Sterol composition of dwarf and tall Pisum sativum seedlings in relation to gibberellic-acid-enhanced shoot growth

Gibberellic acid (GA₃) stimulated shoot elongation in both dwarf and tall cultivars of pea, but more so in the dwarf cultivar. The sterol composition of shoots of both cultivars was similar, with sitosterol being the most abundant compound, followed by stigmasterol and campesterol. Cholesterol could not be detected. Following GA₃ application, levels of free sterols in whole shoots increased whereas glycoside levels tended to fall. The magnitudes of the changes in both classes of sterol were similar in both cultivars. Analyses of stems and leaves separately revealed a greater growth response to GA₃ in the former but no effect of the hormone on the sterol composition of either organ. It is concluded that GA₃ enhancement of shoot growth in pea is not mediated through quantitative changes in cell sterols.     

The early adaptive evolution of calmodulin

Interaction between gene duplication and natural selection in molecular evolution was investigated utilizing a phylogenetic tree constructed by the parsimony procedure from amino acid sequences of 50 calmodulin- family protein members. The 50 sequences, belonging to seven protein lineages related by gene duplication (calmodulin itself, troponin-C, alkali and regulatory light chains of myosin, parvalbumin, intestinal calcium-binding protein, and glial S-100 phenylalanine-rich protein), came from a wide range of eukaryotic taxa and yielded a denser tree (more branch points within each lineage) than in earlier studies. Evidence obtained from the reconstructed pattern of base substitutions and deletions in these ancestral loci suggests that, during the early history of the family, selection acted as a transforming force on expressed genes among the duplicates to encode molecular sites with new or modified functions. In later stages of descent, however, selection was a conserving force that preserved the structures of many coadapted functional sites. Each branch of the family was found to have a unique average tempo of evolutionary change, apparently regulated through functional constraints. Proteins whose functions dictate multiple interaction with several other macromolecules evolved more slowly than those which display fewer protein-protein and protein-ion interactions, e.g., calmodulin and next troponin-C evolved at the slowest average rates, whereas parvalbumin evolved at the fastest. The history of all lineages, however, appears to be characterized by rapid rates of evolutionary change in earlier periods, followed by slower rates in more recent periods. A particularly sharp contrast between such fast and slow rates is found in the evolution of calmodulin, whose rate of change in earlier eukaryotes was manyfold faster than the average rate over the past 1 billion years. In fact, the amino acid replacements in the nascent calmodulin lineage occurred at residue positions that in extant metazoans are largely invariable, lending further support to the Darwinian hypothesis that natural selection is both a creative and a conserving force in molecular evolution.      

Annual Variation in Sterol Levels in Leaves of Taraxacum officinale Weber

Sterol levels in dandelion (Taraxacum officinale Weber) leaves were monitored over a period of 19 months. Sitosterol was the most abundant free sterol, followed by stigmasterol, then campesterol. Cholesterol could not be detected. With the exception of stigmasterol and campesterol, esters were present in greater quantities than were free forms, with 4,4-dimethyl sterol esters being the most abundant type. Glycosides occurred only sporadically. Free 4-demethyl sterols were maximal during the winter months; levels correlated negatively with sunshine and temperature, but proportions did not alter significantly. Sitosterol ester and cycloartenol ester (but not others) showed the opposite response, with levels correlating positively with sunshine and temperature. Relative amounts of 4-demethyl sterol esters remained reasonably constant, but those of cycloartenol ester and 24-methylene cycloartanol ester varied on an annual basis and were negatively correlated with each other.     

Release of growth hormone from ox pituitary slices after pronase treatment

Proteolytic enzymes have been used both to modify properties of the cell membrane and to dissociate cells from many tissues including pituitary (4, 5, 12). Exposure of secretory tissues to pronase can alter their secretory response. Thus incubation of pancreatic islets of Langerhans in the presence of low concentrations of pronase increased the subsequent release of insulin in the presence of stimulatory and nonstimulatory glucose concentrations (7). The purpose of the present investigation was to determine whether low concentrations of pronase have the same stimulatory effect on the release of a pituitary hormone, growth hormone. Such an effect on hormone release could be of some importance in view of the development of dissociated cell systems as models for the study of the control of hormone release (4, 5).     

Steroidal glycoalkaloid content of potato,tomato and their somatic hybrids

Summary Analyses of leaves and ‘tubers’ from somatic hybrids of potato and tomato (‘pomato’ with plastids of potato, ‘topato’ with plastids of tomato) produced by fusion of protoplasts from liquid cultures of dihaploid potato and mesophyll of tomato revealed the presence of the two major potato glycoalkaloids (α-solanine and α-chaconine) as well as the tomato glycoalkaloid (αtomatine). The total alkaloid content of leaves was greater than that of ‘tubers’ and similar to levels in the foliage of parent plants. However, glycoalkaloids were more abundant in hybrid ‘tubers’ than in normal potato tubers by a factor of 5–15. In hybrid foliage, approximately 98% of the alkaloid present was of potato origin whereas in ‘tubers’ the reverse was the case, with tomatine comprising 60–70% of the total alkaloid. The similarities in alkaloid content and ratios between the pomato and the topato lines indicate that plastomes do not influence the biosynthesis and distribution of these alkaloids. The results indicate that major secondary metabolites may prove useful for assessing the hybrid nature of such plants.     

Subcellular localization of steroidal glycoalkaloids in vegetative organs of Lycopersicon esculentum and Solanum tuberosum

Cell fractions from the major vegetative organs of tomato and potato plants were obtained by homogenization and differential centrifugation. In both species, steroidal glycoalkaloids were found to accumulate mainly in the soluble fraction, with smaller amounts in the microsomal fraction. Alkaloids appeared sporadically in the mitochondrial fractions but were only detected in lower fractions as an artefact.