Gary Creissen

Inducible resistance responses play a central role in the defense of plants against pathogen attack. Acquired resistance (AR) is induced alongside defense toward primary attack, providing broad-spectrum protection against subsequent pathogen challenge. The localization and molecular basis of AR in cereals is poorly understood, in contrast with the well-characterized systemic acquired resistance (SAR) response in Arabidopsis. Here, we use Pseudomonas syringae as a biological inducer of AR in barley, providing a clear frame of reference to the Arabidopsis–P. syringae pathosystem. Inoculation of barley leaf tissue with the nonadapted P. syringae pv. tomato avrRpm1 (PstavrRpm1) induced an active local defense response. Furthermore, inoculation of barley with PstavrRpm1 resulted in the induction of broad-spectrum AR at a distance from the local lesion, “adjacent” AR, effective against compatible isolates of P. syringae and Magnaporthe oryzae. Global transcriptional profiling of this adjacent AR revealed similarities with the transcriptional profile of SAR in Arabidopsis, as well as transcripts previously associated with chemically induced AR in cereals, suggesting that AR in barley and SAR in Arabidopsis may be mediated by analogous pathways.

Natural-product effectors of disease resistance in Arabidopsis reveal complementary disabling mechanisms in the pathogen.

Plant aspartic proteases (AP) play key roles in the regulation of biological processes, such as the recognition of pathogens and pests and the induction of effective defense responses. A large number of AP (>400) have been identified in silico in the rice genome. None have previously been isolated and functionally characterized for their involvement in disease resistance. We describe here the isolation and characterization of a gene (OsCDR1) from rice which encodes a predicted aspartate protease. Expression of OsCDR1 was activated upon treatments with benzothiadiazole and salicylic acid, which are signal molecules in plant disease resistance responses. Ectopic expression of OsCDR1 in Arabidopsis and rice conferred enhanced resistance against bacterial and fungal pathogens. The enhanced disease resistance observed in transgenic plants was correlated with induction of pathogenesis-related gene expression and was shown by mutational analysis to be dependent on AP activity of the transgene-encoded product. OsCDR1 accumulates in intercellular fluids (IF) in transgenic plants. Infiltration of IF from transgenic Arabidopsis plants into leaves of wild-type (WT) Arabidopsis induced the systemic defense response. These results demonstrate the conservation of CDR1 function between rice and Arabidopsis during the disease resistance response.

The mutant regulator of APX2 1-1 (rax1-1) was identified in Arabidopsis thaliana that constitutively expressed normally photooxidative stress-inducible ASCORBATE PEROXIDASE2 (APX2) and had ≥50% lowered foliar glutathione levels. Mapping revealed that rax1-1 is an allele of γ-GLUTAMYLCYSTEINE SYNTHETASE 1 (GSH1), which encodes chloroplastic γ-glutamylcysteine synthetase, the controlling step of glutathione biosynthesis. By comparison of rax1-1 with the GSH1 mutant cadmium hypersensitive 2, the expression of 32 stress-responsive genes was shown to be responsive to changed glutathione metabolism. Under photo-oxidative stress conditions, the expression of a wider set of defense-related genes was altered in the mutants. In wild-type plants, glutathione metabolism may play a key role in determining the degree of expression of defense genes controlled by several signaling pathways both before and during stress. This control may reflect the physiological state of the plant at the time of the onset of an environmental challenge and suggests that changes in glutathione metabolism may be one means of integrating the function of several signaling pathways.

In this work we used two different pea cultivars, JI281 is a semidomesticated land race of pea from Ethiopia whereas JI399 is a typical domesticated pea variety. Exposure of pea leaves to excess light (EL) for 1 h caused a reversible photoinhibition of photosynthesis as showed by changes in F v / F m. Although little difference existed between the two pea genotypes with respect to photoinhibition, after 60 min of EL the decline in F v / F m was higher in JI281 than in JI399 leaves. As a consequence of EL, H2O2 increased in both pea cultivars, whereas lipid peroxidation and protein oxidation slightly increased, although differences between cultivars were minimal. The redox state of ascorbate shifted towards its oxidized form under EL stress in both cultivars. Transcript levels of genes coding antioxidant enzymes varied with EL in both cultivars, but the response was more pronounced in JI399. The induction observed during EL was maintained or increased after the stress period, as occurred for cytGR and chlMDHAR . GR protein accumulation and activity correlated with the transcript accumulation in JI399, but not in JI288. In this work, a possible role for H2O2 and redox status of ascorbate in the photoxidative stress signalling is discussed.

We have explored the transient over-expression of Arabidopsis thaliana ascorbate peroxidase 3 (APX3) in Nicotiana benthamiana using a viral vector based on the potato virus X (PVX). Plants infected with a PVX:APX3 hybrid had a similar progression of viral particles compared to control plants infected with a PVX:GFP hybrid, indicating that infection was not affected by the over-expression of heterologous APX3. Our results also showed that in PVX:APX3-infected plants, the hybrid virus directed a high level of APX3 expression and the recombinant protein was functional, as inferred from the higher APX activity compared to mock and PVX:GFP hybrid-infected plants. The PVX recombinant expression system used is a simple and quick method for transient expression of heterologous APXs, which are expected to suffer specific processing in plant cells.

Analysis of the oxidative processes taking place during fruit ripening in a salad tomato variety (Lycopersicon esculentum Mill. cv. Ailsa Craig) revealed changes in oxidative and antioxidative parameters. Hydrogen peroxide content, lipid peroxidation and protein oxidation were measured as indices of oxidative processes and all were found to increase at the breaker stage. The levels of the aqueous-phase antioxidants, glutathione and ascorbate, increased during the ripening process and these increases were associated with significant changes in their redox status, becoming more reduced as ripening progressed. Changes in the activities of superoxide dismutase, catalase and the enzymes involved in the ascorbate-glutathione cycle during ripening indicated that the antioxidative system plays a fundamental role in the ripening of tomato fruits.