Research progress on culture technologies,chemical components,and pharmacological activities of Saussurea involucrata cells / 中国中药杂志

Saussurea involucrata,a traditional Chinese medicinal material,is effective in the treatment of rheumatoid arthritis with cold-dampness blockage syndrome,cold pain in lower abdomen,and menstrual irregularities. However,due to the specific habitat,low natural reproduction rate,slow growth,and overexploitation,it is at the high risk of extinction. S. involucrata cells can be obtained through callus culture,suspension culture,and hairy root culture. This study highlighted the influences of reactor type,culture system,precursor,elicitor type, and light wavelength on the suspension culture of S. involucrate cells. The chemical components of S. involucrata cells mainly include phenylpropanoids,flavonoids,lignans,and steroids,among which phenylpropanoids are the most abundant. S. involucrata cells have multiple pharmacological activities of anti-inflammation,analgesia,activating blood and resolving stasis,immunoregulation,increasing bone density,lowering blood lipids,anti-hypoxia,anti-exercise fatigue,anti-radiation,anti-obesity,and anti-oxidation. Moreover,it has the potential of treating aplastic anemia. This study reviews the cell culture technologies,chemical components,and pharmacological activities of S. involucrata cells,laying a basis for the further research,development,and utilization.

Evaluation of limonoid production in suspension cell culture of Citrus sinensis

ABSTRACTThe use of cell and plant tissue culture techniques to produce economically important active metabolites has been growing. Among these substances are total limonoid aglycones, which are produced by "pera" orange (Citrus sinensis (L.) Osbeck, Rutaceae) and have received considerable attention because of their anticancer actions. The main objective of the present study was to analyze and compare the levels of limonoid aglycones in seeds, callus cultures (originating from seeds), callus cultures (originating from hypocotyls), cell suspensions from hypocotyls cells, and cell suspensions from cotyledons. The cell cultures or C. sinensis were obtained by inoculating two strains of callus in MS medium supplemented with 2.0 µM 2,4-dichlorophenoxyacetic acid, 7.0 µM benzyl aminopurine, and 3% (w/v) sucrose in the dark. The highest concentrations of limonoid aglycone that were obtained were observed in cotyledon cell lines (240 mg/100 g dry weight) that were produced on day 21 of culture and hypocotyl cell lines on day 7 (210 mg/100 g dry weight). Explants of different origins under the same culture conditions had different limonoid aglycone content. The present results may suggest strategies for enhancing the productivity of biologically important limonoid aglycones and investigating the complex pathways of these secondary metabolites in plant tissue cultures.

Bioefficacy of crude extract of Cyperus aromaticus (Family：Cyperaceae) cultured cells, against Aedes aegypti and Aedes albopictus mosquitoes

Objective:To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedes mosquito species during two generations as well. Methods:The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone III, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone III [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedes mosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedes mosquitoes were assessed. Results:Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedes mosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedes mosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex ratio of the adult population either parental or F1 generation of the Aedes mosquito species was not significantly affected by the EI50 dosage of the crude extract of C. aromaticus cultured P4 cells. A significant decrease in the wing length of the treated adult (female and male) of Aedes aegypti as well as the treated female of Ae. albopictus were observed. Longevity of the adult female of the parental generation of both Aedes mosquitoes as well as females of F1 generation of Ae. albopictus were significantly decreased. Conclusions: The present study revealed the potential of the crude extract of C. aromaticus cultured cells in controlling vector mosquito populations in the effort to reduce the transmission of vector borne diseases.

Produção de metabólitos secundários em cultura de células e tecidos de plantas: o exemplo dos gêneros Tabernaemontana e Aspidosperma: [revisão] / Production of plant secondary metabolites in plant cell and tissue culture: the example of Tabernaemontana and Aspidosperma genera: [revision]

Os estudos dos metabólitos secundários de plantas se desenvolveram aceleradamente nos últimos 50 anos. Estes compostos são conhecidos por desempenharem um papel importante na adaptação das plantas aos seus ambientes e também representam uma fonte importante de substâncias farmacologicamente ativas. As técnicas de cultura de células de plantas iniciaramse na década de 1960 como uma possível ferramenta para estudar e produzir os metabólitos secundários de plantas. O uso de cultura de células de planta para a produção de substâncias de interesse contribuiu grandemente para avanços em diversas áreas da fisiologia e bioquímica vegetal. Diferentes estratégias, usando sistemas de cultura in vitro, foram estudadas com o objetivo de aumentar a produção de metabólitos secundários. As plantas dos gêneros Aspidosperma e Tabernaemontana são importantes fontes de alcalóides indólicos biologicamente ativos, sendo que no Brasil existe um número considerável de espécies destes gêneros. As culturas de células de Aspidosperma e Tabernaemontana foram iniciadas há pelo menos 16 anos, as quais produzem um grande número de alcalóides, o que estimulou o desenvolvimento de diversas técnicas para sua produção, extração e identificação.

Studies on plant secondary metabolites have been increasing over the last 50 years. These compounds are known to play a major role in the adaptation of plants to their environment and an important source of active pharmaceuticals. Plant cell culture technologies were introduced at the end of the 1960s as a possible tool for both studying and producing plant secondary metabolites. Different strategies, using in vitro systems, have been extensively studied with the objective of improving the production of secondary plant compounds. The Aspidosperma and Tabernaemontana genera are an important source of biologically active alkaloids and in Brazil there is a considerable number of species of these genera. About 16 years ago cell cultures of Tabernemontana and Aspidosperma were initiated. These cell cultures did produce a number of alkaloids of pharmaceutical interest that stimulated the development of several techniques to production, extraction and identification.

Concentrações de citocinina e carvão ativado na micropropagação de pimenta-do-reino / Cytokynin concentrations and activated charcoal on black pepper micropropagation

A pimenta-do-reino (Piper nigrum) tem grande importância para a economia do Norte do Brasil. A pimenta-do-reino é propagada principalmente por estaquia que, além de ser pouco eficiente, pode possibilitar a propagação da doença fusariose. Dessa forma, testes para verificar a eficiência da micropropagação são pertinentes. O objetivo do trabalho foi testar concentrações de BAP - 6-benzilaminopurina (0,5, 1,5, 3,0 e 4,5mg L-1) e a adição de carvão ativado a 0,2 por cento ao meio de cultura na micropropagação de pimenta-do-reino. O meio de cultura foi composto de sais e vitaminas de Murashige & Skoog, 3 por cento de sacarose e 8g L-1 de ágar. Foram utilizados como explantes ápices caulinares de plântulas da cultivar "Bragantina" germinadas in vitro. Os tratamentos foram avaliados em esquema fatorial 4x2, em delineamento inteiramente casualizado. Após 45 dias no estabelecimento, foi avaliado o número de brotos, mas não foi observada diferença significativa entre os tratamentos, considerando-se os dois fatores e a interação entre eles. Após 45 dias no primeiro subcultivo, foram avaliados o número de brotos e de explantes (segmentos nodais). A interação entre BAP e carvão ativado teve efeito significativo para número de brotos e de explantes, sendo que o carvão influenciou o efeito das concentrações de BAP. A concentração de 0,5mg L-1 de BAP na ausência de carvão ativado gerou maior número de brotos e de explantes. O carvão ativado não é necessário na fase de proliferação de gemas de pimenta-do-reino da cultivar "Bragantina".

Black pepper (Piper nigrum) has great importance for the Northern Brazil’s economy. Black pepper is mainly propagated by mass propagation, which is not very efficient and may spread fusariosis. Thus, micropropagation efficiency tests are pertinent. The objective of this research paper was to test BAP - 6-benzilaminopurine concentrations (0.5, 1.5, 3.0 and 4.5mg L-1) and activated charcoal at 0.2 percent addition to the culture medium for black pepper’s micropropagation. Culture medium was composed of Murashige & Skoog’s salts and vitamins, 3 percent sucrose and 8g L-1 agar. Shoot tips from Bragantina black pepper seedlings germinated in vitro were used as explants. Treatments were evaluated in a 4x2 factorial scheme in a fully randomized design. Forty-five days after establishment, the number of shoots were evaluated, but there was no significative difference between treatments, considering the two factors and the interaction between them. Forty-five days after the first subculture, the number of shoot and explants (nodal segments) were evaluated. Interaction between BAP and activated charcoal had a significative effect on the number of shoots and explants, however, activated charcoal had an influence on BAP concentrations. The concentration of 0.5mg L-1 generated the highest number of shoots and explants. Activated charcoal is not necessary during the Bragantina black pepper shoot bud proliferation phase.