Ancient and modern colonization of North America by hemlock woolly adelgid, Adelges tsugae (Hemiptera: Adelgidae), an invasive insect from East Asia.

Hemlock woolly adelgid, Adelges tsugae, is an invasive pest of hemlock trees (Tsuga) in eastern North America. We used 14 microsatellites and mitochondrial COI sequences to assess its worldwide genetic structure and reconstruct its colonization history. The resulting information about its life cycle, biogeography and host specialization could help predict invasion by insect herbivores. We identified eight endemic lineages of hemlock adelgids in central China, western China, Ulleung Island (South Korea), western North America, and two each in Taiwan and Japan, with the Japanese lineages specializing on different Tsuga species. Adelgid life cycles varied at local and continental scales with different sexual, obligately asexual and facultatively asexual lineages. Adelgids in western North America exhibited very high microsatellite heterozygosity, which suggests ancient asexuality. The earliest lineages diverged in Asia during Pleistocene glacial periods, as estimated using approximate Bayesian computation. Colonization of western North America was estimated to have occurred prior to the last glacial period by adelgids directly ancestral to those in southern Japan, perhaps carried by birds. The modern invasion from southern Japan to eastern North America caused an extreme genetic bottleneck with just two closely related clones detected throughout the introduced range. Both colonization events to North America involved host shifts to unrelated hemlock species. These results suggest that genetic diversity, host specialization and host phylogeny are not predictive of adelgid invasion. Monitoring non-native sentinel host trees and focusing on invasion pathways might be more effective methods of preventing invasion than making predictions using species traits or evolutionary history.

Effects of temperature and photoperiod on the aestivo-hibernal egg diapause of Scymnus camptodromus (Coleoptera: Coccinellidae).

Three sequential studies were conducted on the interacting effects of exposure to low (5°C) temperature for 0, 7, 28, 56, or 84 d followed by incubation at 10, 15, or 20°C on the egg diapause of Scymnus (Neopullus) camptodromus Yu and Liu (Coleoptera: Coccinellidae). This beetle was imported from China as a potential biological control agent for hemlock woolly adelgid, Adelges tsugae (Annand) (Hemiptera: Adelgidae). Very few eggs laid and held at a constant 15 or 20°C showed any indication of development. Only eggs exposed to temperature combinations of 5 and 10°C had >50% hatch. Highest percent hatch and fastest development occurred when eggs were held at 5°C for 56 or 84 d followed by holding at 10°C. A model estimated the lower threshold for postdiapause development to be 2°C. The effect of temperature on egg hatch was similar at photoperiods of 12:12 and 16:8 (L:D) h, suggesting egg development is not governed by photoperiod or light exposure. Collectively these data indicate that S. camptodromus eggs laid in the spring and summer go through an aestivo-hibernal diapause that is maintained by warm temperatures and that development resumes when temperatures drop, in parallel with the development of hemlock woolly adelgid. This concurrent development allows S. camptodromus eggs to hatch while hemlock woolly adelgid is laying eggs. This synchrony between the development of S. camptodromus eggs and the overwintering adelgid suggest this beetle may be a good candidate for the biological control of the hemlock woolly adelgid.

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.

Characterization of terpenoid volatiles from cultivars of eastern hemlock (Tsuga canadensis).

The volatile terpenoid fraction from needles in 13 cultivars of Tsuga canadensis L. (Carriere) was analyzed by gas chromatography with mass spectrometry (GC-MS). The results of this study are considered along with previously reported results for foliar terpenoid levels of the Asian (T. sieboldii, T. chinensis, T. diversifolia), western North American (T. mertensiana, T. heterophylla), and eastern North American species (T. canadensis, T. caroliniana) of hemlock to draw conclusions about the potential of cultivar host resistance to the hemlock woolly adelgid (Adelges tsugae Annand). It is suggested that hemlocks in eastern North America have adapted their terpenoid chemistry for protection against endemic defoliators and that this has made them vulnerable to non-native, sucking pests such as adelgids and scales. Some cultivars of T. canadensis have a terpenoid profile that resembles that of the resistant noneastern North American species and are candidates for biological screening for resistance. Among the cultivars, the variation in terpenoid chemistry did not absolutely correspond with the considerable differences in morphological characters observed, indicating that the terpenoid chemistry is not definitively coupled with hemlock morphology.

Temporal and spatial variation of terpenoids in eastern hemlock (Tsuga canadensis) in relation to feeding by Adelges tsugae.

The terpenoid content of eastern hemlock (Tsuga canadensis) foliage was measured over an annual cycle of development from bud opening, shoot elongation, shoot maturation, to bud-break at the start of the next growing season. The objective was to determine if variation in terpenoid composition is linked with spatial and temporal feeding preferences of the hemlock woolly adelgid (HWA; Adelges tsugae). The HWA has two periods of feeding over the course of 1 yr spanning two complete generations. There are two periods of feeding separated by a nonfeeding period where the adelgid estivates. HWA prefers to feed on mature, rather than young, expanding tissue. Feeding occurs in the leaf cushion at the base of the needle. The needle is the only tissue in hemlock with resin canals that store terpenoids. The needle and leaf cushion of both the current and previous years' growth were analyzed separately over a 1-yr period to examine the variation of terpenoid composition in space and time. Terpenoids were quantified by using headspace solid-phase microextraction/gas chromatography/mass spectrometry (SPME/GC/MS). New growth needles and leaf cushions do not resemble the previous year's growth either visually or in chemical composition until October/November, when the adelgid breaks estivation and begins feeding. Nearly all of the 23 terpenoids present exceeding 0.1% varied significantly either temporally or spatially, usually with complex interactions. Ordination and factor analysis revealed that terpenoids are less variable in mature leaf cushions than in young tissue. By entering a nonfeeding diapause during the late spring and summer, HWA avoids the unstable, variable levels of terpenoids in the immature leaf cushion and needles.

Feeding preference of three lady beetle predators of the hemlock woolly adelgid (Homoptera: Adelgidae).

In a laboratory study, we tested the feeding preferences of three coccinellid predators of hemlock woolly adelgid, Adelges tsugae Annand, an introduced pest of hemlock in the eastern United States. The species tested were Sasajiscymnus tsugae Sasaji & McClure (formerly Pseudoscymnus tsugae) from Japan, Scymnus ningshanensis Yu & Yao from China, and Harmonia axyridis (Pallas), a generalist species introduced from Asia that is currently widespread in eastern hemlock, Tsuga canadensis Carriere, forests. We measured the feeding preference of each beetle species when given the choice of A. tsugae and either 1) Pineus strobi (Hartig) on Pinus strobus L.; 2) Adelges laricis Vallot on Larix decidua Mill.; 3) Adelges cooleyi (Gillette) on Pseudotsuga menziesii (Mirb.) Franco; or 3) Paraprociphilus tessellatus (Fitch) on Alnus serrulata (Ait.) Willd. We evaluated beetle preference for adults, nymphs, and eggs of each prey species. Generally, when adult or nymphal prey stages were compared, S. tsugae preferred A. tsugae adults to P. strobi, A. cooleyi, A. laricis, and P. tessellatus. S. ningshanensis showed less preference between adelgid species, but it did not prefer P. tessellatus nymphs. When preferences for adelgid eggs were assayed, S. tsugae and S. ningshanensis showed no preference between A. tsugae and A. cooleyi or P. strobi, but S. tsugae did prefer A. tsugae to A. laricis. Larvae of S. tsugae were unable to survive on P. tessellatus nymphs. H. axyridis adults readily consumed both A. tsugae and P. tessellatus, but H. axyridis larvae did not complete their life cycle on A. tsugae. Our host range tests suggest that S. ningshanensis and S. tsugae may feed on several species of Adelgidae and that A. tsugae is often preferred.

Analysis of terpenoids from hemlock (Tsuga) species by solid-phase microextraction/gas chromatography/ion-trap mass spectrometry.

A sampling method for determining the volatile terpenoid composition from single needles of seven Tsuga species was developed using headspace solid-phase microextraction (SPME). A reproducible sampling method for the volatile components was generated by examination of sample storage, method of needle cutting, and headspace sampling duration. Following SPME collection of the volatile compounds from the seven Tsuga species, gas chromatography/ion-trap mass spectrometry was used to identify 51 terpenoids present in the needle headspace. A semiquantitative method was devised to express individual terpenoid amounts as a percentage of all of the identified peaks in the chromatogram. The semiquantitative results permitted facile interspecies comparison using principal component analysis. Two components were able to account for 90% of the variance and were interpreted as a "species" component and a "resistance/susceptibility" component. Three interspecies groupings were evident from the principal component analysis: (1) Tsuga canadensis and Tsuga caroliniana; (2) Tsuga chinesnsis, Tsuga diversifolia, Tsuga heterophylla, and Tsuga sieboldii; and (3) Tsuga mertensiana. The finding that T. mertensiana was grouped alone and far removed from the other species adds to the morphological evidence that this species should be segregated from other Tsuga.