Huperzia Selago Research Articles

Characteristics of atypical Huperzia selago subsp. arctica habitats to the south of distribution area

Two localities for <em>Huperzia selago </em>subsp. <em>arctica </em>are recorded from Lithuania, to the south of its known distribution area. The habitats of this subspecies are cutover peatlands whose natural vegetation was disturbed 6-8 years ago during peat exploitation. One of the dominant species of latest vegetation cover is the invasive bryophyte <em>Campylopus introflexus</em>. Characteristics of the habitats of <em>H. selago </em>subsp. <em>arctica </em>and the frequency of this taxon in populations were analysed.

Determination of huperzine a inHuperzia selagoplants from wild population and obtained inin vitroculture by high-performance liquid chromatography using a chaotropic mobile phase

Summary This article is the first report describing a new validated method to determine the content of HupA in Huperzia selago (Huperziaceae) from wild population and obtained in in vitro culture using the chaotropic mobile phase. An aqueous-organic (acetonitrile) mobile phase with an added chaotropic salt (NaPF6) was used. The system of mobile phases ensured very high selectivity and efficiency at up to N = 6683 ± 963 theoretical plates calculated for isocratic mode. A Hypercosil GOLD column, C18 250 × 4.6 mm, and a Hypercosil GOLD precolumn, 5UM 10 × 4 mm, were employed for detection at four wavelengths, 230 nm being analytical. The regression coefficient (R2) of the calibration was 0.9993 over the range 25–1252 μg mL−1. The recovery rates were 98.36–105.1% with RSD <2.9%. The intra- and inter-day precisions, expressed as RSD, ranged from 1.2% to 2.7%. LOD for HupA was 14 ng mL−1 for a signal-to-noise ratio of 3:1. The limit of quantification was 140 ng mL−1. The huperzine A (HupA) content of the plant material ranged from 0.65 mg g−1 dry weight (d.w.) to 1.59 mg g−1 d.w. (material from wild plants) and from 0.44 to 1.10 mg g−1 d.w. (material from in vitro cultures). Interestingly, in our study, plants of H. selago derived from wild population had one of the highest HupA concentrations recorded for a club moss (1.59 mg g−1 d.w.). The findings demonstrate that H. selago, found in Europe and North America, is an alternative source of HupA, richer than H. serrata. In order to confirm that HupA was present in the alkaloid extracts, HPLC-ESI-MS/MS analyses of the patterns were performed in the positive ion mode. The fragmentation quasi-molecular ion of the standard HupA (m/z = 243, [M+H]+) and the ion with m/z = 243 found in the samples were identical, confirming the compound as HupA.

Paraconiothyrium babiogorense sp. nov., a new endophyte from fir club moss Huperzia selago (Huperziaceae)

Paraconiothyrium is a recently established genus within the order Pleosporales. Species from this genus are commonly associated with plants but can also be found in soil samples and be parasitic on fungi. Several isolates of a Paraconiothyrium sp. were obtained from Huperzia selago in Poland. Strains were characterized based on morphological characteristics and molecular data (SSU rDNA, ITS1, 5.8S rDNA, ITS2). Based on its unique morphology and DNA phylogeny, isolates were described as a new species: Paraconiothyrium babiogorense, which represents the first report of a Paraconiothyrium species from fir club moss. Comparison of characters with other Paraconiothyrium species is provided as well as a signature sequence for the new species.

The genusHuperzia(Lycopodiaceae) in the Azores and Madeira

The taxonomy and nomenclature of the genus Huperzia Bernh. in the Azores and Madeira have been reviewed. Plants collected in the Azores and Madeira were characterized morphologically. The independence between two endemic species common to Madeira and the Azores Islands –Huperzia suberecta (Lowe) Tardieu and Huperzia dentata (Herter) Holub – is clearly shown. A clear-cut morphological separation between these taxa and Huperzia selago (L.) Bernh. ex Schrank & Mart. of continental Europe is established. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158, 522–533.

The Gametophyte of Huperzia selago in Culture

ABSTRACT Cultured gametophytes of Huperzia selago are dorsiventral and strap-shaped. They are basically the same as cultured gametophytes of other terrestrial Huperzia species and also essentially the same as gametophytes of H. selago collected from loose soil. Besides shape, the cultured gametophytes have anatomical features found in gametophytes of H. selago from loose soil. These gametophytes are Type III gametophytes as characterized by Bruchmann in 1898. Although both gametangia occur on individual cultured gametophytes, the meristematic groove produces them at different times and they are not intermixed. The number of terrestrial Huperzia species with described gametophytes is still small (six), but that number has tripled as a result of the recent studies on these gametophytes in culture. Because all the terrestrial Huperzia species to date have Type III gametophytes, it would not be unexpected for the undescribed gametophytes of other species in this group to also be Type III gametophytes.

Phylogenetic relationships, host affinity, and geographic structure of boreal and arctic endophytes from three major plant lineages

Although associated with all plants, fungal endophytes (microfungi that live within healthy plant tissues) represent an unknown proportion of fungal diversity. While there is a growing appreciation of their ecological importance and human uses, little is known about their host specificity, geographic structure, or phylogenetic relationships. We surveyed endophytic Ascomycota from healthy photosynthetic tissues of three plant species ( Huperzia selago, Picea mariana, and Dryas integrifolia, representing lycophytes, conifers, and angiosperms, respectively) in northern and southern boreal forest (Québec, Canada) and arctic tundra (Nunavut, Canada). Endophytes were recovered from all plant species surveyed, and were present in <1–41% of 2 mm 2 tissue segments examined per host species. Sequence data from the nuclear ribosomal internal transcribed spacer region (ITS) were obtained for 280 of 558 isolates. Species-accumulation curves based on ITS genotypes remained non-asymptotic, and bootstrap analyses indicated that a large number of genotypes remain to be found. The majority of genotypes were recovered from only a single host species, and only 6% of genotypes were shared between boreal and arctic communities. Two independent Bayesian analyses and a neighbor-joining bootstrapping analysis of combined data from the nuclear large and small ribosomal subunits (LSUrDNA, SSUrDNA; 2.4 kb) showed that boreal and arctic endophytes represent Dothideomycetes, Sordariomycetes, Chaetothyriomycetidae, Leotiomycetes, and Pezizomycetes. Many well-supported phylotypes contained only endophytes despite exhaustive sampling of available sequences of Ascomycota. Together, these data demonstrate greater than expected diversity of endophytes at high-latitude sites and provide a framework for assessing the evolution of these poorly known but ubiquitous symbionts of living plants.

The determination of huperzine A in european Lycopodiaceae species by HPLC‐UV‐MS

A rapid qualitative HPLC-UV-MS method was developed for the detection of huperzine A in Lycopodiaceae species. HPLC coupled with mass spectrometry experiments allowed the identification of the alkaloid and enabled targeted analysis of complex matrices such as herbal extracts. Huperzine A was detected by single ion monitoring of the pseudomolecular ion [M + H]+ at m/z 243.2. The alkaloid was also detected on TLC in a bioautographic enzyme assay with acetylcholinesterase to show the activity of the compound. Four Lycopodiaceae species collected in Switzerland were tested by these methods and Huperzia selago (L.) Bernh. ex Schrank et Martius was the only species found to contain huperzine A.

Somatic embryogenesis and in vitro culture of Huperzia selago shoots as a potential source of huperzine A

In vitro culture of the sporophytes and somatic embryogenesis in Huperzia selago is described together with the results of phytochemical analyses to determine the huperzine A content of H. selago shoots collected from plants growing in vitro and in a natural habitat. The in vitro culture was initiated from shoots of wild plants, which underwent surface and internal sterilization to eliminate the epiphytic and endophytic bacteria and fungi and were next placed on different modifications of the Murashige and Skoog medium (MS). The highest growth frequency was achieved on the MS medium at half-strength mineral salt content (1/2 MS). The same medium was used for the induction of somatic embryogenesis. Cells of the callus, which developed from the apical meristem after 3 months of incubation, transformed into somatic embryos. The stages of their further growth resembled the zygotic embryos of other Lycopodiaceae. The developing embryo had no shoot–root axis, but one that corresponded to the shoot–foot axis with the root growing at its side. The developing somatic embryo smoothly transformed into a young sporophyte with pre-leaves and numerous proper leaves and an embryonal root. HPLC–UV assay demonstrated the highest huperzine A content (3.33 mg g −1 d.w.) in the shoots of plants grown from the somatic embryos. The yield of huperzine A from plants collected from a natural habitat differed from 0.54 (shoots harvested in spring) to 1.27 mg g −1 d.w. (shoots harvested in autumn).

Selagoline, A new Alkaloid from Huperzia Selago

A new natural product, named selagoline, and two known alkaloids, huperzine A and serratidine, were isolated from Huperzia selago (Lycopodiaceae) collected in Iceland. Their structures were determined using 600 and 800 MHz one- and two-dimensional NMR methods supported by Fourier-transform mass spectrometry. Possible role of selagoline as a precursor of 5,15-oxidolycopodane, a component of the classical alkaloids L28 and L31, is discussed.

3- O-Methylrhamnose in lower land plant primary cell walls

Acid hydrolysis of cell wall-rich material derived from the young photosynthetic tissues of lower land plants and charophytes yielded an unusual sugar. This sugar was identified as 3- O-methylrhamnose (trivial name acofriose) by its 1H and 13C NMR spectra. In addition to the usual major monosaccharides (glucose, galactose, mannose, arabinose, xylose, rhamnose, glucuronic acid and galacturonic acid) acofriose was released from cell walls of three charophytic algae ( Chara coralina, Coleochaete scutata and Klebsormidium flaccidium), a hornwort ( Anthoceros caucasicus), leafy and thalloid liverworts, mosses and homosporous lycopodiophytes ( Lycopodium pinifolium, Huperzia selago and Diphasiastrum alpinum). The yield of acofriose approximately equalled that of rhamnose. Acofriose was not detectable in the cell walls of a heterosporous lycopodiophyte, a eusporangiate fern, or any of several leptosporangiate ferns, gymnosperms and angiosperms. Our results show that the primary cell walls of lower land plants and their charophyte ancestors are qualitatively different from those of higher plants: the loss of rhamnose methylation supports the generalisation that major steps in plant evolution have required significant changes in cell wall composition.

Abrupt vegetation and climate changes during the last glacial maximumand last termination in the chilean lake district: a case study from canal de la puntilla (41°s)

ABSTRACT. Multiple overlapping and replicate pollen stratigraphies from Canal de la Puntilla (40°57’09”S, 72°54’18”W, 120 m elevation) reveal that a Nothofagus dombeyi‐type parkland occupied the Valle Central of the Chilean Lake District during the portion of the Last Glacial Maximum between 20,200 and about 14,600 14C yr BP. Dominating this landscape was Nothofagus dombeyi‐type and Gramineae, accompanied by taxa commonly found today in Subantarctic environments and above the Andean tree‐line in the Lake District (Perezia‐type, Valeriana, and Huperzia selago), along with cushion bog taxa characteristic of Magellanic Moorlands (Donatia fascicularis and Astelia pumila). Within this open landscape Nothofagus dombeyi‐type expanded between 20,200 and 15,800 14C yr BP, interrupted by a brief reversal between 19,200 and 18,800 14 C yr BP and followed by a prominent increase in Gramineae pollen between 15,800 and about 14,600 14C yr BP. A major rise of Nothofagus dombeyi‐type began at about 14,600 14C yr BP, followed by decline in non‐arboreal taxa and a remarkable expansion of North Patagonian Rain Forest taxa in pulses centered at 14,200 and 13,000 14C yr BP. Podocarpus nubigena expanded between 12,200 and 9800 14C yr BP, along with increases in Misodendrum and Maytenus disticha‐type between 11,000 and 9800 14C yr BP.Paleovegetation records suggest that mean annual temperature was 6–7°C colder than at present during the coldest episodes between 20,200 and about 14,600 14C yr BP, with twice the modern annual precipitation between 20,200 and 13,000 14C yr BP, suggesting a northward shift and intensification of westerly stormtracks. Slight climate warming occurred between 20,200 and 15,800 14C yr BP, interrupted by cooling events at 19,200 and 15,800 14C yr BP. The initial warming of the last termination started at 14,600 14C yr BP, followed by warming pulses at 14,200 and 13,000 14C yr BP. These events brought glacial conditions to a cool‐temperate climate, slightly cooler and wetter than modern climate, accounting for a total temperature recovery of ≥5°C by about 13,000 14C yr BP. A general reversal in trend is inferred with cooling events at 12,200 and 11,000 14C yr BP.

Germination of Spores of the Lycopodiaceae in Axenic Culture

Spores from four species of Huperzia, three species of Lycopodiella, and three species of Lycopodium were germinated in axenic culture. The ten species included species with subterra- nean mycorrhizal gametophytes and others with surficial photosynthetic gametophytes. The germi- nation of spores from species with mycorrhizal gametophytes was slow, but much faster than pre- viously reported. It occurred in the dark in a few months for most species instead of years as reported by Bruchmann (1910). Although species of Lycopodiella are expected to have rapid germination, spores from two species had slow germination. It appears that not all the species of the Lycopodi- aceae with photosynthetic gametophytes exhibit rapid spore germination as reported by Treub (1884). Germinating the spores of the Lycopodiaceae in axenic culture is difficult. It generally takes a considerable length of time and the percentage of germi- nation is low (Whittier, 1977, 1981; Whittier and Webster, 1986). Efforts over the years to germinate these spores under more natural conditions (i.e., on soil) have demonstrated that spores of the Lycopodiaceae fall into two classes according to how rapidly they germinate (Bierhorst, 1971). The spores of spe- cies with subterranean, mycorrhizal gametophytes germinate slowly (Bruch- mann, 1910; Barrows, 1935) and those of species with photosynthetic, surficial gametophytes germinate more rapidly (DeBary, 1858 (see Chamberlain, 1917, or Barrows, 1935) and Treub, 1884, 1888). Studies on the germination of spores from species of the Lycopodiaceae with mycorrhizal gametophytes provide limited information (Table 1). Bruchmann (1910) germinated the spores of Lycopodium selago L. (Huperzia selago (L.) Bernh. ex Schrank & Mart.), L. annotinum L., and L. clavatum L. in soil cul- tures. Barrows (1935) investigated spore germination with Lycopodium com- planatum L. var. flabelliforme Fernald (L. digitatum Dill. ex A. Br.) and ob- tained 16% germination after one year. More recent attempts to germinate these spores have been carried out in axenic culture (Table 1) but less than 0.1% of the spores germinated. It is probably advantageous for spores forming subterranean gametophytes to germinate slowly and be down in the soil before gametophyte development starts. Indications from earlier studies (Whittier, 1977, 1981; Whittier and Web- ster, 1986) are that spores producing mycorrhizal gametophytes only germinate in the dark, which would stop germination until the spores are covered by soil. However, why these spores of the Lycopodiaceae need long times in the dark before they germinate and why they have such low percentages of germination is not understood. In other pteridophytes with mycorrhizal gametophytes, spore germination is more rapid and can exceed 50% after a few months in axenic culture (Melan and Whittier, 1990; Whittier and Braggins, 1994).

Background levels of heavy metals in plants of different taxonomic groups from a montane rain forest in Sri Lanka

An undisturbed natural reserve area iocated in a tropical montane rain forest at about 1800 m altitude in Sri Lanka served as a study site to investigate and assess the natural background concentration levels of As, Cd, Co, Cu, Ni, Pb and Zn in plants representing different taxonomic groups (divisions) in the plant kingdom. The plants selected were: the lichen,Usnea barbata (old man's beard);Pogonatum sp. (a moss);Lycopodium selago (epiphytic lycopod);Polypodium lanceolatum (epiphytic fern);Bulbophyllum elliae (epiphytic orchid) andActinodaphne ambigua (dicotyledonous large tree). Degree of homogeneity with respect to Cd, Cu, Pb and Zn in homogenised materials of all samples were within acceptable limits, whereasPogonatum sp. showed the highest degree of homogeneity for Pb. In addition to confirming extremely low levels of heavy metals in all plant species, the survey also found that generally the primitive plants,Usnea andPogonatum appear to have a greater tendency to accumulate As, Cd, Co and Pb; in particular,U. barbata appears to be an efficient accumulator for those heavy metals, suggesting its potential use in environmental studies.Actinodaphne ambigua was found to have a specific accumulating ability for nickel. Surface cleaning of theA. ambigua leaves resulted in a substantial decrease in the foliar contents of Cd, Ni, Pb and Zn. Variations in heavy metal contents observed in different plant genera are discussed in terms of their habits and place of growth in the forest. It is anticipated that the background levels presented in this paper from a remote, unpolluted tropical ecosystem will provide useful reference data for comparative environmental studies.

Late‐glacial and Holocene vegetation history of the Flow Country, northern Scotland

The vegetation history of an important conservation area, the Flow Country in northern Scotland, is described using the results of pollen analysis from two closely spaced cores from the Cross Lochs, eastern Sutherland. Buried lake sediments and shallower surface peats yield a continuous record covering the Devensian Late-glacial and the Holocene. The earliest Late-glacial pollen spectra recovered suggest a transition from open habitat to more stable communities, dominated by *Empetrum heath. This Windermere Interstadial vegetation shows some variability, with a phase of increased growth of juniper and birch, followed by deteriorating climate and less stable soil conditions in the transition to the Stadial, with Huperzia selago and Salix. There are poorly defined Loch Lomond Stadial and Stadial-Holocene transition periods, characterized by Rumex/Oxyria, after which much warmer, Holocene conditions occur, with a succession of birch-juniper and birch-hazel woodland. Birch persists in small amounts locally, but the presence of other arboreal pollen is thought to be mostly due to long distance pollen transport. From c. 5500 BP, the landscape is largely treeless, apart from a brief phase of local pine forest c 4500-4000 BP. These changes are compared with other pollen records for northern Scotland. The trends in the Late-glacial vegetation are broadly similar to those displayed by other sites, although many of the changes appear to be less extreme than for areas to the south and west. The lack of Artemisia during the Loch Lomond Stadial in the east Sutherland and Caithness area is particularly noteworthy. This may imply more extensive snow cover. The early Holocene is similar to the rest of northern Scotland, but invasion of Betula was more rapid and the later growth of this genus and hazel was not so dense. The lack of pine in the early Holocene allies the site with eastern Caithness and the Northern Isles, rather than north-west Scotland and the Grampians. It is suggested that the effects of early human populations on such marginal forest areas may have been underestimated in the past. The paradoxical existence of pine macrofossils with no pollen evidence in several areas of Scotland is explained, at least in part, by the lack of temporal precision of pollen diagrams rather than by low pollen productivity, extremely localized occurrences of the species or strong prevailing winds.

The Appalachian Firmoss, a New Species in the Huperzia selago (Lycopodiaceae) Complex in Eastern North America, with a New Combination for the Western Firmoss

The gemmiferous firmosses, containing the type species of Huperzia, Huperzia selago, are distinguished from the other members of Huperzia by concave-sided spores and specialized lateral branches consisting of gemmiphores and detachable gemmae. The species of the group hybridize and the hybrids possess abortive spores with spore fragments and undivided spore mother cells. In the past the systematic treatment of the species in this group has been unsatisfactory because no one has recognized the intermediate sterile hybrids. These can reproduce vegetatively by gemmae, often forming large populations. Undetected hybrid taxa muddle species boundaries by obscuring and combining the already subtle characters that differentiate species. The obviously abortive-spored hybrids have remained undetected by most botanists. Careful field study of sympatric populations of species and hybrids over the range of the taxa, as well as examination of over 6000 herbarium specimens, has resulted in the present alignment of species boundaries and the recognition of a new species, Huperzia appalachiana, in eastern North America. First, we will review the range and characters of the common firmoss species, and then present the range and characters of Huperzia appalachiana. Looking at the three major areas of overlap (the northern Appalachians, the southern Appalachians, and the north shore of Lake Superior), the consistency of characters will be examined as well as of hybrids. The shining firmoss, Huperzia lucidula, is an eastern North American endemic of moist conifer and deciduous forests and swamps, always with some shade. Common in the northern and eastern parts of its range, it extends north of the Great Lakes up into the boreal forest, south in the Appalachians to Georgia and Alabama, west in shaded moist sandstone ravines of Missouri and Arkansas and to the edge of the deciduous woods in Minnesota. It is a robust plant of indeterminate growth habit; the individual stems continue growing indefinitely, becoming decumbent with age and eventually rotting away at their base. The leaves are in annual zones of longer leaves and shorter leaves. The shortest leaves are bud leaves produced at the end of the growing season in the form of winter bud. In H. lucidula, the leaves of the two portions are similar in size and reflexed position. The long leaves are widest beyond the middle with serrate margins and, importantly, have no stomates on the upper surface. The gemmiphores and the gemmae are relatively large. The lateral lobes of the gemmae are broadly obtuse with an apiculate tip. The spores are relatively small, averaging about 25 gm.

Pacific Firmoss (Huperzia miyoshiana) (Lycopodiaceae) in Eastern North America at Gros Morne National Park, Newfoundland

The Pacific Firmoss (Huperzia miyoshiana (Makino) Ching) is an amphi-Pacific element of the Huperzia selago complex (Huperzia selago (L.) Bernh. ssp. miyoshiana (Makino) Calder & Taylor (Beitel, 1986)). It grows on exposed, rocky subalpine slopes with a cool, humid, high precipitation maritime climate (Calder & Taylor, 1968). It is distributed along the coast of eastern Asia northward from Japan and across the Aleutians into southeastern Alaska; from there it ranges south along the coast to Washington state (Fig. 1). An exception to this coastal distribution is the isolated population in the Selkirk Mountains of eastern British Columbia which have a wet, maritimelike climate.

ChemInform Abstract: Alkaloids of Lycopodium selago. Identity of Selagine with Huperzine A and the Structure of a Related Alkaloid.

AbstractSelagine, previously assigned as (I), is found to be identical with huperzine A (II).

Lycopodium hickeyi: A New Species of North American Clubmoss

Recent studies of North American Lycopodiaceae are relatively few, but they have led to some major changes from the taxonomic treatments of the first half of this century (e.g., Eaton, 1890; Fernald, 1950). At the generic level, pteridologists generally agree that the classical genus Lycopodium comprises several distinct genera (0llgaard, 1987), but they do not generally agree as to how many segregate genera to recognize. At the specific level, pteridologists now generally agree that closely related taxa treated by previous workers as varieties of single species are better ranked as distinct species. In North America, examples of this change in rank can be found in the Huperzia selago group (Beitel, unpubl.), the Lycopodiella inundata group (Bruce, 1976), and the Lycopodium complanatum group (Wilce, 1965). This paper deals with the question of rank for a taxon in the L. obscurum group, commonly referred to as the ground pines or tree clubmosses. Over the years, pteridologists have disagreed as to what rank members of this group should receive. In 1803, a half century after Linnaeus described L. obscurum, Michaux distinguished a close relative, L. dendroideum. The former was colloquially referred to as the flat-branched tree clubmoss, the latter as the tree clubmoss due to their different phyllotaxies, orientation, and relative development of the leaves of different ranks. In 1890, D. C. Eaton placed L. dendroideum as a variety of L. obscurum without comment. This placement was followed by nearly all flora writers and writers of popular fern books during the first half of the 1900s. The situation changed when, from 1974 to 1978, Dr. R. James Hickey worked on the taxonomy of the L. obscurum group in North America and eastern Asia (Hickey, 1977, 1978). After detailed studies of geography, ecology, habit, and leaves of the central and lateral axes, he concluded that three species should be recognized: L. obscurum of eastern North America, L. dendroideum of northern North America and eastern Asia, and L. juniperoideum Sw. of eastern Asia. In addition, he concluded that a new variety of L. obscurum should also be recognized: var. isophyllum. The new variety resembled L. dendroideum by its equally spreading and equal-sized leaves which impart a cylindrical aspect to the branchlets. But the new variety more closely resembled L. obscurum by its phyllotaxy and its appressed leaves on the main erect stem below the first lateral branches. In essence, the new taxon was a round-branched variety of the flatbranched tree clubmoss. Hickey (1978) used the varietal category for his new

Étude au microscope électronique à balayage de la bulbille de Huperzia selago (L.) Bernh. (Lycopodium) : méristème apical, initiation foliaire et phyllotaxie

The shoot apex of the bulbil of Huperzia selago (L.) Bernh. (Lycopodium) has been examined by scanning electron microscope. From an ontogenic point of view, changes in shape and size have been observed in the apical meristem. Successive minimal and maximal areas of this meristem are described during a plastochron. Area and cell number are also quantified with the scanning electron microscope. Only one leaf is initiated at the beginning of each plastochron, and the two contact parastichies observed in the shoot apex of the mature bulbil are two foliar helices.

The Chromosomes of Lycopodium lucidulum

Mart.), intermediate sterile hybrids are occasionally found with either or both parents. These hybrids can be distinguished from L. lucidulum by their abortive spores and the presence of stomates on both surfaces of the leaves. The chromosome number of Lycopodium lucidulum Michx. has been reported several times from North America and India. The counts from India (Mehra & Verma, 1957; Ninan, 1958) based on Lycopodium lucidulum sensu Clark (1880), however, are actually from a distinct species, Lycopodium herterianum Kimmerle (Huperzia herteriana (Kumm.) Sen & Sen, L. sikkimense Herter 1909 non K. Muell. 1861), and not the nearctic Lycopodium lucidulum Michx. Love and Love (1958) first reported 2n = 264 for Lycopodium lucidulum from Quebec, but without an accompanying photograph, a drawing, locality data, or a voucher specimen citation. Later (1965), they reported chromosome counts from Mt. Washington, New Hampshire (without citation of voucher) of Lycopodium lucidulum and L. selago (which they made into subspecies of Huperzia selago) remarking . . it has been at last possible to make exact counts of the chromosomes of material of ssp. selago with appressed and patent leaves from Mt. Washington and also ssp. lucidula