On merging Acer sections Rubra and Hyptiocarpa: Molecular and morphological evidence.

In this study, we expanded Acer sect. Rubra Pax to include A. sect. Hyptiocarpa Fang. Traditionally, section Rubra comprises two iconic species, Acer rubrum Linnaeus (red maple) and A. saccharinum Linnaeus (silver maple), of eastern North American forests as well as the rare Japanese montane species, A. pycnanthum K. Koch. Section Hyptiocarpa consists of A. laurinum Hasskarl and A. pinnatinervium Merrill, which occur in subtropical and tropical regions of southwestern China to southeast Asia. Here, we confirm prior phylogenetic results showing the close relationship between sects. Rubra and Hyptiocarpa, and we use scanning electron microscopy to demonstrate that leaves of species within these sections have similar arrangements of cuticular waxes, which account for the silvery color of their abaxial surfaces. We describe that the sections also share labile sex expression; inflorescences that range from compound racemose thyrses, to racemes or umbels and that may have undergone evolutionary reduction; and several features of their fruits, such as seed locules without keels, basal portion of wings straight, acute attachment angle between mericarps, and production of some mericarps that are seedless and partially developed at maturity. Our expansion of sect. Rubra to include sect. Hyptiocarpa better elucidates the biogeographic and evolutionary history of these species. Additionally, we show that A. laurinum and A. pinnatinervium have intergrading morphology and are probably synonymous, but we note that further studies are required to conclude their taxonomic status.

Use of Mycelium and Detached Leaves in Bioassays for Assessing Resistance to Boxwood Blight.

Boxwood blight caused by Calonectria pseudonaviculata is a newly emergent disease of boxwood (Buxus spp. L.) in the United States that causes leaf drop, stem lesions, and plant death. A rapid and reliable laboratory assay that enables screening hundreds of boxwood genotypes for resistance to boxwood blight is needed to enable breeding and selection of resistant cultivars. Using eight boxwood cultivars with differing susceptibilities, we examined parameters for a screening assay comparing whole plant inoculation with detached leaf inoculation, use of mycelium versus spores as the inoculum, comparison of times of the year for inoculation, and comparison of two leaf inoculation methods. Inoculation of detached leaves gave comparable results to inoculation of whole plants when compared across genotypes, although the detached leaf assay resulted in greater percentages of symptom expression. The time of year of plant inoculation (spring, summer, or winter) did not affect the relative expression of symptoms among the most resistant and susceptible genotypes. Inoculating plants with mycelium was as effective as spore inoculation for causing disease symptoms and allowed us to distinguish the more resistant genotypes, yet mycelium inoculation was much easier to prepare in large quantities for multiple assays.

Development of polymorphic genic-SSR markers by cDNA library sequencing in boxwood, Buxus spp. (Buxaceae).

PREMISE OF THE STUDY: Genic microsatellites or simple sequence repeat (genic-SSR) markers were developed in boxwood (Buxus taxa) for genetic diversity analysis, identification of taxa, and to facilitate breeding. • METHODS AND RESULTS: cDNA libraries were developed from mRNA extracted from leaves of Buxus sempervirens 'Vardar Valley' and sequenced using the Illumina MiSeq system. Approximately 11.9 million base pairs of sequence data were examined and 845 genic-SSRs were identified, including 469 dinucleotide, 360 trinucleotide, seven tetranucleotide, one pentanucleotide, and eight hexanucleotide repeats. Primer pairs were designed for 71 selectively chosen genic-SSRs containing trinucleotide repeat motifs and were used to amplify the corresponding loci in 18 diverse boxwood accessions. Twenty-three primer pairs amplified polymorphic loci, with two to 10 alleles per locus. • CONCLUSIONS: These novel polymorphic genic-SSR markers will aid in evaluating genetic diversity of boxwood germplasm and allow verification of hybrids and cultivars for breeding programs.

Ulmus americana (Ulmaceae) is a polyploid complex.

PREMISE OF THE STUDY: Exotic diseases are threatening many North American tree species, and management of diseases requires understanding the biology of the host as well as the pathogen. Ulmus americana is a widespread dominant tree of eastern North America that has been widely planted as an ornamental and shade tree. Populations of the species have suffered heavy mortality from Dutch elm disease, caused by an introduced fungus. Ulmus americana is generally reported to be tetraploid, but the discovery of triploid trees in cultivation suggested that lower ploidy levels may exist in the wild, so the species was surveyed for nuclear DNA content. METHODS: Ploidy level was estimated by flow cytometry for 81 individuals from wild populations of U. americana from throughout the range of the species and for four cultivated trees of interest. KEY RESULTS: Most specimens were tetraploid, as previously reported for the species, but 21% of the wild trees sampled were diploid, a ploidy level not previously confirmed for the species. Tetraploids are found throughout the range of the species. Diploids are most common on the Atlantic coastal plain, Cumberland Plateau, and in southern Ohio, but isolated diploids were also found in central Texas, Oklahoma, and eastern Missouri. Diploids and tetraploids grew in proximity in several areas, but no wild triploids were found in the course of this survey. CONCLUSIONS: The species is genetically heterogeneous, but further research is needed to understand the origin and relations of the different ploidy levels. Understanding the ploidy situation in U. americana will be important in the search for further genotypes that are resistant to Dutch elm disease.

Evaluation of hemlock (Tsuga) species and hybrids for resistance to Adelges tsugae (Hemiptera: Adelgidae) using artificial infestation.

Hemlock (Tsuga) species and hybrids were evaluated for resistance to the hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae). The adelgid was accidentally introduced from Asia to the eastern United States, where it is causing widespread mortality of the native hemlocks, Tsuga canadensis (L.) Carrière and Tsuga caroliniana Engelm. These two native species plus the Asian species Tsuga chinensis (Franch.) E. Pritz and T. dumosa (D.Don) Eichler and Tsuga sieboldii Carrière, and the hybrids T. chinensis x T. caroliniana and T. chinensis x T. sieboldii, were artificially infested with the crawler stage of A. tsugae in the early spring 2006 and 2007. After 8 or 9 wk-when the spring (progrediens) generation would be mature--counts were made of the adelgid. In both years, the density of A. tsugae was highest on T. canadensis, T. caroliniana, and T. sieboldii; lowest on T. chinensis; and intermediate on the hybrids. On T. chinensis and the T. chinensis hybrids, fewer adelgids settled, fewer of the settled adelgids survived, and the surviving adelgids grew slower. Thus, the nature of the host resistance is both nonpreference (antixenosis) and adverse effects on biology (antibiosis). Tree growth (height) was associated with resistance, but no association was found between time of budbreak and resistance that was independent of the taxa. Many of the hybrids grow well, have attractive form, and are promising as resistant landscape alternatives for the native hemlocks.