchego cheese, and the degree of homogeneity of cheeses made under the Manchego Appellation of Origin. The data gathered in this study indicate that sensory and instrumental analysis are useful tools for detecting changes in Manchego cheese during ripening. These changes were first detected by the instrumental analysis (2 months). The panelists detected differences after 4 months' ripening in all the factories. With physicochemical analysis, on the other hand, longer ripening times (6–8 months) are required before such changes become appreciated.     

Analysis of enamel-line length as a morphometric parameter and its application to Quaternary arvicolids

This paper proposes a methodology for taxonomic, phylogenetic and ecological analyses based on enamel distribution in the crown of the tooth. Enamel units (EU) and morphological units (MU) are defined, which can be used to identify both genera and species, and also morphotypes in populations closely located in time. The biological character of the enamel units allows us to propose hypotheses on the genetic characteristics of fossil populations. Mammals, herbivores, Arvicolidae, enamel, methodology, taxonomy, biostratigraphy.     

Enhancement of root hydraulic conductivity by methyl jasmonate and the role of calcium and abscisic acid in this process

The role of jasmonic acid in the induction of stomatal closure is well known. However, its role in regulating root hydraulic conductivity (L) has not yet been explored. The objectives of the present research were to evaluate how JA regulates L and how calcium and abscisic acid (ABA) could be involved in such regulation. We found that exogenous methyl jasmonate (MeJA) increased L of Phaseolus vulgaris, Solanum lycopersicum and Arabidopsis thaliana roots. Tomato plants defective in JA biosynthesis had lower values of L than wild‐type plants, and that L was restored by addition of MeJA. The increase of L by MeJA was accompanied by an increase of the phosphorylation state of the aquaporin PIP2. We observed that MeJA addition increased the concentration of cytosolic calcium and that calcium channel blockers inhibited the rise of L caused by MeJA. Treatment with fluoridone, an inhibitor of ABA biosynthesis, partially inhibited the increase of L caused by MeJA, and tomato plants defective in ABA biosynthesis increased their L after application of MeJA. It is concluded that JA enhances L and that this enhancement is linked to calcium and ABA dependent and independent signalling pathways.