Browsing by Author "Dickinson, Christopher C."
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- Accession-Level Differentiation of Urushiol Levels, and Identification of Cardanols in Nascent Emerged Poison Ivy SeedlingsLott, Aneirin A.; Baklajian, Emily R.; Dickinson, Christopher C.; Collakova, Eva; Jelesko, John G. (MDPI, 2019-11-20)Poison ivy (Toxicodendron radicans (L.) Kuntze) shows accession-level differentiation in a variety of morphometric traits, suggesting local adaptation. To investigate whether the presumed defense compound urushiol also demonstrates accession-level accumulation differences, in vitro nascent germinated poison ivy seedlings from geographically isolated populations were germinated in vitro and then assayed for known urushiol congener accumulation levels. Significant accession-level differences in the accumulation levels of total C15- and C17-, total C15-, total C17-, specific C15 congeners, and specific C17 congeners of urushiol were identified. In addition, hereto novel C15- and C17-urushiol isomers were identified as well. Cardanols are assumed to be the penultimate metabolites giving rise to urushiols, but this assumption was not previously empirically validated. C15-cardanol congeners and isomers corresponding to expected substrates needed to produce the observed C15-urushiol congeners and isomers were identified in the same poison ivy seedling extracts. Total C15-cardanol and C15-cardanol congeners also showed significant accession-level differences. Based on the observed C15-cardanol congeners in poison ivy, the penultimate step in urushiol biosynthesis was proposed to be a cardanol-specific hydroxylase activity.
- Habitat Suitability and Establishment Limitations of a Problematic LianaDickinson, Christopher C.; Jelesko, John G.; Barney, Jacob (MDPI, 2021-01-29)The US native liana, poison ivy (Toxicodendron radicans), responsible for contact dermatitis in humans, is a competitive weed with great potential for expansion in disturbed habitats. To facilitate a better understanding of this threat, we sought to evaluate habitat suitability, population demography, and biotic interactions of poison ivy, using a series of complementary field studies in the two habitats where it most commonly occurs—forest interiors and edges. Of the 2500 seeds planted across both habitats, poison ivy initially colonized forest interiors (32% emergence) at a higher rate than edge habitats (16.5% emergence). However, forest interior seedlings were less likely to survive (interior n = 3; edge n = 15), which might be attributed to herbivore pressure when the seedlings were smaller in the less competitive forest interior. Once established, the poison ivy seedlings appeared to be more tolerant of herbivory, except that of large grazers such as deer. The early life stage of seedling emergence, survival, and establishment are critical in poison ivy success, with biotic pressure, especially from plant competition and deer, limiting recruitment. A suitable habitat of this expanding native liana would increase with increasing forest fragmentation, but might be buffered by the expanding deer population.
- Poison ivy hairy root cultures enable a stable transformation system suitable for detailed investigation of urushiol metabolismLott, Aneirin A.; Freed, Catherine P.; Dickinson, Christopher C.; Whitehead, Susan R.; Collakova, Eva; Jelesko, John G. (Wiley, 2020)Poison ivy (Toxicodendron radicans) is best known for causing exasperating allergenic delayed-contact dermatitis symptoms that last for weeks on persons who have contacted the plant. Urushiols are alkylcatechols produced by poison ivy responsible for causing this dermatitis. While urushiol chemical structures are well known, the metabolic intermediates and genes responsible for their biosynthesis have not been experimentally validated. A molecular genetic characterization of urushiol biosynthesis in poison ivy will require stable genetic transformation and subsequent regeneration of organs that retain the capacity synthesize urushiol. To this end, Agrobacterium rhizogenes was used to generate hormone-independent poison ivy hairy root cultures. Optimal conditions for hairy root formation were skotomorphic poison ivy hypocotyls prick-inoculated with A. rhizogenes, and preferential propagation of cultures with an atypical clumpy hairy root growth habit. The origin of the poison ivy accession used for A. rhizogenes prick-inoculation did not affect the initial formation of calli/ hairy root primordia, but rather significantly influenced the establishment of longterm hormone-independent hairy root growth. A. rhizogenes harboring a recombinant T-DNA binary plasmid with an intron-containing Firefly Luciferase gene produced stable transgenic hairy root lines expressing luciferase activity at high frequency. Poison ivy hairy root lines produced significantly lower steady-state urushiol levels relative to wild-type roots, but higher urushiol levels than a poison ivy undifferentiated callus line with undetectable urushiol levels, suggesting that urushiol biosynthesis requires intact poison ivy organs. The lower urushiol levels in poison ivy hairy root lines facilitated the first identification of anacardic acid metabolites initially in hairy roots, and subsequently in wild-type roots as well. This study establishes a transformation hairy root regeneration protocol for poison ivy that can serve as a platform for future reverse-genetic studies of urushiol biosynthesis in poison ivy hairy roots.