Electrical fusion of protoplasts from sycamore mutants and hybrid selection on hypoxanthine-aminopterin-thymidine (HAT) medium

Summary An electrical fusion method has been used to form somatic hybrids between protoplasts of two mutant cell lines of sycamore tissue culture cells. Both mutants will not grow in a hypoxanthine-aminopterin-thymidine (HAT) medium. It was possible to select the fused hybrids from homospecific fusion products and nonfused protoplasts by the use of HAT medium. In this way the viability and regeneration of the fused cells during the first few weeks of culture could be evaluated. An electron microscopic examination of the fusion process showed that it occurred at a series of points along the surface of the plasmalemma. Cytoplasmic bridges between the two cells were formed separated by vesicles which later dispersed to give complete cytoplasmic continuity between the cells.     

Differential expression within a family of novel wound-induced genes in potato

Summary Wounding in higher plants leads to an increased synthesis of specific messenger RNAs. A cDNA clone complementary to a wound-induced message from potato tubers was used to isolate a lambda clone from a genomic library of Salanum tuberosum var. Maris Piper. DNA sequence analysis has shown that this single genomic clone contains two novel wound-induced genes, called win1 and win2, organised in close tandem array. The coding sequences of these two genes are highly homologous and are interrupted by a single intron. However, the sequences of the introns and flanking regions have diverged widely. Win1 and win2 encode cysteine-rich proteins of 200 and 211 amino-acids, respectively, which show striking homologies to several chitin-binding proteins. Southern analysis of genomic DNA has shown that win1 and win2 are members of a small multi-gene family which is estimated to have a minimum of five members per haploid genome of Maris Piper and appears to be conserved within the Solanaceae. We have shown by Northern analysis and S1 mapping that the two genes exhibit differential organ-specific expression after the wounding of a potato plant.     

Resource utilization of sympatric and experimentally allopatric cutthroat trout and Dolly Varden charr

Summary Resource utilization by cutthroat trout (CT) and Dolly Varden charr (DV) was studied 8 years after experimental transfers from sympatry had established reproducing allopatric populations in two nearby fishless lakes. Allopatric DV significantly increased their utilization of shallow-dwelling zoobenthos, and increased their vertical distribution in comparison to that in sympatry. In contrast, allopatric CT showed little change in the proportions of major prey types utilized, and, if anything, restricted their vertical distribution in comparison to that in sympatry. The results can be explained by the hypothesis that the resource use of DV is strongly influenced by interspecific competition from CT, whereas CT largely remains unaffected by this interaction. An alternative hypothesis, that lake differences can explain the differences in resource use between sympatry and allopatry, was evaluated by comparing food resource availability and other biotic and abiotic characteristics of the three study lakes. None of these could account for the shift in resource use by DV between sympatry and allopatry, but lake differences may explain why allopatric CT showed a restricted habitat use in comparison with their sympatric donor stock. The results of this whole-lake transfer experiment are consistent with earlier reported field and laboratory studies, and suggest that the aggressive dominance of CT is the most important mechanism by which DV are displaced from littoral and near-surface habitats in sympathy with CT.     

Optimal age at sexual maturity of sympatric and experimentally allopatric cutthroat trout and Dolly Varden charr

Summary Reproductive potentials of transplanted curthroat trout (Salmo clarki) and Dolly Varden charr (Salvelinus malma) and of their donor stocks were estimated from life history data. We found good agreement between observed and predicted age at maturity in all populations, and cannot reject the hypothesis that the fish matured at the age maximizing the overall lifetime reproductive potential ( ). Our estimates were insensitive to probable variations in female fecundity, adult mortalityrate and maximum body length. Small changes in either juvenile mortality-rate or individual growth-rate had marked effects on the estimations, as did changes in the Malthusian parameter (r). Three alternative mechanistic explanations of how age at maturity is determined could be rejected. We suggest that fish are able to adjust the maturity age non-genetically to changes in growth-rate, and that temporal variations in juvenile survival-rate allow coexistence of genotypes coding for different ages at maturity at the same growth-rate.     

Cell-cell recognition of host surfaces by pathogens. The adsorption of maize (Zea mays) root mucilage by surfaces of pathogenic fungi.

The adsorption of radioactive mucilage by pathogenic fungi was shown to be dependent upon time, the composition of mucilage, the type of fungal surface (conidia, hyphae, hyphal apices), fungal species, pH and bivalent cations. All fungal adhesins were inactivated by either proteinase or polysaccharase treatments. Adsorption was not inhibited by the numberous mono-, di- and oligo-saccharides that were tested individually, but it was inhibited absolutely by several polysaccharides. This suggested that adsorption of mucilage by pathogens involved conformational and ionic interactions between plant and fungal polymers but not fungal lectins bound to sugar residues of mucilage. Several fractionation schemes showed that pathogens bound only the most acidic of the variety of polymers that comprise mucilage. There was not any absolute distinction between ability to bind radioactive mucilage and type of pathogen or non-pathogen. However, there were notable differences in characteristics of adsorption between two types of pathogen. Differences were revealed by comparison of the adsorption capacities of conidia and germinant conidia and chromatography of radioactive mucilage on germinant conidia. An ectotrophic root-infecting fungus (a highly specialized pathogen) bound a greater proportion of mucilage than did a vascular-wilt fungus (of catholic host and tissue range) with more than one class of site for adsorption. In contrast with the vascular-wilt fungus, sites for adsorption on the specialized pathogen were present solely on surfaces formed by germination.     

The action of exogenous gibberellic acid on polysome formation and translation of mRNA in germinating castor-bean seeds

The amount of protein synthesis in germinating castor-bean seeds has been estimated by the quantitative and qualitative exmainatin of polysomes from the seeds in the presence and absence of gibberellic acid (GA₃). Careful optimisation of polysome extraction procedures was required to minimise the ribonuclease activity in the extracts. Ribonuclease activity in seed extracts increased fourfold over the first 5 d of germination. Gibberellic acid stimulated polysome formation about twofold during the first 4 d of germination. It also stimulated the amount of mRNA associated with polysomes by about twofold during the first 3 d of germination. Between days 1 and 5 of germination, polysome formation was primarily limited by mRNA availability. During the period 0–24 h, polysome formation was independent of mRNA levles. The increase in enzyme activities stimulated by GA₃ was probably the result of an increase in the amount of cellular mRNA. No evidence was obtained for an action of GA₃ on translation other than on the increased production of RNA. Examination of the recruitment of isocitrate-lyase mRNA into polysomes showed that GA₃ did not specifically stimulate production of this enzyme.     

The relationship of root-cap slimes to proteins

1. The patterns of incorporation of radioactivity from d-[U-(14)C]glucose into the pectic components of sections of sycamore roots changed so that sections nearer the tip incorporated relatively more label into arabinose and galactose compared with uronic acid. 2. Radioactive maize root-cap slime was prepared and found to contain three water-soluble component polymers which were electrophoretically (i) neutral, (ii) weakly acidic and (iii) strongly acidic at pH6.5. The neutral component was a glucan. The other components, which could be degraded by trans-elimination, consisted of an acidic backbone chain composed of galacturonic acid and glucose, attached to which were different proportions of neutral sugars. Arabinose, galactose and fucose, the main neutral sugars of the weakly and strongly acidic materials, were absent from the neutral fraction. 3. Fucose was a major sugar in maize-root slime and in a slime of similar composition synthesized by a maize callus of shoot origin. Only trace amounts were found in sycamore, pea and wheat root tips, and in pectin prepared from maize roots and coleoptiles. A high proportion of fucose is therefore a chemical characteristic of maize slime, and slime synthesis indicated a state of differentiation of the tissue. 4. The similarity between the slime and pectin is discussed; slime is a form of pectin modified in such a way as to provide a hydrated protective coating around the root tip.     

Pectic polysaccharides of growing plant tissues

1. The polysaccharide compositions of the cell walls of sycamore cambium and sycamore callus tissue have been analysed and found to be directly comparable. 2. Electrophoretic analyses of the whole pectins prepared from actively growing callus and cambial tissue have shown that these preparations contain, in addition to the neutral and weakly acidic components present in apple fruit, a strongly acidic polygalacturonic acid component. 3. The weakly acidic component of all the pectins was directly comparable with that of the pectinic acid of apple fruit. 4. The components of the whole pectin of sycamore callus tissue have been partially purified and analysed. The neutral and weakly acidic components also found in apple fruit were isolated. 5. The pattern of the composition of the neutral sugars present in the pectins of actively growing tissues of cambium and callus has been compared with those present in apple-fruit pectinic acid. 6. The presence of rhamnose linked as galacturonosyl-(1-->2)-rhamnose has been found in sycamore whole pectin. 7. The difference in the pectins of callus, cambium and fruit appears not to be that of species difference but is more characteristic of the nature of the growth and growth conditions of the cells. This is discussed in relation to the problems of the control and mechanism of plant-cell growth and differentiation.     

THE FINE STRUCTURE AND DEVELOPMENT OF THE COMPANION CELL OF THE PHLOEM OF ACER PSEUDOPLATANUS

At maturity the companion cell of the phloem of the sycamore Acer pseudoplatanus has a large nucleus, simple plastids closely sheathed with rough endoplasmic reticulum, and numerous mitochondria. The cytoplasm contains numerous ribosomes, resulting in a very electron-opaque cytoplasm after permanganate fixation. Bodies similar to the spherosomes of Frey-Wyssling et al. (4) are collected in clusters and these also contain bodies of an unidentified nature similar to those found by Buttrose (1) in the aleurone cells of the wheat grain. The pores through the wall between the companion cell and sieve tube are complex and develop from a single plasmodesma. Eight to fifteen plasmodesmata on the companion cell side communicate individually with a cavity in the centre of the wall which is linked to the sieve tube by a single pore about twice the diameter of an individual plasmodesma. This pore is lined with material of an electron opacity equivalent to that of material bounding the sieve plate pores. The development of the cell organelles, the possible role played in the phloem tissue by the companion cell, and the function of the complex pores contained in its wall are discussed.     

Partial characterization of the polyisoprenoid carrier of N-acetylglucosamine in Glycine max (soya bean).

Radiolabelled anhydrotrypsin was bound by alpha 2M (alpha 2-macroglobulin) sufficiently tightly to resist separation during gel electrophoresis; 2 mol of anhydrotrypsin were bound/mol of alpha 2M, but the interaction differed in important respects from that between active proteinases and alpha 2M. Anhydrotrypsin was bound by the electrophoretically 'fast' form of alpha 2M, although much less effectively than by the 'slow' form. The inactive enzyme was displaced from alpha 2M by trypsin inhibitor, the order of effectiveness being aprotinin > soya-bean trypsin inhibitor > benzamidine. Saturation of alpha 2M with anhydrotrypsin did not prevent subsequent binding and inhibition of active trypsin by the alpha 2M, and the anhydrotrypsin was not displaced during this reaction. Anhydrotrypsin bound by alpha 2M retained its ability to react with antibodies against trypsin, whereas bound trypsin did not.