Matthew H. Collier

Diminished Plant Richness and Abundance Below Lonicera maackii, an Invasive Shrub

Abstract The Asian shrub Lonicera maackii (Amur honeysuckle) is now common in many secondary forests in southwestern Ohio and adjacent states. We found lower species richness and abundance in plots (0.5 m2) placed below crowns of L. maackii than in plots placed away: all species (53% lower richness and 63% lower cover), tree seedlings with canopy potential (−41% richness and −68% density) and seed + bud bank (−34% richness and −33% density). Moreover, most individual taxa had lower abundance below L. maackii: 86% of herbs, 100% of trees and 56% of seed + bud bank taxa. In addition, richness of all species and richness and density of tree seedlings decreased in forests with longer residence time of L. maackii.

Histological analysis of spermatogenesis and the germ cell development strategy within the testis of the male Western Cottonmouth Snake, Agkistrodon piscivorus leucostoma

Cottonmouth (Agkistrodon piscivorus leucostoma) testes were examined histologically to determine the germ cell development strategy employed during spermatogenesis. Testicular tissues from Cottonmouths were collected monthly from swamps around Hammond, Louisiana. Pieces of testis were fixed in Trumps fixative, dehydrated in ethanol, embedded in Spurrs plastic, sectioned with an ultramicrotome, and stained with toluidine blue and basic fuchsin. Spermatogenesis within Cottonmouths occurs in two independent events within a single calendar year. The testes are active during the months of March-June and August-October with spermiation most heavily observed during April-May and October. To our knowledge, this is the first study that describes bimodal spermatogenesis occurring in the same year within the subfamily Crotalinae. During spermatogenesis, no consistent spatial relationships are observed between germ cell generations. Typically, either certain cell types were missing (spermatocytes) or the layering of 3-5 spermatids and/or spermatocytes within the same cross-section of seminiferous tubule prevented consistent spatial stages from occurring. This temporal pattern of sperm development is different from the spatial development found within birds and mammals, being more reminiscent of that seen in amphibians, and has now been documented within every major clade of reptile (Chelonia, Serpentes, Sauria, Crocodylia). This primitive-like sperm development, within a testis structurally similar to mammals and birds, may represent an intermediate testicular model within the basally positioned (phylogenetically) reptiles that may be evolutionarily significant.

Comparative water balance profiles of Orchidaceae seeds for epiphytic and terrestrial taxa endemic to North America

The Orchidaceae have dust-like seeds that use wind currents for long-distance dispersal. Lacking endosperm, orchid embryos consume free-living, mycorrhizal fungi as a carbon source (mycotrophy) after settling on a substrate. Few studies have investigated orchid seed morphology as it relates to ecology, but conceivably variations in seed size and testa characteristics could be linked to water loss rates aimed at maximizing germination in a particular habitat. Seeds of 2 epiphytic, 1 aquatic, and 7 terrestrial orchids native to North America were compared with respect to water balance profiles: Cleistes bifaria, Encyclia tampensis, Epidendrum nocturnum, Habenaria repens, Isotria medeoloides, Liparis elata, L. hawaiensis, Platanthera holochila, P. integrilabia, and P. leucophaea. Water content, water loss rate, activation energy, and equilibrium humidity were assessed for each species. Seeds of epiphytic orchids were smaller, lighter, more porous, and had higher water loss rates compared to terrestrials. No active mechanism for water absorption exists in seeds of either group. Water loss appears to be a species-specific phenomenon that may be linked to the ecological niches these species occupy.

Continuous spermatogenesis and the germ cell development strategy within the testis of the Jamaican Gray Anole, Anolis lineatopus

Testicular tissues from Anolis lineatopus were examined histologically to determine testicular structure, germ cell morphologies, and the germ cell development strategy employed during spermatogenesis. Anoles (N=36) were collected from southern Jamaica from October 2004 to September 2005. Testes were extracted and fixed in Trumps fixative, dehydrated, embedded in Spurrs plastic, sectioned, and stained with basic fuchsin/toluidine blue. The testes of Jamaican Anoles were composed of seminiferous tubules lined with seminiferous epithelia, similar to birds and mammals, and were spermatogenically active during every month of the year. However, spermatogenic activity fluctuated based on morphometric data for February, May and June, and September-December. Sequential increases for these months and decreases in between months in tubular diameters and epithelial heights were due to fluctuations in number of elongating spermatids and spermiation events. Cellular associations were not observed during spermatogenesis in A. lineatopus, and three or more spermatids coincided with mitotic and meiotic cells within the seminiferous epithelium. Although the germ cell generations were layered within the seminiferous epithelium, similar to birds and mammals, the actual temporal development of germ cells and bursts of sperm release more closely resembled that reported recently for other reptilian taxa. All of these reptiles were temperate species that showed considerable seasonality in terms of testis morphology and spermatogenesis. The Jamaican Gray Anole has continuous spermatogenesis yet maintains this temporal germ cell development pattern. Thus, a lack of seasonal spermatogenesis in this anole seems to have no influence on the germ cell development strategy employed during sperm development.

Temporal germ cell development strategy during spermatogenesis within the testis of the Ground Skink, Scincella lateralis (Sauria: Scincidae).

Ground Skink (Scincella lateralis) testes were examined histologically to determine the testicular organization and germ cell development strategy employed during spermatogenesis. Testicular tissues were collected from 19 ground skinks from Aiken County, South Carolina during the months of March-June, August, and October. The testes consisted of seminiferous tubules lined with germinal epithelia in which germ cells matured in close association with Sertoli cells. As germ cells matured, they migrated away from the basal lamina of the epithelia towards the lumina of the seminiferous tubules. The testes were spermatogenically active during the months of March, April, May, June, and October (largest seminiferous tubule diameters and epithelial heights), but entered a quiescent period in August (smallest seminiferous tubule diameter and epithelial height) where only spermatogonia type A and B and early spermatocytes were present in low numbers within the seminiferous epithelium. Although the testicular organization was similar to other amniotes, a temporal germ cell development strategy was employed during spermatogenesis within Ground Skinks, similar to that of anamniotes. Thus, this skinks germ cell development strategy, which also has been recently reported in all other major reptilian clades, may represent an evolutionary intermediate in terms of testicular organization between anamniotes and birds and mammals.

Temporal Germ Cell Development Strategy during Mixed Spermatogenesis within the Male Mediterranean Gecko, Hemidactylus turcicus (Reptilia: Gekkonidae)

Abstract The testes of Hemidactylus turcicus are composed of seminiferous tubules lined with continuous germinal epithelia in which multiple germ cell morphologies can be found during the active months of sperm development. Spermatogenesis is quiescent during September, with only spermatogonia A and spermatogonia B present in the seminiferous epithelia and minimal mitotic activity is observed. Recrudescence begins in October and the early stages of spermatogenesis progress through November. The onset of spermiation is observed in December and continues through August with the heaviest sperm release occurring in June and July. Multiple generations of late elongated spermatids are found in association with early mitotic and meiotic cells during the months of December–August. This temporal germ cell development strategy is similar to that described in other squamates and anamniotes and is different from the spatial development exhibited by birds and mammals, in which germ cell populations collectively progress through the stages of spermatogenesis. The reptilian temporal model of germ cell development within a structurally amniotic testis leads to two hypotheses in character evolution: birds and mammals exhibit convergence of germ cell development strategy, or the spatial development strategy is a synapomorphy in amniotes, and reptiles represent an evolutionary reversal to the strategy employed by anamniotes. These findings, along with present and future data, may allow for more concrete phylogenetic analyses by creating more characters for phylogenetic matrices and may prove to be useful in future histopathological studies.

Temporal germ cell development strategy during continuous spermatogenesis within the montane lizard, Sceloporus bicanthalis (Squamata; Phrynosomatidae)

Sceloporus bicanthalis is a viviparous lizard that lives at higher elevations in Mexico. Adult male S. bicanthalis were collected (n = 36) from the Nevado de Toluca, Mexico (elevation is 4200 m) during August to December, 2007 and January to July, 2008. Testes were extracted, fixed in Trumps, and dehydrated in a graded series of ethanol. Tissues were embedded, sectioned (2 μm), stained, and examined via a light microscope to determine the spermatogenic developmental strategy of S. bicanthalis. In all months examined, the testes were spermiogenically active; based on this, plus the presence of sperm in the lumina of seminiferous tubules, we inferred that S. bicanthalis had year-round or continuous spermatogenesis, unlike most reptiles that occupy a temperate or montane habitat. It was recently reported that seasonally breeding reptiles had a temporal germ cell development strategy similar to amphibians, where germ cells progress through spermatogenesis as a single population, which leads to a single spermiation event. This was much different than spatial development within the testis of other derived amniotes. We hypothesized that germ cell development was temporal in S. bicanthalis. Therefore, we wanted to determine whether reptiles that practice continuous spermatogenesis have a mammalian-like spatial germ cell development, which is different than the typical temperate reptile exhibiting a temporal development. In the present study, S. bicanthalis had a temporal development strategy, despite its continuous spermatogenic cycle, making them similar to tropical anoles.

Clonal variation in floral stage timing in the common dandelion Taraxacum officinale (Asteraceae)

We investigated the hypothesis that dandelion clones (Taraxacum officinale Weber, sensu lato; Asteraceae) differ in their floral stage timing characteristics under a constant set of environmental conditions. To test this hypothesis, plants representing nine different dandelion clones (identified by DNA fingerprinting) were grown in groups of five (N = 45) in a growth chamber for a period of 8 mo, with chamber settings similar to environmental conditions at peak dandelion flowering time for their population sites. Five flowering phenology parameters were monitored daily for a total of 301 buds developing during this time: (1) time to bud; (2) time to full opening and inflorescence maturation (i.e., first anthesis); (3) time to re-closure of an inflorescence; (4) time to fruit (full re-opening of the inflorescence); and (5) total flowering time. Scape length at the appearance of a fully expanded infructescence was also measured for each individual. Significant differences in mean time to inflorescence, mean time to re-closure, mean time to fruit, and mean total flowering time were revealed among some dandelion clones (Kruskal-Wallis, P ≤ 0.0005). No differences in mean number of inflorescence buds per plant (P = 0.2217), mean time to bud (P = 0.2396), or mean scape length (P = 0.3688) were detected among the nine clones. These results suggest that differences in floral stage timing may in part involve varying genotypic environmental response characteristics and that these differences may have potential fitness effects. Further research is needed to determine if such clonal differences are observed under a broader range of uniform environmental conditions.

Productivity differences between dandelion ( Taraxacum officinale ; Asteraceae) clones from pollution impacted versus non-impacted soils

Common dandelions (Taraxacum officinale Weber, sensu lato; Asteraceae) introduced to North America form an assemblage of asexual (agamospermous), clonal lineages derived from Eurasian mixed sexual and asexual populations. We investigated whether selection for more pollution tolerant clonal lineages occurs at polluted sites and selection for more pollution intolerant lineages occurs at unpolluted sites. We tested the above hypothesis by performing reciprocal greenhouse productivity experiments in which unique dandelion clones (12 clones, identified by DNA fingerprinting, from each site type) sampled from two unpolluted and two polluted (moderately enhanced Cu, Pb and Zn soil concentrations) sites were grown pairwise in both unpolluted (nutrient solution only) and polluted (nutrient solution + Cu, Pb and Zn) media (n = 48 paired tests for each media type). Dandelion clones from polluted sites produced fewer and smaller leaves, shorter roots and smaller root diameters, reduced shoot and root dry weights, and reduced total biomass compared to clones from unpolluted sites when clones were grown in unpolluted-media (P ≤ 0.05). In contrast, clones taken from unpolluted sites were shown to produce significantly fewer and shorter leaves, shorter roots and smaller root diameters, reduced shoot and root dry weights, reduced total biomass, a reduced shoot : root biomass ratio, and have much lower survival compared to clones from polluted sites when both were grown in polluted-media (P ≤ 0.05). These results reveal that there was increased selection against unpolluted-site clonal lineages in polluted-media and against polluted-site clonal lineages in unpolluted-media. Across all treatments, clones from unpolluted sites growing in unpolluted-media had the highest proximate measures of fitness. Overall, these findings provide insight into the relationships among anthropogenic environmental contamination and the consequent effects of selective forces acting on dandelion clones and their population genetic architecture.

Reduced seed germination after pappus removal in the North American Dandelion (Taraxacum officinale; Asteraceae).

Abstract This study examined seed ultrastructure in relation to germination of North American dandelion seeds. Based on laboratory rearing observations, it was thought that the design of the pappus acts as a conduit facilitating water entry into the seed. It was hypothesized that seeds without a pappus would yield fewer seedlings and require more time to germinate than seeds with an intact pappus. Seed ultrastructure was investigated using scanning electron microscopy, while relative humidity and fungal association were explored as factors that may confer an advantage to intact seeds. Results indicate that germination for seeds lacking a pappus is 31% lower than control seeds (with an intact pappus) and that the seeds lacking a pappus require more time to germinate. Relative humidity did not differentially affect germination, and while a fungus Cladosporium cladosporioides was recovered internally, its presence neither enhanced germination nor decreased time to germination when tested by antimycotic removal. Electron micrographs revealed that (1) the pappus is hollow and (2) the pericarp of the fruit fuses with and partially encloses the pappus. Fusion of the pappus with the fruit suggests that this structure acts as a device to regulate seed hydration. Nomenclature: North American dandelion, Taraxacum officinale G. H. Weber ex Wiggers; Cladosporium cladosporioides.