Preliminary results on the expression of Chrysanthemum Yellows Phytoplasma genes in Arabidopsis thaliana (Abstract/Poster in atti di convegno)

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  • Preliminary results on the expression of Chrysanthemum Yellows Phytoplasma genes in Arabidopsis thaliana (Abstract/Poster in atti di convegno) (literal)
Anno
  • 2012-01-01T00:00:00+01:00 (literal)
Alternative label
  • Pacifico D., Palmano S., Firrao G., Abbà S., Bertin S., Marzachì C. (2012)
    Preliminary results on the expression of Chrysanthemum Yellows Phytoplasma genes in Arabidopsis thaliana
    in 19th Congress of the International Organisation for Mycoplasmology, Toulouse, France, 15-20 luglio 2012
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Pacifico D., Palmano S., Firrao G., Abbà S., Bertin S., Marzachì C. (literal)
Note
  • Poster (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • Istituto di Virologia Vegetale - CNR, Torino Dipartimento di Biologia e Protezione delle Piante, Università degli Studi di Udine Dipartimento di Valorizzazione e Protezione delle Risorse Agroforestali, Università degli Studi di Torino (literal)
Titolo
  • Preliminary results on the expression of Chrysanthemum Yellows Phytoplasma genes in Arabidopsis thaliana (literal)
Abstract
  • Microarrays analysis suggests that phytoplasma genome expression in the plant can be modulated according to the infection stage (Oshima et al., 2011). A Real Time PCR protocol was set up to study the expression profile of different \"Candidatus Phytoplasma asteris\" Chrysanthemum yellows isolate (CY) genes during infection of Arabidopsis thaliana. Target genes were selected among secreted proteins, known effectors, general metabolism and specific CY genes, obtaining following Illumina sequencing. Genes coding three putative secreted peptides, antigenic membrane protein (Amp), immunodominant membrane protein (Imp), four general transporters (multidrug, mechano-sensitive channel, SecY translocon component and oligopeptide), two specific transporters (arginine, Zn++), one protein involved in phospholipid metabolism and the 30S rRNA gene were selected from the CY genome. Specific primers were designed for each selected gene, and the appropriate Real Time PCR protocols were optimized. Amplification efficiencies for each target were determined by amplification of five dilutions of cDNA obtained from pooled RNA infected plants. CY phytoplasma was transmitted to A. thaliana by infective Macrosteles quadripunctulatus Kirschbaum. Leaf samples were collected at different times post infection, DNA was purified and used as template in diagnostic PCR and in Real Time PCR to determine the phytoplasma titer. Total RNA was extracted, treated with DNase, reverse transcribed and analyzed by quantitative Real Time PCR. Yellowing, leaf deformation, and stunting appeared two weeks after inoculation, and 15% of plants were CY infected five days post inoculation (dpi). At 25 dpi (last sampling date) 83% of plants were infected. The expression level of each target gene was correlated to the actual titer of phytoplasma in each infected A. thaliana to provide a detailed study on gene expression of an unculturable obligate parasite in a model plant. (literal)
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