Maria Apparecida Esquibel

Essential oil composition of Melissa officinalis L. in vitro produced under the influence of growth regulators

It was investigated the effects of indole-3-acetic acid (11.42 µmol L-1), benzylaminopurine (8.87 µmol L-1) on essential oil composition and on the growth of Melissa officinalis in vitro plants. In vitro plantlets developed on MS media, showed 1.4 times in the proportion of nerol and 4.1 of geraniol, when compared with ex vitro plants. Treatments with 11.42 µmol L-1 indole-3-acetic acid plus 8.87 µmol L-1 benzylaminopurine led to 1.7 and 2.2 fold in proportion of nerol and geraniol, respectively in 60-day-old whole plants. These increases might be associated with the action of growth regulators wich stimulate plant growth (shoot organogenesis and elongation) and delaying the alcohol oxidation to aldehydes.

Developmental effects of additional ultraviolet a radiation, growth regulators and tyrosine in Alternanthera brasiliana (L.) Kuntze cultured in vitro

Tissue cultures of Alternanthera brasiliana (L.) Kuntze were treated with different growth regulators (Kinetin and 2,4-D), tyrosine and Ultraviolet A radiation (UV-A; 320 - 400 nm) to investigate their effects on development and pigment production. Nodal segments of plantlets grown from seeds were inoculated in all tested media and different light conditions. After eight weeks, this material was used to evaluate biomass, chlorophyll and betacyanin production. The Murashige and Skoog (MS) + kinetin medium resulted in development of approximately four shoots/explant. This medium plus white light was the best combination for micropropagation with the highest rooting percentage and betacyanin production. Plantlets grown under UV-A illumination reduced biomass accumulation and worse Chlorophyll a / Chlorophyll b ratio. Addition of 2,4-D resulted in inhibition of pigment production and growth of plantlets.

Antinociceptive effects of Abarema cochliacarpos (B.A. Gomes) Barneby & J.W.Grimes (Mimosaceae)

In this study, we investigated the analgesic activity of crude aqueous and methanol extracts obtained from Abarema cochliacarpos bark in mice, and analyzed its phytochemical profile. All the extracts exhibited analgesic properties against the writhing test in mice, but the aqueous and methanol extracts were more active, and more potent than two known analgesic and anti-inflammatory drugs used as reference. They were also active against the capsaicin-model, but inactive when evaluated in the hot-plate test. Phytochemical studies revealed the presence of saponins, catechins, tannins, phenols and anthraquinones.

In vitro propagation of the alkaloid producing plant Datura insignis Barb. Rodr.

A method is presented for the in vitro propagation of Datura insignis Barb. Rodr. Nodal explants were cultured on Murashige and Skoog medium supplemented either with BA alone or in combination with 2,4-D or IAA. Shoot multiplication and elongation were obtained in various growth regulator concentrations. Best results were obtained in a medium with 1.0 mgl-1 of BA. Individual shoots were excised and transfered to growth regulator-free medium for rooting. Additional multiplication was obtained by single-node culture using explants from these in vitro rooted shoots. Rooted plantlets were successfully grown in soil.

Standardized production of Phyllanthus tenellus Roxb. by plant tissue culture

Exigencies as ethic plant raw material are part of the needs of modern phytotherapy. Micropropagation offers opportunities to obtain mass propagation of superior genotypes in short time. This study aimed to develop a protocol of direct and indirect organogenesis of Phyllanthus tenellus Roxb. Nodal segments from plantlets obtained by in vitro germination were subcultured in modified Murashige and Skoog medium added with different plant growth regulators: IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), GA 3 (3-giberelic acid) and KIN (kinetin). The highest proliferation rate was obtained using the combinations: IBA, KIN + GA 3 (3.5 mg L -1 ) and IBA + KIN (2.4 mg L -1 ). Rooting was intensified after 40 days, reaching 100% for all media with indole-3-butyric acid. Addition of 2,4 dichlorophenoxyacetic acid (2,4D) provided the best results for production of friable calli. Acclimatization was 100% effective for plantlets cultured in control medium, with decrease in survival rate in grown plantlets from media added with growth regulators.

Solid phase microextraction (SPME) analysis of volatile compounds produced by in vitro shoots of Lantana camara L. under the influence of auxins and cytokinins

on growth, development and volatile compounds production of Lantana camara L. in vitro plants were investigated. The volatiles were extracted by solid phase microextraction (SPME) and the quantification was performed using GC-MS. A higher content of myrcene, α-phellandrene, α-copaene, trans-caryophyllene and β-gurjunene, and the lower levels of α-pinene and β-pinene were observed in in vitro plants cultivated on different supplementation media than in donor plants.

Growth stimulation produced by methylene blue treatment in sweet potato

Methylene blue as an alternative treatment to gamma rays to stimulate growth in sweet potato tissue cultures, was applied in two different ways:– pre-incubation of nodal explants with methylene blue for 1 h using urea as a permeabilizer;– methylene blue directly incorporated into the culture medium.Both treatments stimulated growth, but the better performance being obtained with the second treatment, which had no toxic effect. The activity and electrophoresis pattern of peroxidase after treatment ofIpomoea batatas plantlets with methylene blue or gamma rays did not show similar results for the two treatments. Peroxidase activity was greater in leaves of gamma ray treated plants compared to the non-treated control. The results obtained with the Methylene blue treatment did not significantly change the peroxidase activity relative to the control.

The presence of concanavalin A and canatoxin in Canavalia ensiformis DC tissue culture

Isolated embryos, cotyledons and embryos plusa fragment of cotyledon from seeds of Canavalia ensiformis (jack bean) were cultured in vitro. Concanavalin A and canatoxin cross-reactive material were detected by double immunodiffusion tests. Canatoxin was detectable until 30 days in cultures of embryos, embryos plus cotyledons and hypocotyls. Concanavalin A was also present in all cultures being detected until 90 days in cultures treated with 6-benzylaminopurine. No concanavalin A was detected in root cultures. Concanavalin A was present in cell suspensions until 45 days of culture; the culture medium contained neither concanavalin A nor canatoxin. Tissue cultures thus can produce Con A and CNTX and will be an important research tool for studying the biosynthesis of such substances.

Developmental Changes Induced by γ Irradiation in Ipomoea batatas L. Lam (Sweet Potato)

Storage roots of Ipomoea batatas L. Lam were exposed to gamma irradiation at 90, 180, and 240 Gy. The two highest doses caused a delay in the initiation of rooting and especially of shooting; they also caused formation of rosette-like, stemless shoots without apical meristems. The storage roots irradiated with 90 Gy did form stems, but 50% of them had no apical meristem. The irradiated storage roots produced absorbent roots which developed shoots after 70 days of growth. Nonirradiated storage roots did not differentiate shoots from the absorbent roots. When grown in vitro, phytomers, stem segments with leaves and an axillary meristem, separated from the shoots of irradiated storage roots and exhibited growth aberrations, very intense rooting, and a delay in shooting. Phytomers from nonirradiated normal plants were irradiated with 10, 20, 30, 40, and 90 Gy and grown in a hormone-free medium. The 40- and 90-Gy doses delayed shooting as well as rooting. Only phytomers exposed to 40 and 90 Gy differentiated shoots from the absorbent roots. A stimulation of growth revealed in the accumulation of dry mass was found in the shoots of phytomers irradiated with 10 to 30 Gy. The long after-effects of irradiation as well as possible causes of growth stimulation are discussed.

Solasodine accumulation in regenerated plants of Solanum torvum Sw

RESUMO: Acumulo de solasodina em plantas micropropagadas de Solanum torvum Sw. A regeneracao de Solanum torvum e a avaliacao do conteudo de solasodina foram os objetivos de cultura de segmentos nodais. A influencia de auxinas (acido 3-indolacetico, acido naftalenoacetico) e de 6-benzilaminopurina no crescimento de S. torvum na micropropagacao foi investigado. Cultura de segmentos nodais foi iniciada por sementes germinadas em meio basico MS acrescido de GA 3 e cultivadas em diferentes concentracoes de AIA, AIA + BAP e ANA + BAP. Plantas da cultura in vitro com 60 dias foram coletadas, congeladas e liofilizadas e o metodo de alaranjado de metila foi utilizado para quantificacao de solasodina para o ensaio espectrofotometrico. Os melhores resultados para regeneracao vegetal e acumulo de solasodina foram alcancados no meio MS sem adicao de reguladores de crescimento havendo, porem grande producao de calos de base friaveis. Esses resultados mostram que a partir desses calos, cultura de celulas pode ser iniciada com aplicacao de outras auxinas e citocininas para o aumento da producao de solasodina alem de diferentes eliciadores, intensidades luminosas e concentracoes de sacarose. Palavras-chave: Solanaceae, jurubeba, auxina, citocinina, alcaloide esteroidal