Profiling and quantifying differential gene transcription in Phytophthora infestans prior to and during the early stages of potato infection

Avrova, Anna O.; Venter, Eduard; Birch, Paul R. J.; Whisson, Stephen C.

doi:10.1016/s1087-1845(03)00063-x

Cited by 94 publications

(96 citation statements)

References 37 publications

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“…Bintje 12,24,33,48,56, and 72 h postinoculation (hpi). Expression of each gene was compared with the ActA gene from P. infestans, which has been used as a constitutively expressed endogenous control (21,22). Although PCR products of expected sizes were generated for Pex147-2 and Pex147-3 from genomic DNA template, no products were detected from cDNA templates in the case of Pex147-3, and expression was only weakly detected in the case of Pex147-2 and only in cDNA from axenic cultured, nonsporulating mycelium.…”

Section: Resultsmentioning

confidence: 99%

“…21 and 22. For real-time RT-PCR, material was prepared, RNA extracted, and experiments conducted as described previously by using the actA gene from P. infestans as a constitutively expressed endogenous control (21). Real-time RT-PCR primers for Pex147 were 5Ј-CGCCATAAACTTTG-CAACCA-3Ј and 5Ј-TGCCGGCTGAATCGTGTAT-3Ј (amplicon size ϭ 92 bp), primers for Pex147-2 were 5Ј-GGTGGCAG-CACAAGAGGC-3Ј and 5Ј-GCAGCTATTGTAGATCCG-ATTGTATC-3Ј (amplicon size ϭ 104 bp), and primers for Pex147-3 were 5Ј-TTGGTGGCAGCACAATGGT-3Ј and 5Ј-CCCAGGTGCATCAGGTAGCT-3Ј (amplicon size ϭ 120 bp).…”

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confidence: 99%

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An ancestral oomycete locus contains late blight avirulence gene Avr3a , encoding a protein that is recognized in the host cytoplasm

Armstrong

Whisson

Pritchard

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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417

386

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The oomycete Phytophthora infestans causes late blight, the potato disease that precipitated the Irish famines in 1846 and 1847. It represents a reemerging threat to potato production and is one of >70 species that are arguably the most devastating pathogens of dicotyledonous plants. Nevertheless, little is known about the molecular bases of pathogenicity in these algae-like organisms or of avirulence molecules that are perceived by host defenses. Disease resistance alleles, products of which recognize corresponding avirulence molecules in the pathogen, have been introgressed into the cultivated potato from a wild species, Solanum demissum, and R1 and R3a have been identified. We used association genetics to identify Avr3a and show that it encodes a protein that is recognized in the host cytoplasm, where it triggers R3a-dependent cell death. Avr3a resides in a region of the P. infestans genome that is colinear with the locus containing avirulence gene ATR1 NdWsB in Hyaloperonospora parasitica, an oomycete pathogen of Arabidopsis. Remarkably, distances between conserved genes in these avirulence loci were often similar, despite intervening genomic variation. We suggest that Avr3a has undergone gene duplication and that an allele evading recognition by R3a arose under positive selection.microsynteny ͉ Phytophthora infestans ͉ hypersensitive response ͉ linkage disequilibrium ͉ hemibiotroph

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

An ancestral oomycete locus contains late blight avirulence gene Avr3a , encoding a protein that is recognized in the host cytoplasm

Armstrong

Whisson

Pritchard

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

417

386

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“…Potato (Solanum tuberosum) infection for real-time RT-PCR analysis was performed as described elsewhere (Avrova et al, 2003). Leaves were harvested before inoculation (B0) and at 12 hpi (B12), 24 hpi (B24), 33 hpi (B33), 48 hpi (B48), 56 hpi (B56), and 72 hpi (B72) as described by GrenvilleBriggs et al (2005).…”

Section: Methodsmentioning

confidence: 99%

“…Template cDNA was derived from mycelium grown in rye broth, sporangia, zoospores, germinating cysts, and germinating cysts with appressoria, as well as potato leaves before inoculation (B0) and at 12 hpi (B12), 24 hpi (B24), 33 hpi (B33), 48 hpi (B48), 56 hpi (B56), and 72 hpi (B72). The actA gene from P. infestans was used as a constitutively expressed endogenous control, and the expression of each gene in mycelium was determined relative to actA as described by Avrova et al (2003). Expression of all genes in different life cycle stages was compared with the level of their expression in a calibrator sample, which was cDNA from mycelium.…”

Section: Sybr Green Real-time Rt-pcr Assaysmentioning

confidence: 99%

Cellulose Synthesis inPhytophthora infestansIs Required for Normal Appressorium Formation and Successful Infection of Potato

et al. 2008

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Cellulose, the important structural compound of cell walls, provides strength and rigidity to cells of numerous organisms. Here, we functionally characterize four cellulose synthase genes (CesA) in the oomycete plant pathogen Phytophthora infestans, the causal agent of potato (Solanum tuberosum) late blight. Three members of this new protein family contain Pleckstrin homology domains and form a distinct phylogenetic group most closely related to the cellulose synthases of cyanobacteria. Expression of all four genes is coordinately upregulated during pre-and early infection stages of potato. Inhibition of cellulose synthesis by 2,6-dichlorobenzonitrile leads to a dramatic reduction in the number of normal germ tubes with appressoria, severe disruption of the cell wall in the preinfection structures, and a complete loss of pathogenicity. Silencing of the entire gene family in P. infestans with RNA interference leads to a similar disruption of the cell wall surrounding appressoria and an inability to form typical functional appressoria. In addition, the cellulose content of the cell walls of the silenced lines is >50% lower than in the walls of the nonsilenced lines. Our data demonstrate that the isolated genes are involved in cellulose biosynthesis and that cellulose synthesis is essential for infection by P. infestans.

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“…Actin A (accession number M59715.1) (35,36), actin B (accession number M59716.1) (35), and elongation factor 2 (accession number EEY57133.1) (36, 37) were employed as internal references. Primers were designed by Primer3Plus (http://www .bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi) (38).…”

Section: Methodsmentioning

confidence: 99%

Phytophthora infestans Cholinephosphotransferase with Substrate Specificity for Very-Long-Chain Polyunsaturated Fatty Acids

Chen

yun

Meesapyodsuk

et al. 2013

Appl Environ Microbiol

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The effective flux between phospholipids and neutral lipids is critical for a high level of biosynthesis and accumulation of verylong-chain polyunsaturated fatty acids (VLCPUFAs), such as arachidonic acid (ARA; 20:4n-6), eicosapentaenoic acid (EPA; 20: 5n-3), and docosahexaenoic acid (DHA; 22:6n-3). Here we describe a cDNA (PiCPT1) from Phytophthora infestans, a VLCPUFAproducing oomycete, that may have a role in acyl trafficking between diacylglycerol (DAG) and phosphatidylcholine (PC) during the biosynthesis of VLCPUFAs. The cDNA encodes a polypeptide of 393 amino acids with a conserved CDP-alcohol phosphotransferase motif and approximately 27% amino acid identity to the Saccharomyces cerevisiae cholinephosphotransferase (ScCPT1). In vitro assays indicate that PiCPT1 has high cholinephosphotransferase (CPT) activity but no ethanolaminephosphotransferase (EPT) activity. Substrate specificity assays show that it prefers VLCPUFA-containing DAGs, such as ARA DAG and DHA DAG, as substrates. Real-time PCR analysis reveals that expression of PiCPT1 was upregulated in P. infestans organisms fed with exogenous VLCPUFAs. These results lead us to conclude that PiCPT1 is a VLCPUFA-specific CPT which may play an important role in shuffling VLCPUFAs from DAG to PC in the biosynthesis of VLCPUFAs in P. infestans.

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Profiling and quantifying differential gene transcription in Phytophthora infestans prior to and during the early stages of potato infection

Cited by 94 publications

References 37 publications

An ancestral oomycete locus contains late blight avirulence gene Avr3a , encoding a protein that is recognized in the host cytoplasm

An ancestral oomycete locus contains late blight avirulence gene Avr3a , encoding a protein that is recognized in the host cytoplasm

Cellulose Synthesis inPhytophthora infestansIs Required for Normal Appressorium Formation and Successful Infection of Potato

Phytophthora infestans Cholinephosphotransferase with Substrate Specificity for Very-Long-Chain Polyunsaturated Fatty Acids

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