@article{kurt_matallana-ramirez_kohlway_whetten_frampton_2020, title={A fast, flexible and inexpensive protocol for DNA and RNA extraction for forest trees}, volume={29}, ISSN={["2171-9845"]}, DOI={10.5424/fs/2020292-16730}, abstractNote={Aim of the study: DNA and RNA extraction are still one of the most important and challenging steps of many molecular genetics applications such as Next-Generation Sequencing technologies. In this study, traditional laboratory preparation protocols and commercially available nucleic acids extraction kits’ features were combined into a procedure suitable for extraction of either DNA or RNA in 96-well plate format at high throughput.Area of study: The study covers forest tree species from the United States of America.Materials and methods: The DNA and RNA protocol were tested on 27 species, including especially recalcitrant forest tree species, from five angiosperm and three gymnosperm families. DNA was also extracted from stored (from 2 to 6 years) silica-dried samples of 11 species of Pinaceae.Main results: The spectrophotometric analysis of DNA and RNA showed that gymnosperms yielded lower quantity, but higher quality nucleic acids than angiosperms which have variable results among species. The quantity and quality of DNA from stored samples were generally lower than fresh silica-dried samples. The RNA results showed high-enough yield (6.6 to 8.8 RIN) for downstream analyses.Research highlights: It was demonstrated that high quality and high molecular weight nucleic acids for Next-Generation Sequencing applications can be isolated from hundreds of samples from a wide range of taxonomic groups. The new protocol has features similar to both traditional laboratory and commercial extraction kits; is easy to set up in any molecular research laboratory, can be applied to a large number of samples (hundreds) in a working day, uses inexpensive reagents and supplies, and is compatible with automation.Key words: Angiosperms; gymnosperms; isolation protocol; nucleic acids.}, number={2}, journal={FOREST SYSTEMS}, author={Kurt, Yusuf and Matallana-Ramirez, Lilian and Kohlway, William and Whetten, Ross and Frampton, John}, year={2020} }
 @article{kohlway_whetten_benson_frampton_2017, title={Response of Turkish and Trojan fir to Phytophthora cinnamomi and P. cryptogea}, volume={32}, ISSN={0282-7581 1651-1891}, url={http://dx.doi.org/10.1080/02827581.2017.1280076}, DOI={10.1080/02827581.2017.1280076}, abstractNote={ABSTRACT Phytophthora root rot, primarily caused by the oomycete Phytophthora cinnamomi, is a large problem for the Christmas tree industry in North Carolina. Fraser fir (Abies fraseri) has no known innate resistance to this pathogen while some exotic fir species, such as Trojan (Abies equi-trojani) and Turkish (Abies bornmuelleriana) fir display varying amounts of resistance. A Phytophthora-resistance screening trial was completed with 1600 seedlings from 12 Turkish and Trojan fir families and Fraser and momi fir (A. firma). Seedlings from each family or species were inoculated with each of eight Phytophthora isolates, six P. cinnamomi and two Phytophthora cryptogea, in an effort to describe variability in isolate aggressiveness. Mortality was assessed as percent shoot necrosis bi-weekly for 16 weeks. Overall, rankings of resistance in fir species confirmed previous single-isolate-based results; momi fir was the most resistant, followed by Turkish, Trojan, and Fraser fir. P. cinnamomi isolates were generally more aggressive than P. cryptogea isolates. The two P. cryptogea isolates resulted in 5.6% and 0.8% mortality on Turkish fir, and 10.9% and 6.7% mortality on Trojan fir, the first reported resistance screen of these host-pathogen combinations. Pearson’s correlation testing identified a high degree of correlation between most isolates and overall mean mortality. Turkish and Trojan fir families appear to possess resistance to Phytophthora species common in North Carolina.}, number={5}, journal={Scandinavian Journal of Forest Research}, publisher={Informa UK Limited}, author={Kohlway, W. H. and Whetten, R. W. and Benson, D. M. and Frampton, J.}, year={2017}, month={Feb}, pages={406–411} }
 @article{pettersson_frampton_rönnberg_shew_benson_kohlway_escanferla_cubeta_2016, title={Increased diversity of Phytophthora species in Fraser fir Christmas tree plantations in the Southern Appalachians}, volume={32}, ISSN={0282-7581 1651-1891}, url={http://dx.doi.org/10.1080/02827581.2016.1265144}, DOI={10.1080/02827581.2016.1265144}, abstractNote={ABSTRACT Phytophthora root rot (PRR) disease afflicts significant economic losses to the Fraser fir Christmas tree industry. In previous surveys conducted in 1972 and from 1997 to 1998 in North Carolina, the incidence of PRR was ∼9.5% with Phytophthora cinnamomi identified as the predominant causal species isolated from infected roots of Fraser fir. Due to increased use of out-of-state planting stock since 2000, we suspected increased diversity of Phytophthora species. During 2014, we surveyed Fraser fir Christmas tree plantations in the Southern Appalachians of North Carolina, Tennessee and Virginia to determine the occurrence of pathogenic root-rotting species of Phytophthora. A weighted sampling strategy based on Christmas tree acreage was deployed to collect symptomatic Fraser fir roots from 103 commercial production fields in 14 counties. Six species of Phytophthora were isolated from infected roots sampled from 82 sites in 13 counties. Phytophthora cinnamomi, P. cryptogea and P. pini represented 70.3%, 23.1% and 1.1% of the 91 isolates. Phytophthora citrophthora, P. europaea and P. sansomeana accounted for the remaining 5.5% of the isolates and have not been identified in previously published Fraser fir surveys conducted in the region. The pathogenicity of P. citrophthora on Fraser fir was confirmed based on completion of Koch’s postulates.}, number={5}, journal={Scandinavian Journal of Forest Research}, publisher={Informa UK Limited}, author={Pettersson, M. and Frampton, J. and Rönnberg, J. and Shew, H. D. and Benson, D. M. and Kohlway, W. H. and Escanferla, M. E. and Cubeta, M. A.}, year={2016}, month={Dec}, pages={412–420} }