Interactions between fungi and bacteria influence microbial community structure in the Megachile rotundata larval gut

Recent declines in bee populations coupled with advances in DNA-sequencing technology have sparked a renaissance in studies of bee-associated microbes. Megachile rotundata is an important field crop pollinator, but is stricken by chalkbrood, a disease caused by the fungus Ascosphaera aggregata. To test the hypothesis that some gut microbes directly or indirectly affect the growth of others, we applied four treatments to the pollen provisions of M. rotundata eggs and young larvae: antibacterials, antifungals, A. aggregata spores and a no-treatment control. We allowed the larvae to develop, and then used 454 pyrosequencing and quantitative PCR (for A. aggregata) to investigate fungal and bacterial communities in the larval gut. Antifungals lowered A. aggregata abundance but increased the diversity of surviving fungi. This suggests that A. aggregata inhibits the growth of other fungi in the gut through chemical or competitive interaction. Bacterial richness decreased under the antifungal treatment, suggesting that changes in the fungal community caused changes in the bacterial community. We found no evidence that bacteria affect fungal communities. Lactobacillus kunkeei clade bacteria were common members of the larval gut microbiota and exhibited antibiotic resistance. Further research is needed to determine the effect of gut microbes on M. rotundata health.     

Disturbances on a Wooded Raised Bog—How Windthrow, Bark Beetle and Fire Affect Vegetation and Soil Water Quality?

Pinus rotundata dominated peatbog (?ofinka Nature Reserve) in the T?eboň Basin, Czech Republic, was affected by “natural” disturbances: wind damage (1984), followed by a bark beetle attack, and fire (1994, 2000). Phytosociological relevés were used to document vegetation. Soil water chemistry was compared in three differently affected stands: (1) an undisturbed Pinus rotundata bog forest, (2) a windthrow – bark beetle affected stand and (3) a site burned by wildfire in 2000. The species composition of the windthrow – bark beetle affected sites and the undisturbed P. rotundata bog forest differed mainly in the shrub and tree layers. Burned sites were partly colonized by anemochorous species (e.g. Taraxacum sp. div.) that disappeared within two or three years after colonization. Bare peat was colonized by bryophytes (e.g. Marchantia polymorpha and Funaria hygrometrica) typical of the disturbed sites, and by Polytrichum sp. div. and Aulacomnium palustre. Most plant species characteristic of the P. rotundata bog forest occurred at the burned sites eight years after the fire, but in different abundances. The edificator of the former community—P. rotundata—was mostly absent. Compared with windthrow followed by the bark beetle attack, fire promoted rapid expansion of Molinia caerulea. Soil water in both the undisturbed P. rotundata bog forest and the windthrow – bark beetle affected sites had a similar composition: very low pH values, high P concentrations, low concentrations of cations (Ca²⁺, Mg²⁺and K⁺) and inorganic nitrogen. The concentrations of soluble reactive phosphorus (SRP) and ${\text{NH}}_4^ + - {\text{N}}$ were negatively correlated with the groundwater table. Total P, SRP and ${\text{NH}}_4^ + - {\text{N}}$ concentrations in the soil water at the burned site were by one order of magnitude higher than those in the P. rotundata bog forest, while concentrations of K⁺, Mg²⁺ and Ca²⁺ were only about two times higher. High concentrations of P and N in the soil water found three years after the fire indicated a long-term elevated nutrient content in the soil water.     

An Orphan Protist Quadricilia rotundata Finally Finds Its Phylogenetic Home in Cercozoa

Quadricilia rotundata is a heterotrophic flagellate with four flagella. However, because this species has no clear morphological characteristics or molecular data affiliating it with any known group, Q. rotundata has been treated as a protist incertae sedis, for a long time. Here, we established a clonal culture of Q. rotundata and sequenced its 18S rDNA sequence. Molecular phylogenetic analysis successfully placed Q. rotundata in an environmental clade within Cercozoa, which contributes to expand the morphological and species diversity within Cercozoa. We also discuss morphological evolution within Cercozoa based on this finding.     

Lumicella,a new genus of the tribe Empoascini (Hemiptera,Cicadellidae, Typhlocybinae) from China

Lumicella rotundata gen. et sp. n. is described based on specimens from Fujian Province, China. Habitus photos and illustrations of male genitalia of this new species are provided. Differences between the new genus and closely related genera are discussed.     

Antifungal phenanthrenes in yam tubers

An extract from the peel of yams (Dioscorea rotundata) showed anti-fungal activity towards both Cladosporium cladoporioides and a variety of ya     

Growth patterns in some indo-west-pacific seagrasses

The plastochrone interval of the rhizomes (PIR) was determined in the monopodially branching seagrasses Cymodocea rotundata Ehrenb. et Hempr. ex Aschers. (1981), Cymodocea serrulata (R.Br.) Aschers. et Magnus (1981), Syringodium isoetifolium (Aschers.) Dandy (1982), Halodule uninervis (Forssk.) Aschers. (1982) and Halophila ovalis (R.Br.) Hook.f. (1982). The growth rate during 1981 was in all species, including Thalassia hemprichii (Ehrenb.) Aschers., constant during the year and the PIR amounted to 6.7 days (Cymodocea serrulata) and 3.9 days (Cymodocea rotundata). However, the PIR observed in 1982 showed in all species a unimodal pattern and a decrease during June, July and August. The PIR was 6.3 days in Halodule uninervis while an interval of 5.2 days was calculated in Syringodium isoetifolium.The growth rates of the horizontal and vertical axes (rhizomes and shoots) were correlated. A regression between the number of leaves on the shoot and the number of nodes on the rhizome resulted for most species in a linear correlation. The slopes of the calculated correlation were rather similar in the tree species with a comparable morphology: 0.44 in Cymodocea serrulata, 0.43 in Cymodocea rotundata and 0.49 in Halodule uninervis. In Syringodium isoetifolium, the longevity of the leaves increased during ageing of the shoot.The proliferation ratio (the ratio shoots/rhizome apices) was measured in all species. A maximum of 0.41 was observed in Thalassia hemprichii. This species and Cymodocea rotundata showed a unimodal pattern. In the last species, the ratio ranged from 0.03 to 0.13. The ratio was constant during the year in Cymodocea serrulata and Syringodium isoetifolium. Proliferation by means of monopodially branching of the rhizome was of little importance in Halodule uninervis. The pattern of proliferative and reiterative branching in the Cymodoceoideae is illustrated by an image of 123 days of growth in the fastest growing specimen.Most species showed growth in fronts. The expansion of monospecific seagrass fringes was monitored by repetitive mapping. The pattern of succession in subtidal meadows was determined.     

Salinity influence on the ecology of phytoflagellate blooms in lower new york bay and adjacent waters

The possible rôle of salinity in phytoflagellate blooms in New York Harbor estuarine and oceanic waters was examined by culture studies of the dominant species. Massartia rotundata (Lohmann) Schiller (=Katodinium rotundatum (Lohmann) Loeblich III) grew best in the range 24–30‰, Olisthodiscus luteus Carter in the range 10–36‰, and Prorocentrum micans Ehrenberg in the range 27–36‰. The optimum ranges for all three species sufficiently match the characteristic bay-ocean range (17–32‰) to eliminate salinity tolerance as a major factor in bloom development. Laboratory simulation of the change from brackish-river to ocean water salinities, by culture preconditioning at 20‰ or at 30‰ before inoculation into a salinity gradient, shows stress effects on growth, especially in Massartia rotundata and Olisthodiscus luteus. It is speculated that salinity stress may temporarily affect the development of particular blooms of these two species.     

Effects of Fungicide and Adjuvant Sprays on Nesting Behavior in Two Managed Solitary Bees,Osmia lignaria and Megachile rotundata

There is a growing body of empirical evidence showing that wild and managed bees are negatively impacted by various pesticides that are applied in agroecosystems around the world. The lethal and sublethal effects of two widely used fungicides and one adjuvant were assessed in cage studies in California on blue orchard bees, Osmia lignaria, and in cage studies in Utah on alfalfa leafcutting bees, Megachile rotundata. The fungicides tested were Rovral 4F (iprodione) and Pristine (mixture of pyraclostrobin + boscalid), and the adjuvant tested was N-90, a non-ionic wetting agent (90% polyethoxylated nonylphenol) added to certain tank mixtures of fungicides to improve the distribution and contact of sprays to plants. In separate trials, we erected screened cages and released 20 paint-marked females plus 30–50 males per cage to document the behavior of nesting bees under treated and control conditions. For all females in each cage, we recorded pollen-collecting trip times, nest substrate-collecting trip times (i.e., mud for O. lignaria and cut leaf pieces for M. rotundata), cell production rate, and the number of attempts each female made to enter her own or to enter other nest entrances upon returning from a foraging trip. No lethal effects of treatments were observed on adults, nor were there effects on time spent foraging for pollen and nest substrates and on cell production rate. However, Rovral 4F, Pristine, and N-90 disrupted the nest recognition abilities of O. lignaria females. Pristine, N-90, and Pristine + N-90 disrupted nest recognition ability of M. rotundata females. Electroantennogram responses of antennae of O. lignaria females maintained in the laboratory did not differ significantly between the fungicide-exposed and control bees. Our results provide the first empirical evidence that two commonly used fungicides and a non-ionic adjuvant can disrupt nest recognition in two managed solitary bee species.     

Aspects of production and biomass of four seagrass species (Cymodoceoideae) from Papua New Guinea

Biomass and production data of the seagrasses Cymodocea serrulata (R. Brown) Aschers. and Magnus, Cymodocea rotundata Ehrenb. et Hempr. ex Aschers., Halodule uninervis (Forssk.) Aschers. and Syringodium iksoetifolium (aschers.) Dandy were collectede in monospecific stands in Bootless Inlet, Papua New Guinea. Cymodocea serrulata and Cymodocea rotundata were studied from November 1980 to November 1981. Total annual mean biomass was 354 and 201 g ADW m⁻², respectively. The largest proportion of these biomass values was contributed by the rhizomes (49 and 36%, respectively) and leaf biomass was ± 30% for both species. Halodule uninervis was studied at an intertidal and a subtidal site. The highest total annual mean biomass (600 g ADW m⁻²) was recorded at the intertidal site, of which 85% was found below ground. The largest proportion of the biomass, at both sites, was contributed by the below-ground vertical axes of the shoots. The biomass of the rhizomes was relatively low (9–12%) for Halodule uninervis. Proportionally, the largest above-ground biomass (40%) was recorded for Syringodium isoetifolium, of which the annual mean biomass was 481 g ADW m⁻².Total production (above and below ground) was 4.9 and 3.0 g ADW m⁻² day⁻¹ for Cymodocea serrulata and Cymodocea rotundata, respectively. Approximately 70% was production of leaves. Total production amounted to 6.0 and 4.0 g ADW m⁻² day⁻¹ for Halodule uninervis at the intertidal and subtidal sites, respectively. The maximum production was recorded for Syringodium isoetifolium, 60% of the 9.0 g ADW m⁻² day⁻¹ was contributed by the leaves. All species reached the maximum production during February and March, when the water temperatures were highest and water was retained above all sites, at all times. The increase of leaf production was mainly due to the increase in biomass of the mature leaves. Significant changes in the plastochrone interval of the leaves were not observed during this period.     

LC–MS/MS analysis of okadaic acid analogues and other lipophilic toxins in single-cell isolates of several Dinophysis species collected in Hokkaido,Japan

Quantification of diarrhetic shellfish poisoning (DSP) toxins (okadaic acid analogues), and other lipophilic toxins in single-cell isolates of the dinoflagellates Dinophysis fortii, D. acuminata, D. mitra, D. norvegica, D. tripos, D. infundibulus and D. rotundata, collected in coastal waters Hokkaido, Japan in 2005, was carried out by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Okadaic acid (OA), dinophysistoxin-1 (DTX1), 7-O-palmitoyldinophysistoxin-1 (DTX3), pectenotoxin-1 (PTX1), pectenotoxin-11 (PTX11), pectenotoxin-2 (PTX2), pectenotoxin-6 (PTX6), pectenotoxin-2 seco-acid (PTX2sa), yessotoxin (YTX) and 45-hydroxyyessotoxin (45-OHYTX) were quantified by LC–MS/MS. PTX2 was the dominant toxin in D. acuminata, D. norvegica and D. infundibulus whereas both DTX1 and PTX2 were the principal toxins in D. fortii. None of the toxins were detected in D. mitra, D. rotundata and D. tripos. These results suggest that D. fortii is the most important species responsible for DSP contamination of bivalves in Hokkaido. This is the first finding of PTX2 in D. infundibulus, and confirms the presence of PTX2 in Japanese D. acuminata and D. norvegica collected from natural seawater.     

Quantitative and dynamic aspects of a mixed seagrass meadow in Papua New Guinea

Aspects of production and biomass were studied from November 1981 to November 1982 in six seagrass species which together from the mixed seagrass meadows in Papua New Guinea. These species, viz. Thalassia hemprichi (Ehrenb.) Aschers., Cymodocea serrulata (R.Br.) Aschers. et Magnus, Cymodocea rotundata Ehrenb. et Hempr. ex Aschers., Syringodium isoetifolium (Aschers.) Dandy, Halodule uninervis (Forssk.) Aschers. and Halophila ovalis (R.Br.) Hook. f. have been previously studied in monospecific seagrass beds. Thalassia hemprichii was the dominant species, followed by Syringodium isoetifolium. These two species were present in all samples and evenly distributed. Cymodocea serrulata and C. rotundata were recorded in 91 and 86%, respectively, of the quadrats sampled. The density, however, varied considerably. Shoots of the remaining two species were found in < 50% of the samples. The percentage presence increased when below-ground plant parts were taken into account.Significant differences in the shoot density were only found in Syringodium isoetifolium. The distribution of the five other species remained unchanged during the year. Annual mean shoot density amounted to 860 for Thalassia hemprichii, 2100 for Syringodium isoetifolium, 200 for Cymodocea serrulata, 250 for C. rotundata and 54 for both Halodule uninervis and Halophila ovalis. All species reached their maximum density from September to November. The mean aboveground production was 3.9 g ash-free dry weight (ADW) m⁻² day⁻¹, of which 64% was contributed by Thalassia hemprichii. Syringodium isoetifolium, which had the highest shoot density, contributed only 17%.The plastochrone interval of the leaves (PIL) was constant in all species and the mean ranged from 10.1 to 11.1 days. The PIL was virtually the same in this mixed meadows as in monospecific seagrass beds. Furthermore, the above-ground relative growth rate was constant during the year. Thalassia hemprichii was the most productive seagrass (mean 0.043 day⁻¹), whereas the lowest mean relative production was observed for Syringodium isoetifolium (0.030 day⁻¹). Total mean production was 6.4 g ADW m⁻² day⁻¹, of which 39% was contributed by the vertical axes, the rhizomes and the roots. The caloric production efficiency of the meadows was 0.58% of the total insolation at the water surface.Thalassia hemprichii was, because of its morphology, the stable element in the meadow. All other species were present at all times and exhibited a continuative process of recolonization.     

Dihydrostilbene phytoalexins from Dioscorea rotundata

2′,3-Dihydroxy-5-methoxybibenzyl (Batatasin IV), its demethyl derivative and 3,5-dihydroxybibenzyl (dihydropinosylvin) were isolated only from flesh of Dioscorea rotundata infected with Botryodiplodia theobromae and may therefore be considered phytoalexins. These compounds were found to be antifungal using bioassays with Cladosporium cladosporioides, Botryodiplodia theobromae, Aspergillus niger and Penicillium schlerotgenum. Dihydro-pinosylvinin also exhibited strong antibacterial activity against Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli.     

The distribution of seagrasses of Yap,Micronesia, with relation to low tide conditions

On the shallow coral shelves of Yap, the distribution of Cymodocea rotundata Ehrenb. & Aschers., Thalassia hemprichii (Ehrenb.) Aschers. and Enhalus acoroides (L.) Royle reflects their tolerance of low tide conditions that include mid-day water temperatures exceeding 40°C and water salinity during heavy rains to 2‰. The restriction of Cyamodocea serrulata (R.Br.) Aschers. & Magnus and Syringodium isoetifolium (Aschers.) Dandy to deeper sites reflects less tolerance to low tide conditions.     

Next-generation sequencing based genotyping,cytometry and phenotyping for understanding diversity and evolution of guinea yams

Key message

Genotyping by sequencing (GBS) is used to understand the origin and domestication of guinea yams, including the contribution of wild relatives and polyploidy events to the cultivated guinea yams.

Abstract

Patterns of genetic diversity within and between two cultivated guinea yams (Dioscorea rotundata and D. cayenensis) and five wild relatives (D. praehensilis, D. mangenotiana, D. abyssinica, D. togoensis and D. burkilliana) were investigated using next-generation sequencing (genotyping by sequencing, GBS). Additionally, the two cultivated species were assessed for intra-specific morphological and ploidy variation. In guinea yams, ploidy level is correlated with species identity. Using flow cytometry a single ploidy level was inferred across D. cayenensis (3x, N = 21), D. praehensilis (2x, N = 7), and D. mangenotiana (3x, N = 5) accessions, whereas both diploid and triploid (or aneuploid) accessions were present in D. rotundata (N = 11 and N = 32, respectively). Multi-dimensional scaling and maximum parsimony analyses of 2,215 SNPs revealed that wild guinea yam populations form discrete genetic groupings according to species. D. togoensis and D. burkilliana were most distant from the two cultivated yam species, whereas D. abyssinica, D. mangenotiana, and D. praehensilis were closest to cultivated yams. In contrast, cultivated species were genetically less clearly defined at the intra-specific level. While D. cayenensis formed a single genetic group, D. rotundata comprised three separate groups consisting of; (1) a set of diploid individuals genetically similar to D. praehensilis, (2) a set of diploid individuals genetically similar to D. cayenensis, and (3) a set of triploid individuals. The current study demonstrates the utility of GBS for assessing yam genomic diversity. Combined with morphological and biological data, GBS provides a powerful tool for testing hypotheses regarding the evolution, domestication and breeding of guinea yams.     

Pathogenicity of Pratylenchus coffeae,Scutellonema bradys,Meloidogyne incognita,and Rotylenchus reniformis on Dioscorea rotundata

Low populations (200 specimens per plant) of Pratylenchus cofl''eae, Scutellonema bradys, Meloidogyne incognita, and Rotylenchulus reniformis stimulated the development of tops, roots, and tubers of Dioscorea rotundata "Guinea" yam. We demonstrated experimentally that P. coffeae was responsible for the deterioration in quality of the yam tuber in Puerto Rico, a condition known as a dry rot of yam. Initial populations of 600 P. coffeae, S. bradys, or M. incognita, and populations of 1,000 P. coffeae or S. bradys per plant were high enough to induce dry rot of the yam tubers. P. coffeae and S. bradys were pathogenic to yam cultivar Guinea, but M. incognita and R. reniformis did not cause necrosis or cracking of the tuber cortex in our experiments.     

Novel microsatellite loci reveal high genetic diversity yet low population structure for alfalfa leafcutting bees in North America

The alfalfa leafcutting bee, Megachile rotundata (ALCB) is an economically important pollinator necessary for seed production of the critical forage crop alfalfa, Medicago sativa. The pollinator was accidentally introduced to North America from Europe approximately 70 years ago, and it is primarily produced in Canada and shipped to the United States annually en masse for seed field pollination. We investigate how the large-scale commercial movement of this bee affects the genetic structure of populations in the North American seed growing system and compare the genetic diversity and structure of introduced North American bees with two native European populations. Using 16 newly developed microsatellite loci, we describe the North American population structure of this bee. ALCBs collected from alfalfa seed farms have a degree of genetic variability similar to one native European population, but lower than the second. Considering that the species was accidentally introduced into North America, we anticipated more signature of a founder effect. Despite the level of genetic variability, we found little, if any, genetic structuring across North America, other than that the North American populations were distinct from the European populations sampled. While we detected some sub-structure in North American populations using Bayesian methods, the structuring was without geographic pattern, and we propose it is the result of the intense human management and movement of these bees. The trade and movement of these bees by humans has created a nearly panmictic M. rotundata population across the continent, which has implications relevant to the preservation and conservation of other bee pollinators.     

Immunological relatedness of cytoplasmic and membrane-bound acid phosphatases from Dioscorea cayenensis rotundata

Antibodies were raised against one cytoplasmic and two membrane-bound acid phosphatases purified from yam tubers (Dioscorea cayenensis rotundata). Experiments of immunoinactivation and immunoelectrophoresis revealed cross-immunological reactions between the cytoplasmic enzyme (acid phosphatase A) and one of two membrane-bound counterparts (acid phosphatase B) suggesting that these molecules share common antigenic determinants. The antibodies raised against the other membrane-bound enzyme (acid phosphatase C) only inhibited and precipitated this enzyme.     

The seagrass and associated macroalgal vegetation of Gazi Bay (Kenya)

The seagrass and macroalgal vegetation of Gazi Bay (at approximately 50 km south of Mombasa) have been studied by means of 88 relevés along 7 transects. Correlation between the distribution of the seagrasses and some abiotic factors (particle size fractions, chemical composition of the substrate) is not well marked. Nevertheless a general zonation and succession of seagrasses could be established:

1. A transition zone between the mangal and the seagrass beds is covered byBoodleopsis pusilla;
2. the pioneer associationHalophila ovalis +Halodule wrightii forms low sandy bumps at the upper limit of the seagrass beds, but also occurs in the whole midlittoral where sandlayers have recently been accumulated (e.g. on coral platforms);
3. the climax vegetation of the intertidal zone seems to beThalassia hemprichii which sometimes is associated withCymodocea rotundata andC. serrulata, certainly in deeper pools and close to low water mark;Halimeda opuntia,Gracilaria salicornia andG. corticata are also frequent in this vegetation type;
4. from low water at neap tide downwards patches of monospecificEnhalus acoroides vegetation can also occur;
5. from mean low water down to approximately ?1 m mixed meadows ofThalassia, C. serrulata, C rotundata andHalodule uninervis are well developed; the seaweedsHalimeda macrooloba andAvrainvillea obscura are also typical for this zone; locally patches ofSyringodium isoetifolium grow on small bumps andHalophila stipulacea grows as a pioneer on bare sand;
6. from ?1 m downwards the whole lagoon is covered by homogeneous, monospecificThalassodendron ciliatum meadows, locally replaced byE. acoroides.

     

Characterization of glutathione S-transferases in a solitary bee,Megachile rotundata (Fab.) (hymenoptera: megachilidae) and inhibition by chalcones,flavone, quercetin and tridiphane-diol

• 1.1. The glutathione S-transferases of Megachile rotundata (Fab.) were characterized eletrophoretically and spectrophotometrically.
• 2.2. Differences were found between sexes with respect to number of isozymes and activity with age.
• 3.3. Inhibition patterns of chalcone, seven of its synthetic derivatives, flavone, quercetin, and tridiphanediol differed with respect to sex and substrate.
• 4.4. Comparisons are made with the honey bee, Apis mellifera L.

     

Evidence for allelopathy as a mechanism of community composition change by an invasive exotic shrub, Chrysanthemoides monilifera spp. rotundata

Chemical interference is increasingly suggested as a mechanism facilitating exotic plant invasion and plant community composition. In order to explore this further, we employed a comprehensive extract-bioassay technique that facilitated detection and demarcation of phytotoxicity, direct allelopathy and indirect allelopathy of bitou bush (Chrysanthemoides monilifera spp. rotundata) compared to an indigenous dominant of the invaded system, acacia (Acacia longifolia var. sophorae). Extracts of the leaves and roots of both species exhibited phytotoxic effects against five indigenous plant species. Evidence for allelopathy between co-evolved indigenous plants was detected between acacia and Isolepis nodosa. Allelopathy between bitou bush and four indigenous plant species was also detected. Therefore we propose that both the acacia and bitou bush have the potential to chemically inhibit the establishment of indigenous plants. Eventual dominance of bitou bush is predicted, however, based on more ubiquitous effects on seedling growth.