Evaluation of promoters and visual markers for transformation of eastern white pine

This report serves to evaluate possible promoters for use in theproduction of transgenic eastern white pine (Pinus strobus L.).Embryogenic cultures of eastern white pine were bombarded with goldparticles coated separately with a variety of gene constructs containingthe UidA -glucuronidase (GUS) or green fluorescentprotein (GFP) reporter gene. Transient expression of the UidAgene, driven by a novel algal virus adenine methyl transferase genepromoter, as well as five other promoters used in angiospermtransformation, were evaluated. The maize alcohol dehydrogenase promoterwas not effective in eastern white pine cultures. The construct with thedoubled Cauliflower Mosaic Virus 35S promoter plus Alfalfa Mosaic Virusenhancer showed the highest levels of expression. GUS expression wasdetected within 24 hours, but decreased after 5 days and was notdetectable 15 days after bombardment. Expression of GUS activity wasrecorded mainly in somatic embryonal heads of various stages ofdevelopment and occasionally in suspensor cells. Similar to GUSexpression, modified green fluorescent protein (GFP) was detected in theembryonal head cells 24 hours after bombardment. GFP-expressingsuspensor cells were both more infrequent and difficult to detect, astheir highly vacuolate nature rendered the GFP presence less visibleagainst the yellow background autofluorescence.     

Phylogenetic Relationships of Xylella fastidiosa Strains Isolated from Landscape Ornamentals in Southern California

ABSTRACT Xylella fastidiosa is an insect-borne, xylem-limited pathogenic bacterium that has been associated with a rise in incidence of diseased landscape ornamentals in southern California. The objective of this study was to genetically characterize strains isolated from ornamental hosts to understand their distribution and identity. Strains of X. fastidiosa isolated from ornamentals were characterized using a multiprimer polymerase chain reaction (PCR) system, random amplified polymorphic DNA (RAPD)-PCR, and sequence analysis of the 16S-23S rDNA intergenic spacer region (ISR). Based on RAPD-PCR and 16S-23S rDNA ISR, strains isolated from daylily, jacaranda, and magnolia clustered with members of X. fastidiosa subsp. sandyi and caused oleander leaf scorch but not Pierce's disease symptoms in glasshouse assays on oleander and grape, respectively. This demonstrated both that our groupings based on genetic characterization were valid and that strains of X. fastidiosa subsp. sandyi are present in hosts other than oleander. Strains isolated from Spanish broom, cherry, and one strain isolated from western redbud clustered with X. fastidiosa subsp. fastidiosa members. Strains isolated from purple-leafed plum, olive, peach, plum, sweetgum, maidenhair tree, crape myrtle, and another western redbud strain clustered with members of X. fastidiosa subsp. multiplex. All strains isolated from mulberry and one from heavenly bamboo formed a separate cluster that has not yet been defined as a subspecies.     

Geographic variation in marine turtle fibropapillomatosis.

We document three examples of fibropapillomatosis by histology, quantitative polymerase chain reaction (qPCR), and sequence analysis from three different geographic areas. Tumors compatible in morphology with fibropapillomatosis were seen in green turtles from Puerto Rico and San Diego (California) and in a hybrid loggerhead/ hawksbill turtle from Florida Bay (Florida). Tumors were confirmed as fibropapillomas on histology, although severity of disease varied between cases. Polymerase chain reaction (PCR) analyses revealed infection with the fibropapilloma-associated turtle herpesvirus (FPTHV) in all cases, albeit at highly variable copy numbers per cell. Alignment of a portion of the polymerase gene from each fibropapilloma-associated turtle herpesvirus isolate demonstrated geographic variation in sequence. These cases illustrate geographic variation in both the pathology and the virology of fibropapillomatosis.