In vitro conservation strategy for endemic and endangered Himalayan liverwort Stephensoniella brevipedunculata Kashyap (Marchantiophyta)

2018 ◽  
Vol 4 (02) ◽  
pp. 81-87
Author(s):  
A. K. Asthana ◽  
S. D. Tewari ◽  
Vishwa Jyotsna Singh ◽  
Isha Pathak ◽  
Vinay Sahu

During the present study an effort has been made to propagate (in-vitro) the endangered and endemic Himalayan liverwort Stephensoniella brevipedunculata Kashyap using different culture media under controlled Laboratory conditions. Axenic cultures of the taxon have been established using tubers as explants. Seven combinations of media with Full Strength Knop’s macronutrients; Half-strength Knop’s macronutrients; Half-strength Knop’s macronutrients + Vitamins; Halfstrength Knop’s macronutrients + 0.2 mg L-1 IBA + 0.1 mg L-1 BAP; Half-strength Knop’s macronutrients + 0.1 mg L-1 Kinetin + 0.1 mg L-1 2,4D; Half-strength Knop’s macronutrients + 0.1 mg L-1 IBA + 0.2 mg L-1 BAP and Hoagland no. 2 basal salt mixture were used for culture. The best growth was observed in the Hoagland no. 2 basal salt mixture medium, in which dichotomously branched young thalli were successfully formed. Subsequently healthy population of culture grown plants has been raised on soil in pots for the first time.

1970 ◽  
Vol 18 (2) ◽  
pp. 131-137 ◽  
Author(s):  
S. Mederos-Molina

Culture medium requirements for micropropagation of Isoplexis chalcantha was achieved for the first time after high degree of contamination and phenolic exudates were detected and solved. Cultures were established from axillary shoots using juvenile branches collected from this medicinal plant. Most satisfying results were obtained using a solidified and a modified MS medium (NO3- : NH4+ ratios) enriched with ascorbic acid or soluble PVP plus GA3, BAP and NAA. Explants (nodal segments) were used for in vitro shoots multiplication and best results were achieved with modified MS plus BAP and auxins. Vigorous shoots rooted without symptoms in the half-strength modified MS enriched with low concentration of IBA. Key words: Isoplexis chalcantha, axillary shoots, contamination, phenolic exudates, culture media, NO3- : NH4+ ratios D.O.I. 10.3329/ptcb.v18i2.3395 Plant Tissue Cult. & Biotech. 18(2): 131-137, 2008 (December)


Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2301 ◽  
Author(s):  
Federica De Castro ◽  
Michele Benedetti ◽  
Giovanna Antonaci ◽  
Laura Del Coco ◽  
Sandra De Pascali ◽  
...  

The novel [Pt(O,O′-acac)(γ-acac)(DMS)], Ptac2S, Pt(II) complex has recently gained increasing attention as a potential anticancer agent for its pharmacological activity shown in different tumor cell lines, studied both in vitro and in vivo. The mechanism of action of Ptac2S, operating on non-genomic targets, is known to be very different from that of cis-[PtCl2(NH3)2], cisplatin, targeting nucleic acids. In this work, we evaluated the cytotoxicity of Ptac2S on the cisplatin resistant Epithelial Ovarian Carcinoma (EOC), SKOV-3 cells, by the MTT assay. A 1H-NMR metabolomic approach coupled with multivariate statistical analysis was used for the first time for Ptac2S to figure out the biological mechanisms of action of the complex. The metabolic variations of intracellular metabolites and the composition of the corresponding extracellular culture media were compared to those of cisplatin (cells were treated at the IC50 doses of both drugs). The reported comparative metabolomic analysis revealed a very different metabolic profile between Ptac2S and cisplatin treated samples, thus confirming the different mechanism of action of Ptac2S also in the Epithelial Ovarian Carcinoma (EOC), SKOV-3 cells line. In particular, higher levels of pyruvate were observed in Ptac2S treated, with respect to cisplatin treated, cells (in both aqueous and culture media). In addition, a very different lipid expression resulted after the exposure to the two drugs (Ptac2S and cisplatin). These results suggest a possible explanation for the Ptac2S ability to circumvent cisplatin resistance in SKOV-3 cells.


Nativa ◽  
2018 ◽  
Vol 6 (1) ◽  
pp. 27
Author(s):  
Marcos Vinícius Marques Pinheiro ◽  
Ana Cristina Portugal Pinto De Carvalho ◽  
Fabrina Bolzan Martins

No intuito de elevar as taxas de sobrevivência durante a etapa de aclimatização e posterior plantio a campo, avaliou-se o enraizamento in vitro de bananeira cv. Pacovan, em diferentes concentrações de sais MS e de sacarose. Utilizou-se DIC, esquema fatorial (6x2x3), com seis meios de cultura [sendo três concentrações de nutrientes do meio MS (100%; 50% de macronutrientes; e 50% dos sais macro e micronutrientes), e duas concentrações de sacarose (1,5/3,0%)], dois fotoperíodos (12/16 h) e três tempos de cultivo (21, 28 ou 35 dias) e seis repetições/tratamento. Analisaram-se: altura da planta, número de folhas/planta, massas frescas e secas das partes aérea e radicular. Para altura da planta, massa fresca da parte aérea e radicular, o meio MS 50% dos sais + sacarose (1,5%) com fotoperíodo de 16 h e tempo de cultivo de 35 dias foi satisfatório. Para massa seca da parte aérea foi MS 50% de sais + sacarose (3%), e para massa seca da parte radicular, MS 100% + sacarose (3%) (em 12hs/28 dias e 16hs/21 dias). Para o alongamento/enraizamento in vitro da bananeira cv. Pacovan sugere-se MS 50% de sais (macro e micronutrientes), redução ou manutenção de sacarose (1,5 ou 3%) em 16h/35 dias de cultivo.Palavra-chave: Musa spp., propagação in vitro, sistema radicular. CHANGES IN CULTURE MEDIUM, PHOTOPERIOD AND TIME OF CULTIVATION AFFECT THE IN VITRO ELONGATION AND ROOTING OF BANANA CV. PACOVAN ABSTRACT:In order to achieve high rates of survival during the acclimatization and later planting in the field, was evaluated the in vitro of banana cv. Pacovan plants under different concentrations of sucrose and MS basal salt mixture. The experiment was assembled in a DIC, in 6x2x3, six different culture media [three different MS salt mixture concentrations (100%; 50% of macronutrients; and 50% of macro/micronutrients) and two sucrose concentrations (1.5/3%)], two photoperiods (12/16 hours) and three cultivation times (21, 28 or 35 days). Each treatment was composed by 6 replicates. Plant height, number of leaves/plant, fresh and dry weight of roots and shoots, were analyzed. Satisfactory results for plant height and shoot and root fresh biomass were observed in MS with macro/micronutrients (50%) + sucrose (3%), 16 hours/35 days. The highest values of shoot dry weight were observed in MS with macro/micronutrients (50%) + sucrose (3%); the highest root dry weight was achieved with MS 100% + sucrose (3%) (12hs/28 and 16hs/21 days). The suggested medium for the in vitro elongation and rooting stage of banana cv. Pacovan is the MS with 50% of salts (macro and micronutrients), reduction or maintenance of sucrose (1.5 or 3%) in 16h/35 days of cultivation.Keywords: Musa spp., in vitro propagation, root system. DOI:


HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.


2020 ◽  
Vol 42 ◽  
pp. e52940
Author(s):  
Simone Sacramento dos Santos Silva ◽  
Everton Hilo de Souza ◽  
Fernanda Vidigal Duarte Souza ◽  
Cristina Ferreira Nepomuceno ◽  
Maria Angélica Pereira de Carvalho Costa

Alcantarea nahoumii (Leme) J. R. Grant is a species native to the Atlantic Forest that stands out for ornamental purposes. The objective of this work was to evaluate the in vitro germination of A. nahoumii seeds and establish a micropropagation protocol for production of seedlings so as to minimize the effects of predatory extractivism and develop an in vitro conservation system. Mature seeds were disinfested, established in three culture media (MS, MS½ and MS⅓) and incubated at four temperatures (20, 25, 30 and 35ºC) in a germination chamber. In the micropropagation experiment, stem segments were introduced in MS medium supplemented with 0.5 μM of 1-naphthaleneacetic acid (NAA) and 0.0, 2.2, 4.4 and 6.6 μM of 6-benzylaminopurine (BAP). For the in vitro conservation, plantlets were established in MS or MS½ medium supplemented with 15 g L-1 or 30 g L-1 of sucrose. The plants were acclimated with commercial substrate. The highest seed germination percentages were promoted by temperature conditions of 20 and 25ºC, with MS culture medium. The highest multiplication rate of shoots was obtained from the treatment without addition of the growth regulator or when combined with 2.2 μM of BAP + 0.5 μM of NAA. The acclimation of the plants occurred with high survival rate. The species can be conserved in vitro under slow growth condition for 24 months when incubated in MS medium supplemented with 30 g L-1 of sucrose.


Rodriguésia ◽  
2019 ◽  
Vol 70 ◽  
Author(s):  
Monique Cristine R. Juras ◽  
Jackeline Jorge ◽  
Rosete Pescador ◽  
Wagner de Melo Ferreira ◽  
Vivian Tamaki ◽  
...  

Abstract Cattleya xanthina is a Neotropical orchid endemic to the Brazilian Atlantic Rainforest, at high risk of extinction. In this paper, we investigated the effects of different culture media on C. xanthina as well as on their endogenous nitrogen status. Culture media studied: Knudson C (KC), Vacin and Went (VW), and Murashige and Skoog (MS), the latter used at two different concentration (full and half-strength; MS/2). After 180 days, plants were transferred to MS medium with different NAA and BA concentrations. In each treatment, biometric parameters were measured and the endogenous levels of photosynthetic pigments, total protein, nitrate and ammonium ions were quantified. Plants grown on KC medium had the lowest concentration of nitrogen but exhibited the greatest shoot development, production of photosynthetic pigments and total protein. Results of growth regulators showed that the highest concentration of auxin stimulated root development and the production of photosynthetic pigments, and that a higher concentration of cytokinin promoted protein synthesis and the development of shoots. Most successful acclimatization was obtained when a mixture of Sphagnum and Pinus bark was used as the substrate.


Cells ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 103
Author(s):  
Lluis Rodas ◽  
Sonia Martínez ◽  
Aina Riera-Sampol ◽  
Hannah J. Moir ◽  
Pedro Tauler

Immune system functionality has been commonly assessed by a whole-blood or isolated-cell stimulation assay. The aim of this study was to determine whether cytokine production in whole-blood-stimulated samples is influenced by age, sex, and smoking. A descriptive cross-sectional study in 253 healthy participants aged 18–55 years was conducted. Whole blood samples were stimulated for 24 h with LPS and concentrations of IL-6, IL-10, and TNF-α were determined in the culture media. Among parameters considered, statistical regression analysis indicated that smoking (change in R2 = 0.064, p < 0.001) and sex (change in R2 = 0.070, p < 0.001) were the main predictors for IL-10 production, with higher values for women and non-smokers. Age was also found to be a significant predictor (change in R2 = 0.021, p < 0.001), with higher values for younger ages. Age (change in R2 = 0.089, p = 0.013) and smoking (change in R2 = 0.037, p = 0.002) were found to be negative predictors for IL-6 production. Regarding TNF-α-stimulated production, age (change in R2 = 0.029, p = 0.009) and smoking (change in R2 = 0.022, p = 0.022) were found to be negative predictors. Furthermore, sex (change in R2 = 0.016, p = 0.045) was found to be a significant predictor, with lower values for women. In conclusion, sex, age, and smoking were found to be independent determinants of stimulated cytokine production. While female sex is associated with higher IL-10 and lower TNF-α production, aging and smoking are associated with lower IL-6, IL-10, and TNF-α production.


Author(s):  
Meena Choudhary ◽  
Inder Dev Arya ◽  
Sarita Arya

The main aim of present study was to overcome the problems associated with the in vitro culture initiation in Terminalia arjuna. The micropropagation of tree species is not easy as shrubs and herbs. Many problems encountered from explant collection to in vitro culture establishment. The problems that have been occurred during T. arjuna micropropagation were culture contamination, phenolic exudation, bud growth inhibition, shoots yellowing and leaf fall. All these problems have been solved by applying certain treatments prior to explant collection and inoculation. The mother tree was lopped in November months (six months prior to explant collection) to remove any inhibitory substance and release bud growth. Different sterilizing agents were used to minimize the bacterial and fungal contamination. Some modification in culture media (use of different concentration of NH4NO3 and KNO3 salts and adenine sulphate) was done. Surface sterilization of nodal explants collected from lopped branches with 0.1% HgCl2 for 8 min., treatment with chilled antioxidant solution (Ascorbic acid, Citric acid and PVP) and half strength of NH4NO3 and KNO3 salts of MS medium supported 100% bud break response with proliferation of green and healthy in vitro shoots. Removing these hurdles already in the initial stage of micropropagation is very important and maximize mass in vitro propagation of this medicinally important Arjun tree. 


2015 ◽  
Vol 23 (2) ◽  
pp. 27-38 ◽  
Author(s):  
Rosen S. Sokolov ◽  
Bistra Y. Atanassova ◽  
Elena T. Iakimova

AbstractIn this study, the effects of compounds providing Fe in chelated (NaFeEDTA and Fe(III)AC) and non-chelated (FeSO4·7H2O) forms as components of culture media, onin vitroshoot multiplication and rooting ofMagnolia soulangeana‘Alexandrina’,Magnolia grandifloraandPrunus cerasifera‘Nigra’ were comparatively evaluated. Each of the tested chemicals was used as a single Fe source in the basal salt medium. In the stages of shoot multiplication and rooting plant response was scored by biometrical indices (number of shoots, leaves and roots, shoot and root length, percent of rooted plants and root hairs). The occurrence of physiological disorders was estimated by visual observations. In presence of FeSO4, symptoms of chlorosis, hyperhy-dricity, early senescence and specific morphology of roots, suggesting Fe deficiency, were observed. These deteriorations were entirely prevented at the application of Fe chelates of which, in this experimental systems, Fe(III)AC was tested for the first time. The addition of Fe(III)AC positively affected the plant quality to extent comparable to that of NaFeEDTA. The obtained data suggest that both applied Fe chelates are more appropriate than non-chelated Fe form and can be alternatively used in the optimization of nutrient media for micropropagation ofMagnoliaandPrunus cerasiferagenotypes.


Parasitology ◽  
1962 ◽  
Vol 52 (3-4) ◽  
pp. 441-457 ◽  
Author(s):  
J. D. Smyth

1. A technique is described for the preparation of axenic cultures of protoscolices of Echinococcus granulosus.2. Protoscolices removed aseptically from hydatid cysts are treated with 0·02% crystalline pepsin at pH 2·0 for 10 min. at 38°C. in order to remove (a) daughter cyst germinal membranes, and (b) dead or degenerating protoscolices.3. Protoscolices may be maintained for more than 100 days in a variety of media. These include media containing various proportions of some of the following: chick embryo extract, beef embryo extract, hydatid fluid, bovine amniotic fluid, Parker 199, bovine serum.4. 10–20 ml. of culture media was used at 38°C., either in roller tubes or shaken in a water-bath. Media were renewed every 48 hr.5. Protoscolices developed into cysts by two routes, either (a) by becoming vesicular, gradually growing in size, and finally becoming enclosed in a laminated envelope, or (b) by forming a ‘posterior bladder’ which became relatively enormous and ultimately absorbed the scolex region, the whole forming a cyst within an envelope as in (a).6. In the majority of media, after about 31–48 days' culture, a thin envelope was secreted which developed later into a thick laminated membrane.7. Cysts with laminated membranes ultimately underwent fatty degeneration and became cytologically abnormal.8. In the most successful experiments, cells or clusters of cells formed within the cysts and these may have represented the anlagen of the protoscolices.9. In no case was segmentation, or any other sign of strobilar development, obtained.10. These results show that the laminated membrane in E. granulosus is initially of parasite origin and not of host origin as believed by some workers to be the case.11. Since there is evidence from the literature that E. multilocularis does not form such an envelope in vitro, a clear developmental difference appears to exist between the two species.12. It is suggested that this difference offers an explanation for the formation of unilocular or multilocular cysts. Since E. granulosus secretes a laminated envelope, its development is limited to a single or unilocular type of cyst. In the case of E. multilocularis no such limitation occurs, so that the invasive multilocular type of cyst could more easily develop.


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