Mari Narusaka

Crosstalk in the responses to abiotic and biotic stresses in Arabidopsis: analysis of gene expression in cytochrome P450 gene superfamily by cDNA microarray.

From Arabidopsis full-length cDNA libraries, we collected ca. 7000 (7K) independent full-length cDNAs to prepare a cDNA microarray. The 7K cDNA collection contains 49 cytochrome P450 genes. In this study, expression patterns of these cytochrome P450 genes were analyzed by a full-length cDNA microarray under various treatments, such as hormones (salicylic acid, jasmonic acid, ethylene, abscisic acid), pathogen-inoculation (Alternaria brassicicola, Alternaria alternata), paraquat, rose bengal, UV stress (UV-C), heavy metal stress (CuSO_4), mechanical wounding, drought, high salinity and low temperature. Expression of 29 cytochrome P450 genes among them was induced by various treatments. Inoculation with A. brassicicola and A. alternata as biotic stresses increased transcript levels of 12 and 5 genes in Arabidopsis plants, respectively. In addition, some of the genes were also expressed by abiotic stresses. This suggests crosstalk between abiotic and biotic stresses. The promoter sequences and cis-acting elements of each gene were studied on the basis of full-length cDNA sequences. Most cytochrome P450 genes induced by both abiotic and biotic stresses contained the recognition sites of MYB and MYC, ACGT-core sequence, TGA-box and W-box for WRKY transcription factors in their promoters. These cis-acting elements are known to participate in the regulation of plant defense. The response of each gene to multiple stresses is strictly regulated.

Cytological and molecular analyses of non-host resistance of Arabidopsis thaliana to Alternaria alternata

SUMMARY When challenged with the necrotrophic fungal pathogen Alternaria alternata Japanese pear pathotype, all tested ecotypes of Arabidopsis plants failed to show hypersensitive cell death, accumulation of detectable levels of reactive oxygen species or accumulation of phytoalexin. We operationally define A. alternata as a non-host pathogen for Arabidopsis plants and show that the protection against A. alternata demonstrated in this study is a non-host penetration resistance. To characterize non-host penetration resistance, we examined the expression patterns of c. 7000 genes by cDNA microarray analysis in Arabidopsis Col-0 plants after inoculation with A. alternata. After inoculation with A. alternata, the transcript levels of 48 genes increased in Col-0 plants. The expression of genes associated with hypersensitive reaction was induced in the non-host penetration resistance to A. alternata, despite the fact that A. alternata had no visible effect on the plants. The non-host penetration resistance to A. alternata was clearly associated with activation of the jasmonate- and ethylene-signalling pathways. In addition, analysis using histochemical staining of GUS activity suggests that defence reactions in non-host penetration resistance are activated locally. The characterization of non-host pathosystem involving Arabidopsis and A. alternata offers an overview of non-host penetration resistance.