Expression pattern of four storage xyloglucan mobilization-related genes during seedling development of the rain forest tree Hymenaea courbaril L.

During seedling establishment, cotyledons of the rain forest tree Hymenaea courbaril mobilize storage cell wall xyloglucan to sustain growth. The polysaccharide is degraded and its products are transported to growing sink tissues. Auxin from the shoot controls the level of xyloglucan hydrolytic enzymes. It is not yet known how important the expression of these genes is for the control of storage xyloglucan degradation. In this work, partial cDNAs of the genes xyloglucan transglycosylase hydrolase (HcXTH1) and beta-galactosidase (HcBGAL1), both related to xyloglucan degradation, and two other genes related to sucrose metabolism [alkaline invertase (HcAlkIN1) and sucrose synthase (HcSUS1)], were isolated. The partial sequences were characterized by comparison with sequences available in the literature, and phylogenetic trees were assembled. Gene expression was evaluated at intervals of 6 h during 24 h in cotyledons, hypocotyl, roots, and leaves, using 45-d-old plantlets. HcXTH1 and HcBGAL1 were correlated to xyloglucan degradation and responded to auxin and light, being down-regulated when transport of auxin was prevented by N-1-naphthylphthalamic acid (NPA) and stimulated by constant light. Genes related to sucrose metabolism, HcAlkIN1 and HcSUS1, responded to inhibition of auxin transport in consonance with storage mobilization in the cotyledons. A model is proposed suggesting that auxin and light are involved in the control of the expression of genes related to storage xyloglucan mobilization in seedlings of H. courbaril. It is concluded that gene expression plays a role in the control of the intercommunication system of the source-sink relationship during seeding growth, favouring its establishment in the shaded environment of the rain forest understorey.

Polyamines, IAA and ABA during germination in two recalcitrant seeds: Araucaria angustifolia (Gymnosperm) and Ocotea odorifera (Angiosperm)

Background and Aims Plant growth regulators play an important role in seed germination. However, much of the current knowledge about their function during seed germination was obtained using orthodox seeds as model systems, and there is a paucity of information about the role of plant growth regulators during germination of recalcitrant seeds. In the present work, two endangered woody species with recalcitrant seeds, Araucaria angustifolia (Gymnosperm) and Ocotea odorifera (Angiosperm), native to the Atlantic Rain Forest, Brazil, were used to study the mobilization of polyamines (PAs), indole-acetic acid (IAA) and abscisic acid (ABA) during seed germination. Methods Data were sampled from embryos of O. odorifera and embryos and megagametophytes of A. angustifolia throughout the germination process. Biochemical analyses were carried out in HPLC. Key Results During seed germination, an increase in the (Spd + Spm) : Put ratio was recorded in embryos in both species. An increase in IAA and PA levels was also observed during seed germination in both embryos, while ABA levels showed a decrease in O. odorifera and an increase in A. angustifolia embryos throughout the period studied. Conclusions The (Spd + Spm) : Put ratio could be used as a marker for germination completion. The increase in IAA levels, prior to germination, could be associated with variations in PA content. The ABA mobilization observed in the embryos could represent a greater resistance to this hormone in recalcitrant seeds, in comparison to orthodox seeds, opening a new perspective for studies on the effects of this regulator in recalcitrant seeds. The gymnosperm seed, though without a connective tissue between megagametophyte and embryo, seems to be able to maintain communication between the tissues, based on the likely transport of plant growth regulators.

Protection of spruce seedlings against pine weevil attacks by treatment of seeds or seedlings with nicotinamide, nicotinic acid and jasmonic acid

Sustainable methods are required to protect newly planted tree seedlings from insect herbivore attack. To this end, here Norway spruce (Picea abies (L.) Karst.) seeds were treated with 2.5 mM nicotinamide (NIC), 2.5 mM nicotinic acid (NIA), 3 mM jasmonic acid (JA) or 0.2 mM 5-azacytidine (5-Aza), and 6-month-old seedlings grown from these seeds were planted at a reforestation area in central Sweden. Attack by pine weevils (Hylobius abietis) was reduced by 50 per cent by NIC treatment, 62.5 per cent by JA treatment and 25 per cent by 5-Aza treatment, when compared with seedlings grown from untreated seeds. Watering 18-month-old spruce seedlings with 2 mM NIC or 2 mM NIA did reduce attack during the first season in the field by 40 and 53 per cent, respectively, compared with untreated plants. Girdling was also reduced by the different treatments. Analysis of conifer seedlings treated with 5-Aza points at a possible involvement of epigenetic mechanisms in this defensive capacity. This is supported by a reduced level of DNA methylation in the needles of young spruce seedlings grown in a greenhouse from NIC-treated seeds. Seed treatment for seedling defense potentiation is simple, inexpensive and also a new approach for forestry with many potential applications.

Dispersion of Seeds of Tree Species by the Leaf-Cutting Ant Acromyrmex subterraneus molestans (Hymenoptera: Formicidae) in Vicosa, Minas Gerais State, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Can Eucalyptus invade native forest fragments close to commercial stands?

Some Eucalyptus species are widely used as a plantation crop in tropical and subtropical regions. One reason for this is the diversity of end uses, but the main reason is the high level of wood production obtained from commercial plantings. With the advancement of biotechnology it will be possible to expand the geographical area in which eucalypts can be used as commercial plantation crops, especially in regions with current climatic restrictions. Despite the popularity of eucalypts and their increasing range, questions still exist, in both traditional planting areas and in the new regions: Can eucalypts invade areas of native vegetation, causing damage to natural ecosystems biodiversity?The objective of this study it was to assess whether eucalypts can invade native vegetation fragments in proximity to commercial stands, and what factors promote this invasive growth. Thus, three experiments were established in forest fragments located in three different regions of Brazil. Each experiment was composed of 40 plots (1 m(2) each one), 20 plots located at the border between the forest fragment and eucalypts plantation, and 20 plots in the interior of the forest fragments. In each experimental site, the plots were paired by two soil exposure conditions, 10 plots in natural conditions and 10 plots with soil exposure (no plant and no litter). During the rainy season, 2 g of eucalypts seeds were sown in each plot, including Eucalyptus grandis or a hybrid of E. urophylla x E. grandis, the most common commercial eucalypt species planted in the three region. At 15, 30, 45, 90, 180, 270 and 360 days after sowing, we assessed the number of seedlings of eucalypts and the number of seedlings of native species resulting from natural regeneration. Fifteen days after sowing, the greatest number of eucalypts seedlings (37 m(-2)) was observed in the plots with lower luminosity and exposed soil. Also, for native species, it was observed that exposed soil improved natural germination reaching the highest number of 163 seedlings per square meter. Site and soil exposure were the factors that have the greatest influence on seed germination of both eucalypt and native species. However, 270 days after sowing, eucalypt seedlings were not observed at any of the three experimental sites. The result shows the inability of eucalypts to adapt to condition outside of their natural range. However, native species demonstrated their strong capacity for natural regeneration in forest fragments under the same conditions where eucalypts were seeded. (C) 2011 Elsevier B.V. All rights reserved.

Attraction of fruit-eating bats with essential oils of fruits: A potential tool for forest restoration

Previous tests with essential oils from ripe chiropterochoric fruits suggested they can be used to attract and capture fruit-eating bats inside forest remnants. Here we evaluated the efficiency of these oils to attract frugivorous bats to open areas. We performed field tests with artificial fruits impregnated with essential oils of the genera Piper or Ficus that were attached to two groups of mist-nets set 50 m outside the border of a forest remnant. One group of artificial fruits received the corresponding oil isolated through hydrodistillation and the other received water only. Fruits with oils attracted significantly more fruit-eating bats, especially Artibeus lituratus that regularly crosses open habitats to reach other forest remnants. The highly significant attraction of A. lituratus by the oil of Piper was unexpected, since this bat is a specialist on Ficus fruits. We hypothesize that in habitats with no fruit available it is possible to attract frugivorous bats with the odor of several ripe fruit species. Furthermore, we verified that almost half of the individuals captured defecated seeds, indicating that the oils also attract recently fed bats, even when their preferred food is available nearby. This technique potentially may increase seed rain at specific locations, being particularly promising to restoration projects.

Forest fragment size and microhabitat effects on palm seed predation

The establishment of plant species depends crucially on where the seeds are deposited. However, since most studies have been conducted in continuous forests, not much is known about the effects of forest fragmentation on the maintenance of abiotic and biotic characteristics in microhabitats and their effects on seed survival. in this study, we evaluated the effects of forest fragmentation on the predation upon the seeds of the palm Syagrus romanzoffiana in three microhabitats (interior forest, forest edge and gaps) in eight fragments of semi-deciduous Atlantic forest ranging in size from 9.5 ha to 33,845 ha in southeastern Brazil. Specifically, we examined the influence of the microhabitat structure, fauna and fragment size on the pattern of seed predation. Fragments < 100 ha showed similar abiotic and biotic characteristics to those of the forest edge, with no seed predation in these areas. Forest fragments 230-380 ha in size did not present safe sites for S. romanzoffiana seed survival and showed high seed predation intensity in all microhabitats evaluated. In fragments larger than 1000 ha, the seed predation was lower, with abiotic and biotic differences among gaps, interior forests and forest edges. In these fragments, the survival of S. romanzoffiana seeds was related to squirrel abundance and interior forest maintenance. Based on these results, we concluded that there are no safe sites for S. romanzoffiana seed establishment in medium- and small-sized fragments as result of the biotic and abiotic pressure, respectively We suggest that on these forest fragments, management plans are needed for the establishment of S. romanzoffiana, such as interior forest improvement and development in small-sized sites in order to minimize the edge effects, and on medium-sized fragments, we suggest post-dispersal seed protection in order to avoid seed predation by vertebrates. our findings also stress the importance of assessing the influence of forest fragmentation on angiosperm reproductive biology as part of the effective planning for the management of fragmented areas. (c) 2006 Elsevier Ltd. All rights reserved.

Seed rain and seed limitation in a planted gallery forest in Brazil

With seeds collected monthly during one year from 53 1-m(2) seed traps, we investigated the seed rain and seed limitation in a gallery forest planted in 1994 in SE Brazil. Contrasting animal- (zoochorous) and wind-dispersed (anemochorous) plants we investigated (1) which aspects of the composition and structure of the vegetation influence the abundance and species richness of the seed rain; (2) if such influences differ between zoochorous and anemochorous seeds; (3) if the abundance and richness of the seed rain sampled under zoochorous and nonzoochorous plant species differ; and (4) if seed limitation (given by the proportion of sites to which seeds were not dispersed) differs between zoochorous and anemochorous plant species, and also between species that have been planted and those that further colonized the area (colonists). Seed rain was intense and dominated by anemochorous species. The overall seed rain was not influenced by the vegetation parameters we analyzed (canopy height and cover, plant size, abundance, and richness) or by the plant species above the seed trap. The abundance and richness of zoochorous seeds in a given spot was influenced by the abundance and richness of zoochorous plants in its immediate vicinity. Seed limitation was higher for anemochorous than zoochorous species and higher for planted than for colonist species. We concluded with recommendations for the initial establishment of a planted forest, including the homogeneous distribution of zoochorous plants to permit a spatially homogeneous zoochorous seedfall, which will likely enhance the chances of survival and successful establishment of seeds.

Germination of Ocotea pulchella (Nees) Mez (Lauraceae) seeds in laboratory and natural restinga environment conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Seed deposition patterns and the survival of seeds and seedlings of the palm Euterpe edulis

The seed deposition pattern created by a seed disperser is one of the components of the efficiency of a species as seed disperser, and ultimately may influence the recruitment of a plant species. In this study, we used the seeds of a bird-dispersed forest palm, Euterpe edulis, to investigate the effects of two distinct seed deposition patterns created by birds that defecate (clumped pattern) and regurgitate seeds (loose-clumped pattern) on the survival of seeds experimentally set in an E. edulis-rich site, and of seedlings grown under shade-house conditions. The study was conducted in the lowland forest of Parque Estadual Intervales, SE Brazil. Clumped and loose-clumped seeds were equally preyed upon by rodents and insects. Although clumped and isolated seedlings had the same root weight after 1 year, the isolated seedlings survived better and presented more developed shoots, suggesting intraspecific competition among clumped seedlings. Our results indicate that animals that deposit E. edulis seeds in faecal clumps (e.g. cracids, tapirs) are less efficient seed dispersers than those that regurgitate seeds individually (e.g. trogons, toucans). Intraspecific competition among seedlings growing from faecal clumps is a likely process preventing the occurrence of clumps of adult palms. (C) 2001 Editions scientifiques et medicales Elsevier SAS.

Granivorous birds as potentially important post-dispersal seed predators in a Brazilian forest fragment

In tropical forests, rodents exert a prominent role as post-dispersal seed predators, while other vertebrates apparently have a minor effect on seed predation. In some forest fragments in southeast Brazil, however, the density of rodents is unusually low, whereas terrestrial granivorous birds are abundant. In this study, we used seeds of a second-growth tree (Croton priscus, Euphorbiaceae) to investigate experimentally the relative importance of rodents and birds as seed predators in a forest fragment. We also estimated the density of terrestrial granivorous birds and trapped small mammals. Results indicate that, as a consequence of decreased rodent population and increased bird abundance, the latter are at least as important as the former as the main post-dispersal seed predators of C priscus. We interpret such apparent imbalance in the interaction between C priscus and their seed predators as a possible consequence of forest fragmentation.

Mobilisation of reserves during germination of seeds of Tabebuia roseo-alba (Bignoniaceae)

Tabebuia roseo-alba is a native Brazilian tree species with wide occurrence, particularly in semi-deciduous forest. The tree has lush flowering and is used in landscaping, arboriculture and for reforestation of dry and rocky soils. Its seeds are small, lightweight and winged, and are wind-dispersed. The objective of this study was to investigate the biochemical changes in T. roseo-alba seeds during germination. We determined the content of starch, structural polysaccharides, lipid and soluble protein, and analysed the proteins by SDS-PAGE. There was a rapid consumption of starch, structural polysaccharides, lipids and proteins during germination of T. roseo-alba seeds. The rapid mobilisation of protein during germination, was accompanied by the synthesis of high molecular mass proteins and degradation of low molecular mass proteins.

Morphological and physiological responses of the recalcitrant Euterpe edulis seeds to light, temperature and gibberellins

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversity of functional traits of fleshy fruits in a species-rich Atlantic rain forest

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)