Seedling morphology and development in the epiphytic cactus Epiphyllum phyllanthus (L.) Haw. (Cactaceae: Hylocereeae)

Seedling morphology is relevant in classification, taxonomy, and vegetation studies to understand plant life cycles, germination succession and requirements, and developmental progression. However, most morphological studies of seedlings lack analysis of organ anatomy, impeding the comprehension of series of development and establishment in a particular environment. Here, we have taken a traditional anatomical approach to examine the stages of seedling development in Epiphyllum phyllanthus, a holo-epiphytic cactus of tribe Hylocereeae. The goals were 1) to offer a comprehensive description of growth series in E. phyllanthus seedlings based on morphological and anatomical analysis and 2) to examine the initial growth phases in the life cycle of this species to identify organ development and understand their adaptive significance in relation to seedling establishment. Our results include descriptions of seed morphology, embryonic features, and seedling vascularization pattern in the root, hypocotyl, cotyledons, and epicotyl. The morphological and developmental patterns in E. phyllanthus seedlings have potential phylogenetic and ontogenetic implications in the Cactaceae. Characters such as the presence of mucilage on the seed coat, the lack of seed operculum, and large cotyledons in E. phyllanthus are comparable to basal cacti, but the root anatomy is more similar to columnar relatives. At the familial level, there is an apparent trend in decreasing number of phloem and xylem poles in the stele of primary root, correlated with degree of specialization and advanced phylogenetic position: tetrarch to septarch-octarch in basal lineages, tetrarch Cereus-type in columnar species, to the diarch vascular system in Rhipsalideae and some species with cylindric/globose stem. © Torrey Botanical Club.

Effects of substrate and temperature on the emergence of Phacelia sp. (Boraginaceae) seedling

This study aimed to define the best substrate and temperature for the emergence of Phacelia sp. seedling, annual ornamental flower gardens. The experimental design was entirely randomized in a factorial scheme 37 (three different types of substrates: vermiculite, sand and sphagnum combined with seven temperature conditions: room temperature, constant at 20, 25, 30 and 35C, and alternated at 20-30C and 25-35C) with 4 replications of 100 seeds each. Emergence (%E) and Emergence Rate (ER) were evaluated. The means of the resulting values were then compared by the Tukey test at 5% confidence level. There was a significant interaction amongst substrates and temperatures for all analyzed variables. For seeds sown in vermiculite and sand, the seedlings had higher %E and emerged fastest at 20C, room temperature and 20-30C that did not present statistically significant data. In sphagnum, seedlings showed greater %E in the alternated at 20-30C, room temperature and 20C and emerged quickly at room temperature and 20C that did not present statistically significant data. At room temperature and 20C, the seedlings had higher %E and emerged faster in vermiculite and sand that did not present statistically significant data. At 25, 30 and 25-35C, the seedlings showed better %E and emerged fastest in sand. At 35C, the seedlings showed either in vermiculite and sand were not significantly different in their emergence, but emerged faster in sand. There was not significantly different among substrates in their emergence at 20-30C, but the seedlings emerged faster in vermiculite and sand. It was concluded that the Phacelia sp. seedlings in all substrates showed greater %E at room temperature, 20C and 20-30C. For all temperatures, the seedlings growth parameters were superior when seeds were sown in sand.

Cryopreservation, early seedling development, and genetic stability of Oncidium flexuosum Sims

An efficient cryopreservation protocol was developed for mature seeds of Oncidium flexuosum Sims. Seed morphology, protocorm formation, and early seedling development were also assessed. The effects of phloroglucinol and Supercool X-1000® as cryoprotectant additives in the vitrification solution were investigated. Dehydration using the plant vitrification solution 2 (PVS2) for 60 and 120 min prior to immersion in liquid nitrogen promoted the highest frequency of in vitro seed germination 6 weeks following culture on half-strength Murashige and Skoog (1/2 MS) medium. Mature seeds submitted to vitrification for 120 min in PVS2 and 1 % phloroglucinol at 0 °C enhanced germination by 68 %, whereas in PVS2 and 1 % Supercool X-1000® germination was just moderately enhanced (26 %). In vitro-germinating seedlings developed healthy shoots and roots without the use of plant growth regulators. After 6 months of growth, there were no differences between in vitro- and ex vitro-grown seedlings for various phenotypic characteristics, including shoot length, number of leaves, number and length of roots, and fresh and dry weight. Seedlings were transferred to greenhouse conditions and successfully acclimatized, further developing into normal plants with over 90 % survival. Comparative analysis of seedlings from control and vitrified seeds using flow cytometry indicated that no change in ploidy levels occurred as a result of cryopreservation, therefore maintaining seedlings genetic stability. In this study, vitrification with PVS2 for 120 min with the addition of 1 % phloroglucinol offers a simple, safe, and feasible protocol for cryopreservation of O. flexuosum mature seeds. © 2013 Springer Science+Business Media Dordrecht.

Growth regulators for in vitro propagation of Tillandsia gardneri (Bromeliaceae)

Tillandsia gardneri Lindi, is a herbaceous perennial with ornamental value. However, in Brazil there is no report about this species' cultivation on a commercial scale. The low multiplication rate of T. gardneri (in average one offshoot/plant/year) leads to illegal over-collection in the wild to meet commercial demands. The development of protocols for in vitro propagation of T. gardneri may be useful for increasing multiplication rate, producing enough plants to supply the ornamental market and also to reduce the pressure over plants collection in the wild. The present study evaluated the effect of growth regulators (6-benzylaminopurine-BA alone or in combinations with naphthaleneacetic acid-NAA) on shoots development from seedlings pre-established in vitro, from seed germination on 1/4 MS medium without growth regulators. Seedlings (with about 1.0 cm long) were re-cultured to solid 1/2 MS media supplemented with growth regulators. After 30 days on the induction medium seedlings were re-cultured to MS basal medium. The experiment was conducted in a complete randomized design with four replications and ten treatments: control (free of growth regulators), BA (0.5, 1.0 and 2.0 mg/L), BA (0.5 mg/L) + ANA (0.1 mg/L), BA (1.0 mg/L) + ANA (0.1 mg/L), BA (1.0 mg/L) + ANA (0.5 mg/L), BA (2.0 mg/L) + ANA (0.1 mg/L), BA (2.0 mg/L) + ANA (0.5 mg/L), and BA (2.0 mg/L) + ANA (1.0 mg/L). The outgrowth of shoots did not occur on medium devoid of growth regulators (control). Regression analysis for some evaluated parameters, such as percentage of seedlings responsive to shoot formation and number of shoots/seedling, and regulators concentrations (BA or ANA) were significant, allowing the establishment of the growth regulators concentration for obtaining the best multiplication rate. Some seedlings maintained in media with ANA (0.5 or 1.0 mg/L) were completely converted into callus masses that turned dark brown leading to seedlings death.

Morphology and anatomy of the diaspores and seedling of Paspalum (Poaceae, Poales)

RESUMO O conhecimento relativo ao diásporo e ao desenvolvimento pós-seminal de Paspalum L. é importante para a conservação da biodiversidade dos campos, devido sua importância na representatividade e no melhoramento genético de pastagens. A morfologia do diásporo e do desenvolvimento pós-seminal de Paspalum dilatatum Poir. (rizomatosa); P. mandiocanum Trin. var. subaequiglume Barreto (estolonífera), P. pumilum Nees. (cespitosa decumbente) e P. urvillei Steud. (cespitosa ereta) foi descrita procurando distinguir as espécies com diferentes formas de crescimento, e levantar características úteis para a taxonomia. P. dilatatum se diferencia por apresentar diásporo oval, de maior tamanho que as demais, com cinco nervuras salientes e tricomas; P. urvillei por apresentar diásporo com uma nervura central mais desenvolvida do que as duas nervuras laterais e tricomas; P. mandiocanum var. subaequiglume por apresentar diásporo com tricomas apenas na margem; e P. pumilum por apresentar diásporo glabro. A cariopse envolve a semente que apresenta embrião diferenciado, disposto lateralmente; apresenta hilo elíptico em todas as espécies estudadas e rostelo em P. dilatatum e P. mandiocanum var. subaequiglume. O desenvolvimento pós-seminal é semelhante nas quatro espécies e se inicia com a germinação, que é marcada pela emergência da coleorriza, seguida pelo coleóptilo. Essas características são comuns às demais Poaceae já estudadas, indicando um padrão para a família e não diferenciam as formas de crescimento.

EFFECTS OF EMERGENCY IN BIOSTIMULANT SEEDLING OF PASSION FRUIT ROXINHO OF KENYA

The increase in market demand for fresh fruits along with the high price of passion fruit juice in domestic and international markets has increased the interest in fruit, especially the purple passion fruit in the center south of the country seeking to export. This study aimed to evaluate the effects of plant growth regulator on the emergence and development of seedlings of passion 'Roxinho of Kenya' when propagated sexually. The treatments consisted of concentrations of commercial product Stimulate (R): control (no biostimulant); 6; 12; 18; 24 and 30 mL. kg(-1) of seed. It were evaluated the percentage of seedling emergence, number of leaves, aerial part dry weight, stem and root (g), root length (mm), diameter and stem length (mm), leaf area and chlorophyll 'a' and 'b'. The application of bio-stimulant in doses of 6 and 12 mL. kg(-1) promotes increased percentage of seedling emergence of Passiflora edulis Sims in a shorter time. The use of it also promotes the development of seedlings, with better results for the dose of 12 and 24 mL. kg(-1) treated seeds.

Seedling fate across different habitats: the effects of herbivory and soil fertility

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

Initial growth of bauhinia variegata trees under different colored shade nets and light conditions

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

PHYSIOLOGICAL CHANGES IN SEEDS AND ANTIOXIDANT METABOLISM OF LETTUCE SEEDLING EXPOSED TO THE ACTION OF Zantedeschia aethiopica Spreng. LEAF EXTRACT

This work aimed to evaluate the influence of different concentrations of Zantedeschia aethiopica Spreng. extract on the physiological performance of the seed and on the response of the antioxidant metabolism of lettuce seedlings. The treatments consisted of leaves extracts from Z. aethiopica at concentrations of 0, 6, 12, 25 and 50%. Germination, first germination count, germination speed and index, length of shoot and radicle, seedling total dry mass, chlorophyll content, activity of superoxide dismutase, catalase and ascorbarte peroxidase enzymes, lipid peroxidation, hydrogen peroxide quantification and seedling emergence, length of organs, and total dry mass of seedlings were evaluated. The percentage of germination, the length of the shoot and radicle of seedlings and the total dry mass of seedlings grown in the greenhouse were reduced as the concentration of the extract increased. There were increases of electrical conductivity, of superoxide dismutase, catalase and ascorbate peroxidadase enzymes and the amount of hydrogen peroxide and lipid peroxidation in seedlings with increasing extract concentration. The extract reduced the physiological quality of lettuce seeds and induced an increased production of hydrogen peroxide in seedlings, which increased the activity of antioxidant enzymes that were not effective in tissue detoxification, resulting in cellular damage and increased numbers of abnormal seedlings.

Growth of seedlings Schizolobium amazonicum (Huber ex Ducke) on substrate fertilized with nitrogen, phosphorus and potassium

The nutritional management of seedlings in the nursery is one of the most important practices that influence seedling quality. The aim of this work was to evaluate the effect of nitrogen, phosphorus and potassium on the development of Schizolobium amazonicum seedlings grown in 250 cm(3) containers with a commercial substrate in the North of Mato Grosso State, Brazil. The experimental design was completely randomized design with five treatments and five replications, each replication being represented by 24 seedlings. The treatments were: control (only commercial substrate); nitrogen fertilization (150 g m(-3) N using ammonium sulfate + 1.0 kg of ammonium sulfate dissolved in 100 L of water and applied in coverage); phosphorus fertilization (300 g P2O5 m(-3) using simple superphosphate); potassium fertilization (100 g m(-3) K2O using potassium chloride + 0.3 kg of potassium chloride dissolved in 100 L of water and applied in coverage) and; complete (a mixture of the three nutrients, 150, 300 and 100 g m(-3) N, P2O5 and K2O, respectively + 1.0 kg of ammonium sulfate + 0.3 kg of potassium chloride). The commercial substrate was composted milled pine bark plus vermiculite. Evaluations of the seedlings were performed at 90 days after sowing. The complete treatment (NPK) gave the highest values for biometric and best plant indices, which express the quality. When analyzing nutrients in isolation; potassium had the lowest effect. Based on these results it can be recommended to fertilize Schizolobium amazonicum seedlings in nurseries with 150, 300 and 100 g m(-3) of N, P2O5 and K2O, respectively, plus 1.0 kg of sulfate ammonium and 0.3 kg of potassium chloride applied in coverage.

Effects of experimental nitrogen additions on plant diversity in an old-growth tropical forest

Response of plant biodiversity to increased availability of nitrogen (N) has been investigated in temperate and boreal forests, which are typically N-limited, but little is known in tropical forests. We examined the effects of artificial N additions on plant diversity (species richness, density and cover) of the understory layer in an N saturated old-growth tropical forest in southern China to test the following hypothesis: N additions decrease plant diversity in N saturated tropical forests primarily from N-mediated changes in soil properties. Experimental additions of N were administered at the following levels from July 2003 to July 2008: no addition (Control); 50 kg N ha−1 yr−1 (Low-N); 100 kg N ha−1 yr−1 (Medium-N), and 150 kg N ha−1 yr−1 (High-N). Results showed that no understory species exhibited positive growth response to any level of N addition during the study period. Although low-to-medium levels of N addition (≤100 kg N ha−1 yr−1) generally did not alter plant diversity through time, high levels of N addition significantly reduced species diversity. This decrease was most closely related to declines within tree seedling and fern functional groups, as well as to significant increases in soil acidity and Al mobility, and decreases in Ca availability and fine-root biomass. This mechanism for loss of biodiversity provides sharp contrast to competition-based mechanisms suggested in studies of understory communities in other forests. Our results suggest that high-N additions can decrease plant diversity in tropical forests, but that this response may vary with rate of N addition.

Canopy gaps promote selective stem-cutting by small mammals of two dominant tree species in an African lowland forest: the importance of seedling chemistry

Small mammals can impede tree regeneration by injuring seedlings and saplings in several ways. One fatal way is by severing their stems, but apparently this type of predation is not well-studied in tropical rain forest. Here, we report on the incidence of 'stem-cutting' to new, wild seedlings of two locally dominant, canopy tree species monitored in 40 paired forest understorey and gap-habitat areas in Korup, Cameroon following a 2007 masting event. In gap areas, which are required for the upward growth and sapling recruitment of both species, 137 seedlings of the long-lived, light-demanding, fast-growing large tropical tree (Microberlinia bisulcata) were highly susceptible to stem-cutting (83% of deaths) - it killed 39% of all seedlings over a c. 2-y period. In stark contrast, seedlings of the more shade-tolerant, slower-growing tree species (Tetraberlinia bifoliolata) were hardly attacked (4.3%). In the understorey, however, stem-cutting was virtually absent. Across the gap areas, the incidence of stem-cutting of M. bisulcata seedlings showed significant spatial variation that could not be explained significantly by either canopy openness or Janzen-Connell type effects (proximity and basal area of conspecific adult trees). To examine physical and chemical traits that might explain the species difference to being cut, bark and wood tissues were collected from a separate sample of seedlings in gaps (i.e. not monitored for stem-cutting). These analyses suggested that, compared with T. bifoliolata, the lower stem density, higher Mg and K and fatty acid concentrations in bark, and fewer phenolic and terpene compounds in M. bisulcata seedlings made them more palatable and attractive to small-mammal predators, likely rodents. We conclude that selective stem-cutting is a potent countervailing force to the current local canopy dominance of the grove-forming M. bisulcata by limiting the recruitment and abundance of its saplings. Given the ubiquity of gaps and ground-dwelling rodents in pantropical forests, it would be surprising if this form of lethal browsing was restricted to Korup.

Postgerminative growth and lipid catabolism in oilseeds lacking the glyoxylate cycle

The glyoxylate cycle is regarded as essential for postgerminative growth and seedling establishment in oilseed plants. We have identified two allelic Arabidopsis mutants, icl-1 and icl-2, which lack the glyoxylate cycle because of the absence of the key enzyme isocitrate lyase. These mutants demonstrate that the glyoxylate cycle is not essential for germination. Furthermore, photosynthesis can compensate for the absence of the glyoxylate cycle during postgerminative growth, and only when light intensity or day length is decreased does seedling establishment become compromised. The provision of exogenous sugars can overcome this growth deficiency. The icl mutants also demonstrate that the glyoxylate cycle is important for seedling survival and recovery after prolonged dark conditions that approximate growth in nature. Surprisingly, despite their inability to catalyze the net conversion of acetate to carbohydrate, mutant seedlings are able to break down storage lipids. Results suggest that lipids can be used as a source of carbon for respiration in germinating oilseeds and that products of fatty acid catabolism can pass from the peroxisome to the mitochondrion independently of the glyoxylate cycle. However, an additional anaplerotic source of carbon is required for lipid breakdown and seedling establishment. This source can be provided by the glyoxylate cycle or, in its absence, by exogenous sucrose or photosynthesis.

The role of nutrients for improving seedling quality in drylands

Forest plantations have been extensively used to combat desertification. In drylands, harsh climate conditions and unfertile soils often preclude seedling establishment. The improvement in seedling quality by manipulating nutrient availability could contribute to increase planting success. However, morpho-functional traits defining optimum seedling quality in drylands, and the fertilization schemes to achieve them, are still under discussion. Several studies suggest that well fertilized seedlings may perform better than nutrient limited seedlings in these environments. However, recent works have shown opposite results. In this review, we discuss the concept of seedling quality in drylands based on an evaluation of the effects of nutrient manipulation on seedling morpho-functional traits and field performance. According to existing data, we hypothesize that nutrient-limited small seedlings may be better adapted to arid environments and unfavorable microsites, where access to water is uncertain and a conservative water use strategy may be advantageous. In contrast, in dry sub-humid areas, areas with deep soils, protected from excess radiation, and areas where irrigation is feasible, well-fertilized big seedlings with high root growth potential may have more chances of success. We discuss this theory in the context of the multiple objectives of dryland restoration and the environmental constrains posed by these areas, and identify knowledge gaps that should be targeted to test our hypothesis.

Water and nutrient availabilities do not affect lignotuber growth and sprouting ability of three eucalypt species of south-eastern Queensland

Shoot biomass and lignotuber size of seedlings of three eucalypt species, Eucalyptus acmenoides Schauer, E. siderophloia Benth. and Corymbia variegata [syn. E. maculata (F. Muell.)K. D. Hill and L. A. S. Johnson], were measured for glasshouse-grown seedlings established under two water and nutrient regimes. Seedlings were subjected to shoot removal (clipping) at ages from 9 to 19 weeks, and transferred to the high water treatment for a further 8 weeks to assess shoot emergence from lignotubers. Seedling shoot biomass was greater in both the high than the low nutrient and water treatments, but lignotuber diameter was not affected significantly. C. variegata seedlings had the largest lignotuber diameters, followed by E. siderophloia and E. acmenoides, respectively. Although growth of shoots was influenced by nutrient availability, results suggest that species differences in the growth of lignotubers was less affected. It is suggested that lignotuber growth was strongly influenced by genotype. More than 70% of C. variegata seedlings clipped at 9 weeks sprouted, compared with only 5 and 10% of seedlings of E. siderophloia and E. acmenoides, respectively. All C. variegata seedlings sprouted after being clipped at 19 weeks, but < 80% of E. siderophloia and < 60% of E. acmenoides sprouted when clipped at the same age. It was concluded that seedlings forming part of the regeneration stratum in dry sclerophyll forests need to be protected from damage for at least 4 months (for C. variegata) or at least 6 months (for E. siderophloia and E. acmenoides) if they are to survive by sprouting from lignotubers.