Transformation of Stylosanthes spp. using Agrobacterium tumefaciens

Tumours were incited on leaf sections of Stylosanthes humilis, S. hamata, S. guianensis and S. scalra following infection by Agrobacterium tumefaciens. The suitability of 2 binary vectors (pGA472, BIN6) for gene transfer in S. humilis was tested and kanamycin-resistant tumour tissue was obtained from infected leaf pieces. The presence and expression of the neomycin phosphotransferase (NPT II) gene in the plant cells was demonstrated by hybridization of the coding region of the NPT II gene of the transposon Tn5 to DNA and RNA of kanamycin resistant tumours and by detection of significant NPT II activity in tissue extracts. Tumours also produced teratomatous shoots expressing the NPT II gene, but these could not be rooted.     

Consistent variation in seed germination across an environmental gradient in a Neotropical savanna

Environmental conditions experienced by mother plants determine germination patterns. Here, we investigated the germination ecology of Miconia albicans (Melastomataceae), a widespread apomictic shrub, along a soil fertility gradient in the Brazilian Cerrado (a Neotropical savanna). The mosaic of vegetations in the Cerrado ranges from grasslands to woodlands, which present different conditions for seedling establishment. Cerrado grasslands are a more unpredictable environment because they are more prone to disturbances such as fires and prolonged droughts than closed woodlands. We expect lower reproductive investment but increasing germinability, germination speed and synchrony in areas with more dense vegetation, due to prolonged droughts in grasslands and higher soil properties in woodlands. Seeds of M. albicans were collected in grasslands, shrublands and woodlands for three years in a Cerrado area in southeastern Brazil. Seeds collected in all vegetations were set to germinate under identical experimental conditions. According to our prediction, we found a consistent pattern of increasing germinability, mean germination time and synchrony in areas with more closed vegetation. Both slower and asynchronous germination in seeds from grassland plants are likely to increase seedling survival under a more unpredictable environment. On the other hand, seeds from plants in the woodlands germinated at higher rates and germination was more synchronous, thus increases the chances of survival under a more competitive light-limited environment. This pattern was found in all three years of study. Our study suggests that stressful environmental conditions experienced by mother plants in the Cerrado may act as cues signalizing unfavourable conditions for seed germination and help explain functional divergence in germination traits within species. In conclusion, we showed for the first time a temporally consistent pattern of intraspecific variation in the germination traits of M. albicans, with different germination strategies at the extremes of the ecological gradient.     

Evolutionary Dynamics of wAu-Like Wolbachia Variants in Neotropical Drosophila spp.

Wolbachia bacteria are common intracellular symbionts of arthropods and have been extensively studied in Drosophila. Most research focuses on two Old Word hosts, Drosophila melanogaster and Drosophila simulans, and does not take into account that some of the Wolbachia associations in these species may have evolved only after their fast global expansion and after the exposure to Wolbachia of previously isolated habitats. Here we looked at Wolbachia of Neotropical Drosophila species. Seventy-one lines of 16 Neotropical Drosophila species sampled in different regions and at different time points were analyzed. Wolbachia is absent in lines of Drosophila willistoni collected before the 1970s, but more recent samples are infected with a strain designated wWil. Wolbachia is absent in all other species of the willistoni group. Polymorphic wWil-related strains were detected in some saltans group species, with D. septentriosaltans being coinfected with at least four variants. Based on wsp and ftsZ sequence data, wWil of D. willistoni is identical to wAu, a strain isolated from D. simulans, but can be discriminated when using a polymorphic minisatellite marker. In contrast to wAu, which infects both germ line and somatic tissues of D. simulans, wWil is found exclusively in the primordial germ line cells of D. willistoni embryos. We report on a pool of closely related Wolbachia strains in Neotropical Drosophila species as a potential source for the wAu strain in D. simulans. Possible evolutionary scenarios reconstructing the infection history of wAu-like Wolbachia in Neotropical Drosophila species and the Old World species D. simulans are discussed.     

Microbial colonization in the seagrass Posidonia spp. roots

The pattern of colonization by microorganisms on root surfaces from three species of seagrass belonging to the genus Posidonia was assessed. Microbial abundance on roots was measured by two electronic microscope techniques. Trends in microbial colonization between species and root order were defined. In addition, eutrophication status of the sampling sites and physiological status of Posidonia oceanica (L.) Delile roots have been taken into account. Our results show high microbial abundance in the Mediterranean species P. oceanica, in comparison with the low rates of colonization found in the Australian species P. australis Hook f. and P. sinuosa Cambridge et Kuo. Microbial density tended to decrease as root order increased, and living roots always showed higher microbial abundance than dead ones. Colonization of P. oceanica roots at the three sites with different environmental status follows different trends according to root order. It is suggested that root age influences the rate of microbial colonization of seagrass roots and that colonization of root surface by microorganisms is associated with organic exudates from the roots rather than with decaying root tissues.     

Histological Comparisons of Parasitism by Schistonchus spp. (Nemata: Aphelenchoididae) in Neotropical Ficus spp.

Syconia (enclosed infructescences) infested with host-specific species of Schistonchus (Aphelenchoididae) were collected from six species of Ficus (Moraceae) native to Florida or Panama. They were sectioned and histologically examined to assess the effects of parasitism. Parasitism by Schistonchus spp. was associated with hypertrophied cells, tissue necrosis, and the presence of an exudate in all species. Occasional hypertrophy of the outer epidermal cells occurred on seed florets, wasp florets, and on the endothecial cells of male florets in F. aurea (subgenus Urostigma) from Florida. Aberrations of the inner mesocarp occurred under the hypertrophied cells on seed florets. In F. laevigata (subgenus Urostigma) from Florida, Schistonchus sp. infested immature male florets and was associated with hypertrophy of endothecial cells, epidermal cells of the anther filaments, and anthers. Schistonchus sp. also caused aberrations of the anther filament, anthers, and pollen. Ficus poponoei (subgenus Urostigma) and F. glabrata (subgenus Pharmacosycea), both from Panama, had hypertrophied outer epidermal cells on seed florets. Ficus poponoei also had Schistonchus sp. within the pedicel of an aborted floret, with hypertrophy of the cortical parenchyma. Ficus trigonata (subgenus Urostigma) from Panama had hypertrophy of the outer epidermis of seed florets. When the outer epidermis on these florets was missing, the inner mesocarp was hypertrophied. Ficus maxima (subgenus Pharmacosycea) from Panama had hypertrophy on the outer epidermis of seed and aborted florets. Schistonchus spp. were not found in wasp larvae or pupae in any of the Ficus spp. examined. Hypertrophy was never observed in the absence of Schistonchus spp.     

Leaf functional traits of Neotropical savanna trees in relation to seasonal water deficit

The seasonal savannas (cerrados) of Central Brazil are characterized by a large diversity of evergreen and deciduous trees, which do not show a clear differentiation in terms of active rooting depth. Irrespective of the depth of the root system, expansion of new foliage in deciduous species occurs at the end of the dry season. In this study, we examined a suite of leaf traits related to C assimilation, water and nutrients (N, P) in five deciduous and six evergreen trees that were among the dominant families of cerrado vegetation. Maximum CO₂ assimilation on a mass basis (A_mass) was significantly correlated with leaf N and P, and specific leaf area (SLA; leaf area per unit of leaf mass). The highest leaf concentrations of both nutrients were measured in the newly mature leaves of deciduous species at the end of the dry period. The differences in terms of leaf N and P between evergreen and deciduous species decreased during the wet season. Deciduous species also invested less in the production of non-photosynthetic leaf tissues and produced leaves with higher SLA and maintained higher water use efficiency. Thus, deciduous species compensated for their shorter leaf payback period by maintaining higher potential payback capacity (higher values of A_mass) and lower leaf construction costs (higher SLA). Their short leafless period and the capacity to flush by the end of the dry season may also contribute to offset the longer payback period of evergreen species, although it may involve the higher cost of maintaining a deep-root system or a tight control of plant water balance in the shallow-rooted ones.     

The relevance of ants as seed rescuers of a primarily bird-dispersed tree in the Neotropical cerrado savanna

Sex ratios can influence mating behaviour, population dynamics and evolutionary trajectories; yet the causes of natural sex ratio variation are often uncertain. Although secondary (birth) sex ratios in guppies (Poecilia reticulata) are typically 1:1, we recorded female-biased tertiary (adult) sex ratios in about half of our 48 samples and male-biased sex ratios in none of them. This pattern implies that some populations experience male-biased mortality, perhaps owing to variation in predation or resource limitation. We assessed the effects of predation and/or inter-specific resource competition (intraguild predation) by measuring the local catch-per-unit-effort (CPUE) of species (Rivulus killifish and Macrobrachium prawns) that may differentially prey on male guppies. We assessed the effects of resource levels by measuring canopy openness and algal biomass (chlorophyll a concentration). We found that guppy sex ratios were increasingly female-biased with increasing CPUE of Macrobrachium, and perhaps also Rivulus, and with decreasing canopy openness. We also found an interaction between predators and resource levels in that the effect of canopy openness was greatest when Macrobrachium CPUE was highest. Our study thus also reveals the value of simultaneously testing multiple environmental factors that may drive tertiary sex ratio variation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Plant community structure, soil properties and microbial characteristics in revegetated quarries

Quarries are an important type of degraded land in southern China requiring ecological improvement and rehabilitation. In this study, plant community structure, soil properties, and microbial biomass and community function were examined at different rehabilitated phases in three quarries, namely Turret Hill Quarry, Lam Tei Quarry and Shek O Quarry, in Hong Kong. Results show that plant species richness and the percentage of native species increased with rehabilitated ages in the three quarries. The highest coverage of woody species was found at older phases, while the lowest woody coverage occurred at younger phases. Soils were strongly to moderately acidic in reaction, and more acidic soils were found in the older than in the younger sites. Organic C as well as total N and P accumulated in soil along with secondary succession in the three quarries, which were positively correlated with woody species richness. Older phases had higher total microbial biomass C and N which were positively correlated with soil organic C, total N and extractable NO₃-N, as well as woody species coverage and native species richness as shown by the biplot of redundancy analysis. Diversity of utilized carbons suggested that metabolic abilities developed gradually with rehabilitation ages in Shek O Quarry, but Turret Hill Quarry and Lam Tei Quarry had similar patterns of carbon source utilization. Principal component analysis further revealed consistent differences in metabolic diversity. Woody coverage and native species richness were significantly correlated with carbon source utilization and functional diversity.     

Additional Fallisia spp. (Apicomplexa: Plasmodiidae) of Neotropical lizards

Fallisia biporcati n. sp. parasitises thrombocytes and lymphocytes of Anolis biporcatus and A. lionotus in Panama. Round or oval schizonts average 13.3 × 11.5 (10.5–16 × 9–13) m, with LW 153.8 (94–208) m², and produce 38.3 (28–60) merozoites. Gametocytes are variably shaped, from round or oval to nearly triangular or rectangular, and average 12.6 × 9.0 (10–15 × 6–12) m, with LW 113.1 (82–150) m² and L/W ratio 1.43 (1.0–2.2). Thrombocytes and lymphocytes of A. poecilopus in Panama are parasitised by F. poecilopi n. sp. Schizonts, oval to elongate in shape, 7.7 × 4.7 (5.5–9 × 3–6) m, with LW 36.5 (22–54) m², are filled with 31.0 (20–51) tiny nuclei or merozoites. Gametocytes are 10.1 × 8.0 (7.5–14 × 6–11) m, with LW 82.0 (45–132) m², round to elongate with L/W ratio 1.27 (1.0–1.6). F. thecadactyli n. sp. parasitises thrombocytes and lymphocytes of Thecadactylus rapicaudus in Panama and Venezuela. Oval, oblong, or triangular schizonts average 10.3 × 8.0 (7–13 × 5–12) m, with LW 86.6 (37–156) m², and produce 40.2 (26–61) merozoites. Gametocytes are round, oval, triangular or elongate, 10.4 × 7.0 (7–15 × 5–11) m, with LW 74.8 (40–154) m² and L/W ratio 1.51 (1.1–2.2). F. dominicensis n. sp. parasitises thrombocytes of A. cybotes on Hispaniola. Schizonts, 6.0 × 4.8 (4–8 × 3–7) m, with LW 29.1 (12–56) m², round, oval, elongate, oblong or lentiform in shape, produce 12.4 (8–22) merozoites. Gametocytes are 6.6 × 5.0 (5–9 × 4–7) m , with LW 33.8 (20–56) m², round, oval or elongate, and L/W ratio of 1.34 (1.1–2.0).     

Plant and lichen colonization in an asbestos mine: Spontaneous bioattenuation limits air dispersion of fibres

Abstract

Plant and lichen phytosociological studies describe the different dynamic stages of colonization. This approach has been applied to the Balangero asbestos mine, the most important of Western Europe up to 1990, which is a potential source for the air-dispersion of carcinogenic fibres. Vegetation relevés and maps have shown that plants and lichens develop spontaneously on the asbestos-rich substrates. Early colonization stages with low-covering hyperaccumulators, such as Thlaspi sylvium and Minuartia laricifolia, and with Thymus alpestris and T. cfr. humifusus, are followed, some decades later, by mature plant communities, which completely cover the asbestos-rich debris, thereby limiting the dispersion of fibres. Lichen colonization, even when limited to low covering early stages with pioneer crustose species, such as Scoliciosporum umbrinum and Candelariella vitellina, and young foliose thalli of Xanthoparmelia tinctina, is important whenever asbestos veins are exposed on serpentinite blocks and walls. Plants and lichens can be considered as bioattenuation (spontaneous bioremediation) agents in ecological recovery.     

Ontogeny and characterization of aerenchymatous tissues of Melastomataceae in the flooded and well-drained soils of a Neotropical savanna

Lavoisiera bergii Cogn., Macairea radula (Bonpl.) DC. and Trembleya parviflora (D. Don) Cogn. are shrubby species of the family Melastomataceae. They occur in the flooded area of the Vereda Grande palm swamp in the Águas Emendadas Ecological Station (a protected area in the midwest of Brazil). M. radula and T. parviflora also occur in the well-drained soil of the cerrado sensu stricto that surrounds the palm swamp. Root and shoot anatomy of the three species show aerenchyma in plants growing in the waterlogged palm swamp area. In the cortex of roots of L. bergii and T. parviflora this aerenchyma develops in a schizo-lysigenous way. Aerenchymatous spaces occur also in the stems. During secondary growth, the phellogen produces an aerenchymatous polyderm. This latter is formed by two cell types, braciform and compact cells, and shows deposition of suberin in the cell walls. In the emerging organs the polyderm does not form gas spaces. The primary aerenchyma is constitutive and occurs also in the root of M. radula and T. parviflora, even when these species are growing in the well-drained soil of cerrado sensu stricto.     

Seasonal gas exchange and water relations in juveniles of two evergreen Neotropical savanna tree species with contrasting regeneration strategies

Colonization dynamics of woody species into grasslands in Neotropical savannas are determined by two main factors: plant-available moisture and fire. Considering seasonality of precipitation and high fire frequency in these ecosystems, vegetative reproduction has been suggested as the main regeneration strategy in woody species. This study examined seasonal variations in water relations and photosynthesis in juveniles of two tree species with contrasting regeneration strategies: Palicourea rigida (sexual reproduction) and Casearia sylvestris (asexual reproduction). The studied species showed similar transpiration rates to deep-rooted adult evergreen tree species during the rainy period, suggesting little water availability limitations on surface soil layers. P. rigida juveniles significantly decreased their leaf water potentials from wet to dry seasons. In C. sylvestris resprouts, there were no seasonal differences in their predawn water potentials and gas exchange parameters, indicating a water deficit avoidance characteristic derived from their connections to deep-rooted adult counterparts allowing access to moist soil at depth even during the drought period. P. rigida rely on strict control of water losses and turgor maintenance through elastic cell walls during the dry season. The iso-hydric behavior of gas exchange and most water relations parameters in C. sylvestris enable turgor maintenance during the dry season which also gives the possibility to achieve foliar expansion under water-stressed conditions for shallow-rooted plants. Nevertheless, in absence of water deficits, P. rigida had the advantage to be physiologically independent individuals, showing an equal or even superior photosynthetic performance that eventually could be translated into a more favorable whole-plant carbon balance and higher growth rates in wet habitats.     

利用柱花草为受体表达口蹄疫病毒外壳蛋白VP1的研究

将口蹄疫病毒外壳蛋白VP1基因克隆到植物表达载体pBI121,并转化到根癌农杆菌(Agrobacteriumtumefaciens)菌株LBA4404中,采用叶盘转化法转化柱花草(Stylosanthesspp.)栽培品种热研二号柱花草(S.guianensiscv.ReyanⅡ),获得了转基因植株,经PCR、PCR-Southern blot和Southern blot分析表明VP1基因已整合到转基因柱花草植株的核基因组中。经RT-PCR、Northern blot分析表明VP1基因已在转基因柱花草中获得转录。     

Effects of fire and defoliation on the life history of native and invader C4 grasses in a Neotropical savanna

African grasses, introduced into Neotropical savannas to improve forage quality, have spread successfully and displaced native plants. To understand their competitive relationships, we compared biomass production and allocation, plant architecture and phenology, net photosynthesis (Pn), water relations, and nutrient content under fire and simulated herbivory between two C₄ grasses, the native Trachypogon plumosus and the introduced Hyparrhenia rufa from a seasonal savanna in Venezuela. All variables were strongly influenced by the rainfall regime. Hyparrhenia produced bigger plants (in mass and size) with a large proportion of mass (>75%) allocated to leaves and culms. Its biomass production was more affected by fire than by defoliation. In contrast, Trachypogon was more affected by defoliation than by fire which promoted a flush of leaf growth even in the dry season. Fire caused up to 85% mortality in Hyparrhenia but none in Trachypogon where it increased inflorescence production. However, fire promoted abundant seed germination and fast seedling growth in Hyparrhenia, enabling it to colonize new areas. During the growing season Trachypogon had higher Pn and lower leaf water potential (Ψ) than Hyparrhenia but differences among treatments were not significant for either grass. Pn of Trachypogon ceased at a lower Ψ (−3.0 MPa) than in Hyparrhenia (−2.0 MPa), indicating its higher tolerance to water stress. During the dry season, Trachypogon leaves remained alive and retained low Pn. Leaf nutrient content was higher during the rainy season in both species. Differences in Pn could not explain the higher seasonal biomass production of Hyparrhenia. However, its water stress evasion strategy, larger biomass allocated to leaves, abundant germination and fast seedling growth appeared to be responsible for the success of Hyparrhenia as an invader of Neotropical savannas. Received: 17 August 1998 / Accepted: 3 March 1999     

Native bird breeding in a chronosequence of revegetated sites

Restoration of degraded landscapes through replantings of native vegetation has been proceeding in response to habitat loss and fragmentation and plummeting biodiversity. Little is known about whether the investments in ecological restoration have resulted in biodiversity benefits. We evaluated the potential of restored sites to support populations by assessing bird breeding activity. We surveyed 21 revegetated sites of various ages (9–111 years) in the box–ironbark region of Victoria, Australia. Sites differed in landscape context, patch features and in-site characteristics. The latter, including whether sites were grazed, amounts of fallen timber and numbers of remnant trees, were most important in affecting overall bird breeding activity. Patch-configuration (e.g., shape, area) was of secondary importance. Landscape context appeared to have little effect on bird breeding except for one species. While these results suggest that in-site habitat structure is the predominant driver, we caution against dismissing the importance of patch characteristics and landscape context for two reasons. First, the available sites covered a relatively small range of areas (<54 ha), and we could not provide a broad range of landscape-contextual contrasts given that we could only use existing plantings. Second, much of the breeding activity was by bird species known to be tolerant of smaller woodland areas or of the open countryside. We show that there is very little breeding activity in replantings by species that have declined dramatically in rank abundance between large ‘reference’ areas and fragmented landscapes. It seems likely that most replantings provide habitat configurations unsuited for dealing with declines of species most vulnerable to habitat loss and fragmentation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Temporal variation in extrafloral nectar secretion in different ontogenic stages of the fruits of Alibertia verrucosa S. Moore (Rubiaceae) in a Neotropical savanna

In this study, we evaluate the temporal variation in extrafloral nectaries (EFNs) secretion in different ontogenic stages of Alibertia verrucosa (Rubiaceae) fruits in a Neotropical savanna. We observe greater nectar secretion rate in fruits of intermediate size compared with young or ripe fruit, indicating that they are possibly more protected by ants. In addition, the nectar secretion was higher at night, a pattern that could be associated with an increase of herbivore pressure and water stress during the daylight hours. In fact, due to the high temperature and low humidity during the day in savannas, most herbivores display strong nocturnal activity, and plants can avoid nectar secretion in this period. Our results indicate that A. verrucosa can change the ants' attraction according to EFN secretion during the ontogenic stages of the fruits, probably secreting more nectar when the biotic defense are more necessary for the protection of the fruits and the plant as a whole.     

Plant regeneration from protoplast fusion inPassiflora spp.

Summary Protoplasts were isolated from leaf explants ofPassiflora edulis var.flavicarpa (the yellow passion fruit) and from cell suspensions of fivePassiflora species. Chemical fusion was performed using polyethylene glycol and the microcolonies obtained were transferred to growth medium to produce calli. Electrophoresis of soluble proteins and analysis of isoenzymes from calli produced from the fusion experiments were performed to select somatic hybrids. Specific polypeptide bands allowed the identification of somatic hybrids betweenP. edulis var.flavicarpa (+)P. alata, P. edulis var.flavicarpa (+)P. amethystina, P. edulis var.flavicarpa (+)P. cincinnata, P. edulis var.flavicarpa (+)P. giberti andP. edulis var.flavicarpa (+)P. coccinea. An average of 3 to 5% hybrid calli were obtained. With the exception of theP. edulis var.flavicarpa (+)P. coccinea, whole plants were recovered from all hybrids. These somatic hybrids showed 4n=36 chromosomes, which represents a further evidence of their hybridity.