Phytotoxin

Abstract: A new allele of the coronatine-insensitive locus (COI1) was isolated in a screen for Arabidopsis thaliana mutants with enhanced resistance to the bacterial pathogen Pseudomonas syringae. This mutant, designated coi1-20, exhibits robust resistance to several P. syringae isolates but remains susceptible to the virulent pathogens Erisyphe and cauliflower mosaic virus. Resistance to P. syringae strain PstDC3000 in coi1-20 plants is correlated with hyperactivation of PR-1 expression and accumulation of elevated levels of salicylic acid (SA) following infection, suggesting that the SA-mediated defense response pathway is sensitized in this mutant. Restriction of growth of PstDC3000 in coi1-20 leaves is partially dependent on NPR1 and fully dependent on SA, indicating that SA-mediated defenses are required for restriction of PstDC3000 growth in coi1-20 plants. Surprisingly, despite high levels of PstDC3000 growth in coi1-20 plants carrying the salicylate hydroxylase (nahG) transgene, these plants do not exhibit disease symptoms. Thus resistance to P. syringae in coi1-20 plants is conferred by two different mechanisms: (i) restriction of pathogen growth via activation of the SA-dependent defense pathway; and (ii) an SA-independent inability to develop disease symptoms. These findings are consistent with the hypotheses that the P. syringae phytotoxin coronatine acts to promote virulence by inhibiting host defense responses and by promoting lesion formation.

Abstract: The present invention provides for a method to circumvent the problem of using antibiotic resistant selectable markers. In particular, target plants are transformed using a plastid vector which contains heterologous DNA sequences coding for a phytotoxin detoxifying enzyme or protein. The selection process involves converting an antibiotic-free phytotoxic agent by the expressed phytotoxin detoxifying enzyme or protein to yield a nontoxic compound. The invention provides for various methods to use antibiotic-free selection in chloroplast transformation.

Abstract: Thaxtomin A is a phytotoxin produced by Streptomyces scabies and other Streptomyces species, the causative agents of common scab disease in potato and other taproot crops. At nanomolar concentrations, thaxtomin causes dramatic cell swelling, reduced seedling growth, and inhibition of cellulose synthesis in Arabidopsis. We identified a mutant of Arabidopsis, designated txr1, that exhibits increased resistance to thaxtomin as a result of a decrease in the rate of toxin uptake. The TXR1 gene was identified by map-based cloning and found to encode a novel, small protein with no apparent motifs or organelle-targeting signals. The protein, which has homologs in all fully sequenced eukaryotic genomes, is expressed in all tissues and during all developmental stages analyzed. Microarray transcript profiling of some 14,300 genes revealed two stomatin-like genes that were expressed differentially in the txr1 mutant and the wild type. We propose that TXR1 is a regulator of a transport mechanism.

Abstract: (1) Experimental plant populations were grown at varying densities and varying levels of phytotoxins in the soil to investigate the potential interacting influences of allelopathy and resource competition on plant response and yield-density relationships. (2) Experiments were performed with (a) bahiagrass, Paspalum notatum, grown in soil treated with gallic acid and hydroquinone, putative inhibitors produced by Polygonella myriophylla and (b) tomato, Lycopersicon esculentum, grown in soils from under and around black walnut, Juglans nigra. (3) Phytotoxicity decreased as plant density increased. This was attributed to plant 'dilution' of phytotoxins, i.e. the sharing of the available phytotoxin among many plants at high densities such that each received a small sub-lethal dose. (4) Phytotoxins caused characteristic deviations from expected yield-density relationships. Low to moderate phytotoxin concentrations caused a decrease in the slope of the log yield-log density relationship. Contrary to the expected consequences of increased density and resource competition, the presence of high phytotoxin concentrations may cause a reversal in the slope of predicted log yield-log density relationships at low plant densities, such that maximum individual plant weight occurs at an intermediate density. (5) Demonstration of either decreasing phytotoxicity with increasing plant density or a reversal in slope of the predicted log yield-log density relationship is proposed as an indication of the presence of toxic substances in soil. (6) The ecological implications of these results are discussed.