The presence of partially 3-O-methylated mannogalactan from the fruit bodies of edible basidiomycetes Pleurotus ostreatus 'florida' Berk. and Pleurotus ostreatoroseus Sing

The partially 3-O-methylated mannogalactans were isolated from the fruiting bodies of edible basidiomycetes Pleurotus ostreatus 'florida' Berk. and Pleurotus ostreatoroseus Sing. They were obtained via successive aqueous extraction, freeze thawing, and precipitation with Fehling solution and then investigated using (13)C- and (1)H-nuclear magnetic resonance spectroscopy (including COSY, TOCSY and HMQC techniques), methylation analysis and Smith degradation. The main chain consisted of (1-->6)-linked alpha-D-galactopyranosyl residues containing 3-O-Me-alpha-D-galactopyranoses, a part of these units being substituted in the position O-2 with beta-D-mannopyranose residues. The heteropolysaccharides found were similar with differences only in the levels of the 3-O-Me-alpha-D-galactopyranoses residues. The presence of partially 3-O-methylated mannogalactan appears to be typical of Pleurotus spp.     

Inhibition of plant-pathogenic fungi by the barley cystatin Hv-CPI (gene Icy) is not associated with its cysteine-proteinase inhibitory properties

The recombinant barley cystatin Hv-CPI inhibited the growth of three phytopathogenic fungi (Botrytis cinerea, Colletotrichum graminicola, and Plectosphaerella cucumerina) and the saprotrophic fungus Trichoderma viride. Several mutants of barley cystatin were generated by polymerase chain reaction approaches and both their antifungal and their cysteine-proteinase inhibitory properties investigated. Point mutants R38-->G, Q63-->L, and Q63-->P diminished their capacity for inhibiting papain and cathepsin B, retaining their antifungal properties. However, mutant C68-->G was more active for papain and cathepsin B than the wild type. These results indicate that in addition to the consensus cystatin-reactive site, Q63-V64-V65-A66-G67, the A37-R38-F39-A40-V41 region, common to all cereal cystatins, and the C68 residue are important for barley cystatin activity. On the other hand, the K92-->P mutant is inactive as a fungicide, but still retains measurable inhibitory activity for papain and cathepsin B. Against B. cinerea, the antifungal effect of Hv-CPI and of its derived mutants does not always correlate with their activities as proteinase inhibitors, because the Q63-->P mutant is inactive as a cystatin, while still inhibiting fungal growth, and the K92-->P mutant shows the reciprocal effects. These data indicate that inhibition of plant-pathogenic fungi by barley cystatin is not associated with its cysteine-proteinase inhibitory activity. Moreover, these results are corroborated by the absence of inhibition of intra- and extramycelia-proteinase activities by barley cystatin and by other well-known inhibitors of cysteine-proteinase activity in the fungal zymograms of B. cinerea.     

Homoeologous chromosomal location of the genes encoding thionins in wheat and rye

Summary Thionins are high sulphur basic polypeptides present in the endosperm of Gramineae. In wheat there are three thionins encoded by genes located in the long arms of chromosomes 1A, 1B and 1D. Rye has one thionin encoded by a gene which has been assigned to chromosome 1R after analysis of the Imperial-Chinese Spring rye-wheat disomic addition lines. Commercial varieties and experimental stocks with a 1B/1R substitution carry the thionin from rye ( _R) instead of the _B thionin from wheat. The _R thionin gene is not located in the large chromosomal segment representing most of the short arm of chromosome 1R.     

Differential Allelic Expression at a Locus Encoding an Endosperm Protein in Tetraploid Wheat (TRITICUM TURGIDUM)

Two hydrophobic endosperm proteins, designated CM3 and CM3', have been purified from appropriate cultivars of tetraploid wheat (T. turgidum) and characterized. They are inherited as though encoded by alleles at a single locus, designated Cm3a and Cm3b, respectively. The net amount of protein molecules has been measured for each of the alleles at one, two and three doses. The amount of CM3' is 50%–65% of that found for CM3. For both, there is a linear gene dosage response. These effects were observed not only in the parental material and the reciprocal F₁ generations, but also in the segregating F₂ generation, indicating that the quantitative difference depends on differences in the structural gene or is controlled by regulatory or modifier gene(s) linked to it.     

Gene expression in alloploids: genetic control of lipopurothionins in wheat

Lipopurothionins are complexes of basic polypeptides and polar lipids found in petroleum ether extracts of wheat endosperm. Location of the structural genes for the protein moiety and of genes probably controlling the lipid moiety has been achieved by analysis of compensated nulli-tetrasomic and ditelosomic lines of Triticum aestivum L. cv. Chinese Spring, as well as of other genetic stocks. There are two electrophoretic variants of the apoprotein designated α and β purothionins. Structural genes for α purothionins are located in the long arm of chromosomes 1B and 1D, and for the β variant in the long arm of 1A. These genes have been tentatively designated Pur-A1, Pur-B1, and Pur-D1. The aminoacid composition of purified α and β purothionins from Triticum aestivum (genomes AABBDD) and T. durum (AABB), and of β purothionin from T. monococcum (AA) is also consistent with this conclusion and suggests that the α purothionin encoded by gene Pur-B1 probably differs from that encoded by gene Pur-D1 in at least three positions of the aminoacid sequence. A gene (or genes) located in the short arm of chromosome 5D markedly affects the level of lipopurothionin but does not affect apoprotein synthesis. It is concluded that they control the lipid moiety which is required for solubility in petroleum ether.     

Comparative phylogenetic analysis of cystatin gene families from arabidopsis,rice and barley

The plant cystatins or phytocystatins comprise a family of specific inhibitors of cysteine proteinases. Such inhibitors are thought to be involved in the regulation of several endogenous processes and in defence against pests and pathogens. Extensive searches in the complete rice and Arabidopsis genomes and in barley EST collections have allowed us to predict the presence of twelve different cystatin genes in rice, seven in Arabidopsis, and at least seven in barley. Structural comparisons based on alignments of all the protein sequences using the CLUSTALW program and searches for conserved motifs using the MEME program have revealed broad conservation of the main motifs characteristic of the plant cystatins. Phylogenetic analyses based on their deduced amino acid sequences have allowed us to identify groups of orthologous cystatins, and to establish homologies and define examples of gene duplications mainly among the rice and barley cystatin genes. Moreover, the absence of a counterpart between the two monocots, as well as strong variations in the motifs that interact with the cysteine proteinases, may be related to a species-specific evolutionary process. This cystatin classification should facilitate the assignment of proteinase specificities and functions to other cystatins as new information is obtained.Electronic Supplementary Material Supplementary material is available for this article at