大车前（Plantago major L. "Giant Turkish."）体外优化再生体系的建立及Na+/H+逆向转运蛋白基因SeNHX1 和TtNHX1 的功能鉴定

大车前（Plantago major L. "Giant Turkish."）不仅有很高的药用价值，在生态学研究方面也是重要模式植物。大车前的组织培养工作，目前报道很少。对其组织培养体系的建立，为筛选大车前耐盐突变体和基因转化建立高效的体外再生系统和实验平台体系。通过愈伤组织诱导和直接不定芽再生途径， 建立了大车前（Plantago major L. "Giant Turkish."）的快速高效再生系统。叶片外植体在含有1.0 mg/L NAA的MS培养基中培养3周后，形成愈伤组织，愈伤组织在含4.0 mg/L 6-BA的MS培养基中成功再生，得到完整植株。种子外植体在含0.2 mg/L IAA和1.0 mg/L TDZ的MS培养基中培养4周后产生大量的丛生芽，对9株再生植株进行RAPD检测表明，部分植株在DNA水平上发生了变异。 植物抵御盐胁迫的一个重要机制是在液泡中积累Na+，从而使细胞质内Na+保持在较低水平，并且降低细胞渗透势。Na+运输到液泡是由液泡Na+/H+逆向转运蛋白完成的。本实验室已从盐生植物盐角草（Salicornia europaea）和番杏（Tetragonia tetragonioides）中分别克隆得到SeNHX1和TtNHX1基因。本文研究了SeNHX1和TtNHX1基因在酵母突变体里的作用。TtNHX1和SeNHX1蛋白在缺陷型酵母菌株里的表达能够提高这些菌株对NaCl、LiCl和潮霉素的抗性，提高到与野生型相当的抗性水平。说明TtNHX1和SeNHX1有着与酵母ScNHX1相似的细胞定位和作用机制，是ScNHX1的功能类似蛋白。

In situ nutrient removal from aquaculture wastewater by aquatic vegetable Ipomoea aquatica on floating beds

Nutrient-rich effluents caused rising concern due to eutrophication of aquatic environment by utilization of a large amount of formula feed. Nutrient removal and water quality were investigated by planting aquatic vegetable on artificial beds in 36-m(2) concrete fishponds. After treatment of 120 days, 30.6% of total nitrogen (TN) and 18.2% of total phosphorus (TP) were removed from the total input nutrients by 6-m(2) aquatic vegetable Ipomoea aquatica. The concentrations of TN, TP, chemical oxygen demand (COD) and chlorophyll a in planted ponds were significantly lower than those in non-planted ponds (P<0.05). Transparency of water in planted ponds was much higher than that of control ponds. No significant differences in the concentration of total ammonia nitrogen (TAN), nitrate nitrogen (NO3-N) and nitrite nitrogen (NO2--N) were found between planted and non-planted ponds. These results suggested that planting aquatic vegetable with one-sixth covered area of the fishponds could efficiently remove nutrient and improve water quality.

Identification of Phenylethanoid Glycosides in Plant Extract of Plantago asiatica L. by Liquid Chromatography-Electrospray Ionization Mass Spectrometry

The present work describes a liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method for rapid identification of phenylethanoid glycosides in plant extract from Plantago asiatica L. By using a binary mobile phase system consisting of 0.2% acetic acid and acetonitrile under gradient conditions, a good separation was achieved on a reversed-phase C-18 column. The [M-H](-) ions, the molecular weights, and the fragment ions of phenylethanoid glycosides were obtained in the negative ion mode using LC-ESI-MS. The identification of the phenylethanoid glycosides (peaks 1-3) in the extract of P. asiatica L. was based on matching their retention time, the detection of molecular ions, and the fragment ions obtained by collision-induced dissociation (CID) experiments with those of the authentic standards and data reported in the literature.

Biocaumulação de alumínio em rebentos micropropagados de Plantago sp.

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2009

Germination requirements of Plantago algarbiensis seeds, an endangered species endemic to the Algarve region (South of Portugal)

The ecdysteroid, 20-hydroxyecdysone or beta-ecdysone, is a steroid hormone which plays a crucial role in molting, metamor- phosis and reproduction of arthropods. This ecdysteroid and its analogues have high potential to be used as insecticides. Previous studies in our laboratory have demonstrated that Vitex glabrata R.Br. (commonly known as Khai-Nao), an indigenous herbaceous plant of Thailand, synthesized and accumulated high quantity of 20-hydroxyecdysone. Therefore, the aim of this study was to investigate the effect of precursor and elicitors feeding on cell growth and 20-hydroxyecdysone production of V. glabrata suspension cultures. Plant cells were cultured in half strength MS medium containing 30 g/l glucose and supplemented with 2.0 mg/l 6- benzylaminopurine (BAP) and 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Cells were incubated on a rotary shaker at 120 rpm under continuous light of 2000 lux at 25 °C. Sterilized cholesterol (5 and 10 mg/l) as precursor was added to the cell cultures on the day of inoculation, while chitosan (50, 100 and 200 mg/l) and methyl jusmonate (100 and 200 mM) as elicitors were added to the cell cultures on day 6 after cultivation.

Los hipoglicemiantes orales y psyllium plantago en el control del paciente diabético (por) Rita Bertha Ramírez Quintero

Tesis (Maestría en Ciencias, con especialidad en Medicina Familiar) U.A.N.L.

Glomus intraradices and Gigaspora margarita arbuscular mycorrhizal associations differentially affect nitrogen and potassium nutrition of Plantago lanceolata in a low fertility dune soil

Two controlled microcosm experiments aimed at a critical re-assessment of the contributions of divergent arbuscular mycorrhizal (AM) fungi to plant mineral nutrition were established that specifically targeted Plantago lanceolata–Glomus intraradices (B.B/E) and –Gigaspora margarita (BEG 34) symbioses developed in a native, nutrient limited, coastal dune soil. Plant tissue nitrogen (N), phosphorus (P) and potassium (K) status as well as plant growth parameters and levels of mycorrhizal colonization were assessed at harvest. In addition to the general well-established mycorrhizal facilitation of P uptake, the study was able to demonstrate a G. intraradices-specific contribution to improved plant nitrogen and potassium nutrition. In the two respective experiments, G. intraradices-inoculated plants had 27.8% and 40.8% more total N and 55.8% and 23.3% more total K when compared to Gi. margarita inoculated counterparts. Dissimilar overall contribution of the two isolates to plant nutrition was identified in AM-genus specific differences in plant tissue N:P:K ratios. G. intraradices inoculated and non-mycorrhizal plants generally exhibited N:P:K ratios indicative of P limitation whereas for Gi.margarita mycorrhizal plants, corresponding ratios strongly implied either N or K limitation. The study provides further evidence highlighting AM functional biodiversity in respect to plant nutrient limitation experienced by mycorrhizal P. lanceolata in an ecologically relevant soil system.

Arbuscular mycorrhizal modulation of diazotrophic and denitrifying microbial communities in the (mycor)rhizosphere of Plantago lanceolata

Impacts of divergent arbuscular mycorrhizal (AM) fungi, Glomus intraradices and Gigaspora margarita, on denitrifying and diazotrophic bacterial communities of Plantago lanceolata in nutrient-limited dune soil were assessed. We hypothesized AM species-related modifications that were confirmed in respective bacterial nirK and nifH sequence polymorphism -based community clustering and community variance allocation. The denitrifying community appeared more responsive to AM fungi than the nitrogen-fixing community. Nevertheless, the main explanatory variable, in both cases, was plant age. We conclude that AM fungi can modify N-cycling microbial rhizosphere communities and future work should aim to verify the functional significance and mechanistic basis.

Olfactory selection of Plantago lanceolata by snails declines with seedling age

Background and Aims Despite recent recognition that (1) plant–herbivore interactions during the establishment phase, (2) ontogenetic shifts in resource allocation and (3) herbivore response to plant volatile release are each pivotal to a comprehensive understanding of plant defence, no study has examined how herbivore olfactory response varies during seedling ontogeny. Methods Using a Y-tube olfactometer we examined snail (Helix aspersa) olfactory response to pellets derived from macerated Plantago lanceolata plants harvested at 1, 2, 3, 4, 5, 6 and 8 weeks of age to test the hypothesis that olfactory selection of plants by a generalist herbivore varies with plant age. Plant volatiles were collected for 10 min using solid-phase microextraction technique on 1- and 8-week-old P. lanceolata pellets and analysed by gas chromatography coupled with a mass spectrometer. Key Results Selection of P. lanceolata was strongly negatively correlated with increasing age; pellets derived from 1-week-old seedlings were three times more likely to be selected as those from 8-week-old plants. Comparison of plant selection experiments with plant volatile profiles from GC/MS suggests that patterns of olfactory selection may be linked to ontogenetic shifts in concentrations of green leaf volatiles and ethanol (and its hydrolysis derivatives). Conclusions Although confirmatory of predictions made by contemporary plant defence theory, this is the first study to elucidate a link between seedling age and olfactory selection by herbivores. As a consequence, this study provides a new perspective on the ontogenetic expression of seedling defence, and the role of seedling herbivores, particularly terrestrial molluscs, as selective agents in temperate plant communities.

Avaliação toxicológica pré-clínica do fitoterápico contendo Aristolochia cymbifera, Plantago major, Luehea grandiflora, Myrocarpus frondosus, Piptadenia colubrina (Cassaú composto) em ratos wistar.

Avaliou-se a segurança de um fitoterápico, constituído de extratos fluidos de Aristolochia cymbifera (“cassaú”), Plantago major L.(“transagem”), Luehea grandiflora Mart.(“açoita-cavalo”), Myrocarpus frondosus Allemão (“cabreúva”), Piptadenia colubrina Benth (“angico”) (Cassaú Composto® ), através de estudos de toxicidade aguda e subcrônica, tendo como base a resolução Nº 90, de 16 de março de 2004 da ANVISA (Agência Nacional de Vigilância Sanitária). Para o teste de toxicidade aguda, ratos Wistar de ambos os sexos foram tratados por via oral com uma única dose de 26 ml/kg, correspondendo a 20 vezes a dose terapêutica indicada pelo fabricante para seres humanos adultos. Os resultados revelaram haver sinais de toxicidade sistêmica com o aparecimento de ataxia, porém de forma transitória e reversível, não causando interferência no desenvolvimento ponderal dos animais, nos consumos de água e ração, nas produções de urina e fezes, bem como alterações macroscópicas nos órgãos dos animais. Avaliou-se também a exposição a doses repetidas do fitoterápico (toxicidade subcrônica). Constituiram-se 4 grupos experimentais (10 animais/sexo/dose), onde administrou-se por via oral a ratos Wistar, durante 30 dias, doses diárias de 1,3 ml/kg, 6,5 ml/kg e 13 ml/kg, respectivamente a dose terapêutica indicada pelo fabricante para seres humanos adultos, 5 vezes, e 10 vezes a dose terapêutica, além de um grupo controle, onde administrou-se o veículo do fitoterápico. Os resultados revelaram ausência de toxicidade sistêmica, fundamentados na ausência de alterações hematológicas e bioquímicas sangüíneas, bem como peso e análises histopatológicas dos órgãos, nos diferentes grupos. As flutuações nos consumos de água e ração, bem como produções de urina e fezes, não influenciaram de maneira negativa o desenvolvimento ponderal dos animais. Concluiu-se portanto, que a utilização do fitoterápico nas doses e períodos referidos pode ser considerado segura.