Influence of the nitrogen level on root growth and morphology of two potato varieties differing in nitrogen acquisition

Two potato (Solanum tuberosum L.) cultivars (Astrid and Bodenkraft) differing in their nitrogen acquisition from the soil (Hunnius, 1981) were used in nutrient solutions to study the effect of increasing concentrations of nitrate (0.05; 0.5; 5.0 mol m^-3) particularly on root growth and morphology. In each variety increasing nitrogen concentrations stimulated shoot growth more than root growth. At all nitrate concentrations, the variety with higher nitrogen acquisition (Astrid) had a significantly larger root system. The larger root system of Astrid compared to Bodenkraft was particularly evident when surface area and total length of the roots, instead of root dry weight were used as parameters. The results stress the importance of root length and surface area for nitrogen acquisition from soils.     

Microbial degradation of dehydrodiconiferyl alcohol,a lignin substructure model

Bacteria, yeasts, and molds which grew in a medium containing a synthetic lignin — a dehydrogenation polymer (DHP) of coniferyl alcohol — as a sole carbon source, were isolated from soil. One fungus, Fusarium solani M-13-1, was found to degrade the DHP most vigorously among the isolated organisms. It was shake-cultured in a medium containing dehydrodiconiferyl alcohol (DHCA) (I), an important lignin model compound, and the following six metabolic products were isolated and identified: 1) Phenylcoumaran--aldehydic (II) and -carboxylic compounds, 2) phenylcoumaran--aldehydic compound (IV), formed by release of a 2-carbon fragment from the phenylcoumaran--carboxylic compound, 3) 5-acetylvanillyl alcohol (V), formed by cleavage of the coumaran ring and reduction of the -aldehyde group, 4) 5-carboxyvanillyl alcohol (VI), formed by subsequent oxidation of the acetyl group, and 5) the -ether of DHCA (VII), considered to be a by-product. A degradation pathway for DHCA was proposed on the basis of these metabolic products.Non-Standard Abbreviations DHP dehydrogenation polymer - DHCA dehydrodiconiferyl alcohol - DDQ dichlorodicyano-p-benzoquinone - DDHQ dichlorodicyano-p-hydroquinone - Ar aromatic - TLC thin layer chromatography - GC-MS gas chromatography-mass spectrometry     

Industrial applications of a cloned neutral protease gene in Bacillus subtilis

Summary Genes for the -amylase and neutral protease were cloned from an industrial Bacillus isolate, Bsl, onto two separate plasmids and introduced into a B. subtilis strain. Both plasmids were stably maintained in this strain. Analysis of the extracellular proteins showed that the plasmidcarrying strain produced predominantly the Bsl -amylase and neutral protease with few contaminating B. subtilis exoenzymes. The presence of high levels of protease enabled the strain to produce considerably more -amylase when grown on a complex industrial medium rich in protein.     

The NGF complex from the African ratMastomys natalensis

The 7S NGF complex from the male mouse submaxillary gland consists of the , and subunits in the ratio ₂₂. The (NGF) subunit contains all the known biolocial activity of 7S NGF. The and subunits are both members of glandular kallikrein gene family, yet only subunit has protease activity. The subunit plays a role in the processing of preproNGF to its mature form, while the role of the subunit is not yet understood. Despite the fact that 7S NGF has been extensively characterized, no other NGF complex has been characterized, nor have the or subunits been observed in tissues which express NGF. We have therefore purified and characterized the NGF complex from the submaxillary glands of the ratMastomys natalensis in order to more fully understand the roles of the and subunits. The NGF complex from M. natalensis contains subunits similar to those found in mouse 7S NGF. Although similar, there are significant differences between mouse and M. natalensis NGF complexes, especially in the degree of post-translational modification of the and NGF subunits, the expression of esterase activity and the ease with which the complexes dissociate. Evidence is presented that suggests that the NGF complex from M. natalensis may consist of subunits in the ratio ₂. The amino acid sequence of the M. natalensis NGF suggests some, but not all, ways in which these differences arise.Special issue dedicated to Dr. Lawrence Austin     

Molecular cloning of the T4 genome: organization and expression of the tail fiber gene cluster 34--38

Summary T4 derivatives that carry T4 tail fiber genes 34–38 have been isolated and characterized by genetic, structural and functional analysis. 32 T4 recombinants were identified by a marker rescue screen of 310 T4 clones generated by restriction of partial cytosine-containing T4 DNA with either HindIII or EcoRI and ligation into appropriately cleaved vectors. These tests defined 15 recombinant classes with respect to the contiguous stretches of genome recovered. Restriction enzyme structural analysis identified 7 HindIII fragments and 7 EcoRI fragments, established a restriction map covering about 11 kb, and indicated the orientation of the DNA inserts within the vectors. The cloned tail fiber genes are expressed efficiently from promoters and complement in vivo T4 phage carrying amber mutations in the tail fiber genes. Polypeptides corresponding to gp34-gp38 have been detected by SDS polyacrylamide gel electrophoresis of ³⁵S-labeled extracts of appropriate T4 recombinant infected UV-treated host cells. The genetic, structural and functional maps of the T4 tail fiber gene cluster have been correlated, and provide a rational approach to genetically directed DNA sequence analysis of genes 34–38 and their mutant variants that affect the assembly, structure and function of the tail fibers.     

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.     

Functional characterisation of two cytochrome<Emphasis Type="Italic"> b</Emphasis><Subscript>5</Subscript>-fusion desaturases from<Emphasis Type="Italic"> Anemone leveillei</Emphasis>: the unexpected identification of a fatty acid Δ<Superscript>6</Superscript>-desaturase

The Ranunculaceae are known to accumulate a wide range of unusual fatty acids in their seed lipids, and this variability has been advocated as a taxonomic marker. The Anemone species, Anemone leveillei L. and Anemone rivularis Buch.-Ham., have previously been reported to accumulate ⁵-desaturated fatty acids in their seed tissue [K. Aitzetmüller (1995) Plant Syst Evol 9:229–240]. Two cDNAs, AL1 and AL2, with similarity to plant cytochrome b₅-fusion "front-end" desaturases were isolated from developing seeds of A. leveillei and their function identified by expression in Saccharomyces cerevisiae. AL2 was characterised as a sphingolipid long-chain-base ⁸-desaturase, while AL1 acted as a fatty acid desaturase. However, AL1 did not produce ⁵-desaturated fatty acids as expected; instead, when expressed in transgenic S. cerevisiae or Arabidopsis thaliana this enzyme was functionally characterised as a ⁶-desaturase. Northern analysis confirmed the expression of this gene in seed tissue and leaf tissue of A. leveillei, though ⁶-desaturated fatty acids were found to accumulate only in the leaf tissue. The unexpected characterisation of a ⁶-desaturase in A. leveillei has implications for the use of fatty acids in chemotaxonomic studies. This is also the first report of a higher-plant ⁶-desaturase from a family other than the Boraginaceae.Abbreviations ALA -linolenic acid - DMOX 4,4-dimethyloxazoline - EDA eicosadienoic acid - FAME fatty acid methyl ester - GLA -linolenic acid - LA linoleic acid - LCB long chain base - ORF open reading frame - OTA octadecatetraenoic acid     

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.     

Averaging methods in the delayed-logistic equation

The delayed logistic equation is analyzed using the averaging method. Using the transformation of coordinates v=ln N/K it is shown that the first order term in perturbation theory yields N=K exp(r ^* cos t/2) when the delay time T exceeds some critical value T _c. The amplitude r^* is equal to (40/3 – 2)^1/2 and is an expansion parameter that is proportional to (T – T_c). Comparison of the exponential solution of N and numerical results for the ratio N _maximum/N _minimum provides a good fit for values of larger than the results using the N coordinate as the perturbed coordinate.     

Solvent production from starch: effect of pH on α-amylase and glucoamylase localization and synthesis in synthetic medium

Summary The fermentation of starch by Clostridium acetobutylicum ATCC 824 has been reviewed in an optimised synthetic medium. A progressive increase of pH from 4.4 to 5.2 led to a higher production of extracellular -amylase whereas glucoamylase was poorly affected. A portion of these enzymes was cell-associated and on increasing the pH from 4.4 to 5.8 a decrease was noted in cell-bound enzymes. The association was higher for the glucoamylase than for the -amylase. The highest rate of starch consumption was at pH 5.2 whereas due to the earlier shift to solvent production at low pH, the highest solvent production was at pH 4.4. This study suggested that the level of -amylase and then the rate of starch hydrolysis was the limiting step of sugar catabolism in C. acetobutylicum ATCC 824. Correspondence to: P. Soucaille     

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.     

Die Brutzeiten einiger Gänsearten und ihrer Bastarde in identischen Bedingungen

Zusammenfassung An Hand von 229 Brutbeginn-Daten von freilebenden Gänsen, die während der Jahre 1956–1966 in Seewiesen (Obb.) (48°N, 11°11E) brüteten, wurden die mittleren Brutbeginn-Daten von 5 Gänsearten und von Artbastarden bestimmt. Es zeigte sich, daß die untersuchten Arten unter diesen Bedingungen in derselben Reihenfolge brüteten, wie ihre Artgenossen in freier Wildbahn. Die mittleren Brutbeginn-Termine wurden allerdings um so mehr vorverlegt, je später die Art normalerweise brütet (Abb. 1). , die mit artfremden verpaart waren, brüteten zur selben Zeit wie ihre Artgenossen, die mit artgleichen verpaart waren (Abb. 1). GraugansxSchneegans-Bastard-, die mit Schneegantern verpaart waren, begannen meist nach den Graugänsen, aber stets vor den Schneegänsen zu brüten (Abb. 1, 2). Das intermediäre Brüten dieser wird als starkes Argument für die Richtigkeit der Hypothese gewertet, nach welcher die artspezifisch verschiedenen Brutzeiten wenigstens zum Teil genetisch bedingt sind. In der Diskussion wird die Frage kritisch erörtert, wie weit schon allein die Tatsache, daß die verschiedenen Arten über Generationen hinweg in derselben Reihenfolge wie ihre wildlebenden Artgenossen zu brüten beginnen, als Beweis für derartige genetische Unterschiede angesehen werden kann.

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Summary In 229 cases onset of breeding was recorded from free-living geese of 5 species and of some hybrids of these species, kept in Seewiesen/Obb. (48° N, 11° 11E) from 1956 to 1966. It was found that the species under these conditions bred in the same seasonal sequence as did wild birds. The mean breeding times, however, were found to be advanced in relation to the onset of breeding in the wild (Fig. 1). This was especially evident in the case of late-breeding species. paired with of another species came into breeding condition at the same time as paired with of the same species (Fig. 1). GraylegxSnowgoose hybrid paired with Snowgoose in most cases started to breed later than Greyleg geese but always earlier than the mean breeding time for Snowgeese (Fig. 1, 2). This intermediate breeding time is taken as a strong argument for the hypothesis that the species specific differences in breeding times are, at least in part, genetic in origin. The question as to the extent to which the differences in breeding times alone, persisting for generations in the same sequence as those of wild birds, can be attributed to genetic differences between the species, is critically discussed.

     

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).     

The occurrence and identification of intracellular polyglucose storage granules inMethylococcus NCIB 11083 grown in chemostat culture on methane

The accumulation of intracellular storage granules (0.03–0.5 m) byMethylococcus NCIB 11083 when grown under conditions of ammonia limitation with methane as the sole source of carbon and energy was inversely proportional to the dilution rate. The isolated material was composed entirely of glucose residues and the infra-red spectrum exhibited characteristic absorption bands at 925 cm^-1, 845 cm^-1 and 745±4 cm^-1, indicating the presence of (14) glycosidic linkages. The polymer dissolved in hot water to give an opalescent solution that formed a violet iodine complex with an absorption maximum at 550 nm, identical to that observed with reference amylopectin. The percentage of the polysaccharide released as maltose by the action of - and -amylases was 55–64% and 80–90% respectively, values very similar to those obtained by the action of these enzymes on reference amylopectin and glycogen. Methylation analysis indicated that the average interior and exterior chain lengths of the polymer were 2.7 and 10.0 glucose units respectively and confirmed that theMethylococcus polyglucose is a branched polymer composed of units joined by 14 and 16 linkages. The number average molecular weight of the polymer is 2–4.5×10⁵. The stored polymer was metabolised by the organism and its metabolism resulted in the synthesis of protein.Abbreviations GLC gas liquid chromatography - MS mass spectroscopy - PAAN peracetylated aldononitriles     

Testing for non-randomness in sizes and habitats of West Indian lizards: choice of species pool affects conclusions from null models

Summary Several null models are proposed for testing whether size or habitat differences in West IndianAnolis lizards are greater than expected by chance. The models differ primarily in choice of the pool from which species are sampled to form random communities. Regardless of choice of pool, size differences in the Lesser Antilles are greater than null models predict; the pool using species on the known source (Puerto Rico) gives a greater variance in ratios but about the same mean ratio (for males), or a greater mean ratio (for females), compared with the pool composed of species on the islands being tested (the Stronget al., 1979, Galápagos procedure). On satellite islands of the Greater Antilles, sizes do not differ more than expected from null models. Pools composed of mainland-source species give null communities with more small or more large ratios than those composed of island species, depending upon whether four-species islands are included or excluded, respectively. Colwell and Winkler's unmodified Narcissus hypothesis is contradicted by these results in procedures where species not likely to be able to occur on small islands are included in the species pool. Using the most biologically reasonable, but not other, choices of source pool, species on satellite islands of the Greater Antilles differ more in structural habitat then expected by chance. In contrast to some of the results on size, here mainland-source pools are more likely to produce a statistically significant difference between real and random communities, as predicted by the Narcissus hypothesis. However, exclusion of structural habitat categories not found on satellite islands is necessary to achieve this significance.     

Protein kinases expressed by interstitial cells of Cajal

Interstitial cells of Cajal (ICC) are involved in the generation of electrical rhythmicity of intestinal muscle and in the transduction of neural inputs in the gut. Although the expression of receptors for neurotransmitters and hormones and some second messengers have been investigated in ICC, the protein kinases present in these cells have not been well documented. This study has demonstrated the immunohistochemical localisation of PKA, PKC and PKC in ICC that were identified by the known ICC marker, c-Kit, in the guinea-pig gut. Other PKCs, PKC , , , , , and , and Ca²⁺-calmodulin-dependent protein kinase II were not localised in ICC. Double labelling studies were conducted on longitudinal muscle–myenteric plexus and external muscle–myenteric plexus preparations of the oesophagus, stomach (fundus, corpus and antrum), duodenum, distal ileum, caecum, proximal and distal colon, and rectum. The three protein kinases were detected in c-Kit-immunoreactive ICC at the level of the myenteric plexus (IC-MY), in the muscle (IC-IM) and at the level of the deep muscular plexus (IC-DMP) in the small intestine. PKA was found in over 90% of IC-IM in all regions examined, and in over 90% of IC-MY in the gastric body and antrum and throughout the small and large intestines. PKC was in the majority of ICC in the gastric body and antrum and in the small intestine, but was largely absent from ICC in the oesophagus, proximal stomach and large intestine. PKC occurred in the majority of ICC in all regions except the rectum. The intensity of staining was greatest for PKA, with PKC giving comparatively weak labelling of ICC. PKA was also detected in myenteric neurons, smooth muscle, macrophages and fibroblast-like cells. PKC labelling occurred in large, multipolar neurons throughout the small and large intestine, as well as in lymph vessels and in capillaries. It is concluded that PKA, PKC and PKC are all present in ICC, with the differences in their localisations suggesting specific roles for each in ICC function.     

Pigment-pigment interactions and secondary structure of reconstituted algal chlorophyll a/b-binding light-harvesting complexes of Chlorella fusca with different pigment compositions and pigment-protein stoichiometries

Earlier we have shown by in vitro reconstitution experiments that the pigment composition of the chlorophyll alb-binding light-harvesting complex of the green alga Chlorella fusca could be altered in a relatively broad range (Meyer and Wilhelm 1993). In this study we used these reconstituted complexes of different pigment loading to analyze the excitonic interactions between the pigment molecules and the secondary structure by means of circular dichroism spectra in the visible and the far UV spectral regions, respectively. We found that, in contrast to the expectations, the pigment composition and pigment content hardly affected the circular dichroism spectra in the visible spectral region. Reconstituted complexes, independent of their pigment composition, exhibited the most characteristic circular dichroism bands of the native light-harvesting complex, even if one polypeptide bound only 3 chlorophyll a, 3 chlorophyll b and 1–2 xanthophyll molecules. Full restoration of the protein secondary structure, however, could not be achieved. The -helix content depended significantly on the pigment composition as well as on the pigment-protein ratio of the reconstituted complexes. Further binding of pigments resulted in restoration of the minor excitonic circular dichroism bands, the amplitudes of which depended on the pigment content of the reconstituted complexes. These data suggest that in the reconstitution of light-harvesting complexes a central cluster of pigment molecules plays an important role. Further binding of pigments to the peripheral binding sites appeared also to stabilize the protein secondary structure of the reconstituted complexes.Abbreviations CD- circular dichroism - LHC- chlorophyll a/b light-harvesting complex(es) - LHC II- light-harvesting complex(es) of Photosystem II of higher plants - LHCP- light-harvesting Chl a/b-binding protein(s) - PP- polypeptide(s)     

The Expression of Transforming Growth Factor Alpha Receptor Protein and its Activation in Chicken Ovarian Granulosa Cells of Maturing Follicles

Epidermal growth factor (EGF) and transforming growth factor alpha (TGF-) are structurally related growth factors that exert their biological actions by binding to the same cell-surface receptor, EGF receptor. However, in chicken cells, human EGF binds with approximately 100-fold lower affinity than human TGF-. In a previous study, we localized EGF/TGF- receptor immunohistochemically in the granulosa and theca of the developing follicles of laying hens. We have also shown that TGF- binds to cell-surface receptors of the granulosa cells. The present study characterizes the nature of the EGF/TGF- receptor. Immunoprecipitation of receptor proteins from cultured granulosa cells with an anti-EGF receptor antibody (12E) shows the expression of a 170-kDa receptor protein. The expression of the receptor protein decreases with follicular enlargement between the F3 and F1. Incubation of the cells with [125I]TGF- followed by crosslinking with bis(sulphosuccinimidyl)suberate showed that TGF- binds a similar (170 kDa) receptor protein immunoprecipitated with the 12E anti-EGF receptor antibody. The binding of TGF- to granulosa cells caused receptor protein oligomerization, yielding the monomeric (170 kDa) and dimeric (340 kDa) protein forms. Oligomerization seemed to favour the formation of the dimeric rather than the monomeric form. Culturing granulosa cells with luteinizing hormone or follicle-stimulating hormone increased the expression of both monomer and dimer forms of the receptor proteins compared with the control. Western blotting analysis with anti-phosphotyrosine antibody revealed that the lysates of TGF--stimulated cells express phosphotyrosine-containing receptor proteins of 170 kDa and 340 kDa. The results show that chicken granulosa cells express the 170-kDa EGF=TGF- receptor protein, which dimerizes on binding to TGF-, suggesting that the receptor protein may be involved in the signal transduction of TGF- actions in the chicken granulosa cells.     

Variations regarding Susto causality among the cakchiquel of Guatemala

George Foster's model of personalistic and naturalistic disease theories is employed in the present analysis of fright-sickness among Cakchiquel villagers in highland Guatemala. Field data from Panajachel and San Antonio Aguas Calientes suggest that pronounced intrasocietal competition favors personalistic interpretation, with sorcery cited as the ultimate source, rather than naturalistic interpretation, which emphasizes chance or destiny. Village differences in subsistence ecology and internal competition apparently underlie variations in both the social function and assumed etiology of fright-sickness.