Development of high frequency multiple shoot formation in Persian clover (<Emphasis Type="Italic">Trifolium resupinatum</Emphasis> L.)

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1M BA and 1M IBA or 14.1M BA and 1M IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8M IBA.     

Betrachtungen zum Problem der Frostresistenzzüchtung von Tomaten

Zusammenfassung Zweijährige Selektion auf Frostresistenz bei einer Kreuzung der argentinischen Tomatensorte Platense mit der nordamerikanischen Pearl Harbour unter Freilandbedingungen erwies sich bei Überprüfung in der Kühlkammer im Vergleich zu frostempfindlichen Kultursorten als ergebnislos. Es wird daher der Schluß gezogen, daß auch bei Tomaten eine erfolgreiche Züchtung auf Frostresistenz nur durch Einkreuzung mit frostresistenten Wildarten möglich ist, die auch vorhanden sind. Unter den verfügbaren Wildarten wurde Frostresistenz beiL. peruvianum undL. hirsutum gefunden.     

Pulse schemes for the measurement of3 JC′Cγ and3 JNCγ scalar couplings in 15N,13C uniformly labeled proteins

Pulse sequences are presented for the measurement of³J_CC and³J_NC scalar couplings for allC containing residues in¹⁵N,¹³C uniformly labeled proteins. The methodsdescribed are based on quantitative J correlation spectroscopy pioneered byBax and co-workers [Bax et al. (1994) Methods Enzymol., 239, 79–105].The combination of ³J_CC and³J_NC scalar coupling constants allows theassignment of discrete rotameric states about the ₁ torsion angle in cases where such states exist or, alternatively,facilitates the establishment of noncanonical ₁conformations or the presence of rotameric averaging. The methods areapplied to a 1.5 mM sample of staphylococcal nuclease.     

Unequal specific uptake rates of α- and β-glucose by yeast andE.coli

The specific uptake rate of -glucose by yeast (S. cerevisiae) and recombinant yeast is higher than that of -glucose. On the other hand, the specific uptake rate of -glucose byE.coli and recombinantE.coli is less than that of -glucose. These results imply that the unequal specific uptake rates of optical isomers of glucose should be included in modeling and optimizing high-cell density fermentations, fed batch fermentations, and immobilized cell systems, where cell density is maintained high and glucose concentration is low.     

Aspects of Reproductive Biology of the Scalloped Hammerhead Shark, Sphyrna Lewini, off Northeastern Brazil

Ninety four scalloped hammerhead sharks, Sphyrna lewini (53 females and 41 males) ranging in size from 121 to 321cm total length (TL), were collected from surface gillnetters operating off northeastern Brazil and throughout the southwestern equatorial Atlantic Ocean between January and December 1996. A common regression for TL and eviscerated weight (EW) was calculated as, logEW = –11.786 + 2.889 logTL. Females and males were categorised into reproductive stages (4 and 2, respectively) according to morphological changes in their gonads. Size at sexual maturity for females was estimated to be 240cm, while males appeared to mature at between 180 and 200cm. Gravid females had between 2 and 21 embryos or pups, varying in TL from 3 to 38cm. There was no relationship between maternal length and size of litter. Copulation and parturition appear to occur outside the sampled area and possibly closer to the coast. With the exception of slightly lower uterine and ovarian fecundities, the results support the few existing data on the reproductive cycle of S. lewini in other areas.     

Reproductive Ecology of Pterapogon kauderni, an Endemic Apogonid from Indonesia with Direct Development

Pterapogon kauderni does not have secondary sexual dimorphism, and has an equal sex ratio (50.8% males:49.2% females). Females first mature at a length of about 34mm. Data from both laboratory-bred and wild-collected eggs and embryos are not consistent with the presence of differential reproductive allocation in this species. Analysis from clutches brooded by wild-collected males shows a significant intra-clutch and among-clutch variation in egg size. Mean individual egg size (diameter, volume, or weight) is not correlated with the size (standard length, head length, or lower jaw length) of the males brooding those eggs. P. kauderni has a lunar reproductive cycle, with a major spawning peak during full moon and a smaller peak during the last quarter. Similarly, it has a major juvenile release (settlement) peak at full moon, with a minor peak at new moon. However, one population, which is located in a site with distinct environmental conditions and which was sampled a year earlier, shows a different reproductive pattern, last quarter being the phase where most of spawn and release events take place.     

Genetics of the sex ratio anomaly in Drosophila hybrids of the Virilis group

Summary A study was made of the effect of genotype and temperature (25 and 17°C) on sex ratio in the hybrids D. virilis Sturt. X D. littoralis Sokolov. A genetic system has been found controlling sex-differential viability. In the F₁ of the reciprocal hybrids D. virilis X D. littoralis the sex ratio is normal, though at 17°C females are slightly excessive. The abnormal sex ratio is observed only in the progeny of test crosses.The major gene causing the death of female progeny of the cross [ (, D. virilis x , D. littoralis) x D. virilis] x D. littoralis is located on chromosome 2 of D. virilis. It is expressed as a lethal if chromosome 5 is heterogeneous virilis-littoralis. Chromosome 3 of D. virilis bears a modifier-enhancer and chromosome 5, a suppressor, of this lethal found in chromosome 2. This genetic system has a maternal effect and functions at 25°C, interacting with the X-chromosome of D. littoralis. If the maintainance temperature is lowered to 17°C, the progeny of the cross hybrid FB₁ x D. littoralis is predominantly female. Partial death of males is accounted for by a disturbance in the interaction between the genes of X-chromosome in certain combinations with the D. virilis autosomes and the Y-chromosome of the paternal species D. littoralis.Sex-differential mortality in the hybrids D. virilis x D. littoralis is one of the isolating factors between these species which does not appear to act until the second and subsequent F₁ generations due to the formation of the recombination load.     

Genetic control of β-diketone and hydroxy-β-diketone synthesis in epicuticular waxes of barley

Summary Five eceriferum, (cer) mutants in barley which influence -diketone and hydroxy--diketone synthesis in spike and internode epicuticular waxes have been characterized. The mutation cer-u ⁶⁹ blocks the synthesis of hydroxy--diketones and leads to a compensatory increase in the amount of -diketones, indicating that -diketones are precursors of the hydroxy--diketones. Furthermore, highly lobed wax plates were observed for the first time on barley lemmas, in addition to the characteristic wax tubes. Both diketone classes are selectively and proportionally reduced in the spike wax of cer-i ¹⁶, which has shorter wax tubes. The three mutants cer-c ³⁶, -q ⁴², and -c,u ¹⁰⁸ synthesize neither diketone class and form no wax tubes. In contrast to the variable composition of most individual barley wax classes, only a single -diketone was identified, namely hentriacontan-14,16-dione.     

Identification of beta subunit of the rhesus monkey chorionic gonadotropin (rmCG)

Chorionic gonadotropin (CG) is a placental derived hormone that plays a crucial role in successful implantation and establishment of early pregnancy in the primates. The rhesus monkey was chosen as a model to understand the feasibility of developing human DNA immuno-contraceptive. The coding region of rhesus monkey CG -subunit (rmCG) was isolated by the TDRT-PCR method. The nucleotide sequence including the leader peptide was 499 nucleotide long and encoded 166 amino acids. In comparing with the previous known primates CG -subunits, the rmCG was the highest degree of homology with baboon CG -subunit at the deduced amino acid sequence (94%), 79.5% homology with human CG -subunit and 70.4% homology with marmoset monkey CG -subunit. The eukaryotic expression vector pCMV4-rmCG inserted full-coding cDNA sequence of rmCG was constructed, and the expression of rmCG -subunit in HeLa cells transient expressing system in vitro and BALB/c mice in vivo was determined. The results demonstrated that the recombinant PCMV4-rmCG eukaryotic expression vector could express rmCG -subunit in vitro and in vivo.     

The de novo origin of the quiescent center regenerating root apices of Zea mays

Summary In roots from which the root cap and quiescent center have been removed new apical tissues regenerated in line with the main axis of the root. Regeneration of these tissues occurred from the region of the proximal meristem, which extends for no more than 350 m from the cut surface. Accompanying the regeneration of new apical tissues is a change in the architecture of the root apex and initiation and enlargement of a new quiescent center. A possible role for the quiescent center in the establishment of pattern at the apex is considered. Regeneration of the original apex failed to occur in those roots from which the root cap, quiescent center and proximal meristem were excised.Abbreviation QC quiescent center     

Beta-1,3-glucanase from unfertilized eggs of the sea urchin Strongylocentrotus intermedius. Comparison with beta-1,3-glucanases of marine and terrestrial mollusks

-1,3-Glucanase (Lu) was isolated from unfertilized eggs of the sea urchin Strongylocentrotus intermedius. A comparative study of some properties of -1,3-glucanase Lu and -1,3-glucanases with different action types—endo--1,3-glucanase from crystalline style of the marine mollusk Spisula sachalinensis (LIV) and exo--1,3-glucanase from the terrestrial snail Eulota maakii (LII)—was performed. It was found that -1,3-glucanase Lu hydrolyzes laminaran with a high yield of glucose in the reaction products. The enzyme hydrolyzes substrates with retention of the glycosidic bond configuration, is able to cleave modified substrates, and exhibits transglycosylating activity. All properties of -1,3-glucanase from S. intermedius were more similar to those of the endo--1,3-glucanase from the marine mollusk (LIV) than exo--1,3-glucanase LII from the terrestrial snail. The differences in the effect of LIV and Lu on laminaran are probably related to the functions of -1,3-glucanase Lu from sea urchin eggs (which, in contrast to LIV, is not a digestive enzyme).     

Fine structure of sensory tubes on the antennule of Conchoecia spinirostris (Ostracoda,Crustacea)

Summary In addition to setae, the first antennae of Conchoecia spinirostris also bear soft sensory tubes (4 tubes + 1 seta; 2 tubes + 3 setae). These tubes were examined electron microscopically. Each tube is divided into 4 regions: the stem, the bulbous region, the main region, and the tip. A tube contains 40–60 multiciliated dendrites, some hypodermal cells, and nonneuronal cells, and it has a specialized cuticle. Each dendrite develops within the tube, on the terminal 5–8 m of its inner dendritic segment, approx. 25 cilia in a 9 × 2 + 0 pattern, whose rootlets are absent or only poorly developed. Each cilium splits up into 9 ramifications which extend into the tip. These ramifications partly take a spirallike course and form a ring in the distal main part beneath the cuticle. Their membranes often dilate into spindleshaped swellings. In the center of the middle and distal parts of the main region approx. 7 dendrites without cilia are located, one of them reaches into the tip. The poreless cuticle is extremely delicate and electron lucid. In contrast to the cuticle of the setae it is elastic and soft. Special substructures are described. The tubes are completely covered by a filamentous surface coat. Because of the structure and the thin walled nature of the cuticle, permeability for dissolved substances is assumed. The ciliary ramifications are likely to represent the receptive apparatus. The sensory tubes are interpreted as chemoreceptors. They can best be compared with the chemoreceptors of certain crustaceans, but differ strongly from the types of sensilla found in insects.Supported by project 3540 of the Fonds zur Förderung der wissenschaftlichen Forschung in Österreich. The author is deeply indebted to Prof. Gamulin (Dubrovnik) for his support and to Prof. R. Riedl and Dr. W. Klepal for helpful discussions     

Existence and uniqueness of a sharp travelling wave in degenerate non-linear diffusion Fisher-KPP equations

In this paper we use a dynamical systems approach to prove the existence of a unique critical value c ^* of the speed c for which the degenerate density-dependent diffusion equation u _ct = [D(u)u _x ] _x + g(u) has: 1. no travelling wave solutions for 0 < c < c ^*, 2. a travelling wave solution u(x, t) = (x - c ^* t) of sharp type satisfying (– ) = 1, () = 0 ^*; '(^*–) = – c ^*/D'(0), '(^*+) = 0 and 3. a continuum of travelling wave solutions of monotone decreasing front type for each c > c ^*. These fronts satisfy the boundary conditions (– ) = 1, '(– ) = (+ ) = '(+ ) = 0. We illustrate our analytical results with some numerical solutions.     

A posteriori time-varying filtering of averaged evoked potentials

The problem of estimating an unknown transient signal, given an ensemble of waveforms, in which this signal appears as a nonrandom component in the presence of additive noise is considered. This problem is solved by generalizing the method of a posteriori Wiener filtering. In the new method, the ensemble average is filtered by a time-varying system which is based on estimated time-varying power spectra of signal and noise. The nature of this system, and the computational procedures involved, are discussed in detail. A software package for time-varying filtering is briefly described. Application of the method is illustrated by a simulation example, which also provides a comparison to time-invariant a posteriori Wiener filtering.     

Upstream non-coding region which confers polar expression to Ri plasmid root inducing gene rolB

Summary Root differentiation could be elicited on carrot discs by transformation with the agropine Ri plasmid rolB gene cloned in the binary vector Bin19, provided two conditions were met. Firstly, an adequate auxin supply had to be provided. This was achieved by co-inoculation with a strain carrying only the auxin synthetic genes of the T_R-DNA. Most of the resulting roots were then shown to harbour only rolB and no aux genes. Secondly, an extended non-coding region (1200 bp) at the 5 end of rolB had to be included in the construction. A shorter (300 bp) 5 region, including TATA and CCAAT boxes, was not sufficient to trigger root differentiation. Both the extended (B1185) and reduced (B310) 5 regions of rolB were then cloned upstream of the -glucuronidase (GUS) reporter gene and infections carried out both on the apical and on the basal side of carrot discs. Strong expression of GUS, visualized histochemically as an intense blue colouring of transformed cells was observed with B1185-GUS constructions on the apical side of the discs. Only occasionally could coloured cells be observed on the basal side of the discs with B1185-GUS and on both apical and basal sides with B310-GUS constructions. Strong GUS expression was, on the contrary, achieved on cells of both auxin-rich (apical) and auxin-depleted (basal) sides of the discs with the strong constitutive viral promoter, CaMV35S. These results indicate the presence of an upstream regulatory region which confers polar expression to the rolB gene and suggest a role for auxin in its activation.     

Leaf water potential,component potentials and relative water content in a xeric grass,Agropyron dasystachyum (Hook.) Scribn.

Summary Leaf water potential ( _l ), osmotic potential ( _s ), pressure potential ( _p , turgor pressure), relative water content (R) and their interrelationships were determined for a xeric grass (Agropyron dasystachyum) found in the grasslands of Canada. Thermocouple psychrometers were used to measure _l and _s ; _p was obtained by subtraction. _l dropped from near 0 bars to about-28 bars as R went from 90% to 75%. R greater than 90% was not observed, perhaps because of a systematic error in determination of turgid water content. R remained relatively high in A. dasystachyum, even at low _l . The slope of the _l -R relationship was similar to other species which are generally considered to be drought tolerant. _p as high as 14 bars was observed. Most of the decrease in _l was accounted for by a decline in _p . The ability of A. dasystachyum to adjust to fluctuating water stress over the growing season is probably as much related to changes in tissue structure and turgor relationships as to simple changes in osmotic potential.     

The genetic defect in the various types of human β-galactosidase deficiency

Summary Gene localization studies revealed the presence of two structural -galactosidase (GAL) loci on the human chromosomes 3 and 22 (de Wit et al., 1979). To determine the function of these genes, proliferating hybrid cell lines were isolated following fusion of fibroblasts from two different patients with a GAL deficiency and Chinese hamster cells. The hybrids were analyzed electrophoretically and immunologically.Fibroblasts from a patient with an adult type of GAL deficiency associated with a neuraminidase deficiency were used for the first fusion. No evidence for a structural GAL mutation was found in these hybrids. The absence of a structural GAL mutation is consistent with a primary defect in neuraminidase in this adult patient.Fibroblasts from a patient with the infantile type 1 G_M1-gangliosidosis were used for the second fusion. It is concluded that the human determinants present in the isolated hybrid lines occur in heteropolymeric man-Chinese hamster molecules. The heteropolymeric isoenzyme in (+3–22) hybrids is very labile and is sensitive to neuraminidase treatment. Therefore it is concluded that the infantile type 1 patient is mutated in the structural GAL gene on chromosome 3. Because this patient has a primary defect in G_M1-GAL, the GAL gene on chromosome 3 is apparently a G _M1-GAL gene. Interaction of the two GAL loci results in an additional band of GAL activity on electrophoresis. This suggests that the gene on chromosome 22 is also a structural G _M1-GAL gene.     

Evolution of a protein superfamily: Relationships between vertebrate lens crystallins and microorganism dormancy proteins

Summary A search of sequence databases shows that spherulin 3a, an encystment-specific protein ofPhysarum polycephalum, is probably structurally related to the - and -crystallins, vertebrate ocular lens proteins, and to Protein S, a sporulation-specific protein ofMyxococcus xanthus. The - and -crystallins have two similar domains thought to have arisen by two successive gene duplication and fusion events. Molecular modeling confirms that spherulin 3a has all the characteristics required to adopt the tertiary structure of a single -crystallin domain. The structure of spherulin 3a thus illustrates an earlier stage in the evolution of this protein superfamily. The relationship of - and -crystallins to spherulin 3a and Protein S suggests that the lens proteins were derived from an ancestor with a role in stressresponse, perhaps a response to osmotic stress.     

A Cytogenetic Study of the Leaf Beetle Genus Cyrtonus (Coleoptera, Chrysomelidae)

The chromosomes of ten species of Cyrtonus and the genome sizes of six are surveyed. Among the total of 15 chromosomally studied species, 11 have 2n=28 chromosomes and a 13+Xy_p male meioformula, three have 2n=40 and 19+Xy_p and one 2n=46 and 22+Xy_p. All but one species with 28 chromosomes show only metacentric or submetacentric chromosomes, whereas the species with 40 and 46 chromosomes display some telocentrics or subtelocentrics, that are probably derived from the former by centric fissions. However, since the number of major chromosome arms is strikingly higher in these latter species (NF=70 and 78) than in the 28-chromosome species (mostly NF=56), other chromosomal rearrangements such as pericentric inversions or heterochromatin accretions could also be involved. The genome sizes display a narrow range, from 1C=0.6–1.22pg, and they are not significantly correlated with the chromosome numbers. Some possible factors implied in the rough chromosomal evolution of Cyrtonus are discussed in relation to a few other genera of the subfamily Chrysomelinae.