Collection periods in the in vitro establishment of raspberry tree cultivars

The raspberry tree, a species from the Rosaceae family and the Rubus genus, is included in the group of small fruit that has been drawing the attention of the global consuming market for carrying antioxidant properties and substances capable of fighting free radicals, bringing health benefits. The objective of this work was to evaluate the effect of collection periods on the in vitro establishment of raspberry trees (Rubus idaeus L.). Explants were collected in different seasons of the year (winter, spring, summer and autumn), and were inoculated in a MS culture medium. The experimental design was totally randomized, and arranged in a factorial scheme. At 28 days, the following variables were evaluated: oxidation, fungal and bacterial contamination survival and establishment. There was less phenolic oxidation in raspberry tree explants collected in winter. Explants contamination by endogenic bacteria occurred in higher percentages in winter. Although summer was the season of the year with higher percentage of explants’ phenolic oxidation for both cultivars, the highest in vitro establishment percentages for the Heritage cultivar occurred in summer and fall. However, for the Fallgold cultivar, spring and winter were the seasons with greater in vitroestablishment.

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DESINFESTAÇÃO E MEIO DE CULTURA PARA O ESTABELECIMENTO In Vitro DE SEGMENTOS NODAIS DE Liquidambar styraciflua

FLORESTA ◽

10.5380/rf.v40i3.18916 ◽

2010 ◽

Vol 40 (3) ◽

Author(s):

Gilvano Ebling Brondani ◽

Fabricio Augusto Hansel ◽

Leonardo Ferreira Dutra ◽

Ivar Wendling

Keyword(s):

Sodium Hypochlorite ◽

Bacterial Contamination ◽

Culture Medium ◽

Morphological Characteristics ◽

Liquidambar Styraciflua ◽

Nodal Segments ◽

Hydroponic System ◽

Randomized Design ◽

In Vitro Establishment

O objetivo deste trabalho foi testar a desinfestação e meios de cultura para o estabelecimento in vitro de segmentos nodais de Liquidambar styraciflua L. Os explantes foram coletados de minicepas propagadas pelo processo de estaquia e manejadas em minijardim clonal sob sistema semi-hidropônico em leito de areia. O experimento foi conduzido em delineamento inteiramente casualizado no arranjo fatorial (3x2x2), sendo os fatores constituídos por três clones (L 26, L 35 e L 63), duas metodologias de desinfestação (A1 - hipoclorito de sódio (NaOCl) durante 10 minutos a 2,5% v/v de cloro ativo e A2 - explantes mergulhados durante 40 minutos em solução a base de benomyl à 1% p/v) e dois meios de cultura (WPM e MS), com quatro repetições. Os clones não diferiram em relação à assepsia e meio de cultura, obtendo-se média de 4% de explantes isentos de contaminação. Contudo, cerca de 80% dos explantes apresentaram contaminação bacteriana, indicando a necessidade do desenvolvimento de um protocolo de desinfestação mais eficiente. Embora tenham ocorrido poucas diferenças entre os meios de cultura testados, o meio de cultura MS apresentou superioridade em relação ao WPM para a maioria das características morfológicas estudadas.Palavras-chave: Micropropagação; assepsia; contaminação bacteriana; hipoclorito de sódio; benomyl. AbstractDisinfestation and culture medium for the in vitro establishment of Liquidambar styraciflua nodal segments. The objective of this research was to test the culture medium sterilization for the in vitro establishment of Liquidambar styraciflua L. nodal segments. Ministumps, from which the explants were collected, were propagated by cutting process and managed in clonal mini garden under semi-hydroponic system in a sand bed. The experiment was conducted in a completely randomized design under factorial arrangement (3x2x2); the factors were: three clones (L 26, L 35 and L 63), two sterilization methods (A1 - sodium hypochlorite (2.5% v/v of active chlorine) during 10 minutes; A2 - explants immersed during 40 minutes in a solution of benomyl 1% w/v) and two culture mediums (WPM and MS), with four replications. The clones did not differ in relation to the asepsis and culture medium. In average, 4% of the explants were free of contamination. However, almost 80% of the material presented bacterial contamination, what indicated the necessity of developing a more efficient sterilization protocol. Although only a little difference between the culture mediums had been observed, the MS medium showed superiority in relation to the WPM regarding the majority of the morphological characteristics studied.Keywords: Micropropagation; asepsis; bacterial contamination; sodium hypochlorite; benomyl.

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In vitro establishment of blackberry (Rubus sp.) cultivar 'Xavante'

Ciência Rural ◽

10.1590/0103-8478cr20140988 ◽

2016 ◽

Vol 46 (9) ◽

pp. 1542-1545

Author(s):

Tânia Regina Pelizza ◽

Fabiane Nunes Silveira ◽

Roberta Sabatino Ribeiro ◽

Bruno Dalazen Machado ◽

Leo Rufatto ◽

...

Keyword(s):

Bacterial Contamination ◽

Ms Medium ◽

Lower Survival ◽

In Vitro Establishment

ABSTRACT: The objective of this study was to evaluate the in vitro establishment of 'Xavante' blackberry under different conditions of plant luminosity and concentrations of salts of MS medium. The maintenance of 'Xavante' blackberry plants in the absence of light, with the use of MS medium with salt concentrations of 75% and 100% reduce bacterial contamination and the percentage of explants oxidized and provide the highest percentage of surviving and established explants. The 50% salt dilution in MS medium is not recommended when plants are subjected to the absence of light due to the high percentage of oxidized explants and consequently lower survival and establishment.

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Clonal bamboo production based on in vitro culture

Bioscience Journal ◽

10.14393/bj-v36n4a2020-48169 ◽

2020 ◽

Vol 36 (4) ◽

Author(s):

Anatálya dos Santos Ribeiro ◽

Alexssandra Jéssica Rondon de Figueiredo ◽

Gabriela Cristina Rech Tormen ◽

André Luís Lopes da Silva ◽

Wellington Ferreira Campos ◽

...

Keyword(s):

Activated Carbon ◽

Large Scale ◽

Culture Medium ◽

Clonal Plant ◽

Adventitious Rooting ◽

Clonal Plants ◽

Ex Vitro ◽

Bambusa Vulgaris ◽

In Vitro Establishment

Bamboo species are an alternative for the composition of forest plantations. However, their potential has not been explored due to the hard time in producing large-scale clonal plants. Thus, the aim this work was to evaluate the in vitro establishment, bud multiplication and ex vitro rooting of Bambusa vulgaris. The first experiment tested different systemic and contact fungicide solutions, based on exposure time, during the establishment phase. Established explants were subjected to evaluation of residual fungicide effect on subcultures during the multiplication and elongation phases. The second experiment evaluated the influence of activated carbon on ex vitro survival and on adventitious rooting. Explant immersion in liquid culture medium added with 1.0 mL of fungicide for 120 hours has favored the in vitro establishment and reduced fungal contamination. On the other hand, it favored the shoot emission of shoots per explant during the multiplication phase. Both rooting induction culture medium and mini-incubator system use were effective in enabling adventitious root formation. The presence of activated carbon in the rooting induction culture medium resulted in a higher clonal plant survival rate.

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In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bactéria

Pesquisa Agropecuária Tropical ◽

10.1590/1983-40632019v4953673 ◽

2019 ◽

Vol 49 ◽

Author(s):

Ana Paula de Azevedo Pasqualini ◽

Gabriela Xavier Schneider ◽

Hugo Pacheco de Freitas Fraga ◽

Luiz Antonio Biasi ◽

Marguerite Quoirin

Keyword(s):

Molecular Identification ◽

Culture Medium ◽

Bacterial Isolate ◽

Fungal Growth ◽

Endophytic Bacterium ◽

Liquid Culture Medium ◽

Rdna Sequencing ◽

In Vitro Establishment ◽

Bambusa Oldhamii

ABSTRACT In plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test.

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Antioxidant properties of cystic fibrosis sputum

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00349.2004 ◽

2005 ◽

Vol 288 (5) ◽

pp. L903-L909 ◽

Cited By ~ 8

Author(s):

Nurlan Dauletbaev ◽

Jens Rickmann ◽

Klaus Viel ◽

Holger Diegel ◽

Christian von Mallinckrodt ◽

...

Keyword(s):

Cystic Fibrosis ◽

Glutathione Peroxidase ◽

Culture Medium ◽

Antioxidant Properties ◽

Sputum Induction ◽

Protein Thiols ◽

Fetal Bovine ◽

Heme Peroxidases ◽

In Cells

Oxidative stress is a likely contributor to the pathogenesis of cystic fibrosis (CF) lung disease. However, hydrogen peroxide (H2O2), a physiological oxidant, is not elevated in CF exhalates. H2O2may be neutralized by antioxidants in CF airway secretions. The H2O2-detoxifying capacity of CF airway secretions, obtained via sputum induction, was studied in an in vitro H2O2cytotoxicity model. 16HBE14o- cells were exposed to H2O2in culture medium containing either 0 or 10% fetal bovine serum (FBS) or 10% CF sputum supernatant (extracted without use of dithiothreitol). The efficiency of H2O2neutralization was estimated by measuring intracellular oxidant levels (dihydrorhodamine 123) after 2 h and cell viability (propidium iodide) after 24 h of H2O2exposure. Furthermore, the presence of reduced thiols (DTNB assay) and reduced glutathione (recycling assay) in CF sputum samples was evaluated. CF sputum extracts completely prevented intracellular oxidant accumulation seen in cells incubated with H2O2in both control media (i.e., 0 or 10% FBS). Furthermore, CF sputum abolished cell death in 16HBE14o- cells exposed to up to 1 mM H2O2. In contrast, there was 100% cytotoxicity in cells exposed to 600 μM H2O2in both control media. The H2O2-detoxifying potential of CF sputum was sustained after catalase and heme peroxidases were inactivated by sodium azide, which does not affect glutathione peroxidase. In addition, reduced protein thiols were found in abundance in CF sputum. In conclusion, CF sputum is capable to neutralize H2O2and abundant reduced thiols and/or glutathione peroxidase are fully sufficient to detoxify H2O2.

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A METHOD FOR THE PHYSIOLOGICAL STUDY OF TISSUES IN VITRO

Journal of Experimental Medicine ◽

10.1084/jem.38.4.407 ◽

1923 ◽

Vol 38 (4) ◽

pp. 407-418 ◽

Cited By ~ 70

Author(s):

Alexis Carrel

Keyword(s):

Nutrient Medium ◽

Bacterial Contamination ◽

Culture Medium ◽

Residual Activity ◽

Physical Factors ◽

Dynamic Changes ◽

Fluid Medium ◽

Continuous Growth ◽

Pure Cultures

1. A method has been developed which allows the continuous growth of pure strains of fibroblasts, epithelium, and leucocytes in a medium which undergoes but slight spontaneous deterioration. 2. The principle of the method is to leave the tissues undisturbed while the medium is changed. This was realized by special containers allowing the change of the medium without bacterial contamination and by the simultaneous use of a solid and a fluid medium. 3. The curve of growth of pure cultures of fibroblasts and epithelial cells in a nutrient medium is a parabola; in a non-nutrient medium, it is S-shaped and expresses the residual activity of the tissues. Leucocytes invade the culture medium progressively, as do bacteria, but never aggregate in a tissue. 4. The method is used for the study of the morphological and dynamic changes occurring in tissues under the influence of chemical and physical factors.

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Effects of a commercial biocide, kasugamycin and consistency of the culture medium on the in vitro establishment of bamboo1

Pesquisa Agropecuária Tropical ◽

10.1590/1983-40632019v4955435 ◽

2019 ◽

Vol 49 ◽

Author(s):

Daniela Werner Ribeiro dos Santos ◽

Théo Piucco Rocker ◽

Thiago Sanches Ornellas ◽

Miguel Pedro Guerra

Keyword(s):

Liquid Culture ◽

Culture Medium ◽

Liquid Culture Medium ◽

Bambusa Vulgaris ◽

Phyllostachys Bambusoides ◽

In Vitro Establishment ◽

Dendrocalamus Asper

ABSTRACT The contamination by microorganisms and oxidation of explants in the in vitro establishment of bamboo are recurrent problems for its micropropagation. In the present study, effects of the biocide Plant Preservative Mixture (PPM™), the antibiotic kasugamycin and the consistency of the culture medium were evaluated in the in vitro establishment of Bambusa vulgaris,Phyllostachys bambusoides and Dendrocalamus asper. The presence of PPM™ in the culture medium had a significant effect using 2 mL L-1 or 4 mL L-1 concentrations, as well as in the liquid culture medium, increasing the plants established in the autumn. Kasugamycin promoted variable responses for all the three species, depending on the season. There was interaction among the factors, demonstrating that higher rates of viable plants can be obtained by combining different strategies to reduce the oxidation and contamination. For the in vitro establishment of B. vulgaris,P. bambusoides and D. asper, it is recommended to add 2 mL L-1 or 4 mL L-1 of PPM™ to the liquid culture medium.

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EFFECT OF 6- BENZYLAMINOPURINE AND THE CULTURE MEDIUM MS (1962) ON THE IN VITRO ESTABLISHMENT OF Prosopis pallida (WILLD.) KUNTH

REBIOL ◽

10.17268/rebiol.2019.39.02.03 ◽

2019 ◽

Vol 39 (2) ◽

pp. 30-40

Author(s):

Diego Campos ◽

Claudia Chávez ◽

Segundo Lopéz ◽

Armando Gil- Rivero ◽

Angélica Lopéz -Zavaleta ◽

...

Keyword(s):

Culture Medium ◽

In Vitro Establishment

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Androgenesis of Red Cabbage in Isolated Microspore Culture In Vitro

Plants ◽

10.3390/plants10091950 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1950

Author(s):

Anna Mineykina ◽

Ludmila Bondareva ◽

Alexey Soldatenko ◽

Elena Domblides

Keyword(s):

Culture Medium ◽

Microspore Culture ◽

Doubled Haploids ◽

Antioxidant Properties ◽

F1 Hybrids ◽

Anthocyanin Content ◽

Isolated Microspore Culture ◽

Culture In Vitro ◽

Red Cabbage

Red cabbage belongs to the economically important group of vegetable crops of the Brassicaceae family. A unique feature of this vegetable crop that distinguishes it from other members of the family is its unique biochemical composition characterized by high anthocyanin content, which gives it antioxidant properties. The production mainly uses F1 hybrids, which require constant parental lines, requiring 6–7 generations of inbreeding. Culture of isolated microspores in vitro is currently one of the promising methods for the accelerated production of pure lines with 100% homozygosity. The aim of this study is to investigate the factors and select optimal parameters for successful induction of red cabbage embryogenesis in isolated microspore culture in vitro and subsequent regeneration of DH plants. As a result of research, for the first time, it was possible to carry out the full cycle of obtaining DH plants of red cabbage from the induction of embryogenesis to their inclusion in the breeding process. The size of buds containing predominantly microspores at the late vacuolated stage and pollen at the early bi-cellular stage has to be selected individually for each genotype, because the embryoid yield will be determined by the interaction of these two factors. In the six samples studied, the maximum embryoid yield was obtained from buds 4.1–4.4 mm and 4.5–5.0 mm long, depending on the genotype. Cultivation of microspores was carried out on liquid NLN culture medium with 13% sucrose. The maximum number of embryoids (173.5 ± 7.5 pcs./Petri dish) was obtained on culture medium with pH 5.8 and heat shock at 32 °C for 48 h. Successful embryoid development and plant regeneration by direct germination from shoot apical meristem were achieved on MS culture medium with 2% sucrose and 0.7% agar, supplemented with 6-benzylaminopurine at a concentration of 1 mg/L. Analysis of the obtained regenerated plants, which successfully passed the stage of adaptation to ex vitro conditions by flow cytometry, showed that most of them were doubled haploids (up to 90.9%). A low number of seeds produced by self-fertilization in DH plants was observed.

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In vitro seed germination and multiplication of Calophyllum brasiliense

Pesquisa Florestal Brasileira ◽

10.4336/2016.pfb.36.87.1168 ◽

2016 ◽

Vol 36 (87) ◽

pp. 185

Author(s):

Sheila Susy Silveira ◽

Juliana Degenhardt-Goldbach ◽

Marguerite Germaine Ghislaine Quoirin

Keyword(s):

Bacterial Contamination ◽

Culture Medium ◽

Germination Rate ◽

Nodal Segment ◽

In Vitro Germination ◽

Fungal Contamination ◽

Natural Reproduction ◽

Calophyllum Brasiliense ◽

Free Rate

Calophyllum brasiliense is a tree species with limited natural reproduction. In vitro germination may be an alternative for obtaining high-quality seedlings. Seeds were maintained in water before surface disinfestation and compared with control seeds (i.e. not immersed), without differences between treatments. HgCl2 used during surface-disinfestation reduced contamination rates of cultures. Fungal contamination was reduced with fungicide added to culture medium (23 to 6.4%), although bacterial contamination increased (24 to 36%). In another experiment, seeds were immersed in plant preservative mixture (PPM™) prior to surface disinfestation. By combining immersion for 48 h and 2 mL L-1 in culture medium, contamination was only 6%. Seeds immersion in GA3 prior to surface disinfestation reduced root formation as concentration increased. Germination rate and GSI were reduced, respectively, from 72% and 0.129 (24 h) to 60% and 0.092 (48 h) according to exposure time to GA3. After 90 days in multiplication medium containing benzylaminopurine, average number of shoots per nodal segment was 3.4. In conclusion, in vitro germination of C. brasiliense seeds is feasible in sucrose-free WPM medium and reaches a high contamination-free rate (up to 93.3%).

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