Effects of thionins on β-glucuronidase in vitro and in plant protoplasts

Thionins cause the irreversible inactivation of β-glucuronidase (GUS) in vitro in a dose- and time-dependent manner. The enzyme is also sensitive to externally added thionins when expressed in the cytoplasmic compartment of tobacco protoplasts transformed with the Gus gene under the 35S promoter of the cauliflower mosaic virus. In protoplasts transformed with the Gus gene fused to a signal peptide, where GUS is translocated into the lumen of the endoplasmic reticulum, the activity is significantly increased both by externally-added and by transiently-expressed thionin, suggesting that it interferes with GUS secretio

The barley a-thionin promotor is rich in negative regulatory motifs and directs tissue-specific expression of a reporter gene in tobacco.

The promoter of the barley α-thionin gene (1.6 kb) fused to the β-glucuronidase (GUS) gene directs temporally-controled, tissue-specific expression in the endosperm of transgenic tobacco. The nucleotide sequence of this promoter shows negative regulatory motifs which have been functionally analyzed in other gene

Expression of the alpha-thionin gene from barley in tobacco confers enhanced resistance to bacterial pathogens

Thionins are cysteine-rich, 5 kDa polypeptides which are toxic to plant pathogens in vitro. Expression of the gene encoding alpha-thionin from barley endosperm, under the 35S promoter from cauliflower mosaic virus, conferred to transgenic tobacco enhanced resistance to the bacterial plant pathogens Pseudomonas syringae pv. tabaci 153 and P. syringae pv. syringae. The barley alpha-thionin gene, which has two introns, was correctly spliced in tobacco. The alpha-thionin in transgenic plants had the expected mobility in the gradient, when separated by high- performance liquid chromatography, reacted with monospecific antibodies and showed the expected antibiotic properties in vitr

Characterization and analysis of thionin genes

The general designation of thionins has been proposed for a family of homologous proteins that have been isolated from different tissues in a wide range of plant taxa and have been variously named purothionins, viscotoxins, crambins, etc. (see Garcia-Olmedo et al., 1989). The possible involvement of thionins in plant defense was first suggested, on the basis of their in vitro toxicity to plant pathogens, by Fernandez de Caleya et al., (1972). Those observations had been prompted by earlier reports concerning the antimicrobial properties of these polypeptides (Stuart and Harris, 1942; Balls and Harris, 1944). Work on the thionins, which has been actively pursued over the past half-century, has been recently reviewed in detail (Garcia-Olmedo et al., 1989). For this reason, earlier work will only be partially summarized in the present chapter, which will focus on recent developments concerning thionin genes and their potential role in plant defense mechanisms

The thionins: a protein family that includes purothionins, viscotoxins and crambins

Almost half a century ago, a crystalline protein material was obtained from lipid extracts of wheat endosperm and designated "purothionin" (Trvpwo, wheat; 9 e i w v , sulphur) on account of its high sulphur content (Balls et al., 1942a,b). This material, which was thought to be the oxidized form of a powerful oxidation-reduction system, was found to have baetericidal and fungicidal properties (Stuart and Harris, 1942), to inhibit fermentation of wheat mashes (Balls and Harris, 1944), and to be toxic to laboratory animáis (Coulson et al., 1942)

Trypsin/α-amylase inhibitors and thionins: possible defence proteins from barley

This chapter reviews recent work on the trypsin/α-amylase inhibitor and thionin protein families. The genomic distribution of protein genes in barley and related species, gene expression and in vitro activities are considered. Some of the evidence of a possible defence role against stored products pests for inhibitors and thionins is briefly discusse