Preliminary investigations into a potential ant invader in Kruger National Park,South Africa

The super‐abundance of Lepisiota incisa (Forel) in settlement areas of Kruger National Park, South Africa has raised concerns that it might be exotic and could negatively impact on natural ecosystems. We documented the current distribution of this ant species around the main settlement in Kruger, assessed how ant diversity varies across habitats, and investigated potential mechanisms facilitating dominance by L. incisa. Around the main camp of Skukuza, pitfall traps were set in five habitats differing in anthropogenic influence. Baiting trails were conducted to determine whether L. incisa and native ants differed in numerical and behavioural dominance. Aggression assays were performed on L. incisa to provide information on colony structure. Although L. incisa was found in all habitats, it had a significantly higher abundance in gardens and appears confined to human‐disturbed areas. It was numerically dominant recruiting more workers to food baits than all other ant species combined. Aggression levels were low between most nests of L. incisa indicating a potential supercolony structure. More information is urgently needed on the genetics, physiology and origins of L. incisa, and monitoring of its current distribution is recommended. This species warrants attention because there may be significant potential for overseas invasion.     

Isozyme evidence on the specific distinctness and phylogenetic position of <Emphasis Type="Italic">Vicia incisa</Emphasis> (Fabaceae)

Vicia incisa is a taxonomically controversial species that has been also treated as a subspecies of V. sativa because of a great morphological similarity. The phylogenetic position of V. incisa is uncertain because various DNA markers have provided contradictory results. Isozymes of V. incisa encoded by 15 loci and resolved with the use of polyacrylamide gel electrophoresis (PAGE) are described and compared with those of seven related species belonging to sections Vicia, Sepium, Lathyroides and Pseudolathyrus in order to get new evidence about its taxonomic rank and phylogenetic position. Phylogenetic relationships are analyzed with maximum parsimony and neighbour joining methods. Vicia incisa is shown to differ from all three subspecies of V. sativa including, sativa, cordata and nigra, by alternate variants of ten isozymes out of 15 analysed. Instead, V. incisa has much more similarity to V. grandiflora and V. sepium by sharing eight isozyme variants which differ from the subspecies of V. sativa. The most parsimony and neighbour joining analyses of the isozyme variation placed V. incisa as basally linked to the V. grandiflora and V. sepium species couple in the clade of section Sepium (= sect. Atossa), while the subspecies of V. sativa together with V. lathyroides formed a separate clade of section Vicia. The isozyme data provide further support to the species status of V. incisa.     

Chemical Composition and Antimicrobial Activity of Essential Oils from the Aerial Parts of Asteriscus graveolens (Forssk.) Less. and Pulicaria incisa (Lam.) DC.: Two Asteraceae Herbs Growing Wild in the Hoggar

In recent years, antimicrobial activities of essential oils have been intensively explored, mainly in researching and developing new antimicrobial agents to overcome microbial resistance. The present study investigates the chemical composition and antimicrobial activities of essential oils obtained from two Asteraceae: Asteriscus graveolens (Forssk .) Less . and Pulicaria incisa (Lam .) DC. Chemical analysis was performed using a combination of capillary GC‐FID and GC/MS analytical techniques. The major component of Asteriscus graveolens were cis‐chrysanthenyl acetate (31.1%), myrtenyl acetate (15.1%), and kessane (11.5%), while for Pulicaria incisa the main components were chrysanthenone (45.3%) and 2,6‐dimethylphenol (12.6%). The oils obtained from the aerial parts were tested against sixteen microbial strains by agar well diffusion technique and dilution methods and showed minimum inhibitory concentrations (MIC) in the range of 19 – 1250 μg/ml. A good antibacterial activity against a common nosocomial pathogen, Acinetobacter baumanniiATCC 19606 was observed, especially from Pulicaria incisa essential oil, with a MIC value up to 19 μg/ml. These results give significant information about the pharmacological activity of these essential oils, which suggest their benefits to human health, having the potential to be used for medical purposes.     

Effects of light intensity and nitrogen starvation on growth, total fatty acids and arachidonic acid in the green microalga Parietochloris incisa

The effects of light and nitrogen deficiency on biomass, fatty acid content and composition were studied in Parietochloris incisa, the unicellular freshwater chlorophyte accumulating very high amounts of arachidonic-acid-rich triacylglycerols. P. incisa cultures grown on complete nutrient medium and under high light (400 μmol photons m^{− 2} s⁻¹) showed the highest rate of growth in comparison to medium (200 μmol photons m⁻² s⁻¹) and low (35 μmol photons m⁻² s⁻¹) light intensity. Cultures grown under high light (on complete BG-11 medium) attained higher volumetric contents of total fatty acids and arachidonic acid due to greater increase in biomass. Nitrogen starvation brought about a strong increase in the arachidonic acid proportion of total fatty acids. Thus, adjustments to cultivation conditions could serve as an efficient tool for manipulation of yield and relative content of arachidonic acid in P. incisa. The significance of the changes in lipid metabolism for adaptation of P. incisa to high-light stress and nitrogen deficiency is also discussed.     

A multivariate morphological analysis of the Cardamine concatenata alliance (Brassicaceae)

Principal coordinate analysis using Gower’s similarity coefficient and canonical discriminant analysis were performed using morphological data to assess species distinctness in theCardamine concatenata alliance, comprising the eastern North American dentarias. The results of an overall principal coordinate analysis of the alliance using quantitative and qualitative characters revealed distinct and separate clusters that correspond to five previously recognized species (C. angustata, C. concatenata, C. diphylla, C. dissecta, C. maxima). One additional taxon,C. incisa, was found to group closely with theC. diphylla cluster. In a second principal coordinate analysis including only individuals fromC. diphylla andC. incisa, both are supported as distinct morphological entities. Canonical discriminant analysis of only above-ground quantitative characters was less effective in distinguishing previously recognized taxa, in keeping with the emphasis on qualitative characters in previous taxonomic treatments of the group.Cardamine incisa (Eames) Schumann (pro hybr.) is raised to the rank of species. A key to the species of theCardamine concatenata alliance is provided.     

Cytogenetic studies on natural intergeneric hybridization inAster alliances

The karyotype and meiosis of the 12-ploid plants—one of the offspring of the natural F₁ hybrid (Aster ageratoides subsp.ovatus (2n=36) ×Kalimeris incisa (2n=72), 2n=72)—were examined. The 2n=108 chromosomes of the 12-ploids were found to consist of 18 large chromosomes and 90 small chromosomes. In meiosis of the PMCs of the 12-ploid, chromosome configurations of 3_III+46_II+7_I, 2_III+48_II+6_I and 3_III+47_II+5_I were observed. All the univalents and trivalents were small, and among the 46–48 bivalents nine were large and the remaining 37–39 were comparatively small. The large bivalents were found to represent autosyndetic pairings, and the small bivalents and trivalents were probably formed by autosyndetic pairings. The large chromosomes of the 12-ploids were found to coincide with the large chromosomes ofovatus, and the 90 small chromosomes to correspond to small chromosomes ofovatus andK. incisa. The 12-ploids were concluded to have been produced by a fusion of an unreduced gamete of the F₁ plant and a reduced gamete ofK. incisa which was growing in proximity to the F₁s. Thus the 12-ploids were regarded to be an amphidiploid having 36 chromosomes ofovatus and 72 chromosomes ofK. incisa.     

EFFECT OF NITROGEN STARVATION ON OPTICAL PROPERTIES,PIGMENTS, AND ARACHIDONIC ACID CONTENT OF THE UNICELLULAR GREEN ALGA PARIETOCHLORIS INCISA (TREBOUXIOPHYCEAE,CHLOROPHYTA)1

Spectral properties of cell suspensions, individual cells, and extracts of the unicellular green alga Parietochloris incisa (Reisigl) Shin Watan. grown under low light were studied. Long‐term nitrogen (N) deprivation resulted in a decrease of chloroplast volume, appearance of numerous large cytoplasmic oil bodies, and the deposition of triacylglycerols with a high proportion of arachidonic acid. Chlorophylls a and b underwent a synchronous decline, whereas carotenoids (Car) showed a relative increase. Simultaneously, significant qualitative changes in the spectral properties of P. incisa individual cells, cell extracts, and cell suspensions were observed. To a large extent, the spectral changes observed in cell suspension could be attributed to a decrease in overall pigment content, leading to a gradual weakening of the so‐called package effect and accumulation of additional amounts of Car over chl, most probably, in oil bodies. Several optical characteristics of cell suspensions could serve as sensitive indicators of N‐deficiency in P. incisa. Furthermore, the absorption ratios, A₄₇₆/A₆₇₆ and A₆₅₀/A_676, showed close correlations with the Car‐to‐chl ratio and relative arachidonic acid (AA) content, respectively. The latter makes it possible to suggest that the increase in AA percentage in P. incisa proceeds in parallel with a decrease in cell chl content, accounting for the weakening of the package effect. N‐replenishment resulted in complete recovery of cell optical properties. The possible significance of the changes in cell ultrastructure, pigments, lipids, and optical properties is discussed with special reference to the ability of algae to adapt to and survive under conditions of long‐term nutrient deficiency.     

Peroxidases as taxonomic characters in Lophozia incisa (Schrad.) Dum. and Lophozia opacifolia (Culm.) Meyl.

Abstract

Peroxidases in 21 samples of Lophozia incisa from the Tatra Mountains and 14 samples of L. opacifolia from the Tatra Mountains and the Swiss Alps were electrophoretically assayed. Although both taxa proved to be polymorphic, consistent differences were found between them. This indicates that electrophoretic peroxidase phenotypes provide an additional taxonomic character supporting the morphological and ecological differences, a summary of which is provided.     

Carotenoid-to-chlorophyll ratio as a proxy for assay of total fatty acids and arachidonic acid content in the green microalga Parietochloris incisa

The relationships between pigment (carotenoid and chlorophyll) content with accumulation of total fatty acids (TFA) and arachidonic acid (AA) were studied in the green microalga Parietochloris incisa (Trebouxiophyceae, Chlorophyta) grown under different PFDs (35, 200, and 400 μmol photons m⁻² s⁻¹) and nitrogen availabilities. The growth of P. incisa under higher light and nitrogen deficiency was accompanied by accumulation of FA, an increase in carotenoid and a decline in chlorophyll content. It was found that the carotenoid-to-chlorophyll ratio (but not the individual pigment content) correlates closely with the volumetric content of both TFA and AA. Analysis of scattering-compensated absorption spectra of P. incisa suspensions revealed their tight relationship in the blue-green range of the spectrum with the carotenoid-to-chlorophyll ratio, TFA, and AA content. These findings allowed the development of algorithms for the non-destructive assay of TFA and AA in cell suspensions in the ranges of 0.09–3.04 and 0.04–1.7 μg mL⁻¹, with accuracy of 0.06 and 0.01 μg mL⁻¹, respectively, via analytically measured carotenoid-to-chlorophyll ratio and using the ratio of absorption coefficients at 510 and 678 nm, with accuracy of 0.07 and 0.02 μg mL⁻¹, respectively. The feasibility of obtaining essential spectral information concerning the physiological condition of P. incisa using a standard spectrophotometer is also shown.     

Resilience to Freezing in the Vegetative Cells of the Microalga Lobosphaera incisa (Trebouxiophyceae,Chlorophyta)

The chlorophyte microalga Lobosphaera incisa was isolated from the snowy slopes of Mt. Tateyama in Japan. This microalga stores exceptionally high amounts of the omega-6 LC-PUFA arachidonic acid in triacylglycerols, and therefore represents a potent photosynthetic source for this essential LC-PUFA. Assuming that freezing tolerance may play a role in adaptation of L. incisa to specific ecological niches, we examined the capability of L. incisa to tolerate extreme sub-zero temperatures. We report here, that the vegetative cells of L. incisa survived freezing at −20°C and −80°C (over 1 month), without cryoprotective agents or prior treatments. Cells successfully recovered upon thawing and proliferated under optimal growth conditions (25°C). However, cells frozen at −80°C showed better recovery and lower cellular ROS generation upon thawing, compared to those preserved at −20°C. Photosynthetic yield of PSII, estimated by F_v/F_m, temporarily decreased at day 1 post freezing and resumed to the original level at day 3. Interestingly, the thawed algal cultures produced a higher level of chlorophylls, exceeding the control culture. The polar metabolome of the vegetative cells comprised a range of compatible solutes, dominated by glutamate, sucrose, and proline. We posit that the presence of endogenous cryoprotectants, a rigid multilayer cell wall, the high LC-PUFA content in membrane lipids, and putative cold-responsive proteins may contribute to the retention of functionality upon recovery from the frozen state, and therefore for the survival under cryospheric conditions. From the applied perspective, this beneficial property holds promise for the cryopreservation of starter cultures for research and commercial purposes.     

Dynamics of pleustonic ostracod populations in small ponds on the Island of Martín García (Río de la Plata,Argentina)

The biogeography and ecological preferences of Neotropical freshwater ostracods are poorly known, and more so the dynamics of populations and habitat selection of species living in pleustonic environments of temporary ponds. In the present survey we analyze the population changes of ostracods living in pleustonic environments of small freshwater bodies on Martín García Island (Río de la Plata, Argentina). Between June 2005 and June 2007, monthly samples of floating vegetation from eight different ponds on the island were collected, and limnological parameters were measured in situ. The results of multivariate logistic regression showed that the presence of ostracods was significantly related to high dissolved oxygen content and high water temperature. In addition, multivariate regression analysis indicated that, when ostracods were present, their total abundance was negatively related to floating vegetation dry weight. Four ostracod species were found: Strandesia bicuspis, Chlamydotheca incisa, Cypridopsis vidua, and Bradleytriebella trispinosa. The seasonal variation in abundances indicated that populations of the most common species, S. bicuspis and C. incisa, were denser during the summer and autumn months. The results of canonical correspondence analysis showed that individuals of S. bicuspis were more abundant at higher temperatures and lower conductivity than C. incisa. Further research is needed to clarify the observed negative correlation between floating vegetation dry weight and ostracod density and the possible differential thermal preference of the two species studied.     

The effects of ultraviolet radiation on photosynthetic performance,growth and sunscreen compounds in aeroterrestrial biofilm algae isolated from building facades

The effects of artificial ultraviolet radiation [UVR; 8 W m⁻² ultraviolet-A (UVA), 0.4 W m⁻² ultraviolet-B (UVB)] on photosynthetic performance, growth and the capability to synthesise mycosporine-like amino acids (MAAs) was investigated in the aeroterrestrial green algae Stichococcus sp. and Chlorella luteoviridis forming biofilms on building facades, and compared with the responses of two green algae, from soil (Myrmecia incisa) and brackish water (Desmodesmus subspicatus). All species exhibited decreasing quantum efficiency (F _v/F _m) after 1–3 days exposure to UVR. After 8–12 days treatment, however, all aeroterrestrial isolates exhibited full recovery under UVA and UVA/B. In contrast, D. subspicatus showed only 80% recovery after treatment with UVB. While Stichococcus sp. and C. luteoviridis exhibited a broad tolerance in growth under all radiation conditions tested, M. incisa showed a significant decrease in growth rate after exposure to UVA and UVA/B. Similarly D. subspicatus grew with a reduced rate under UVA, but UVA/B led to full inhibition. Using HPLC, an UV-absorbing MAA (324 nm-MAA) was identified in Stichococcus sp. and C. luteoviridis. While M. incisa contained a specific 322 nm-MAA, D. subspicatus lacked any trace of such compounds. UV-exposure experiments indicated that all MAA-containing species are capable of synthesizing and accumulating these compounds, thus supporting their function as an UV-sunscreen. All data well explain the conspicuous ecological success of aeroterrestrial green algae in biofilms on facades. Biosynthesis and accumulation of MAAs under UVR seem to result in a reduced UV-sensitivity of growth and photosynthesis, which consequently may enhance survival in the environmentally harsh habitat.     

ENTOMOPHILOUS,INTRAFLORAL POLLINATION IN PHYLA INCISA

Experimental pollination studies indicate that probing into the corolla tube by insects is a requirement for autogamous reproduction in Phyla incisa. Insect-mediated self-fertilization combines the anthecology of allogamous reproduction with the genetic consequences of autogamy.     

Helminth parasites of the Eurasian badger (<Emphasis Type="Italic">Meles meles</Emphasis> L.) in the Basque Country (Spain)

The helminthological study of 26 Eurasian badgers (Meles meles L.) in the Basque Country (northern Spain) revealed the presence of two trematode (Euryhelmis squamula and Brachylaima sp.), three cestode (Atriotaenia incisa, Mesocestoides sp. and Taenia sp.) and five nematode (Aonchotheca putorii, Physaloptera sp., Molineus patens, Uncinaria criniformis and Strongyloides sp.) species. All 15 individuals analysed for Trichinella sp. were negative. The coprological analysis (flotation and migration) revealed the excretion of strongylid, capillariid and trematode eggs, and Crenosoma sp. and Angiostrongylus sp. larvae. No sex- or age-related differences were found in the parasite burden or egg and larvae excretion. A. incisa and M. patens abundances were positively correlated, both species being negatively correlated with badger weight. The presence of adults of U. criniformis and strongylid eggs in faeces was not independent. Badgers excreting Angiostrongylus larvae were in poorer condition than those not excreting.     

Fertilizer impacts on the understorey of a regenerating Mountain Ash (Eucalyptus regnans F. Muell.) forest,Victoria

Abstract We examined the effects of adding fertilizers on nutrient concentrations in foliage, and on species composition and density of the understorey in a 10 year old Mountain Ash (Eucalyptus regnans F. Muell.) forest. Nutrient concentrations in foliage of three woody understorey species (Acacia dealbata, Pomaderris aspera and Olearia argophylla) showed no significant response to the addition of phosphorus and nitrogen either alone, in combination or combined with other nutrients. The phosphorus concentration in foliage and stems of two herbaceous species (Australina pusilla and Urtica incisa) was significantly increased by applying phosphorus. The addition of fertilizers had no significant effect on species composition and density of the understorey.     

NITROGEN STARVATION INDUCES THE ACCUMULATION OF ARACHIDONIC ACID IN THE FRESHWATER GREEN ALGA PARIETOCHLORIS INCISA (TREBUXIOPHYCEAE)1

The Chlorophyte Parietochloris incisa comb. nov (Trebuxiophyceae) was found to be the richest plant source of the pharmaceutically valuable long‐chain polyunsaturated fatty acid (PUFA), arachidonic acid (20:4ω6, AA). Over 90% of total AA are deposited in triacylglycerols (TAG). Under nitrogen starvation, the fatty acid content constituted over 35% of dry weight and the proportion of AA exceeded 60% of total fatty acids. Consequently, we obtained an AA content of over 20%. This is, to the best of our knowledge, the highest reported content of any PUFA in algae. Increasing the biomass concentration resulted in an enhancement of both the proportion of AA and the fatty acid content. We hypothesize that one of the roles of TAG in P. incisa is to serve as a reservoir of AA that can be used for the construction of membranal lipids.     

Life cycle and morphology ofMelampsora yezoensis onSalix serissaefolia

Uredinial and telial states of aMelampsora species occurring on the leaves ofSalix serissaefolia were for the first time recorded. Field observations and inoculation experiments showed that the spermogonial and aecial states of the fungus were formed onChelidonium majus var.asiaticum andCorydalis incisa. The fungus was identified asMelampsora yezoensis based on the morphological observation of all the spore states. Urediniospores were able to infectS. erocarpa andS. pierotii as well asS. serissaefolia andS. jessoensis. Salix serissaefolia, S. eriocarpa, andS. pierotii are new uredinial and telial hosts ofM. yezoensis, andC. majus var.asiaticum andC. incisa are new spermogonial and aecial hosts of the fungus. Contribution No. 124, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry University of Tsukuba.     

Identification of four distinct ‘Candidatus Phytoplasma’ species in pomegranate trees showing witches' broom,little leaf and yellowing in Jordan,and preliminary insights on their putative insect vectors and reservoir plants

During field surveys conducted in northern Jordan from June to November 2020, phytoplasma-like symptoms, including leaf yellowing/reddening and rolling, little leaf and witches' broom were observed in pomegranate. Disease incidence in 22 surveyed orchards ranged from 30% to 65%. Nested PCR-based amplification of 16S rRNA gene detected phytoplasmas in 17% of collected symptomatic pomegranate trees. Amplicon nucleotide sequence analyses allowed attributing the detected phytoplasmas to ‘Candidatus Phytoplasma solani’, ‘Ca. P. aurantifolia’, ‘Ca. P. asteris’ and ‘Ca. P. ulmi’. These phytoplasmas were found in plants showing specific symptoms and differentially distributed in the considered locations. Additionally, three cicadellids (Macrosteles sexnotatus, Cicadulina bipunctata and Psammotettix striatus) and two non-crop plants (Plantago major and Capsicum annuum) resulted hosting ‘Ca. P. asteris’ strains, and one cicadellid (Balclutha incisa) was carrying a ‘Ca. P. solani’ strain. A new pomegranate disease complex associated with multiple phytoplasmas, including ‘Ca. P. aurantifolia’ and ‘Ca. P. ulmi’, never reported before in this host plant, is described here. Moreover, preliminary indications are provided on its possible epidemiology in Jordan, involving two putative insect vectors (M. sexnotatus, B. incisa) first reported in the Country.