American chestnut (Castanea dentata [Marsh.] Borkh.) was once the dominant hardwood species in Eastern North America before an exotic fungal pathogen, Cryphonectria parasitica (Murrill) Barr, functionally eliminated it across its range. One promising approach toward restoring American chestnut to natural forests is development of blight‐tolerant trees using genetic transformation. However, transformation and related processes can result in unexpected and unintended phenotypic changes, potentially altering ecological interactions. To assess unintended tritrophic impacts of transgenic American chestnut on plant–herbivore interactions, gypsy moth (Lymantria dispar L.) caterpillars were fed leaf disks excised from two transgenic events, Darling 54 and Darling 58, and four control American chestnut lines. Leaf disks were previously treated with an LD50 dose of either the species‐specific Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) or the generalist pathogen Bacillus thuringiensis subsp. kurstaki (Btk). Mortality was quantified and compared to water blank controls. Tree genotype had a strong effect on the efficacies of both pathogens. Larval mortality from Btk‐treated foliage from only one transgenic event, Darling 54, differed from its isogenic progenitor, Ellis 1, but was similar to an unrelated wild‐type American chestnut control. LdMNPV efficacy was unaffected by genetic transformation. Results suggest that although genetic modification of trees may affect interactions with other nontarget organisms, this may be due to insertion effects, and variation among different genotypes (whether transgenic or wild‐type) imparts a greater change in response than transgene presence.
Polyethylene glycol (PEG) was evaluated for its influence on hardening of in vitro-propagated `Fern' strawberries (Fragaria ×ananassa) when applied just before transplanting. Strawberries were micropropagated via shoot tips and grown in vitro until roots were well developed. Plantlets were then transferred onto filter paper bridges in liquid medium with 15% (w/v) of PEG-8000. After treatment in the medium for various periods, the plants were compared to the control (no PEG) for water loss from detached leaves, stomatal aperture, and survival rates after transplanting. Leaf epicuticular wax was also quantified. Overall, the in vitro PEG treatment was not successful in significantly increasing hardiness and survivability of the strawberry plants after transplanting from in vitro conditions to a soil medium. Osmotic stress was created, but apparently not for the time needed to increase survival. Further tests are needed to pinpoint the proper exposure time required to increase hardiness and survivability after transplanting plantlets. To increase survival, the time exposed to PEG should be 15, 18, or possibly 21 days.
Callus induction and regeneration of alkaligrass (Puccinellia distans) was developed in our laboratory for use in transformation studies of turfgrass. Particle bombardment of the embryogenic callus is being evaluated using a helium particle inflow gun constructed at Colorado State Univ., according to the design of Philippe et al. (Ohio State Univ., 1993). Its utility in delivering DNA to plant cells is being tested by measuring the frequency of transient gene expression of a reporter gene (GUS pBI121) in embryogenic callus of alkaligrass. Varying pressure of helium and the distance of the calli in the chamber are also being evaluated for efficiency in transformation.
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