Transformation of Phytophthora capsici with genes for green and red fluorescent protein for use in visualizing plant-pathogen interactions

Dunn, Amara R.; Fry, B. A.; Lee, T. Y.; Conley, Kathleen; Balaji, Vasudevan; Fry, William E.; McLeod, A.; Smart, Christine D.

doi:10.1007/s13313-013-0222-2

Cited by 23 publications

(26 citation statements)

References 48 publications

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“…A pepper isolate of P. capsici (designated “Yolo-1,” from Yolo County, CA, United States; also pathogenic on tomato) was used for most experiments. A P. capsici isolate transformed with the green fluorescent protein (GFP) was a gift of Christine Smart and William Fry, Cornell University ( Dunn et al, 2013 ). Wild-type and transformant P. capsici strains were maintained on V8 juice agar plates or V8 juice amended with 100 mg/L geneticin (G418; Gibco), respectively.…”

Section: Methodsmentioning

confidence: 99%

Abscisic Acid as a Dominant Signal in Tomato During Salt Stress Predisposition to Phytophthora Root and Crown Rot

et al. 2018

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Salt stress predisposes plants to Phytophthora root and crown rot in an abscisic acid (ABA)-dependent manner. We used the tomato–Phytophthora capsici interaction to examine zoospore chemoattraction and assessed expression of pathogenesis-related (PR) genes regulated by salicylic acid (SA) and jasmonic acid (JA) following a salt-stress episode. Although salt treatment enhances chemoattraction of tomato roots to zoospores, exudates from salt-stressed roots of ABA-deficient mutants, which do not display the predisposition phenotype, have a similar chemoattraction as exudates from salt-stressed, wild-type roots. This suggests that ABA action during predisposing stress enhances disease through effects on plant responses occurring after initial contact and during ingress by the pathogen. The expression of NCED1 (ABA synthesis) and TAS14 (ABA response) in roots generally corresponded to previously reported changes in root ABA levels during salt stress onset and recovery in a pattern that was not altered by infection by P. capsici. The PR genes, P4 and PI-2, hallmarks in tomato for SA and JA action, respectively, were induced in non-stressed roots during infection and strongly suppressed in infected roots exposed to salt-stress prior to inoculation. However, there was a similar proportional increase in pathogen colonization observed in salt-stressed plants relative to non-stressed plants in both wild-type and a SA-deficient nahG line. Unlike the other tomato cultivars used in this study that showed a strong predisposition phenotype, the processing tomato cv. ‘Castlemart’ and its JA mutants were not predisposed by salt. Salt stress predisposition to crown and root rot caused by P. capsici appears to be strongly conditioned by ABA-driven mechanisms in tomato, with the stress compromising SA-and JA-mediated defense-related gene expression during P. capsici infection.

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Section: Methodsmentioning

confidence: 99%

Abscisic Acid as a Dominant Signal in Tomato During Salt Stress Predisposition to Phytophthora Root and Crown Rot

et al. 2018

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“…Samples were treated with 10 μl crude extract solution or solvent controls, and inoculated with 20 μl of 1x10 5 zoospores ml -1 suspension of either P . capsici isolate OP97 or NY0664-1 expressing red fluorescent protein (RFP) ([ 26 ]; kindly provided by C. Smart, Cornell University) as described above. The inoculated plates were incubated at 25°C with a 16h light/ 8h dark cycle for 72 hours.…”

Section: Methodsmentioning

confidence: 99%

Exocarp Properties and Transcriptomic Analysis of Cucumber (Cucumis sativus) Fruit Expressing Age-Related Resistance to Phytophthora capsici

et al. 2015

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Very young cucumber (Cucumis sativus) fruit are highly susceptible to infection by the oomycete pathogen, Phytophthora capsici. As the fruit complete exponential growth, at approximately 10–12 days post pollination (dpp), they transition to resistance. The development of age-related resistance (ARR) is increasingly recognized as an important defense against pathogens, however, underlying mechanisms are largely unknown. Peel sections from cucumber fruit harvested at 8 dpp (susceptible) and 16 dpp (resistant) showed equivalent responses to inoculation as did whole fruit, indicating that the fruit surface plays an important role in defense against P. capsici. Exocarp from 16 dpp fruit had thicker cuticles, and methanolic extracts of peel tissue inhibited growth of P. capsici in vitro, suggesting physical or chemical components to the ARR. Transcripts specifically expressed in the peel vs. pericarp showed functional differentiation. Transcripts predominantly expressed in the peel were consistent with fruit surface associated functions including photosynthesis, cuticle production, response to the environment, and defense. Peel-specific transcripts that exhibited increased expression in 16 dpp fruit relative to 8 dpp fruit, were highly enriched (P<0.0001) for response to stress, signal transduction, and extracellular and transport functions. Specific transcripts included genes associated with potential physical barriers (i.e., cuticle), chemical defenses (flavonoid biosynthesis), oxidative stress, penetration defense, and molecular pattern (MAMP)-triggered or effector-triggered (R-gene mediated) pathways. The developmentally regulated changes in gene expression between peels from susceptible- and resistant- age fruits suggest programming for increased defense as the organ reaches full size.

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“…Although ATMT has not been widely used in oomycetes ( Judelson and Ah-Fong, 2009 ), it has been shown to be superior to other methodologies in fungi because the production of protoplasts may affect the fitness of the pathogen ( Dunn et al, 2013 ; Keller et al, 1990 ). Additionally, the probability of single copy transformants is much higher in comparison to other methodologies ( Frandsen, 2011 ).…”

Section: Discussionmentioning

confidence: 99%

“…Genetic transformation of Phytophthora spp. was developed for diverse applications, including the quantification of resistance levels in potato cultivars ( Kamoun et al, 1998 ), evaluation of chemical inducers of disease resistance ( Si-Ammour et al, 2003 ), gene silencing for functional characterization ( Blanco and Judelson, 2005 ; Bos et al, 2010 ; van West et al, 1999a ), cell biology studies of pathogen structures ( Ah-Fong and Judelson, 2011 ; Bozkurt et al, 2011 ; Chaparro-Garcia et al, 2011 ; Dunn et al, 2013 ; Schornack et al, 2010 ) and more recently, genome editing using the CRISPR/cas9 system ( Fang and Tyler, 2016 ).…”

mentioning

confidence: 99%

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

Ochoa¹,

Herrera²,

Navia³

et al. 2019

Plant Pathol J

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Transformation of Phytophthora capsici with genes for green and red fluorescent protein for use in visualizing plant-pathogen interactions

Cited by 23 publications

References 48 publications

Abscisic Acid as a Dominant Signal in Tomato During Salt Stress Predisposition to Phytophthora Root and Crown Rot

Abscisic Acid as a Dominant Signal in Tomato During Salt Stress Predisposition to Phytophthora Root and Crown Rot

Exocarp Properties and Transcriptomic Analysis of Cucumber (Cucumis sativus) Fruit Expressing Age-Related Resistance to Phytophthora capsici

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

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