Spectral differentiation of oak wilt from foliar fungal disease and drought is correlated with physiological changes

Hyperspectral reflectance tools have been used to detect multiple pathogens in agricultural settings and single sources of infection or broad declines in forest stands. However, differentiation of any one disease from other sources of tree stress is integral for stand and landscape-level applications in mixed species systems. We tested the ability of spectral models to differentiate oak wilt, a fatal disease in oaks caused by Bretziella fagacearum ``Bretz'', from among other mechanisms of decline. We subjected greenhouse-grown oak seedlings (Quercus ellipsoidalis ``E.J. Hill'' and Quercus macrocarpa ``Michx.'') to chronic drought or inoculation with the oak wilt fungus or bur oak blight fungus (Tubakia iowensis ``T.C. Harr. & D. McNew''). We measured leaf and canopy spectroscopic reflectance (400–2400 nm) and instantaneous photosynthetic and stomatal conductance rates, then used partial least-squares discriminant analysis to predict treatment from hyperspectral data. We detected oak wilt before symptom appearance, and classified the disease with high accuracy in symptomatic leaves. Classification accuracy from spectra increased with declines in photosynthetic function in oak wilt-inoculated plants. Wavelengths diagnostic of oak wilt were only found in non-visible spectral regions and are associated with water status, non-structural carbohydrates and photosynthetic mechanisms. We show that hyperspectral models can differentiate oak wilt from other causes of tree decline and that detection is correlated with biological mechanisms of oak wilt infection and disease progression. We also show that within the canopy, symptom heterogeneity can reduce detection, but that symptomatic leaves and tree canopies are suitable for highly accurate diagnosis. Remote application of hyperspectral tools can be used for specific detection of disease across a multi-species forest stand exhibiting multiple stress symptoms.

Development of novel Quercus rubra chloroplast genome CAPS markers for haplotype identification

Our main objective was to generate cost-effective chloroplast (cp) DNA markers that are easy to apply and to score. In combination with already published cpSSR markers they should increase haplotype resolution in populations. To discover new cpDNA markers, we sequenced 87-97 % of the entire chloroplast genome (except the second inverted repeat) of 8 trees representing different regions of the Quercus rubra L. natural range with 4,030X-6,297X coverage and assembled the genome sequences using the publicly available chloroplast genome of Quercus rubra L. as a reference. In total, 118 single nucleotide polymorphisms (SNPs) and 107 insertions or deletions (indels) were detected, and 15 cleaved amplified polymorphic sequence (CAPS) markers were developed for Q. rubra. Using these new markers together with five chloroplast microsatellite or simple sequence repeat (cpSSR) markers, we identified 10 haplotypes in our diversity panel of 19 Q. rubra populations. Specifically, two haplotypes based only on the cpSSR markers could now be separated in five haplotypes. These markers are useful to assess haplotype diversity with high resolution and are also transferable to a closely related species, Quercus ellipsoidalis E. J. Hill.

Experimental evidence for selection against hybrids between two interfertile red oak species

Reproductive isolation between related oak species within one taxonomic section is incomplete. Even though pre- and post­zygotic isolation mechanisms have been described for interfer­tile oak species, natural hybridization is common in contact zones between related oaks. The apparent restriction of inter­specific hybrids between ecologically divergent species to intermediate environments in contact zones suggests postzy­gotic isolation via environmental selection against hybrids in parental environments. Overrepresentation of hybrids in seeds as compared to adult trees provides additional indirect evi­dence for selection against hybrids. Here, we used genetic assignment analyses in progeny obtained from a sympatric stand of Quercus rubra and Quercus ellipsoidalis, two interfertile species with different adaptations to drought, to characterize the number of hybrids and “pure” species in the non-germinated acorns and in seedlings. The occurrence of 43.6 % F1 hybrids and introgressive forms among the non-germinated acorns and their very low frequency in the seedlings (9.3 %) is to our knowledge the first direct evidence for early selection against hybrids in oaks possibly as result of genetic incompatibilities. Additionally, we found a signature of positive selection on EST-SSR PIE200 in Q. rubra which needs further confirmation. These results contribute to our under­standing of reproductive isolation and divergence between interfertile oak species with different ecological adaptations.

Predominant nitidulid species (Coleoptera: Nitidulidae) associated with spring oak wilt mats in Minnesota

Nitidulids are primary vectors of the oak wilt pathogen, Ceratocystis fagacearum (Bretz) Hunt, in the north-central United States. Species of adult nitidulids associated with different ages of oak wilt fungus mats on red oaks (Quercus rubra L. and Quercus ellipsoidalis E.J. Hill) during spring in east-central Minnesota were determined. The exoskeletal surfaces of representative specimens were assayed for the presence and abundance of the pathogen. Six species comprised 94% of 2542 adults, representing at least 12 species, collected between 1994 and 1996. Of these six species, Colopterus truncatus Randall and Epuraea corticina Erichson were the most abundant ones on immature mats (94% of 154 nitidulids). They were also more abundant than the other species on mature mats (77% of 868 nitidulids). Carpophilus sayi Parsons was the most common species (28% of 1134 nitidulids) on aging mats, while Carpophilus sayi and three Glischrochilus species (Glischrochilus sanguinolentus (Oliv.), Glischrochilus fasciatus (Oliv.), and Glischrochilus quadrisignatus Say) were predominant on declining mats (80% of 214 nitidulids). Multiple numbers of individuals of each species were commonly found on the mats. The smaller bodied species, C. truncatus and E. corticina, had the lowest numbers of fungal propagules on their bodies, while the highest incidence and numbers of viable propagules were found for the three largest bodied species (Glischrochilus spp.). These results are important to elucidating the principal nitidulid species involved in successful transmission of the pathogen in Minnesota.