In vitro culture

ContentsBovine demi-embryos were produced by micromanipulation of day 6–7 embryos from superovulated cows. Monolayers of bovine oviduct epithelial cells (BOEC) were produced from slaughterhouse material. Half of the demi-embryos were cultured on a confluent layer of bovine oviduct epithelial cells either in Hams F-10 with 10% NCTC-135 and with 10% FCS or in Earles Medium 199 with 10% FCS for 4–24 hours, followed by freezing of the survivors, while the corresponding other halfs were used for sexing. three 16-Cell, 1 morula, 41 compact morulae, 4 early blastocysts, 7 blastocysts and 2 expanded blaszocysts were besected and cultured. An overall survival rate of 83% among the 63 in vitro cultured demi-embryos was seen.Bovine demi-embryos were produced by micromanipulation of day 6–7 embryos from superovulated cows. Monolayers of bovine oviduct epithelial cells (BOEC) were produced from slaughterhouse material. Half of the demi-embryos were cultured on a confluent layer of bovine oviduct epithelial cells either in Hams F-10 with 10% NCTC-135 and with 10% FCS or in Earles Medium 199 with 10% FCS for 4–24 hours, followed by freezing of the survivors, while the corresponding other halfs were used for sexing. three 16-Cell, 1 morula, 41 compact morulae, 4 early blastocysts, 7 blastocysts and 2 expanded blaszocysts were besected and cultured. An overall survival rate of 83% among the 63 in vitro cultured demi-embryos was seen.Inhalt: In vitro Kultivierung von geteilten Rinderembryonen mit bovinen Ovidukt-Epithel-AmmenzellenSechs bis sieben Tage alte Embryonen von superovulierten Rindern wurden mit Hilfe eines Mikromanipulators geteilt. Monolyer von bovinen Eileiterepithelzellen (BOEC) wurden aus Schlachthofmaterial gerüchtet. Eine Hälfte der Embryonen diente der Geschlechtsbestimmung, die andere Hälfte wurde auf konfluierenden Monolayern entweder mit Hams F-10, 10% NCTC-135 und 10′% FCS oder in TCM 199 rnit 10% FCS für 2 bis 4 Stunden kultiviert. Die überlebenden Stadien wurden eingefroren. Drei 16-Zeller, 1 Morula, 41 kompaktierte Morulae, vier frühe und sieben Blastozysten sowie 2 expandierte Blastocysten wurden geteilt und kultiviert. Insgesamt betrug die Oberlebensrate bei 63 in vitro kultivierten Zwillingsembryonen 83%.Sechs bis sieben Tage alte Embryonen von superovulierten Rindern wurden mit Hilfe eines Mikromanipulators geteilt. Monolyer von bovinen Eileiterepithelzellen (BOEC) wurden aus Schlachthofmaterial gerüchtet. Eine Hälfte der Embryonen diente der Geschlechtsbestimmung, die andere Hälfte wurde auf konfluierenden Monolayern entweder mit Hams F-10, 10% NCTC-135 und 10′% FCS oder in TCM 199 rnit 10% FCS für 2 bis 4 Stunden kultiviert. Die überlebenden Stadien wurden eingefroren. Drei 16-Zeller, 1 Morula, 41 kompaktierte Morulae, vier frühe und sieben Blastozysten sowie 2 expandierte Blastocysten wurden geteilt und kultiviert. Insgesamt betrug die Oberlebensrate bei 63 in vitro kultivierten Zwillingsembryonen 83%.

Infection by human hepatitis C virus (HCV) is the principal cause of post-transfusion hepatitis and chronic liver diseases worldwide. A reliable in vitro culture system for the isolation and analysis of this virus is not currently available, and, as a consequence, HCV pathogenesis is poorly understood. We report here the first robust in vitro system for the isolation and propagation of HCV from infected donor blood. This system involves infecting freshly prepared macrophages with HCV and then transmission of macrophage-adapted virus into freshly immortalized B-cells from human fetal cord blood. Using this system, newly isolated HCV have been replicated in vitro in continuous cultures for over 130 weeks. These isolates were also transmitted by cell-free methods into different cell types, including B-cells, T-cells and neuronal precursor cells. These secondarily infected cells also produced in vitro transmissible infectious virus. Replication of HCV-RNA was validated by RT-PCR analysis and by in situ hybridization. Although nucleic acid sequencing of the HCV isolate reported here indicates that the isolate is probably of type 1a, other HCV types have also been isolated using this system. Western blot analysis shows the synthesis of major HCV structural proteins. We present here, for the first time, a method for productively growing HCV in vitro for prolonged periods of time. This method allows studies related to understanding the replication process, viral pathogenesis, and the development of anti-HCV drugs and vaccines.

The production of strong healthy plant material ex vitro is essential for an efficient seed potato production programs. Current study shows an applicable method for enhancing performance of potato plantlet subsequent in vitro phase. Using of ventilation and omitting sucrose from the nutrient medium (photoautotrophic culture) resulted in strong plantlets with higher fresh and dry weight performed better during acclimatization. Furthermore, Using of culture media contain macronutrients based on potato leaves macronutrients concentrations produced vigorous plantlets. Varietal differences were recorded between the two tested varieties Cara and Diamant. To obtain healthy start material for seed potato program this study recommend the use of in vitro photoautotrophic culture and modified nutrient media based on potato leaf macronutrient concentrations.

The positive effect of ventilation of the culture container on in vitro shoot proliferation and quality was already proven for different species. Hereafter we report on the evolution of the headspace during in vitro culture of plantain in a Temporary Immersion Bioreactor (TIB) on the one hand, and culture on semi-solid medium on the other hand. The CO2 and C2H4 concentration reached a maximum of 12% and 0.45 μl l−1, respectively in the control treatment on semi-solid medium, compared to 5.7% CO2 and 0.06 μl l−1 C2H4 in TIB. The minimal O2-concentration on semi-solid medium was 15.1%, compared to 19.3% in TIB. The multiplication rate was best in TIB, 6.4 compared to 4.3 in semi-solid conditions, and this was also the case for shoot height (4.3 cm compared to 3.3 cm), and leaf number (2.6 compared to 1.6). Moreover shoots produced on semi-solid medium showed distorted leaves. A typical day-night pattern in CO2 and O2 concentration was observed in TIB, as well as on semi-solid medium; this is illustrative for the photosynthetic capacity of the plant material produced in both systems.

ABSTRACT: Cassava is one of the most important food crops in Africa. Meristem culture is an effective method of eliminating viruses and other systemic diseases spread through the vegetative propagation of stems. However, in semi-arid conditions, survival of ex vitro plants in the field is often disappointing. When an increasing range of light regimes in vitro was provided, the fresh and dry masses more than doubled their values between 29 and 369 mmol s−1 m−2 PPFDs. Increases in numbers of senescent leaves and stem thickness were also recorded with increasing PPFD. However, PPFD above 101 mmol s−1 m−2 resulted in 30–70% reduction in plant survival, with the thin plants with the smallest fresh and dry masses being the ones with highest survival rates. High light and temperature levels in the greenhouse were also found to be critical for plant survival. It was also shown that transpirational loss from detached leaves and epicuticular wax deposits were not good indicators for predicting survival of ex vitro cassava plantlets during acclimatisation.

We obtained callus, cell suspension and hairy root cultures of Tropaeolum majus and we demonstrated their ability to produce glucotropaeolin and myrosinase. In hairy roots glucotropaeolin content and myrosinase activity were higher in comparison with callus, cell suspension and leaves of intact plants. In hairy root cultures the highest glucotropaeolin contents were detected on the 9th day of culture. In

Soria-Soto, M., Molina-Fernández, D., Hidalgo-Hidalgo, V., Adroher, F.J., Benítez, R., Malagón, D., 2013. In vitro cultivation of Anisakis simplex, causal agent of the human anisakiasis, in: III Jornadas Internacionales y V Nacionales de Ciencias de la Salud, Facultad de Ciencias de la Salud, Universidad de Granada (Eds.), "Nuevos Retos en Investigación, Docencia y Clínica en Ciencias de la Salud". Granada, pp. 223–227.
Anisakis simplex is a parasite ascaridoid nematode, main etiological agent of the parasitic illness known as human anisakiasis, which affects the gastrointestinal tract. The natural final hosts are cetaceans. The intermediate/paratenic hosts are crustaceans, fishes, and cephalopods, which harbor the third larval stage (L3) of this nematode. When the
human, who acts as an accidental host, consumes these raw, marinated or undercooked hosts harboring L3 of Anisakis, has a high risk of acquiring human anisakiasis.

Mass proliferation from the lower half bulb scales and adventitious bulb regeneration from in vitro cultured leaf explants of Madonna Lilly (Lilium candidum L.) was achieved using various concentrations of BAP-IBA. Regenerated plantlets were transferred to greenhouse for adaptation where they flowered after two years.

The present study was conducted at ICAR-Central Potato Research Institute, Regional Station Gwalior during 2007-08 to 2016-17 for 10 years on micro-propagted material multiplied at the station to identify suitable tissue culture/ hi-tech based seed potato production systems using micro-plants, micro-tubers and mini-tubers and performance of seven popular cultivars viz Kufri Lauvkar, Kufri Chandramukhi, Kufri Surya, Kufri Bahar, Kufri Chipsona-1, Kufri Chipsona-3 and Kufri Sindhuri planted during 10 years under these three hi-tech systems for potato breeder seed production. Variation in performance among different varieties for establishment/emergence, mean tuber weight and seed multiplication rate (both by number and weight), percent

The production of reactive oxygen species (ROS) in plants is a common event in metabolic and physiological processes as well as in the response to biotic and abiotic stress. In this paper we will report that root tissue from axenically grown tomato cultivars and Lycopersicon wild species can be used for the determination of superoxide production. Superoxide generation was evaluated following the treatment of root tissues with two general elicitors of the defence response: laminarin and calcium ionophore A23187. Results demonstrated that elicitor reactivity in terms of superoxide generation of the tomato cultivars and the wild species used was different. This suggested varying levels of competence for non-specific active defence. The proposed technique merges the advantages of in vitro cultures and of whole tissues and also demonstrates that root tissue is a suitable material for evaluating free radical release.

The present study was conducted at ICAR-Central Potato Research Institute, Regional Station Gwalior during 2007-08 to 2016-17 for 10 years on micro-propagted material multiplied at the station to identify suitable tissue culture/ hi-tech based seed potato production systems using micro-plants, micro-tubers and mini-tubers and performance of seven popular cultivars viz Kufri Lauvkar, Kufri Chandramukhi, Kufri Surya, Kufri Bahar, Kufri Chipsona-1, Kufri Chipsona-3 and Kufri Sindhuri planted during 10 years under these three hi-tech systems for potato breeder seed production. Variation in performance among different varieties for establishment/emergence, mean tuber weight and seed multiplication rate (both by number and weight), percent

The present study was conducted at ICAR-Central Potato Research Institute, Regional Station Gwalior during 2007-08 to 2016-17 for 10 years on micro-propagted material multiplied at the station to identify suitable tissue culture/ hi-tech based seed potato production systems using micro-plants, micro-tubers and mini-tubers and performance of seven popular cultivars viz Kufri Lauvkar, Kufri Chandramukhi, Kufri Surya, Kufri Bahar, Kufri Chipsona-1, Kufri Chipsona-3 and Kufri Sindhuri planted during 10 years under these three hi-tech systems for potato breeder seed production. Variation in performance among different varieties for establishment/emergence, mean tuber weight and seed multiplication rate (both by number and weight), percent <3 g tuber (both by number and weight) and tuber yield (m 2) were recorded during the study. Kufri Sindhuri, Kufri Bahar and Kufri Surya under micro-plant based, Kufri Bahar and Kufri Surya and Kufri Chipsona-1 under micro-tuber and mini-tuber based multiplication system performed better than other varieties under G-0 conditions for yield attributing parameters. Seed multiplication rate by number was higher in micro-plant system of seed production (5.50) followed by micro-tuber systems (5.31) and mini-tuber (4.43) based system of seed production. Mean tuber weight was higher for mini-tuber system of seed production (22.46 g) followed by micro-plant (15.59 g) and micro-tuber systems (14.62) and showed higher tuber yield i.e 130 tuber/m 2 and 3.10 Kg/m 2 in mini-tuber based system followed by111 tuber/m 2 and 1.74 kg/m 2 in micro-plant based system and by 86 tuber/m 2 and 1.16 kg/m 2 in micro-tuber based seed potato production system. Hence performance of Kufri Sindhuri, Kufri Bahar and Kufri Surya under micro-plant based, Kufri Bahar and Kufri Surya and Kufri Chipsona-1 under micro-tuber and mini-tuber based seed multiplication system was better in 10 years of the study.