Karyotype evolution in apomictic Boechera and the origin of the aberrant chromosomes

Chromosome rearrangements may result in both decrease and increase of chromosome numbers. Here we have used comparative chromosome painting (CCP) to reconstruct the pathways of descending and ascending dysploidy in the genus Boechera (tribe Boechereae, Brassicaceae). We describe the origin and structure of three Boechera genomes and establish the origin of the previously described aberrant Het and Del chromosomes found in Boechera apomicts with euploid (2n = 14) and aneuploid (2n = 15) chromosome number. CCP analysis allowed us to reconstruct the origin of seven chromosomes in sexual B. stricta and apomictic B. divaricarpa from the ancestral karyotype (n = 8) of Brassicaceae lineage I. Whereas three chromosomes (BS4, BS6, and BS7) retained their ancestral structure, five chromosomes were reshuffled by reciprocal translocations to form chromosomes BS1‐BS3 and BS5. The reduction of the chromosome number (from x = 8 to x = 7) was accomplished through the inactivation of a paleocentromere on chromosome BS5. In apomictic 2n = 14 plants, CCP identifies the largely heterochromatic chromosome (Het) being one of the BS1 homologues with the expansion of pericentromeric heterochromatin. In apomictic B. polyantha (2n = 15), the Het has undergone a centric fission resulting in two smaller chromosomes – the submetacentric Het′ and telocentric Del. Here we show that new chromosomes can be formed by a centric fission and can be fixed in populations due to the apomictic mode of reproduction.     

Competitive interactions between grass and succulent shrubs at the ecotone between an arid grassland and succulent shrubland in the Karoo

Nearest-neighbour analysis was used to examine the competitive interactions between Stipagrostis brevifolia, a C₄ perennial grass, and two leaf succulent shrubs, Ruschia robusta and Leipoldtia pauciflora, at the ecotone between semi-arid grassland and succulent shrubland in the Karoo. The root distribution in the soil was also compared to assess the degree of overlap in the potential use of soil resources. Regressions between the combined sizes of interspecific, nearest-neighbour species and the distance between them showed significant positive correlations for S. brevifolia and R. robusta, which suggest the presence of competition. We infer from individual species regressions that the grass exerted a stronger competitive force on the shrub R. robusta than R. robusta on the grass. There was also evidence for strong intraspecific competitive relationships within S. brevifolia and R. robusta. There was no evidence of competition between S. brevifolia and L. pauciflora or among L. pauciflora individuals. S. brevifolia had the deepest root system, and was recorded at depths of 70 cm. Most of this root mass occurred between 10 and 40 cm. Ruschia robusta roots were recorded as deep as 55 cm, but more than 90% was found in the top 20 cm of the soil, creating a degree of overlap with the vertical root distribution of S. brevifolia. A clear separation in rooting depths occurred between S. brevifolia, and L. pauciflora which had only 3% of the total root mass below 10 cm. The partial overlap in the vertical root distribution between S. brevifolia and R. robusta may account for the observed competitive relationship, but each species dominates in a different layer, potentially minimising the net competition between S. brevifolia and R. robusta. Our findings demonstrate the possibility of a two-layer water-obtaining strategy in a semi-desert ecosystem, where the succulent shrubs seem to be playing the typical “grass” role described in most models of water partitioning between grass and woody plants. The stronger competitive effect of S. brevifolia on R. robusta at all the sites is of significance to species dynamics, and might be related to winter/summer rainfall dynamics at the climatic transition.     

Inter and intra-species-related differences in the regulation of the cardiac autonomic system

The heart rate response to isoproterenol (HR-Iso), density and affinity (k_d) of β-adrenergic (β-AR) and muscarinic (M₂) receptors were compared among three rodents with different generation-life histories of confinement and of high altitude exposure. The European guinea pig (Cavia porcellus) (EGp), a laboratory animal that arrived in Europe after the Spanish Conquest of South America and the Peruvian guinea pig (C. porcellus) (PGp), a semi-wild animal that came from the altiplano to sea level at least 25 generations ago, were used for intra-species comparison. Wistar rats (WR) were used for inter-species comparison as representative of a typical sea level laboratory animal. The HR-Iso was lower in EGp than in the PGp. The PGp showed the highest β-AR density (P<0.0005) and the highest β-AR k_d values (P<0.0005) when compared to both EGp and WR groups (β-AR B_max (fmol mg⁻¹ prot), WR, 19±4; Egp, 34±10; PGp, 74±15. β-AR k_d (pM), WR, 24±10; Egp, 17±7; PGp, 39±14). In contrast, PGp showed lower M₂ receptor density values than the EGp (P<0.0005). The WR had the highest M₂ receptor densities (M₂ B_max (fmol mg⁻¹ prot), WR, 188±15; Egp, 147±9; PGp, 118±6 and M₂ k_d (pM), WR, 65±12; Egp, 67±6; PGp, 92±2). The inter and intra-species differences found may be related to their respective history of confinement rather than to their history of exposure to high altitude.     

Genomic constitution and taxonomy of the Chinese hexaploids Elymus cylindricus and E. breviaristatus (Poaceae: Triticeae)

Elymus cylindricus (2n = 6x = 42) and E. breviaristatus (2n = 6x = 42) are distributed in grasslands and deserts of northern and north‐western China. Their genomic constitution and taxonomic status are unclear. Elymus cylindricus was crossed with E. wawawaiensis J.R.Carlson & Barkworth ( StH ), Roegneria grandis Keng ( StY ) and Campeiostachys dahurica (Turcz. ex Griseb.) B.R.Baum, J.L. Y ang & C. Y en var. dahurica ( StYH ). Meiotic pairing in the hybrids E. cylindricus × E. wawawaiensis ( StH ), E. cylindricus × R. grandis ( StY ) and E. cylindricus × C. dahurica var. dahurica ( StYH ) showed on average 10.00, 11.30 and 20.92 bivalents per cell, respectively. Elymus breviaristatus was crossed with C. dahurica var. dahurica ( StYH ) and E. cylindricus. Chromosome pairing in the hybrids of E. breviaristatus × C. dahurica var. dahurica and E. breviaristatus × E. cylindricus showed on average 19.60 and 19.27 bivalents, respectively. Genomic in situ hybridization (GI SH ) revealed the presence of St , Y and H genomes in E. cylindricus and E. breviaristatus. An intergenomic rearrangement was observed in E. cylindricus using GI SH . Meiotic pairing data and GI SH indicated that both E. cylindricus and E. breviaristatus are allohexaploids containing the StYH genomes. Elymus cylindricus and E. breviaristatus should be treated as Campeiostachys dahurica var. cylindrica and Campeiostachys breviaristata, respectively.     

Sprouty1 and Sprouty2 limit both the size of the otic placode and hindbrain Wnt8a by antagonizing FGF signaling

Multiple signaling molecules, including Fibroblast Growth Factor (FGF) and Wnt, induce two patches of ectoderm on either side of the hindbrain to form the progenitor cell population for the inner ear, or otic placode. Here we report that in Spry1, Spry2 compound mutant embryos (Spry1^−/−; Spry2^−/− embryos), the otic placode is increased in size. We demonstrate that the otic placode is larger due to the recruitment of cells, normally destined to become cranial epidermis, into the otic domain. The enlargement of the otic placode observed in Spry1^−/−; Spry2^−/− embryos is preceded by an expansion of a Wnt8a expression domain in the adjacent hindbrain. We demonstrate that both the enlargement of the otic placode and the expansion of the Wnt8a expression domain can be rescued in Spry1^−/−; Spry2^−/− embryos by reducing the gene dosage of Fgf10. Our results define a FGF-responsive window during which cells can be continually recruited into the otic domain and uncover SPRY regulation of the size of a putative Wnt inductive center.     

Comparative analysis of the eye anatomy in fossorial and surface-living skink species (Reptilia: Scincidae), with special reference to the structure of the retina

We compared the eye anatomy of the scotopic fossorial Acontias orientalis, Acontias rieppeli and Typhlosaurus vermis with that of the photopic surface-living Trachylepis punctatissima, with particular reference to the retina. The findings were compared with published data on gecko species (Röll, 2001), to determine whether similar trends existed. The vestigial eye of T. vermis was not comparable with that of the other three skink species. The findings in A. orientalis, A. rieppeli and T. punctatissima were as follows: (a) A. rieppeli lacked a conus papillaris, (b) A. orientalis, A. rieppeli and T. punctatissima were pure-cone species but lacked a fovea, (c) estimated cone density in A. orientalis and A. rieppeli was lower than that in T. punctatissima, (d) the ellipsoid cone segment was smaller and the paraboloid segment larger in A. orientalis and A. rieppeli with the reverse in T. punctatissima, (e) VCL%, ONL%, OPL% and GCL% in A. orientalis and A. rieppeli were significantly greater than that of T. punctatissima, (f) INL% and IPL% in T. punctatissima was significantly greater, and (g) T. punctatissima had abundant Müller cells and fibres. Findings in the gecko species were congruent with those of the three skink species of the present study.     

Enhancing the flux of D-glucose to the pentose phosphate pathway in Saccharomyces cerevisiae for the production of D-ribose and ribitol

Phosphoglucose isomerase-deficient (pgi1) strains of Saccharomyces cerevisiae were studied for the production of D-ribose and ribitol from D-glucose via the intermediates of the pentose phosphate pathway. Overexpression of the genes coding for NAD⁺-specific glutamate dehydrogenase (GDH2) of S. cerevisiae or NADPH-utilising glyceraldehyde-3-phosphate dehydrogenase (gapB) of Bacillus subtilis enabled growth of the pgi1 mutant strains on D-glucose. Overexpression of the gene encoding sugar phosphate phosphatase (DOG1) of S. cerevisiae was needed for the production of D-ribose and ribitol; however, it reduced the growth of the pgi1 strains expressing GDH2 or gapB in the presence of higher D-glucose concentrations. The CEN.PK2-1D laboratory strain expressing both gapB and DOG1 produced approximately 0.4 g l⁻¹ of D-ribose and ribitol when grown on 20 g l⁻¹ (w/v) D-fructose with 4 g l⁻¹ (w/v) D-glucose. Nuclear magnetic resonance measurements of the cells grown with ¹³C-labelled D-glucose showed that about 60% of the D-ribose produced was derived from D-glucose. Strains deficient in both phosphoglucose isomerase and transketolase activities, and expressing DOG1 and GDH2 tolerated only low D-glucose concentrations (≤2 g l⁻¹ (w/v)), but produced 1 g l⁻¹ (w/v) D-ribose and ribitol when grown on 20 g l⁻¹ (w/v) D-fructose with 2 g l⁻¹ (w/v) D-glucose.     

Phytochemical Study of the Genus Salpichroa (Solanaceae), Chemotaxonomic Considerations,and Biological Evaluation in Prostate and Breast Cancer Cells

Twelve Salpichroa taxa have been phytochemically analyzed. From the aerial parts of S. scandens, four known salpichrolides A, C, I, S, and an unreported withanolide named salpichrolide V ( 1 ), were isolated. In S. dependens, S. gayi, S. glandulosa subsp. glandulosa, S. glandulosa subps. weddellii, S. leucantha, S. micrantha, S. microloba, S. proboscidea, S. ramosissima, S. tristis var. tristis, and S. weberbauerii, no withanolides were found. The chemical content of ca. 85% of the Salpichroa taxa is in agreement with molecular studies, which suggest that Salpichroa and Jaborosa, a genus considered morphologically close to Salpichroa, are distant in the systematic of the Solanoideae subfamily. Moreover, the in vitro cytotoxic activity of a set of natural salpichrolides and derivatives was examined against two prostate carcinoma cell lines (PC3 and LNCaP) and two human breast cancer cell lines (MCF‐7 and T47D). Several compounds showed moderate activity (IC₅₀ = 64.91 – 29.97 μm ).     

Host species and vegetable fruit suitability and preference by the parasitoid wasp Fopius arisanus

Parasitoids that oviposit in a concealed host inside a plant part need to be able to find both the plant and the host. Egg parasitoids of fruit‐infesting Tephritidae need to assess the oviposition site based both on the host egg and the infested fruit. Infestation by Tephritidae fruit flies threatens fruit and vegetable production. Management methods have been implemented including biological control, using Fopius arisanus Sonan (Hymenoptera: Braconidae). The parasitism by F. arisanus in three Tephritidae flies in vegetable fruits was investigated. Laboratory assays were conducted to assess the parasitoid's preference and survival. Zucchini, sweet pepper, and tomato were artificially infested with eggs of Bactrocera dorsalis Hendel, Ceratitis capitata Wiedemann, and Ceratitis cosyra Walker (all Diptera: Tephritidae), then exposed to mated naïve F. arisanus females in a 20:1 egg:parasitoid ratio. Parasitoid behavioral activities (resting, antennating, probing, ovipositing) were observed on the infested fruits. Parasitism rate was determined by dissection of fruit fly eggs under a stereomicroscope. Behavioral activities of F. arisanus differed between all the fruits when infested with B. dorsalis or C. cosyra eggs but differed only between some of the fruits when infested with C. capitata. Fopius arisanus preferred B. dorsalis over C. capitata and C. cosyra, with a parasitism rate 2× higher on B. dorsalis compared to the Ceratitis species. Preference for fruits was dependent on the infesting fruit fly. The emergence of F. arisanus was higher with B. dorsalis than with Ceratitis spp. Although B. dorsalis completed its development earlier than Ceratitis spp., host fly species did not affect the developmental time of F. arisanus. We discuss the significance of F. arisanus preference in relation to naturally occurring Tephritidae infestations. We also discuss whether some fruits might constitute a refuge for Tephritidae flies and whether this will affect the current biological control efforts against B. dorsalis.     

Meso- and bathypelagic distribution and abundance of chaetognaths in the Atlantic sector of the Southern Ocean

We conducted multinet sampling during winter and summer in the Southern Ocean (Atlantic sector) to investigate the effect of water mass, season and water depth on abundance and species composition of meso- and bathypelagic chaetognaths. Eukrohnia hamata (mean 115 ind. 1,000 m⁻³) and Sagitta marri (mean 51 ind. 1,000 m⁻³) were dominant, complemented by E. bathypelagica (mean 19 ind. 1,000 m⁻³) and E. bathyantarctica (mean 19 ind. 1,000 m⁻³) below 1,000 m. A further six species were identified, among them the rare bathypelagic species Heterokrohnia fragilis and the subtropical Eukrohnia macroneura that is new to the Antarctic. Water depth and season were the principal determinants of abundance and species composition patterns, indicating vertical seasonal migration and vertical segregation of species. The life cycles of E. hamata and S. marri were studied additionally. Their maturity stages were vertically segregated and prolonged reproductive periods are suggested for both species.     

Nutrient Retention and the Problem of Hydrologic Disconnection in Streams and Wetlands

Some aquatic systems have disproportionately high nutrient processing rates, and may be important to nutrient retention within river networks. However, the contribution of such biogeochemical hot spots also depends on water residence time and hydrologic connections within the system. We examined the balance of these factors in a comparative study of nitrate (NO₃ ⁻) uptake across stream and flow-through wetland reaches of northern Wisconsin, USA. The experimental design compared NO₃ ⁻ uptake at different levels: the ecosystem level, for reaches (n = 9) consisting of morphologically contrasting subreaches (SLOW, low mean water velocity; REF, reference, or higher mean water velocity); the sub-ecosystem level, for subreaches consisting of morphologically contrasting zones (TS, transient storage zone; MC, main channel zone). SLOW subreaches had 45% lower ecosystem-level uptake rate (K, t⁻¹) on average, indicating reduced uptake efficiency in flow-through wetlands relative to streams. The four largest K values (total n = 24) also occurred in REF subreaches. TS:MC uptake rate varied (range 0.1–6.0), but MC zones consistently accounted for most NO₃ ⁻ uptake by the ecosystem. In turn, TS influence was limited by a tradeoff between TS zone uptake rate and the strength of TS–MC hydrologic connection (α or F _med). Additional modeling of published hydrologic parameter sets showed that strong MC dominance of uptake (>75% of total uptake), at the scale of solute release methods (meters to kilometers, hours to days), is common among streams and rivers. Our results emphasize that aquatic nutrient retention is the outcome of a balance involving nutrient uptake efficiency, water residence time, and the strength of hydrologic connections between nutrient sources and sinks. This balance restricts the influence of hydrologically disconnected biota on nutrient transport, and could apply to diverse ecosystem types and sizes.     

Length–weight relationships of six fish species from the Zengqu River and the Ouqu River,southwest China

This study provides the first report of length–weight relationships (LWRs) for six endemic fish species collected from the Zengqu River and the Ouqu River, two of the tributaries to the upper reaches of the Jinsha River, southwest China. Samples were obtained in May 2013 and September 2013 using various types of fishing gear. A total of 413 specimens belonging to six fish species were analyzed; their LWRs were W = 0.0069L^3.086 (r² = 0.987) for Schizothorax wangchiachii (Fang, 1936), W = 0.0076L^3.029 (r² = 0.980) for Schizothorax dolichonema (Herzenstein, 1889), W = 0.0029L^3.315 (r² = 0.961) for Schizopygopsis malacanthus (Herzenstein, 1891), W = 0.0082L ^2.970 (r² = 0.950) for Ptychobarbus kaznakovi (Nikolsky, 1903), W = 0.0017L^3.464 (r² = 0.957) for Triplophysa stolioczkae (Steindachner, 1866), and W = 0.0037L ^3.190 (r² = 0.956) for Triplophysa orientalis (Herzenstein, 1888). Prior to this study, the LWRs for S. dolichonema, P. kaznakovi and T. orientalis were unknown. In addition, new maximum sizes for four of the species are provided.     

Morphological and functional seed traits of the wild medicinal plant Dioscorea strydomiana,the most threatened yam in the world

• Morphological and functional seed traits have important roles in characterising the species regeneration niche and help to understand the reproductive biology of rare and threatened plants, which can thus support appropriate plant conservation measures.
• Seed morphometric and dispersal kinetics of the critically endangered Dioscorea strydomiana were measured and compared with those of four other Dioscorea species, and seed germination response under constant temperatures (5–35 °C) was compared with that of the congeneric and widespread D. sylvatica.
• Seed mass of D. strydomiana (ca. 14 mg) was twice that of D. sylvatica, but similar to or smaller than the other species examined. Seeds of D. strydomiana have the lowest speed of descent and lowest variability in most of the morphological traits considered, suggesting lower phenotypic plasticity but higher variance in the wing‐loading value. Seeds of D. strydomiana reached maximum germination at 15 °C (ca. 47%), which decreased slightly to ca. 37% at 25 °C and was completely inhibited at 35 °C. D. sylvatica seeds started to germinate at 10 °C (ca. 3%), reached 75–80% germination at 15–20 °C and maximum (ca. 90%) at 25–30 °C. Base temperatures for germination (T_b) were 9.3 and 5.7 °C, for D. strydomiana and D. sylvatica, respectively. Due to the higher germination percentages of D. sylvatica, ceiling and optimum temperatures could also be modelled for this species, suggesting higher sensitivity to high temperature for seeds of D. strydomiana.
• The detected poor seed lot quality of D. strydomiana suggests difficulties in reproduction from seed, highlighting the need for further investigation and conservation actions for this threatened yam species.

     

Pre-steady-state analysis of the turn-on and turn-off of water permeability in the kidney collecting tubule

Summary Water transport across the mammalian collecting tubule is regulated by vasopressin-dependent water channel insertion into and retrieval from the cell apical membrane. The time course of osmotic water permeability (P _f ) following addition and removal of vasopressin (VP) and 8-Br-cAMP was measured continuously by quantitative fluorescence microscopy using an impermeant fluorophore perfused in the lumen. Cortical collecting tubules were subjected to a 120 mOsm bath-to-lumen osmotic gradient at 37°C with 10–15 nl/min lumen perfusion and 10–20 ml/min bath exchange rate. With addition of VP (250 U/ml), there was a 23±3 sec (sem,n=16) lag in whichP _f did not change, followed by a rise inP _f (initial rate 1.4±0.2×10^–4 cm/sec²) to a maximum of 265±10×10^–4 cm/sec. With addition of 8-Br-cAMP (0.01–1mm) there was an 11±2 sec lag. For [8-Br-cAMP]=0.01, 0.1 and 1mm, the initial rate ofP _f increase following the lag was (units 10^–4 cm/sec²): 1.1±0.1, 1.2±0.1 and 1.7±0.3. MaximumP _f was (units 10^–4 cm/sec): 64±4, 199±9 and 285±11. With removal of VP,P _f decreased to baseline (12×10^–4 cm/sec) with aT _1/2 of 18 min; removal of 0.1 and 1mm 8-Br-cAMP gaveT _1/2 of 4 and 8.5 min. These results demonstrate (i) a brief lag in theP _f response, longer for stimulation by VP than by 8-Br-cAMP, representing the transient build-up of biochemical intermediates proximal to the water channel insertion step, (ii) similar initialdP _f /dt (water channel insertion) over a wide range of [8-Br-cAMP] and steady-stateP _f values, and (iii) more rapidP _f decrease with removal of 8-Br-cAMP than with VP. These pre-steady-state results define the detailed kinetics of the turn-on and turn-off of tubuleP _f and provide kinetic evidence that the rate-limiting step for turn-on ofP _f is not the step at which VP regulates steady-stateP _f . If water channel insertion is assumed to be the rate-limiting step in the turn-on ofP _f , these results raise the possibility that water channels must be activated following insertion into the apical membrane.     

Variation in the SSUrDNA of the Genus Protostelium Leads to a New Phylogenetic Understanding of the Genus and of the Species Concept for Protostelium mycophaga (Protosteliida,Amoebozoa)

Members of the genus Protostelium (including P. mycophaga, P. nocturnum, and P. okumukumu) are protosteloid amoebae commonly found in terrestrial habitats on dead plant matter. They, along with the closely allied nominal genus Planoprotostelium, containing the single species Pl. aurantium, all have an amoeboid trophic stage with acutely pointed subpseudopodia and orange lipid droplets in the granuloplasm. These amoebae form stalked fruiting bodies topped with a single, usually deciduous spore. The species are identified based on their fruiting body morphologies except for Pl. aurantium which looks similar to P. mycophaga in fruiting morphology, but has amoebae that can make flagella in liquid medium. We built phylogenetic trees using nuclear small subunit ribosomal DNA sequences of 35 isolates from the genera Protostelium and Planoprotostelium and found that (1) the nonflagellated P. nocturnum and P. okumukumu branch basally in the genus Protostelium, (2) the flagellate, Pl. aurantium falls within the genus Protostelium in a monophyletic clade with the nominal variety, P. mycophaga var. crassipes, (3) the cultures initially identified as Protostelium mycophaga can be divided into at least three morphologically recognizable taxa, P. aurantium n. comb., P. apiculatum n. sp., and P. m. rodmani n. subsp., as well as a paraphyletic assemblage that includes the remainder of the P. mycophaga morphotype. These findings have implications for understanding the ecology, evolution, and diversity of these amoeboid organisms and for using these amoebae as models for other amoeboid groups.     

Anti‐HSV‐1 and HSV‐2 Flavonoids and a New Kaempferol Triglycoside from the Medicinal Plant Kalanchoe daigremontiana

Kalanchoe daigremontiana (Crassulaceae) is a medicinal plant native to Madagascar. The aim of this study was to investigate the flavonoid content of an aqueous leaf extract from K. daigremontiana (Kd), and assess its antiherpetic potential. The major flavonoid, kaempferol 3‐O‐β‐d ‐xylopyranosyl‐(1 → 2)‐α‐l ‐rhamnopyranoside ( 1 ), was isolated from the AcOEt fraction (Kd‐AC). The BuOH‐soluble fraction afforded quercetin 3‐O‐β‐d ‐xylopyranosyl‐(1 → 2)‐α‐l ‐rhamnopyranoside ( 2 ) and the new kaempferol 3‐O‐β‐d ‐xylopyranosyl‐(1 → 2)‐α‐l ‐rhamnopyranoside‐7‐O‐β‐d ‐glucopyranoside ( 3 ), named daigremontrioside. The crude extract, Kd‐AC fraction, flavonoids 1 and 2 were evaluated using acyclovir‐sensitive strains of HSV‐1 and HSV‐2. Kd‐AC was highly active against HSV‐1 (EC₅₀ = 0.97 μg/ml, SI > 206.1) and HSV‐2 (EC₅₀ = 0.72 μg/ml, SI > 277.7). Flavonoids 1 and 2 showed anti‐HSV‐1 (EC₅₀ = 7.4 μg/ml; SI > 27 and EC₅₀ = 5.8 μg/ml; SI > 8.6, respectively) and anti‐HSV‐2 (EC₅₀ = 9.0 μg/ml; SI > 22.2 and EC₅₀ = 36.2 μg/ml; SI > 5.5, respectively) activities, suggesting the contribution of additional substances to the antiviral activity.     

Unravelling the structure and function of the petal appendages in the tribe Schwenckieae (Solanaceae)

• The tribe Schwenckieae (Solanaceae) is characterised by the presence of appendages on the corolla, a diagnostic trait for the group. These appendages constitute a median distal projection of the three‐lobed petal and occur in the genera Melananthus and Schwenckia but are absent in Heteranthia.
• We investigated the micromorphology and anatomical structure of the appendages and lateral petal lobes of Schwenckia americana (two varieties), S. angustifolia, S. curviflora and S. novaveneciana, and Melananthus fasciculatus. We also performed histochemical tests to determine if the appendages are involved in the production of volatiles, acting as a fragrance secretory structure (osmophore).
• The appendages have a uniseriate epidermis, whose cells store phenolics and lipids. The parenchyma is starch‐rich just prior to anthesis in all species studied. The sensory test and anatomical analyses identified scent‐secreting tissues, not only in the appendages, but also in the lateral petal lobes, whose cells are papillose with a sculptured surface. The α‐naphthol p‐phenylenediamine (NADI) reaction detected volatile (essential oils) compounds in S. americana var. americana and S. americana var. angustifolia.
• We demonstrated the secretory tissues and the production of lipids in the corolla appendages of Schwenckia and Melananthus, which indicate their osmogenic function and probable scent emission to attract pollinators.

     

Biochemical characterization of the recombinant human Drosophila homologues Timekeeper and Andante involved in the Drosophila circadian oscillator

The Drosophila clock proteins timekeeper (CK2α^Tik) and andante (CK2β^And) are mutated CK2α and CK2β subunits, respectively.In order to revisit the hypothesis concerning a perturbation of the β/β and/or α/β subunit association, involving the andante mutant we have cloned, expressed and purified the recombinant andante mutant CK2β^And and a CK2 holoenzyme composed of CK2β^And and the wildtype CK2α subunit. Biochemical analyses using gel filtration analysis, inhibitor and heat treatment, as well as urea denaturation studies did not yield significant differences between the wildtype holoenzyme (α₂β₂) and a holoenzyme containing wildtype CK2α and andante CK2β^And.The timekeeper mutant, CK2α^Tik has been reported to show a significant reduction in enzyme activity. In order to closely investigate the reason for this reduction in activity, we have also cloned and expressed the human homologue of Drosophila timekeeper. Using a CK2 holoenzyme containing the human timekeeper mutant and the wildtype CK2β subunit we could confirm a strongly reduced activity towards CK2 substrates, but also a significant reduction in the autophosphorylation of the CK2β in the absence of any substrate. Based on a structure-based model we postulate that the mutation M161K in Drosophila (i.e. M163K in human) is responsible for the drastic loss of activity, where the lysine residue may cause improper binding of the tri-nucleotide.