Pollen grain development and male sterility in the perfect flowers of Maytenus obtusifolia Mart. (Celastraceae).

Perfect flowers of Maytenus obtusifolia have partial sterility of pollen grains, resulting in collapsed and developed free microspores. However, the cellular events resulting in partial male sterility have not been determined. In pistillate flowers of this species, male sterility is related to the premature programmed cell death (PCD) in tapetum and sporogenic cells. The process occurs through autophagy via macroautophagy and massive autophagy and is associated with sporophytic cytoplasmic male sterility (CMS). Here, we characterised the development of pollen grains and investigated the cellular events that result in tapetum cells and free microspores PCD in perfect flowers, using light and transmission electron microscopy combined with the TUNEL (Terminal deoxynucleotidyl transferase mediated dUDP end-Labeling) assay and the ZIO (Zinc iodide-osmium tetroxide) method. Pollen grain development in perfect flowers was divided into eight developmental stages based on the characteristics of the pollen grains. Tapetum cells undergo PCD at the free microspore stage, through a macroautophagic process, by formation of autophagosomes and by autophagosomes giving rise to lytic vacuoles at maturity. In the final stage of PCD, massive autophagy occurs by rupture of the tonoplast. The development of viable and inviable microspores diverges at the vacuolated microspore stage, when PCD occurs in some free microspores, causing interruption of pollen development through necrosis. These events result in the observed partial male sterility. Viable microspores undergo mitosis and develop into tricellular pollen grains. Male sterility in hermaphrodite individuals is here interpreted as gametophytic CMS.

Production of the quinone-methide triterpene maytenin by in vitro adventitious roots of Peritassa campestris (Cambess.) A.C.Sm. (Celastraceae) and rapid detection and identification by APCI-IT-MS/MS.

Establishment of adventitious root cultures of Peritassa campestris (Celastraceae) was achieved from seed cotyledons cultured in semisolid Woody Plant Medium (WPM) supplemented with 2% sucrose, 0.01% PVP, and 4.0 mg L⁻¹ IBA. Culture period on accumulation of biomass and quinone-methide triterpene maytenin in adventitious root were investigated. The accumulation of maytenin in these roots was compared with its accumulation in the roots of seedlings grown in a greenhouse (one year old). A rapid detection and identification of maytenin by direct injection into an atmospheric-pressure chemical ionization ion trap tandem mass spectrometer (APCI-IT-MS/MS) were performed without prior chromatographic separation. In vitro, the greatest accumulation of biomass occurred within 60 days of culture. The highest level of maytenin--972.11 µ g·g⁻¹ dry weight--was detected at seven days of cultivation; this value was 5.55-fold higher than that found in the roots of seedlings grown in a greenhouse.

Capacidade de enraizamento de estacas de Maytenus muelleri Schwacke com a aplicação de ácido indol butírico relacionada aos aspectos anatômicos / Rooting capacity of Maytenus muelleri Schwacke cuttings with indolebutyric acid application related to anatomical aspects

A espinheira-santa (Maytenus muelleri - Celastraceae) é a planta medicinal nativa do Sul do Brasil, cujas folhas são tradicionalmente utilizadas pela medicina popular para o tratamento de úlceras e outros problemas gástricos. Existem poucos trabalhos publicados sobre a produção de mudas e técnicas de propagação vegetativa da espécie. A propagação de espinheira-santa por estaquia poderia ser um método eficiente para obtenção de material homogêneo, com características genéticas desejáveis, produzido a partir de plantas matrizes selecionadas. O presente trabalho teve por objetivo estudar os efeitos da aplicação de ácido indol butírico (AIB), em solução e em pó, no enraizamento de estacas de espinheira-santa coletadas nas quatro estações do ano (abril/2005 a janeiro/2006), bem como averiguar, por meio de análises anatômicas e histoquímicas das estacas, a presença de possíveis impedimentos à iniciação do enraizamento adventício. Estacas provenientes de ramos de plantas matrizes de seis anos cultivadas da Estação Experimental do Canguiri, Pinhais, PR, foram coletadas e tratadas com AIB (0, 1500, 3000 mg L-1 ou mg kg-1), em solução alcoólica (50 por cento v/v) e em talco. Aos 365 dias foram avaliadas as porcentagens de estacas enraizadas e mortas, número e comprimento médio de raízes formadas por estaca. Análises anatômicas e histoquímicas com lugol e cloreto férrico foram realizadas. A estação mais promissora para o enraizamento foi o verão/2006 com 62,50 por cento para o tratamento controle, devido à menor lignficação dos ramos no período de intenso crescimento vegetativo. O número médio de raízes formadas por estaca foi de 6,94 (solução) e o comprimento médio de raízes formadas/estaca chegou a 4,82 cm nesta mesma estação. As concentrações de AIB aplicadas não foram eficientes na indução radicial, independentemente do modo de aplicação. Foi detectada a presença de uma camada quase contínua de fibras e braquiesclereídes, a qual constitui barreira anatômica à indução radicial. Os testes histoquímicos revelaram a presença de amido e de compostos fenólicos nas estacas, em todas as estações do ano. A dificuldade ou demora no enraizamento não pode ser justificada pela falta de reservas de amido nos tecidos das estacas, mas pode ser justificada pela presença de compostos fenólicos, possivelmente do grupo dos monofenóis, que causam a degradação do AIA, interferindo negativamente na indução do enraizamento.

"Espinheira-santa" (Maytenus muelleri - Celastraceae) is a medicinal plant native to Southern Brazil, the leaves of which are traditionally used in popular medicine for the treatment of stomach ulcers and other gastric problems. There are few published studies about seedling production and vegetative propagation techniques for this species. The propagation of "espinheira-santa" by cuttings could be an efficient method to obtain homogeneous material, with desirable genetic characteristics, produced from selected mother plants. This paper aimed to study the effects of indolebutyric acid (IBA) application, in solution and in powder, on the rooting of "espinheira-santa" cuttings, collected in four seasons (April/2005 to January/2006), as well as to investigate, by means of cutting anatomical and histochemical analyses, the presence of possible impediments to adventitious rooting initiation. Cuttings from branches of six-year mother plants grown at "Estação Experimental do Canguiri", Pinhais, Paraná State, Brazil, were collected and treated with IBA (0, 1500, 3000 mg L-1 or mg kg-1) in alcoholic solution (50 percent v/v) and in powder. After 365 days, the percentages of rooted and dead cuttings, the number and mean length of roots/cutting were evaluated. Anatomical and histochemical analyses were performed with lugol and ferric chloride. The most promising season for rooting was Summer/2006, with 62.50 percent of rooting for the control treatment, due to the lesser lignification degree of branches in intense vegetative growth period. The mean number of roots/cutting was 6.94 (solution) and the mean length of roots/cutting was 4.82 cm in that same season. The applied IBA concentrations were not efficient in inducing root growth, regardless of the application method. An almost continuous layer of fibers and stone cells was detected, constituting an anatomical barrier for rooting induction. The histochemical tests revealed the presence of starch and phenolic compounds in cuttings, in all seasons. The difficulty or delay in rooting cannot be justified by the absence of starch reserve in the cutting tissues but by the presence of phenolic compounds, possibly of the group of monophenols, which cause IAA degradation, negatively affecting rooting induction.

[Population structure of endangered Monimopetalum chinense and its relationships with environmental factors].

Based on the field investigation data obtained from the typical plots of four community types, i. e. , secondary shrub, Phyllostachys edulis forest, Cunninghamia lanceolata forest, and Pinus massoniana forest, in the Zongli Village of Qimen County in Anhui Province, this paper studied the mean basal diameter and structure of Monimopetalum chinense population, and the effects of environmental factors on the population characteristics. The results showed that the mean basal diameter of M. chinense in the communities was in the order of P. edulis forest > P. massoniana forest > C. lanceolata forest > secondary shrub, and significantly larger in the two former forests than in the others (P < 0.05). The population structure of M. chinense also differed with habits. In secondary shrub and P. massoniana forest, the structure was a aptypical pyramid-like form, suggesting that the population was stable; in P. edulis forest, it was a spindle type, indicating that the population was at the early stage of declining; whereas in C. lanceolata forest, it was a typical pyramid-like form, with most young individuals in the population. The survival curve of the whole population belonged to Deevey II, suggesting that the population was in developing tendency with no declination. M. chinense preferred the sites with low altitude, high soil moisture and organic matter contents, gentle slope, and high coverage of tree layer; while frequent human disturbance decreased its natural regeneration and stability. Based on the results obtained, some preliminary protection suggestions were proposed.

Successful seed germination of the nickel hyperaccumulator Stackhousia tryonii.

BACKGROUND AND AIMS: Stackhousia tryonii, a rare nickel hyperaccumulating herb, is endemic to ultramafic (serpentine) soils of central Queensland, Australia. The effects of eight dormancy-relieving treatments on germination of stored seeds of Stackhousia tryonii were investigated under controlled light and temperature conditions. * METHODS: The treatments were: untreated (control i), leached and dehydrated (primed control ii), treating with gibberellic acid (150 and 300 microM), smoke extract (5 and 10 %, v/v) and potassium cyanide (40 and 80 mM). * KEY RESULTS: Freshly harvested seeds did not germinate. Germination percentage increased with time of storage for up to 18 months (38.3 %). Gibberellin, smoke extract and cyanide treatments did not significantly improve germination. Light did not affect seed germination and there was no interaction between dormancy-relieving treatments and light. A significant inhibition of germination occurred in seeds treated with 5 % (but not 10 %) aqueous smoke extract. Saturated fatty acids, predominantly tridecanoic (C13:0), constituted about 90 % of the total fatty acids in the oil of freshly harvested seeds. In contrast, there was increased accumulation (>75 %) of mono-unsaturated (oleic, c18:1) and poly-unsaturated (linoleic, c18:2; linolenic, c18:3) fatty acids in the oil of stored seeds. * CONCLUSIONS: Seeds of S. tryonii require an after-ripening period for germination.

Rapid in vitro multiplication and restoration of Celastrus paniculatus Willd. sub sp. paniculatus (Celastraceae), a medicinal woody climber.

Nodes, shoot tips, internodes and leaf bases (approximately 1.0 cm) excised from young vines of the flowering woody climber, Celastrus paniculatus WilId. sub. sp. paniculatus (Celastraceae) were cultured in Murashige and Skoog (MS) medium containing agar (0.6%), sucrose (3%) and varied concentrations of 6-benzyl aminopurine (BAP) and kinetin. All the explant types were regenerative and maximum number (3.6) and frequency (94%) of axillary shoot formation of (5.08 cm long) was recorded in the nodes cultured in BAP (1 mg L(-1)) after 6 weeks. Combinations of BAP (1 mg L(-1)) and indole-3-acetic acid/l-naphthalene acetic acid (0.01-1 mg L(-1); IAA/NAA) tested with nodes induced formation of less number (3 and 2.2) of shoots at same frequency (94%). All the explant types viz. node, shoot tip, internode and leaf base of in vitro derived shoots responded earlier and better in lower concentrations of BAP (0.5-2 mg L(-1)) with formation of 8, 3.1, 6.4 and 1.8 shoots respectively during the same period. In spite of the advanced and increased caulogenic responses, differences in cytokinin requirements between different explants observed during culture initiation still persisted with the nodes, shoot tips, internodes and petiole segments responding best at 0.5, 1 and 2 mg L(-1) BAP, respectively. The repeated reculture up to 10 cycles of the nodes from the shoot cultures each at 6-week intervals enabled multiplication and stocking of shoots without decline. Rooting of 3-7 cm shoot cuttings was induced in half-strength MS liquid medium containing IAA (1 mg L(-1)) with formation of 7.25 roots of 2.41 cm length within 6 weeks. Rooted plants were established at 84-96% rate in community pots without hardening, the least value (84%) being obtained with NAA- induced thick and calloid rooted plants. Four month old community potted plants were reintroduced into native forest habitats at 95% efficiency and 8 months after restoration, the plants were uniform in morphological, growth, cytological and peroxidase and esterase isozyme characteristics.