Volatile Composition Screening of Salix spp. Nectar Honey: Benzenecarboxylic Acids,Norisoprenoids, Terpenes,and Others

Salix spp. nectar honey volatiles of Croatian origin were analyzed by headspace solid‐phase microextraction (HS‐SPME) and ultrasonic solvent extraction (USE), followed by gas chromatography and mass spectrometry (GC, GC/MS). Isolated volatiles were found in the honey headspace and extracts with almost exclusive distribution of several abundant compounds (e.g., phenylacetic acid, pinocembrin, 8‐hydroxy‐4,7‐dimethylcoumarin, and 3‐hydroxy‐trans‐β‐damascone in the extracts, or safranal and lilac alcohols in the headspace). Comparison with Croatian Salix spp. honeydew honey revealed similarities regarding distribution of important shikimate pathway derivatives (e.g., high percentage of phenylacetic acid) and several norisoprenoids (α‐isophorone and 4‐oxoisophorone). On the other hand, distinct features of this honey were occurrence of compounds such as pinocembrin, 8‐hydroxy‐4,7‐dimethylcoumarin, phenylacetonitrile, norisoprenoids (major ones: 3‐hydroxy‐trans‐β‐damascenone and trans‐β‐damascone), more pronounced variability of linalool‐derived compounds, as well as the abundance of 3‐methylpropanoic acid, 3‐methylbutanoic acid, 2‐methylpentanoic acid, and 3‐methylpentan‐1‐ol.     

Description of Euroleptochromus gen.n. (Coleoptera,Staphylinidae, Scydmaeninae) from Baltic amber,with discussion of biogeography and mouthpart evolution within Clidicini

A new extinct species of the ant‐like stone beetle supertribe Mastigitae, Euroleptochromus sabathi gen. & sp.n. is described from Eocene Baltic amber. A phylogenetic analysis of Clidicini, with representatives of Leptomastacini and Mastigini as out‐group taxa, provided strong support for a sister‐group relationship between the Neotropical Leptochromus and the new genus. The monophyly of Clidicini is questioned because of an alternative placement of Nearctic Papusus as a sister taxon to Leptomastacini + [Clidicus + (Palaeoleptochromus + (Euroleptochromus + Leptochromus))]. A dispersal‐vicariance analysis provided three alternative scenarios for the evolution of Mastigitae; with Laurasia as the ancestral area of the supertribe, major branching events occurring within either Eurasia or Laurentia and two trans‐Beringia dispersals in Late Cretaceous and Eocene. Euroleptochromus, Palaeoleptochromus and Leptochromus share highly derived structures on postgenae and maxillary palps, probably as part of a specialised feeding or prey capture mechanism. The formation of these modifications in Clidicini is demonstrated to involve a process (traced back to the Campanian, 79 Ma) of elongation and narrowing of maxillary palps and forming a cuticular setal projection from a broadened insertion site of sensory setae.     

Phytochemical Analysis,Biological Activity,and Secretory Structures of Stachys annua (L.) L. subsp. annua (Lamiaceae) from Central Italy

Stachys annua subsp. annua, well‐known in central Italy as ‘stregona annuale’, is an annual, small, slightly‐scented herb, commonly found in fields and uncultivated areas in almost all regions of Italy. In folk medicine, its aerial parts were used as anti‐catarrhal, febrifuge, tonic, and vulnerary. In the present work, the chemical composition of the flowering aerial parts was studied. The hydrodistilled volatile oil, analysed by GC/MS, showed sesquiterpenoids as the major fraction (42.5%); phytol (9.8%), germacrene D (9.2%), and spathulenol (8.5%) were the most abundant constituents. The volatile oil was assayed for antioxidant and cytotoxic activity by DPPH, ABTS, FRAP, and MTT methods. The cytotoxicity results against HCT116, A375, and MDA‐MB 231 human tumor cell lines were significant, with IC₅₀ values of 23.5, 37.2, and 41.5 μg/ml, respectively, whereas the antioxidant power was negligible. The EtOH extract was composed mainly of three glycosidic flavonoids, namely 7‐{[2‐O‐(6‐O‐acetyl‐β‐D ‐allopyranosyl)‐β‐D ‐glucopyranosyl]oxy}‐5,8‐dihydroxy‐2‐(4‐methoxyphenyl)‐4H‐1‐benzopyran‐4‐one ( 1 ), 7‐{[6‐O‐acetyl‐2‐O‐(6‐O‐acetyl‐β‐D ‐allopyranosyl)‐β‐D ‐glucopyranosyl]oxy}‐2‐(3,4‐dihydroxyphenyl)‐5,8‐dihydroxy‐4H‐1‐benzopyran‐4‐one ( 2 ), and 7‐{[6‐O‐acetyl‐2‐O‐(β‐D ‐allopyranosyl)‐β‐D ‐glucopyranosyl]oxy}‐2‐(3‐hydroxy‐4‐methoxyphenyl)‐5,8‐dihydroxy‐4H‐1‐benzopyran‐4‐one ( 3 ). On the contrary, iridoids, considered chemotaxonomic markers of the genus Stachys, were absent in this species. Finally, the morphological and histochemical survey showed that glandular trichomes were composed of two main types, i.e. peltate type A and capitate types B and C giving positive response for both lipids and polyphenols.     

Observations on Purpureofilum apyrenoidigerum gen. et sp. nov. from Australia and Bangiopsis subsimplex from India (Stylonematales, Bangiophyceae, Rhodophyta)

Purpureofilum apyrenoidigerum gen et sp. nov. was obtained from a mangrove habitat in New South Wales, Australia. It had unbranched uniseriate to multiseriate filaments less than 1 mm tall, with a unicellular base. Each cell had a single multilobed parietal chloroplast without a pyrenoid. During reproduction vegetative cells were discharged directly as monospores that remained motile for several hours after release. Spores with long tails moved more slowly (0.053–0.195 μm sχ) than spores without tails (0.43–1.76 μm s′¹). Phylo‐genetic analysis of sequences of the small subunit of the nuclear‐encoded rRNA and plastid‐encoded ribu‐lose bisphosphate carboxylase/oxygenase genes revealed that Purpureofilum is a member of the Stylonematales and is most closely related to the filamentous genus Bangiopsis. Bangiopsis differs from Purpureofilum by having longer (to 5 mm) multiseriate filaments, cells containing a stellate chloroplast, a conspicuous central pyrenoid, and monospores often formed in packets. Monospores of Bangiopsis were also motile. Transmission electron microscopy investigation of Purpureofilum and Bangiopsis revealed that the Golgi complexes are associated only with rough endo‐plasmic reticulum and that the plastid contains a peripheral thylakoid; this combination of features being the same as in all other multicellular members of the Stylonematales. The low molecular weight carbohydrates of Purpureofilum and Bangiopsis were digenea‐side and sorbitol, which were present in most other Stylonematales.     

A NEW AGAROPHYTE SPECIES,GELIDIUM EUCORNEUM SP. NOV. (GELIDIALES,RHODOPHYTA), BASED ON MOLECULAR AND MORPHOLOGICAL DATA1

Gelidium is an economically and ecologically important agar‐producing genus. Although the taxonomy of Gelidium has been the focus of many published studies, there is still a need to reevaluate species‐level diversity. Herein, we describe Gelidium eucorneum sp. nov. based on specimens collected off Geojedo on the southern coast of Korea. G. eucorneum is distinguished by cartilaginous thalli with brush‐like haptera, rhizoidal filaments concentrated in the medulla, and globose cytocarps that are horned with multiple determinate branchlets. The species occurs in wave‐exposed intertidal sites, sometimes in association with other mat‐forming algae. Phylogenetic analyses (rbcL, psaA, and cox1) reveal that G. eucorneum is unique and clearly distinct from other species of the genus. The clade containing Gelidium vagum and Acanthopeltis longiramulosa was resolved as a sister group to G. eucorneum. We suggest that the diverse morphologies of G. eucorneum, G. vagum, and Acanthopeltis developed from a common ancestor in East Asian waters. This study shows that even in well‐studied areas, more agarophyte species are to be added to the world inventory of red algae.     

Phylogenetics and population genetics of the louse fly,Lipoptena mazamae,from Arkansas,U.S.A.

Louse flies, also known as deer keds (Lipoptena mazamae Rondani), infest cervids such as white‐tailed deer, Odocoileus virginianus and vector pathogens such as Anaplasma and Bartonella schoenbuchensis to cattle and humans, respectively. The population genetic structure of 30 L. mazamae collected from white‐tailed deer in four regions of Arkansas, U.S.A., designated by county boundaries, was examined using DNA sequences of a 259‐bp region of the mitochondrial DNA rRNA 16S gene. Of the 259 nucleotide characters, 33 were variable and 6 haplotypes were identified. Two haplotypes occurred only once (haplotype 3 and 4), whereas two other haplotypes occurred in 43% (haplotype 1 in two regions) and 40% (haplotype 6 in three regions) of the samples. Phylogenetic relationships of the six L. mazamae haplotypes were constructed with other Hippoboscid and Glossinid samples and two clades resulted. Clade 1 was located in the north and western Ozarks whereas clade 2 was found in the northern and eastern Ozarks. Results from the present study indicate that Lipoptena may be a polyphyletic genus; consequently, more research into genetic variation within this genus is necessary.     

Aspidistra albopurpurea,A. khangii,A. lubae and A. stellata spp. nov. (Asparagaceae,Convallariaceae s.s.) from Indochina

Aspidistra albopurpurea, A. khangii, A. lubae and A. stellata discovered recently are described and illustrated as species new to science. All novelties are local endemics of northern Vietnam and southeastern Laos. Aspidistra albopurpurea is unique in the genus with a white/red stigma in striking contrast to the large milk‐white tube base. Aspidistra stellata belongs to a group of species with extraordinary long and narrow lineate perigone lobes. Aspidistra khangii and A. lubae belong to a small group of species having erect, vertical shoots. The first species is easily distinguished by its stout stem, down‐curved peduncle and obpyramidal, container‐shaped perianth. Aspidistra lubae has characteristic horizontal, peduncles and urceolate flowers. Besides the type variety, A. lubae includes var. lancifolia, which differs in its erect, lanceolate, immaculate, shortly petiolate leaves. Both varieties of A. lubae were found growing, often intermixed, in the same locality in very similar ecological conditions.     

Identification and typification of Nepenthes blancoi,with N. abalata sp. nov. from the western Visayas,Philippines

Nepenthes blancoi Bl. is neotypified on material conspecific with N. alata Blanco. Material previously associated with N. blancoi is described as N. abalata Jebb & Cheek sp. nov. This is a grassland species seemingly related to N. philippinensis Macfarl. but with similarities to the grassland species of Indo‐China. Its conservation status is assessed as ‘Critically Endangered’ (CR).     

Salvia officinalis L. Essential Oils from Spain: Determination of Composition,Antioxidant Capacity,Antienzymatic, and Antimicrobial Bioactivities

Four essential oils (EO s) from Salvia officinalis L. cultivated in Spain (Murcia Province) were analyzed by gas chromatography coupled with mass spectrometry (GC /MS ) to determine their relative and absolute compositions. The main components were α ‐thujone (22.8 – 41.7%), camphor (10.7 – 19.8%), 1,8‐cineole (4.7 – 15.6%), and β ‐thujone (6.1 – 15.6%). Enantioselective gas chromatography identified (?)‐α ‐thujone and (+)‐camphor as the main enantiomers in all the analyzed EO s. Furthermore, when the EO s were tested to determine their antioxidant activity against free radicals and as ferric reducing and ferrous chelating agents, all were seen to have moderate activity due to the compounds they contained, such as linalool or terpinene. Because of their known relation with inflammatory illnesses and Alzheimer 's disease, respectively, the inhibition of lipoxygenase and acetylcholinesterase was studied using the EO s. Some individual compounds also inhibited these enzymes. In addition, the studied EO s were able to inhibit the growth of Escherichia coli , Staphylococcus aureus , and Candida albicans . The characterization carried out increases our awareness of the possible uses of S . officinalis EO as natural additives in food, cosmetics, and pharmaceuticals.     

Simenoside A,a New Triterpenoid Saponin from Gypsophila simonii Hub.‐Mor.

Saponins are amphiphilic glycoconjugates which give soap‐like foams in H₂O. A new triterpenoid saponin, simenoside A ( 1 ), based on gypsogenin aglycone, and the known saponin 2 were isolated from Gypsophila simonii Hub.‐Mor. The structure of the new saponin was elucidated as 3‐O‐β‐D ‐galactopyranosyl‐(1→2)‐[β‐D ‐xylopyranosyl‐(1→3)]‐β‐D ‐glucuronopyranosylgypsogenin 28‐O‐β‐D ‐glucopyranosyl‐(1→3)‐[β‐D ‐glucopyranosyl‐(1→2)‐β‐D ‐xylopyranosyl‐(1→4)]‐α‐L ‐rhamnopyranosyl‐(1→2)‐β‐D ‐fucopyranosyl ester on the basis of extensive spectral analyses and chemical evidence. Saponins 1 and 2 were isolated from G. simonii for the first time.     

Development of species-specific primers for the ectoparasitic nematode species Xiphinema brevicolle, X. diffusum, X. elongatum, X. ifacolum and X. longicaudatum (Nematoda: Longidoridae) based on ribosomal DNA sequences

The objective of this study was to develop single‐step PCR species‐specific primers that reliably discriminate four economically important Xiphinema species (X. brevicolle, X. elongatum, X. ifacolum and X. longicaudatum) and X. diffusum that is taxonomically very similar to X. brevicolle. Each species‐specific reverse primer was located in the ITS‐1 rDNA region and was used in combination with a universal forward primer located in the 18S rDNA gene. Primer reliability was confirmed by screening seven and 11 populations, respectively of X. diffusum and X. elongatum. Potential species‐specific primers were also identified for X. brevicolle, X. longicaudatum and X. ifacolum, however too few populations of these species were available to thoroughly assess their reliability. For all species‐specific primers, specificity was demonstrated by the absence of cross‐reactions with 14 non‐target Xiphinema species. Multiplex PCR was effective and reproducible for two (X. longicaudatum and X. ifacolum) or three (X. brevicolle, X. diffusum and X. elongatum) of the target nematode species, thus improving the applicability of the diagnostic primers.     

Morphology,Conjugation, and Postconjugational Reorganization of Dileptus tirjakovae n. sp. (Ciliophora,Haptoria)

ABSTRACT. We studied the morphology, conjugation, and postconjugational reorganization of a new haptorid ciliate, Dileptus tirjakovae n. sp., using conventional methods. Dileptus tirjakovae is characterized by two abutting, globular macronuclear nodules and scattered brush kinetids. Conjugation is similar to that in congeners, that is, it is temporary, heteropolar, and the partners unite bulge‐to‐bulge with the proboscis. Some peculiarities occur in the nuclear processes: there are two synkaryon divisions producing four synkaryon derivatives, of which two become macronuclear anlagen, one becomes the micronucleus, and one degenerates. Unlike spathidiids, D. tirjakovae shows massive changes in body shape and ciliary pattern before, during, and after conjugation: early and late conjugants as well as early exconjugants resemble Spathidium, while mid‐conjugants resemble Enchelyodon. These data give support to the hypothesis that spathidiids evolved from a Dileptus‐like ancestor by reduction of the proboscis. Dileptus tirjakovae exconjugants differ from vegetative cells by their smaller size, stouter body, shorter proboscis, and by the lower number of ciliary rows, suggesting one or several postconjugation divisions. Although 83% of the exconjugants have the vegetative nuclear pattern, some strongly deviating specimens occur and might be mistaken for distinct species, especially because exconjugants are less than half as long as vegetative cells.     

Aedes albopictus oviposits with other Aedes species in artificial oviposition cups: a case study in Knox County,Tennessee, U.S.A.

Interspecific associations between two mosquito species can lead to effects such as competition, species displacement, and species stability. To better understand Aedes albopictus (Skuse) and other Aedes species, we monitored eggs in artificial oviposition cups (ovitraps) within Knox County, TN, U.S.A., during the 2016 and 2017 mosquito seasons. In 2016, one black and one white ovitrap were placed at 18 sites for 21 weeks, while in 2017 black and white ovitraps baited with grass‐infused or deionized water were placed at 11 sites for nine weeks. Eggs were identified to species and resulting counts were used to determine the degree of interspecific association using Cole's coefficients (C₇) and the degree of heterogeneity across space and time using generalized linear mixed models (GLMM). Here, Aedes mosquitoes are generally ovipositing in black cups with grass‐infused water, and Ae. albopictus eggs co‐occurred with other Aedes species more often than would be expected. Finding a positive significant interspecific association between Ae. albopictus and other Aedes eggs suggests that methods used to control Ae. albopictus may also control other Aedes mosquitoes. Finding that Ae. albopictus co‐occurs with other Aedes mosquitoes warrants additional research to evaluate outcomes associated with co‐occurrence within the study area.     

Coleifolides A and B,Two New Sesterterpenoids from the Aerial Parts of Scutellaria coleifolia H.Lév.

Coleifolides A and B ( 1 and 2 ), two new sesterterpenoids with a β‐methyl‐α,β‐unsaturated‐γ‐lactone moiety, were isolated from the aerial parts of Scutellaria coleifolia H.Lév . (Lamiaceae), together with three known compounds. Their structures were elucidated by NMR and MS examinations. Coleifolides A and B were concluded to be partially racemic compounds by the HPLC analysis using a chiral column or introduction of chiral derivatizing agents. The absolute configuration of the major isomer was determined by analyses of the CD spectrum as well as NMR data of (R)‐ and (S)‐2‐NMA derivatives. Coleifolides A and B are structurally similar to manoalide derivatives, previously isolated from marine sponges, and appear to be the first examples of this type of compounds being isolated from higher plants.     

Vegetative Characteristics of Recently Reforested Bottomlands in the Lower Cache River Watershed, Illinois, U.S.A.

Abstract Interest in restoring native ecosystems is resulting in conversion of marginal agricultural lands to bottomland hardwood‐dominated forests in the midwestern and midsouthern United States. Growing stock for these efforts typically consists of planted oak (Quercus spp.) and volunteer vegetation. Reports of mixed reforestation success and the lack of post‐establishment tree growth data prompted this evaluation of vegetation characteristics of 5‐ to 7‐year‐old operational restorations in the Lower Cache River Watershed in southernmost Illinois, U.S.A. Fraxinus pennsylvanica (green ash), Acer negundo (box‐elder), and Liquidambar styraciflua (sweetgum) together comprised 77% of all tree stems observed. Full stocking of overstory tree species can be expected to produce a closed canopy stand within 160 m of a forested edge, due primarily to the abundance of rapidly growing volunteer‐origin trees. Planted oaks contributed minimally to total tree stocking but were present in sufficient numbers to eventually improve wildlife habitat, and therefore satisfied restoration objectives. Oak height was 23% greater when in the presence of a non‐oak tree species. Herbaceous cover was dominated by Solidago gigantea (late goldenrod) and Juncus spp. (rushes). Solidago gigantea was associated with poor growth and low density of non‐oak stems, whereas Juncus dudleyi (Dudley's rush) was associated with taller non‐oak stems. These results suggest that the presence of volunteer‐origin trees is crucial for the creation of full stand stocking that will result in rapid development of a closed canopy forest. Improved success of future reforestation efforts will require more intensive methods to establish adequate stocking beyond 160 m of a forest edge. Methods described here could be adapted for agricultural field to forest restorations in other regions to predict critical distances from volunteer seed sources within which supplemental planting would be unnecessary to meet tree stocking objectives.     

THE RECLASSIFICATION OF PFIESTERIA SHUMWAYAE (DINOPHYCEAE): PSEUDOPFIESTERIA,GEN. NOV.1

Pfiesteria shumwayae Glasgow et Burkholder is assigned to a new genus Pseudopfiesteria gen. nov. Plate tabulation differences between Pfiesteria and Pseudopfiesteria gen. nov. as well as a maximum likelihood phylogenetic analysis based on rDNA sequence data warrant creation of this new genus. The Kofoidian thecal plate formula for the new genus is Po, cp, X, 4′, 1a, 6′′, 6c, PC, 5+s, 5′′′, 0p, 2′′′′. In addition to having six precingular plates, P. shumwayae comb. nov. also has a distinctive diamond or rectangular‐shaped anterior intercalary plate. Both Pfiesteria and Pseudopfiesteria gen. nov. are reassigned to the order Peridiniales based on an apical pore complex (APC) with a canal (X) plate that contacts a symmetrical 1′, four to five sulcal plates, and the conservative hypothecal tabulation of 5′′′, 0p, and 2′′′′. These morphological characters and the life histories of Pfiesteria and Pseudopfiesteria are consistent with placement of both genera in the Peridiniales. Based on the plate tabulations for P. shumwayae, P. piscicida, and the closely related “cryptoperidiniopsoid” and “lucy” groups, the family Pfiesteriaceae is amended to include species with the following tabulation: 4‐5′, 0‐2a, 5‐6′′, 6c, PC, 5+s, 5′′′, 0p, and 2′′′′ as well as an APC containing a pore plate (Po), a closing plate (cp), and an X plate; the tabulation is expanded to increase the number of sulcal plates and to include a new plate, the peduncle cover (PC) plate. Members of the family have typical dinoflagellate life cycles characterized by a biflagellated free‐living motile stage, a varying number of cyst stages, and the absence of multiple amoeboid stages.     

Picrorhiza tungnathii sp. nov. from western Himalaya,India

The new species Picrorhiza tungnathii Pusalkar is described and illustrated from the western Himalaya, India. It is distinguished from the allied P. kurrooa Royle ex Benth. by being 10–25 cm tall, having a moderately dense, (10–)15–25‐flowered spike, a zygomorphic, 2‐lipped, glandular‐ciliate corolla that is equaling or slightly exceeding the calyx and partly visible or sub‐exserted between the calyx lobes, a conspicuously long (longer than the corolla lobes), curved corolla tube (1/2–4/5 the length of the calyx), unequal corolla lobes that are ovate‐lanceolate to lanceolate, acute to sub‐acuminate and moderately ciliate, a mid‐lobe of the upper lip that is obliquely erect, galeate with retuse or emarginate apex, lateral corolla lobes that are slightly smaller than the mid‐lobe of the upper lip, a lower corolla lip that is slightly shorter than the lateral lobes, obliquely erect or spreading, didynamous, long‐exserted (2–3 times the corolla) stamens, pollen of the Paederota‐type and a style that is 2–3 times as long as the corolla.     

Apurinic/apyrimidinic endonuclease 1, p53, and thioredoxin are linked in control of aging in C. elegans

Deletions in mitochondrial DNA (mtDNA) accumulate during aging. Expression of the Caenorhabditis elegans apurinic/apyrimidinic endonuclease 1 (APE1) ortholog exo‐3, involved in DNA repair, is reduced by 45% (P < 0.05) during aging of C. elegans. Suppression of exo‐3 by treatment with RNAi resulted in a threefold increase in mtDNA deletions (P < 0.05), twofold enhanced generation of reactive oxygen species (ROS) (P < 0.01), distortion of the structural integrity of the nervous system, reduction of head motility by 43% (P < 0.01) and whole animal motility by 38% (P < 0.05). Suppression of exo‐3 significantly reduced life span: mean life span decreased from 18.5 ± 0.4 to 15.4 ± 0.1 days (P < 0.001) and maximum life span from 25.9 ± 0.4 to 23.2 ± 0.1 days (P = 0.001). Additional treatment of exo‐3‐suppressed animals with a mitochondrial uncoupler decreased ROS levels, reduced neuronal damage, and increased motility and life span. Additional suppression of the C. elegans p53 ortholog cep‐1 in exo‐3 RNAi‐treated animals similarly decreased ROS levels, preserved neuronal integrity, and increased motility and life span. In wild‐type animals, suppression of cep‐1, involved in downregulation of exo‐3, increased expression of exo‐3 without a significant effect on ROS levels, preserved neuronal integrity, and increased motility and life span. Suppression of the C. elegans thioredoxin orthologs trx‐1 and trx‐2, involved in the redox chaperone activity of exo‐3, overrides the protective effect of cep‐1 RNAi treatment on neuronal integrity, neuronal function, mean and maximum life span. These results show that APE1/EXO‐3, p53/CEP‐1, and thioredoxin affect each other and that these interactions determine aging as well as neuronal structure and function.