Identification of potato genes induced during colonization by Phytophthora infestans

Abstract

Summary Suppression Subtractive Hybridization (SSH) was applied in a search for genes induced during the compatible interaction between Phytophthora infestans and potato. Using potato leaves that had been treated with benzo(1,2,3)thiadiazole-7-carbothioic acid S-methylester (BTH) as the control tissue, a low redundancy library with a relatively low frequency of the classic plant Pathogenesis-Related (PR) genes was generated. 288 of the clones were screened for induced sequences using Inverse Northern analysis (hybridizing the arrayed clones with radiolabelled cDNA populations). Of the 75 clones that were detectable by this method, 43 appeared to be induced. Eleven of these clones were then analysed by total RNA blot analysis, and elevation of transcript levels during P. infestans infection was confirmed for 10 of them. Some of the cDNAs analysed by RNA blot analysis have homology to genes already known to be induced during infection, e.g. to beta-1,3-glucanase. Another group of cDNAs have homology to enzymes involved in detoxification: gamma-glutamylcysteine synthetase, cytochrome P450, glutathione S-transferase and an MRP-type ABC transporter. Other infection induced cDNAs encode putative proteins that have not previously been reported to be induced by infection: e.g. the ER-located chaperone BiP, and a homologue of Aspergillus nidulans SudD, which was isolated as a suppressor of a mutation in chromosome disjunction. The differential library therefore presents the opportunity to analyse the metabolic changes occurring during infection, and the disease process itself in more detail.

Large-Scale Gene Discovery in the Oomycete Phytophthora infestans Reveals Likely Components of Phytopathogenicity Shared with True Fungi

“…Tissue from the mating zone was harvested after 6 days. PN, PO (unsubtracted potato interactions)-as described (Beyer et al 2001), zoospores were spot-inoculated onto leaves from cv. Bintje and, after 6 days, tissue was harvested from within and outside the lesion for libraries PN and PO, respectively.…”

“…Bintje and, after 6 days, tissue was harvested from within and outside the lesion for libraries PN and PO, respectively. PU, PW, PX (in vitro potato interactions)-as described (Beyer et al 2001), mycelia were grown in rye broth for 6 days, washed with water, and placed in water or on a potato leaf floating in water to generate control and tester samples, respectively. Both control and tester samples were pooled from 4-, 8-, and 24-h time-points.…”

A novel non-protein-coding infection-specific gene family is clustered throughout the genome of Phytophthora infestans

“…Several approaches exist for isolating such differentially expressed genes (reviewed by Birch et al, 2003). A PCR-based method called suppression subtractive hybridization (SSH) has been used to isolate potato genes upregulated following P. infestans challenge Avrova et al, 1999Avrova et al, , 2004Beyer et al, 2001), and to identify P. infestans genes that are upregulated in the interaction with potato (Beyer et al, 2002). However, despite the ability to enrich for pathogen transcripts, SSH cDNA libraries constructed from host-oomycete interactions still yield a majority proportion of host sequences (Bittner-Eddy et al, 2003).…”

In silico genome-wide identification and phylogenetic analysis of the WRKY transcription factor family in sweet orange (Citrus sinensis)

“…Moreover, studies have shown that WRKYTFs have regulatory roles in signaling and modulation pathways of various molecular and physiological processes like seed germination, pollen development, biomass accumulation, and flowering regulation, and are also involved in the biosynthesis of secondary metabolites (Sun et al, 2003;Rushton et al, 2012;Yu et al, 2013). The WRKY TFs have also shown defense responses to different abiotic stresses, including heat, cold, water deficiency, and salinity (Wu et al, 2009;Jiang and Deyholos, 2009;Niu et al, 2012) and different biotic stresses, such as fungal, and viral, and bacterial infection (Beyer et al, 2001;Huh et al, 2012;Pan and Jiang, 2014).…”