Influencia de reguladores de crecimiento sobre el establecimiento de cultivos in vitro de Salvia hispanica L. ("chía") y sobre su contenido de ácidos grasos / Influence of growth regulators on the establishment of in vitro cultures of Salvia hispanica L. ("chia") and its content of fatty acids

Salvia hispanica L. (Labiateae), comúnmente conocida como "chía", es una especie herbácea anual cuyo cultivo está ampliamente extendido por América del Sur. Es utilizada como cultivo industrial y para la ela-boración de alimentos funcionales por su contenido en proteínas, antioxidantes, fibras y lípidos esenciales. El aceite de sus semillas contiene la mayor proporción (68 %) de omega-3 que cualquier fuente vegetal conocida. El objetivo de este trabajo fue establecer cultivos in vitro de "chía" y analizar la influencia de diferentes reguladores de crecimiento sobre la inducción de callos, sobre el crecimiento de estos y sobre su contenido de ácidos grasos. Se iniciaron cultivos de callos in vitro a partir de explantos de tallos sin nudos y de hojas de plántulas axénicas de 20 días de edad, utilizando 3 tratamientos de reguladores de crecimiento diferentes. El medio Murashige & Skoog modificado (MSRT) con el agregado de ácido 2,4-diclorofenoxia-cético (2,4-D) a una concentración de 2,25 µM y un fotoperíodo de 16 horas fueron las condiciones óptimas para la inducción de callos. Para el mantenimiento de los callos el tratamiento más adecuado resultó ser bencilaminopurina (BAP) a una concentración de 1 µM. La cinética de crecimiento se caracterizó por un período de latencia hasta el día 20 de cultivo, seguido de un período de crecimiento exponencial entre los días 20 y 54. El tratamiento con 2,4-D (2,25 µM) mostró la más alta velocidad específica de crecimiento (0,22 ± 0,01 /día), el tiempo de duplicación más bajo (31,51 ± 1,00 día) y la mayor biomasa máxima (1,46 ± 0,01 g PF). El contenido de ácidos grasos en los callos de "chía" fue 0,83 % en promedio de todos los tratamientos después de 6 meses en cultivo y no mostró variaciones significativas (0,2 % - 0,3 %) entre los tratamientos aplicados (p < 0,05). (AU)

O uso de plantas medicinais por moradores de Quixadá­Ceará / The use of medicinal plants to habitants from Quixada-Ceara

RESUMO O presente trabalho teve por objetivo observar o uso de plantas medicinais por moradores de Quixadá-Ceará. A amostragem adotada foi 5% da população urbana adulta na sede do município, o que resultou em 263 domicílios visitados, onde foi aplicado um questionário semi-estruturado. De acordo com os resultados ainda temos um número expressivo de pessoas que fazem uso de plantas medicinais na cidade de Quixadá-Ceará (57,41%), e que a maioria dos entrevistados (97%) considera que o uso indiscriminado de plantas medicinais não causa nenhum problema à saúde. A maioria dos entrevistados foram mulheres (63,57%) e o maior número de informações e conhecimentos sobre plantas medicinais concentrou-se na faixa de 51-60 anos (24,71%). Com relação ao nível de escolaridade, 34,22% dos entrevistados afirmou ter ensino médio completo. A família botânica mais representativa foi a Lamiaceae (21,05%) e a espécie mais citada foi a erva-cidreira. Dos entrevistados 93,37% consideraram utilizar o chá como medicamento e 6,63% alegaram fazer a utilização do mesmo como alimento. Quanto à frequência do uso de plantas medicinais de forma contínua, 27% alegaram fazer o uso diariamente.

ABSTRACT This study aimed to observe the use of medicinal plants by residents from Quixada-Ceara. The sample used was 5% of the adult urban population in the county seat, which resulted in 263 households visited, where we applied a semi-structured questionnaire. According to the results, there is still a significant number of people who uses medicinal plants in the city of Quixada-Ceara (57.41%), and the majority of participants (97%) consider that the indiscriminate employment of medicinal plants do not cause any health problem. Most respondents were female (63.57%) and the highest number of information and knowledge about medicinal plants were concentrated among 51-60 years old individuals (24.71%). In regard of the educational level, 34.22% of the participants claimed to have completed high school. The botanical family Lamiaceae was the most representative (21.05%) and the most cited specie was the lemongrass. 93.37% of the respondents pledged to use tea as medicine and 6.63% claimed to use it as food. Regarding the employment frequency of the medicinal plants, 27% claimed to take them daily.

Effects of light and temperature on isoprene emission at different leaf developmental stages of eschweilera coriacea in central Amazon / Efeitos da luz e da temperatura sobre a emissão de isopreno em diferentes estádios de desenvolvimento foliar de eschweilera coriacea na Amazônia Central

Isoprene emission from plants accounts for about one third of annual global volatile organic compound emissions. The largest source of isoprene for the global atmosphere is the Amazon Basin. This study aimed to identify and quantify the isoprene emission and photosynthesis at different levels of light intensity and leaf temperature, in three phenological phases (young mature leaf, old mature leaf and senescent leaf) of Eschweilera coriacea (Matamatل verdadeira), the species with the widest distribution in the central Amazon. In situ photosynthesis and isoprene emission measurements showed that young mature leaf had the highest rates at all light intensities and leaf temperatures. Additionally, it was observed that isoprene emission capacity (Es) changed considerably over different leaf ages. This suggests that aging leads to a reduction of both leaf photosynthetic activity and isoprene production and emission. The algorithm of Guenther et al. (1999) provided good fits to the data when incident light was varied, however differences among E S of all leaf ages influenced on quantic yield predicted by model. When leaf temperature was varied, algorithm prediction was not satisfactory for temperature higher than ~40 °C; this could be because our data did not show isoprene temperature optimum up to 45 °C. Our results are consistent with the hypothesis of the isoprene functional role in protecting plants from high temperatures and highlight the need to include leaf phenology effects in isoprene emission models.

O isopreno emitido pelas plantas corresponde em cerca de um terço das emissões globais de compostos orgânicos voláteis anualmente. A maior fonte de emissão de isopreno para a atmosfera global é a Bacia Amazônica. Este estudo objetivou identificar e quantificar a emissão de isopreno e fotossíntese em diferentes níveis de intensidade de luz e temperatura foliar, em três fases fenológicas (folha madura recente, folha madura tardia e folha senescente) de Eschweilera coriacea (Matamatá verdadeira) - a espécie com maior distribuição na Amazônia central. In situ, as medidas de fotossíntese e emissão de isopreno da folha madura recente apresentaram as maiores taxas em todos os níveis de luz e de temperatura. Adicionalmente, a capacidade de emissão de isopreno (ES) mudou consideravelmente entre as diferentes idades foliares, sugerindo que o envelhecimento reduz a atividade fotossintética e a produção/emissão de isopreno. O algoritmo de Guenther et al. (1999) demonstrou bom ajuste para a emissão de isopreno em diferentes intensidades de luz, entretanto, diferenças na ES entre as idades foliares influenciaram no rendimento quântico estimado pelo modelo. Em relação à temperatura foliar, a estimativa do algoritmo não foi satisfatória para as temperaturas acima de ~40 °C; isto provavelmente ocorreu pelo fato dos dados não apresentarem temperatura ótima até 45 °C. Nossos resultados são consistentes com a hipótese do isopreno ter um papel funcional para proteger as plantas de altas temperaturas e apontam a necessidade de incluir os efeitos da fenologia foliar em modelos de emissão de isopreno.

Identificação botânica e química de espécies vegetais de uso popular no Rio Grande do Norte, Brasil / Botanical and chemical identification of plant species of popular use in Rio Grande do Norte, Brazil

Plantas medicinais são utilizadas mundialmente como uma das principais formas de cuidado primário de saúde. No entanto, a literatura indica que muitas espécies podem apresentar composição química variável, toxicidade ou difícil identificação. O objetivo do presente estudo foi obter critérios úteis para o controle de qualidade farmacognóstico das principais espécies vegetais de uso popular no estado do Rio Grande do Norte, utilizando metodologias de análise botânica (morfodiagnose macro e microscópica) e química (triagem fitoquímica e cromatografia em camada delgada), evitando assim adulterações ou uso inadequado dessas plantas medicinais no estado. No total, sete espécies foram analisadas Acmella oleracea, Chenopodium ambrosioides, Lippia alba, Mentha piperita, Ocimum gratissimum, Peumus boldus e Rosmarinus officinalis. Diversos marcadores botânicos e fitoquímicos foram identificados, contribuindo dessa forma para a correta identificação destas espécies de plantas medicinais importantes no estado do Rio Grande do Norte.

Medicinal plants are worldwide used as one of the main forms of primary healthcare. However, the literature indicates that many species may have variable chemical composition, toxicity, or even difficult identification. The aim of this study was to obtain useful criteria for pharmacognostic quality control of the main plant species of popular use in the state of Rio Grande do Norte, using methods of botanical (macro and microscopic morphodiagnosis) and chemical (phytochemical screening and thin-layer chromatography) analysis, thus preventing adulteration or inappropriate use of these medicinal plants in the state. In total, seven species were analyzed Acmella oleracea, Chenopodium ambrosioides, Lippia alba, Mentha piperita, Ocimum gratissimum, Peumus boldus and Rosmarinus officinalis. Several botanical and phytochemical markers were identified, thereby contributing to the correct identification of these important medicinal plant species in the state of Rio Grande do Norte.

Application of DNA labeling technology in forensic botany / 法医学杂志

Forensic botany is a study of judicial plant evidence. Recently, researches on DNA labeling technology have been a mainstream of forensic botany. The article systematically reviews various types of DNA labeling techniques in forensic botany with enumerated practical cases, as well as the potential forensic application of each individual technique. The advantages of the DNA labeling technology over traditional morphological taxonomic methods are also summarized.

Recent advances of amplified fragment length polymorphism and its applications in forensic botany / 法医学杂志

Amplified fragment length polymorphism (AFLP) is a new molecular marker to detect genomic polymorphism. This new technology has advantages of high resolution, good stability, and reproducibility. Great achievements have been derived in recent years in AFLP related technologies with several AFLP expanded methodologies available. AFLP technology has been widely used in the fields of plant, animal, and microbes. It has become one of the hotspots in Forensic Botany. This review focuses on the recent advances of AFLP and its applications in forensic biology.

Rapid in vitro propagation of the threatened endemic orchid, Ipsea malabarica (Reichb.f.) J D Hook through protocorm-like bodies.

Rapid propagation of I. malabarica (Reichb. f.) J D Hook, an endemic and endangered orchid of the Western Ghats of Kerala, India through conversion of axillary buds to protocorm-like bodies (PLBs), and subsequent plant regeneration was achieved. Growth regulators and sugar displayed significant influence in the induction of PLBs. In vitro derived shoots from field grown rhizomes of Ipsea cultured on Murashige and Skoog (MS) medium with 13.3 microM N6-benzyladenine (BA) containing 2% commercial grade sugar turned the axillary buds to PLBs within 25 days, and developed a mean of 33.1 PLBs within 50 days. Kinetin (KIN) did not induce PLBs, but facilitated axillary bud proliferation. Transfer of PLBs on medium having same concentration of BA and sugar facilitated rapid multiplication, and developed a mean of 47.5 PLBs. No decline of PLB proliferation was observed up to 10th subculture. Half strength MS medium with 6.97 microM KIN facilitated conversion of 98% PLBs to plantlets. On this media, a mean of 5.8 roots were also developed per shoot. Shoots developed bulbs during culture were grown to rhizomes. Increase of sugar to 6 or 8% hastened the development of bulbs/rhizomes. Reintroduction of PLB-derived plantlets in the natural habitat i.e. at Vellarimala (at 1300 m height) of the Western Ghats of Kerala was attempted as a means to assist in situ conservation. This is the first report of conversion of axillary buds to PLBs. The protocol enables to surmount the threat of extinction of this endemic and endangered orchid.

Micropropagation of Clerodendrum aculeatum through adventitious shoot induction and production of consistent amount of virus resistance inducing protein.

Rapid micropropagation through adventitious shoot induction from in vitro raised leaf explants of Clerodendrum aculeatum (Verbenaceae), was successfully achieved for the first time. Basal portion of the leaves showed highest regeneration potential when grown on MS medium supplemented with BA (5.0 mg/l) and NAA and IBA (0.5 mg/l of each). Shoots after elongation in growth regulator-free medium, were rooted in MS medium containing 0.5 mg/l of NAA and IBA. Aqueous leaf extract of in vitro raised plants, induced high degree of resistance against viruses in susceptible healthy hosts when applied prior to virus inoculation. Upon purification from leaves of cultured plants, the resistance inducing protein, showed molecular mass of 34 kDa. Amount of resistance inducing protein obtained from leaves of cultured plants, was consistent throughout the year, as compared to the protein isolated from leaves of field grown plants, which showed marked seasonal fluctuation. The purified 34 kDa protein from in vitro raised plants, was serologically related to field grown plants and possessed similar characteristics. The micropropagated plants were successfully established in earthen pots under greenhouse conditions.

Micropropagation of Glandularia perakii Cov. et Schn. (Verbenaceae), a native species with ornamental potential

Glandularia perakii is a perennial species with beautiful violet flowers that grows in the stony soil of Mendocine pedemont. A plentiful and prolonged flowering confers it an important ornamental potential. In this paper, a method of propagation of G. perakii from nodal segments is reported. Proliferating microshoot cultures were obtained by placing nodal segment on Murashige and Skoog medium (MS) supplemented with 20 g.L-1 of sucrose without growth regulators. In this medium multiplication rate after 20 days was 7.9. Rooted plants were acclimatized successfully.

In vitro propagation of Lagerstromia parviflora Roxb. from adult tree.

A micropropagation protocol based on axillary bud proliferation has been developed from mature Lagerstromia parviflora adult tree. Nodal segments cultured on woody plant medium supplemented with 5.0 microl. BAP and 0.25 microm IAA gave maximum (86.9%) morphogenetic response. Proliferated shoots (10.7 per explants) were elongated to 3.9 cm within 6 weeks. In vitro produced micro-shoots were subjected to an IBA treatment (500 ppm for 2 min. dip) and placed under misting conditions for rooting. Misting beds were prepared with sand: soil (3:1) for 80.6% rooting and was acclimatized. Shoot length seems to be important to induce adventitious roots. The highest (91.7%) rooting was recorded on shoots ranging a length between 3.1-4.0 cm. Rooted and hardened plants were later transferred to poly bags and maintained in shadenet house. The protocol has the realizes capacity to produce 260 plants from a single explants within 10 months multiplication cycle.

High frequency in vitro propagation of Phyllanthus amarus Schum. & Thom. by shoot tip culture.

With the aim of micropropagation of Phyllanthus amarus, an important medicinal herb, shoot tips were cultured in Murashige and Skoog's medium supplemented with kinetin/ BAP singly or in combination with IAA. Growth regulators at lower range (0.1-1.0 mg L(-1)) stimulated direct regeneration of shoots. Kinetin was superior to BAP and kinetin-IAA combination was more suitable than kinetin alone. About 15 shoots were yielded per explant after 30 days of culture in the medium containing kinetin and IAA both at 0.1mg L(-1). The cluster of proliferated shoots elongated and rooted simultaneously under the same treatment following another subculture, thus shortening the total time schedule of micropropagation. Shoot tips of regenerated shoots were continuously used to regenerate new shoots with periodic transfer to fresh medium resulting in a steady supply of normal, healthy plants without any deviation in the production rate during a continuous one year culture. Micropropagated plants were successfully established in soil with high survivality (80%).

In vitro propagation of Mussaenda erythrophylla Schum and Thom cv. scarlet through multiple shoot regeneration.

Axillary bud explants were induced to form shoots on Murashige and Skoog's (MS)' basal medium. Best yield (9 shoots per explant) was obtained when the medium was supplemented with adenine sulphate (40 mg/L) and 6-benzylaminopurine (2.25 mg/L). The shoots were rooted on half strength MS basal medium supplemented with indole butyric acid (0.5 mg/L) and having thiamin-HCl (800 mg/L). Regenerated plantlets were successfully acclimatized. This is the first report of micropropagation in the genus Mussaenda without callus intervention.

Citrus tissue culture employing vegetative explants.

Citrus being a number one fruit of the world due to its high nutritional value, huge production of fruits and fruit products, the citrus industry may be considered a major fruit industry. Though citrus orchard area in India is comparable to USA, the produce is far less, while its export is nil. Biotechnology has played an outstanding role in boosting the citrus industry, e.g., in Spain, which is now the biggest exporter of citrus fruit with the application of micrografting. Amongst the fruit trees, perhaps the maximum tissue culture research has been done in citrus during the past four decades, however, the results of practical value are meagre. The shortfalls in citrus tissue culture research and some advancements made in this direction along with bright prospects are highlighted, restricting the review to vegetative explants only. Whilst utilization of nucellar embryogenesis is limited to rootstocks, the other aspects, like, regeneration and proliferation of shoot meristems measuring 200 microm in length--a global breakthrough--of two commercially important scion species, Citrus aurantifolia and C. sinensis and an important rootstock, C. limonia, improvement of micrografting technique, cloning of the same two scion species as well as some Indian rootstock species, employing nodal stem segments of mature trees, of immense practical value have been elaborated. A rare phenomenon of shift in the morphogenetic pattern of differentiation from shoot bud differentiation to embryoid formation occurred during the long-term culture of stem callus of C. grandis. Stem callus-regenerated plants of C. aurantifolia, C. sinensis and C. grandis showed variation in their ploidy levels and a somaclonal variant of C. sinensis, which produced seedless fruits was isolated. Tailoring of rooting in microshoots to a tap root-like system by changing the inorganic salt composition of the rooting medium, resulting in 100% transplant success, and germplasm preservation through normal growth culture of shoots of C grandis without loss of regeneration capacity during 31 years, observed so far, are some other significant results. Plants of C. aurantifolia and C. sinensis raised from shoot meristem and micrografting were grown in a nethouse and those from nodal stem segments in the field along with the in vitro-raised plants of rootstocks, namely, C. jambhiri, C. karna and C. limonia. All the plants showed normal healthy growth. Significantly enough, the meristem regenerated plants of C. aurantifolia attained the reproductive phase just in 1 year of transplantation to soil similar to those raised from nodal stem segments of mature trees, which also produced normal fruits in the subsequent year while growing under field conditions. Thus, a significant fundamental concept of a maturity factor, carried over through as small a shoot meristem as 200 microm in length to cloned plants has been demonstrated. The concept is of far-reaching significance in citrus industry besides production of pathogen-free orchards.

Estudo farmacognóstico e farmacológico de Bredemeyera floribunda Willdenow / Bredemeyera floribunda Willdenow pharmacognostic and pharmacological study

Bredemeyera floribunda Willdenow, espécie vegetal vulgarmente conhecida como botica inteira, ocorre com freqüência nos campos cerrados do país. A botica inteira, cuja parte empregada são as raízes, é utilizada na medicina popular como expectorante, diurética, em problemas circulatórios e para baixar a pressão arterial. O estudo botânico detalhado das partes empregadas comercialmente foi efetuado e fotografias ilustram a tese. As características botânicas importantes na identificação da droga são: 1) características macroscópicas - as raízes possuem forma cilíndrica e coloração castanho-amarelada. Estrias bem evidentes podem ser observadas ao lado de cicatrizes arredondadas em forma de crateras...

In vitro propagation of Acorus calamus Linn.--a medicinal plant.

High frequency in vitro propagation protocol was standardized from rhizome explants of A. calamus. Maximum shoot multiplication frequency was obtained on Murashige and Skoog's media supplemented with 4 mg/l 6-benzyl amino purine and 0.5 mg/l indole-3-acetic acid. Regenerated shoots were rooted in vitro or directly transferred to sterile soil and well developed roots were observed within two weeks. The rooted plants were successfully established in the field.

In vitro multiplication of Nothapodites foetida (Wight.) Sleumer through seedling explant cultures.

In vitro multiplication of Nothapodites foetida (Wight.) Sleumer was achieved using axenic seedling explant cultures. Isolated nodes (1.0-1.2 cm) and shoot tips (1.0-1.5 cm) cultured in Murashige and Skoog's agar medium containing varying concentrations of TDZ, BA and combinations of 2iP and GA3. Single shoot (0.8-1.2 cm) was regenerated in each culture after 6 weeks. Axillary shoots were then excised and recultured for 8 weeks in medium containing TDZ (0.05 mgL-1) which formed shoots (about 4 in no.; 2 cm) from the basal node. Axillary branches (2) which formed on 60% of these shoots after 10-12 weeks of culture were separated and recultured in the same medium for 8 weeks. Three shoots (0.8-1.0 cm) per culture were regenerated. Shoots of 0.8-1.8 cm length were subcultured on a low cytokinin (0.01 mgL-1 TDZ) regime to induce shoot elongation (2.0-3.5 cm) in 4 weeks. Shoot cuttings were rooted (60%) in the medium containing IBA (1.5 mgL-1). Rooted plantlets established in pots (90%) after hardening resumed normal growth in 3 months.

Micropropagation of sweet orange, Citrus sinensis Osbeck. for the development of nucellar seedlings.

Protocol for micropropagation of elite plants of sweet orange (Citrus sinensis) through nucellar embryo culture has been standardized. Three to four nucellar embryos and a zygotic embryo could be excised from a single mature seed and successfully generated as healthy plants in basal MS medium. MS medium supplemented with NAA (1 mg/L) or 2, 4.D (1 mg/L) promoted callus development in both nucellar and zygotic embryos. GA3 (1 mg/L) enriched medium induced plantlets initiation but their growth was very poor. No significant differences were observed between initial growth patterns of nucellar and zygotic seedlings developing from the same ovule. Five to six shoots were obtained from collar region of both category of embryos in MS medium supplemented with BAP (1 mg/L) within 60 days of inoculation. The number of plantlets were almost doubled after their transfer in the same medium and culture for another 30 days. Higher doses of BAP resulted in initiation of callus directly from the embryos. The regenerated shoots (2-3 cm) could be rooted in MS medium supplemented with either only NAA (0.75 mg/L) or NAA (0.50 mg/L) and IBA (2.0 mg/L). A number of plantlets could be obtained from a nucellar embryo grown shoot within a limited time period.

A method for large-scale multiplication of Curculigo orchioides through bulbil formation from leaf explant in shake flask culture.

An efficient method has been developed for large-scale multiplication of Curculigo orchioides (Hypoxidaceae), an endangered medicinal plant, through direct bulbil formation from leaf explants in shake flask cultures. Leaf-segments (7 x 10 mm) were cultured in B5 liquid medium containing KNO3 (200 mgNL-1), (NH4)2SO4 (50 mgNL-1), benzyl adenosine (2.2 microM), adenine (0.11 mM), indole butyric acid (1.0 microM) and polyvinyl pyrrolidone (250 mgL-1). About 95% of explants produced maximum number of bulbils (546/flask at 6 weeks growth) in the medium. Shake flask cultures yielded 2737 bulbils/L medium whereas static cultures yielded 624 bulbils/L medium. Germination of bulbils was maximum (90.62%) on agar-gelled B5 medium containing benzyl adenosine (2.2 microM) and gibberellic acid (3.5 microM). Plantlets developed in vitro were successfully transferred to soil with a high rate of survivability (90%) and were comparable to natural population in growth and vigour.