Species-specific alleles at a <Emphasis Type="Italic">β-</Emphasis>tubulin gene show significant associations with leaf morphological variation within <Emphasis Type="Italic">Quercus petraea</Emphasis> and <Emphasis Type="Italic">Q. robur</Emphasis> populations

Quercus petraea and Q. robur are largely sympatric oak species in western and central Europe and known for their intensive genetic exchange which has made the discovery of species-diagnostic markers a huge challenge. Various natural white oak populations (Q. petraea/Q. robur including mixed stands) were investigated for their variability and differentiation patterns at a β-tubulin gene (qutub8) in a European-wide survey. This gene was chosen as a possible candidate among loci subjected to selection and maintaining integrity between species. Two frequent alleles depicted as indels within qutub8’s first intron showed remarkably high interspecific genetic differentiation, with Weir and Cockerham’s theta per allele values ranging from 0.17 to 0.30 for one allele and from 0.04 to 0.19 for the other allele in such mixed oak stands where the multi-allelic qutub8 locus showed significant interspecific F _ST . For three mixed stands, qutub8’s F _ST significantly departed from the expected neutral differentiation patterns (F _ST ranging from 0.063 to 0.080 for this multi-allelic marker) and thus could be influenced by selection. Significant associations were found between genotypic variation and leaf dimensions as well as leaf structure patterns, after having accounted for species and stand effects. Qutub8 represents a locus that exhibits significant species differentiation and is linked to morphological discriminant traits. Consequently, qutub8 likely contributes to species divergence within the European white oak complex.     

Identification and characterization of polypeptide growth factors secreted by murine embryonal carcinoma cells

Undifferentiated P19 and PC13 murine embryonal carcinoma (EC) cells have been analyzed for their ability to secrete polypeptide growth factors. This has been carried out by a combination of specific bioassays and the use of biochemical and immunological detection methods. Both P19 and PC13 EC cells secrete a platelet-derived growth factor (PDGF)-like growth factor, a type beta transforming growth factor, and insulin-like growth factors. In addition, PC13 EC cells secrete a heparin-binding growth factor functionally related to fibroblast growth factor, while P19 EC cells secrete transforming growth factor-alpha. This is the first demonstration for secretion of transforming growth factor-alpha by an equivalent of early embryonic cells. The possible paracrine growth stimulating effects of these growth factors have been tested on differentiated derivatives of P19 EC cells, corresponding to all three germ layers. The differences in growth factor production by various embryonal carcinoma cells are discussed in relation to the developmental origin of these cell lines.     

Identification of autodigestion target sites in Bacillus subtilis neutral proteinase.

Autocatalytic degradation of purified Bacillus subtilis neutral proteinase was examined under various conditions. At elevated temperatures, under non-inhibitory conditions, mature protein was rapidly degraded, but no accumulation of specific breakdown products occurred. However, by incubating purified neutral proteinase on ice during extended periods of time, specific peptides accumulated. These peptides were analysed by SDS/PAGE and Western blotting, and the N-terminal sequences were determined for the four major peptides, which had sizes of 30, 22, 20 and 15 kDa. Sequence data identified five fission sites in the neutral proteinase, three of which were identical with autodigestion target sites in thermolysin, a thermostable neutral proteinase. Comparison of the identified fission sites of the B. subtilis neutral proteinase with the known substrate-specificity of the enzyme indicated that they were in agreement, showing a preference for the generation of fissions at the N-terminal side of large hydrophobic residues, such as leucine, isoleucine and methionine. These results suggest a high degree of similarity in the three-dimensional structures of B. subtilis neutral proteinase and thermolysin.     

Red light enhancement of the phototropic response of etiolated pea stems

In the subapical third internode of 7-day-old etiolated pea seedlings, the magnitude of phototropic curvature in response to continuous unilateral blue illumination is increased when seedlings are pre-exposed to brief red light. The effect of red light on blue light-induced phototropism becomes manifest maximally 4 or more hours after red illumination, and closely parallels the promotive action of red light on the elongation of the subapical cells. Ethylene inhibits phototropic curvature by an inhibitory action on cell elongation without affecting the lateral transport of auxin. Pretreatment of seedlings with gibberellic acid causes increased phototropic curvature, but experiments using ¹⁴C-gibberellic acid indicate that gibberellic acid itself is not laterally transported under phototropic stimuli. Neither red light nor gibberellic acid treatment has any promotive effect on blue light-induced lateral transport of ³H-indoleacetic acid. Under conditions where phototropic curvature is increased by red light treatment, low concentrations of indoleacetic acid applied in lanolin paste to the apical cut end of the seedling cause an increased elongation response in subapical tissue. This could explain increased phototropic curvature caused by red light treatment.     

PATHS OF TRANSTUBULAR WATER FLOW IN ISOLATED RENAL COLLECTING TUBULES

The cells of perfused rabbit collecting tubules swell and the intercellular spaces widen during osmotic flow of water from lumen to bath induced by antidiuretic hormone (ADH). Ouabain had no influence on these changes. In the absence of net water flow intercellular width was unaffected when tubules were swollen in hypotonic external media. Therefore, during ADH-induced flow widening of intercellular spaces is not a consequence of osmotic swelling of a closed intercellular compartment containing trapped solutes, but rather is due to flow of solution through the channel. Direct evidence of intercellular flow was obtained. Nonperfused tubules swollen in hypotonic media were reimmersed in isotonic solution with resultant entry of water into intercellular spaces. The widened spaces gradually collapsed completely. Spaces enlarged in this manner could be emptied more rapidly by increasing the transtubular hydrostatic pressure difference. In electron micrographs a path of exit of sufficient width to accommodate the observed rate of fluid flow was seen at the base of the intercellular channel. It is concluded that the intercellular spaces communicate with the external extracellular fluid and that water, having entered the cells across the luminal plasma membrane in response in ADH, leaves the cells by osmosis across both the lateral and basilar surface membranes.     

DNA synthesis and division in interkingdom heterokaryons

Summary Drosophila cells and plant protoplasts were fused by polyethylene glycoldimethylsulfoxide treatment. DNA synthesis in plant and animal cells and in their heterokaryons were monitored by [³H]thymidine autoradiography. In appropriate media both the parental cells and the heterokaryons preserved their DNA synthetic activity. Division of the heterokaryons were also observed.     

An explanation of the inhibition of root growth caused by indole-3-acetic Acid

Low concentrations of indole-3-acetic acid inhibit the growth of pea root sections by inducing the formation of the growth regulator, ethylene gas. Ethylene is produced within 15 to 30 minutes after indole-3-acetic acid is applied and roots begin to swell immediately after they are exposed to the gas. Carbon dioxide competitively inhibits ethylene action in roots, impedes their geotropic response, and partially reinstates auxin inhibited growth. It is concluded that ethylene participates in the geotropic response of roots, but not that of stems.