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        Visualization of Phytophthora palmivora Infection in Oil Palm Leaflets with Fluorescent Proteins and Cell Viability Markers

        Ochoa, Juan C.,Herrera, Mariana,Navia, Monica,Romero, Hernan Mauricio The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.1

        Bud rot (BR) is the most devastating disease affecting oil palm (Elaeis guineensis) crops in Colombia. Its causal agent, Phytophthora palmivora, initiates the infection in immature oil palm leaflets producing necrotic lesions, followed by colonization of opportunistic necrotrophs, which increases disease damage. To improve the characterization of the disease, we transformed P. palmivora using Agrobacterium tumefaciens-mediated transformation (ATMT) to include the fluorescent proteins CFP-SKL (peroxisomal localization), eGFP and mRFP1 (cytoplasmic localization). The stability of some transformants was confirmed by Southern blot analysis and single zoospore cultures; additionally, virulence and in vitro growth were compared to the wild-type isolate to select transformants with the greatest resemblance to the WT isolate. GFP-tagged P. palmivora was useful to identify all of the infective structures that are commonly formed by hemibiotrophic oomycetes, including apoplastic colonization and haustorium formation. Finally, we detected cell death responses associated with immature oil palm tissues that showed reduced susceptibility to P. palmivora infection, indicating that these tissues could exhibit age-related resistance. The aim of this research is to improve the characterization of the initial disease stages and generate cell biology tools that may be useful for developing methodologies for early identification of oil palm materials resistant or susceptible to BR.

      • KCI등재

        Visualization of Phytophthora palmivora Infection in Oil Palm Leaflets with Fluorescent Proteins and Cell Viability Markers

        Juan C,Ochoa,Mariana Herrera,,nica Navia,Herná,n Mauricio Romero 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.1

        Bud rot (BR) is the most devastating disease affecting oil palm (Elaeis guineensis) crops in Colombia. Its causal agent, Phytophthora palmivora, initiates the infection in immature oil palm leaflets producing necrotic lesions, followed by colonization of opportunistic necrotrophs, which increases disease damage. To improve the characterization of the disease, we transformed P. palmivora using Agrobacterium tumefaciens-mediated transformation (ATMT) to include the fluorescent proteins CFPSKL (peroxisomal localization), eGFP and mRFP1 (cytoplasmic localization). The stability of some transformants was confirmed by Southern blot analysis and single zoospore cultures; additionally, virulence and in vitro growth were compared to the wild-type isolate to select transformants with the greatest resemblance to the WT isolate. GFP-tagged P. palmivora was useful to identify all of the infective structures that are commonly formed by hemibiotrophic oomycetes, including apoplastic colonization and haustorium formation. Finally, we detected cell death responses associated with immature oil palm tissues that showed reduced susceptibility to P. palmivora infection, indicating that these tissues could exhibit age-related resistance. The aim of this research is to improve the characterization of the initial disease stages and generate cell biology tools that may be useful for developing methodologies for early identification of oil palm materials resistant or susceptible to BR.

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        The NPR1 family of transcription cofactors in papaya: insights into its structure, phylogeny and expression

        Santy Peraza-Echeverria,Jorge M. Santamaría,Gabriela Fuentes,Mariana de los Ángeles Menéndez-Cerón,Miguel Ángel Vallejo-Reyna,Virginia Aurora Herrera-Valencia 한국유전학회 2012 Genes & Genomics Vol.34 No.4

        The NPR1 (non-expressor of pathogenesis related gene 1) gene was initially identified in Arabidopsis as a master regulator of the systemic acquired resistance (SAR). Five additional NPR1 homologues have been identified in Arabidopsis whose function range from regulators of SAR to plant development. In the present study, we characterized the structure, phylogeny and expression of the NPR1 family in papaya (Carica papaya L.), one of the most important tropical fruit crops. We identified four NPR1 homologues in the papaya genome sequence (CpNPR1 to CpNPR4). Overall, the four papaya predicted NPR1 proteins showed the characteristic BTB/POZ and ankyrin domains of the Arabidopsis NPR1 family. Twelve additional open reading frames showing homology to retrotransposon elements or genes involved in different physiological processes were found in close proximity to the papaya NPR1homologues. The phylogenetic analysis revealed that the papaya NPR1 sequences resolved in three clades, each clade containing two Arabidopsis NPR1 homologues involved either in the positive regulation of SAR (clade I), negative regulation of SAR (clade II) or plant development (clade III), suggesting a similar function for the corresponding papaya NPR1homologues. Furthermore, the expression of the four papaya NPR1 homologues was detected in both vegetative and reproductive tissues. The present study has provided the first comparative analysis of the NPR1 family in a tropical fruit crop and expanded our knowledge on this type of genes in dicotyledoneous plants. The identification of the full set of papaya NPR1 homologues will pave the way for their systematic functional analysis and new opportunities for engineering disease resistance in this crop.

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