scholarly journals Germination In Vitro, Micropropagation, and Cryogenic Storage for Three Rare Pitcher Plants: Sarracenia oreophila (Kearney) Wherry (Federally Endangered), S. leucophylla Raf., and S. purpurea spp. venosa (Raf.) Wherry

HortScience ◽  
2012 ◽  
Vol 47 (1) ◽  
pp. 74-80 ◽  
Author(s):  
Cameron Northcutt ◽  
Daniel Davies ◽  
Ron Gagliardo ◽  
Kylie Bucalo ◽  
Ron O. Determann ◽  
...  
Keyword(s):  
Culture Media ◽  
Shoot Multiplication ◽  
Habitat Destruction ◽  
Plant Establishment ◽  
Pitcher Plants ◽  
Tissue Culture Media ◽  
In Vitro Micropropagation ◽  
Seed Cryopreservation

The genus Sarracenia forms a group of carnivorous pitcher plants native to North America. Habitat destruction and overcollection have caused pitcher plants to become rare, including U.S. federally endangered S. oreophila as well as S. leucophylla and S. purpurea spp. venosa (Raf.) Wherry, both listed as endangered in several states. Protocols for in vitro germination, sustainable shoot micropropagation, shoot establishment in soil, and seed cryopreservation are presented. Six-min sulfuric acid scarification treatments coupled with appropriate tissue culture media resulted in germination in vitro within 3 weeks, often reaching greater than 50%. Best germination for S. leucophylla and S. purpurea occurred on one-third strength Murashige and Skoog (MS) salts, whereas S. oreophila germinated best on one-sixth strength MS salts. Adjustment of pH to 4.5 to simulate a bog environment further increased germination for S. leucophylla. Shoot multiplication occurred at optimal levels when explants were placed on media in the presence of a cytokinin without auxin with greatest multiplication on 6-benzylaminopurine (BAP) or trans-zeatin and best shoot quality on trans-zeatin. Plant establishment in soil required both an in vitro rooting treatment and use of shoot clusters resulting in greater than 80% survival in soil. Seed cryopreservation tests with all three species suggest storage in liquid N2 followed by in vitro micropropagation and plant establishment can be used to preserve material long term.

Reproductive fluids, added to the culture media, contribute to minimizing phenotypical differences between in vitro-derived and artificial insemination-derived piglets

2022 ◽  
pp. 1-13
Author(s):  
Evelyne París-Oller ◽  
Cristina Soriano-Úbeda ◽  
Ramsés Belda-Pérez ◽  
Lucía Sarriás-Gil ◽  
Jordana S. Lopes ◽  
...  
Keyword(s):  
Glucose Tolerance ◽  
Culture Media ◽  
Growth Parameters ◽  
Tolerance Test ◽  
Oral Glucose ◽  
Long Term Effects ◽  
Hematological Indices ◽  
Reproductive Techniques

Abstract The addition of reproductive fluids (RF) to the culture media has shown benefits in different embryonic traits but its long-term effects on the offspring phenotype are still unknown. We aimed to describe such effects in pigs. Blood samples and growth parameters were collected from piglets derived from in vitro-produced embryos (IVP) with or without RF added in the culture media versus those artificially inseminated (AI), from day 0 to month 6 of life. An oral glucose tolerance test was performed on day 45 of life. We show here the first comparative data of the growth of animals produced through different assisted reproductive techniques, demonstrating differences between groups. Overall, there was a tendency to have a larger size at birth and faster growth in animals derived from in vitro fertilization and embryo culture versus AI, although this trend was diminished by the addition of RFs to the culture media. Similarly, small differences in hematological indices and glucose tolerance between animals derived from AI and those derived from IVP, with a sex-dependent effect, tended to fade in the presence of RF. The addition of RF to the culture media could contribute to minimizing the phenotypical differences between the in vitro-derived and AI offspring, particularly in males.

AFLP Analysis and Improved Phytoextraction Capacity of Transgenic gshI-Poplar Clones (Populus x canescens L.) for Copper in vitro

2005 ◽  
Vol 60 (3-4) ◽  
pp. 300-306 ◽  
Author(s):  
Gábor Gyulai ◽  
Mervyn Humphreys ◽  
András Bittsánszky ◽  
Kirsten Skøtc ◽  
József Kiss ◽  
...  
Keyword(s):  
Culture Media ◽  
Leaf Disc ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Tissue Culture Media ◽  
Dna Fragment ◽  
High Concentrations ◽  
Cu Uptake ◽  
Concentration Series

Abstract Clone stability and in vitro phytoextraction capacity of vegetative clones of P. x canescens (2n = 4x = 38) including two transgenic clones (ggs11 and lgl6) were studied as in vitro leaf disc cultures. Presence of the gshI-transgene in the transformed clones was detected in PCR reactions using gshI-specific primers. Clone stability was determined by fAFLP (fluorescent amplified DNA fragment length polymorphism) analysis. In total, 682 AFLP fragments were identified generated by twelve selective primer pairs after EcoRIDMseI digestion. Four fragments generated by EcoAGTDMseCCC were different (99.4% genetic similarity) which proves an unexpectedly low bud mutation frequency in P. \ canescens. For the study of phytoextraction capacity leaf discs (8 mm) were exposed to a concentration series of ZnSO4 (10-1 to 10-5 ᴍ) incubated for 21 days on aseptic tissue culture media WPM containing 1 μᴍ Cu. Zn2+ caused phytotoxicity only at high concentrations (10-1 to 10-2 ᴍ). The transgenic poplar cyt-ECS (ggs11) clone, as stimulated by the presence of Zn, showed elevated heavy metal (Cu) uptake as compared to the non-transformed clone. These results suggest that gshI-transgenic poplars may be suitable for phytoremediation of soils contaminated with zinc and copper.

Adaptation to chronic length change in explanted airway smooth muscle

2003 ◽  
Vol 95 (1) ◽  
pp. 448-453 ◽  
Author(s):  
Jahanbakhsh Naghshin ◽  
Lu Wang ◽  
Peter D. Paré ◽  
Chun Y. Seow
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Culture Media ◽  
Length Change ◽  
Functional Change ◽  
Muscle Stiffness ◽  
Airway Narrowing

It has been shown that airway smooth muscle in vitro is able to maintain active force over a large length range by adaptation in the absence of periodic stimulations at 4°C (Wang L, Paré PD, and Seow CY. J Appl Physiol 90: 734–740, 2001). In this study, we show that such adaptation also takes place at body temperature and that long-term adaptation results in irreversible functional change in the muscle that could lead to airway hyperresponsiveness. Rabbit tracheal muscle explants were passively maintained at shortened and in situ length for 3 and 7–8 days in culture media; the length-tension relationship was then examined. The length associated with maximal force generation decreased by 10.5 ± 3.8% (SE) after 3 days and 37.7 ± 8.5% after 7 or 8 days of passive shortening. At day 3, the left shift in the length-tension curve due to adaptation at short lengths was reversible by readapting the muscle at a longer length. The shift was, however, not completely reversible after 7 days. The results suggest that long-term adaptation of airway smooth muscle could lead to increased muscle stiffness and force-generating ability at short lengths. Under in vivo condition, this could translate into resistance to stretch-induced relaxation and excessive airway narrowing.

scholarly journals Micropropagation of Bacopa monnieri using Cyanobacterial Liquid Medium

1970 ◽  
Vol 20 (2) ◽  
pp. 225-231 ◽  
Author(s):  
Meenakshi Banerjee ◽  
Priyanka Modi
Keyword(s):  
Plant Tissue ◽  
In Vitro Propagation ◽  
Liquid Culture ◽  
Culture Media ◽  
Shoot Multiplication ◽  
Axillary Node ◽  
Bacopa Monnieri ◽  
Shoot Initiation ◽  
Aulosira Fertilissima

Hot extract of Aulosira fertilissima (cyanobacterium) added in different proportions to MS as a liquid culture media for the in vitro propagation of Bacopa monnieri (L.) Pennell. Maximum numbers of shoots were induced from axillary node in MS media (40 ml) + Aulosira extract (60 ml) and maximum shoot multiplication was observed when Kn (1.0 mg/l) was added in the shoot initiation media (mentioned above). Surprisingly rooting was also found to be best in the same combination of MS + cyanobacterial extract that was used for initiation and multiplication of shoots. On an average within a period of three subcultures (2 - 3 months) the nodal explants generated 400 shoots.  Rooted plantlets were successfully transferred to the field, after acclimation in the net house.   Key words: Baccopa monnieri, Cyanobacterial extract, Regeneration, Acclimation   D.O.I. 10.3329/ptcb.v20i2.6917   Plant Tissue Cult. & Biotech. 20(2): 225-231, 2010 (December)

scholarly journals In vitro tissue culture and genetic analysis of two high-CBD medical cannabis (Cannabis sativa L.) breeding lines

Genetika ◽  
2020 ◽  
Vol 52 (3) ◽  
pp. 925-941 ◽  
Author(s):  
Spela Mestinsek-Mubi ◽  
Sinja Svetik ◽  
Marko Flajsman ◽  
Jana Murovec
Keyword(s):  
Tissue Culture ◽  
Cannabis Sativa ◽  
Therapeutic Potential ◽  
Culture Media ◽  
Shoot Culture ◽  
Medical Cannabis ◽  
Breeding Lines ◽  
In Vitro Tissue Culture ◽  
Tissue Culture Media

The species Cannabis sativa L. has recently witnessed a resurgence of interest all over the world due to its multipurpose applications and the scientific confirmation of its pharmacological properties. Genotypes with high cannabinoid content are appreciated in the pharmaceutical and cosmetic industries due to their therapeutic potential. These genotypes, with predominantly high cannabidiol (CBD) content, are the subject of research and breeding in several programs, but to date, little data is published on the in vitro tissue culture of cannabis. Our study focused on the establishment of an efficient micropropagation method for two high-CBD breeding lines (MX-CBD-11 and MX-CBD-707) as the basis for advanced biotechnological breeding approaches. The results demonstrated that the in vitro culture of medical cannabis can be initiated on different culture media, that cultured plants can be successfully acclimatized, and that nodal position, and especially the genotype, have a significant influence on the success of shoot culture establishment. They showed that the published tissue culture media optimized for one high-THC strain of Mexican cannabis are not as efficient for other genotypes of (medical) cannabis. We complemented this research with a genetic study of 95 plants of the two breeding lines with 16 microsatellite (SSR) markers which clustered the plants based on breeding line. The results demonstrated that only 8 markers are needed for discrimination of all analyzed plants and their usefulness for clonal identification.

scholarly journals In vitro Regeneration and Callogenesis of Libidibia ferrea

2020 ◽  
pp. 14-24
Author(s):  
Daniel da Silva ◽  
Angela Maria Imakawa ◽  
Kamylla Rosas Vieira Guedes ◽  
Flávio Mauro Souza Bruno ◽  
Paulo de Tarso Barbosa Sampaio
Keyword(s):  
Culture Media ◽  
In Vitro Regeneration ◽  
Shoot Multiplication ◽  
Light Sources ◽  
Multiplication Rate ◽  
Medicinal Species ◽  
Efficient System ◽  
Blue Led ◽  
Friable Callus

Libidibia ferrea (Fabaceae) is a valuable medicinal species in the Amazon, but as it is a protected plant, collection from natural populations is forbidden. Therefore, establishing an efficient system for in vitro regeneration and to improve callogenesis of this species is desirable. To determine the optimal nutritional factors needed for shoot multiplication and callus induction, different culture media, plant growth regulators and LED light sources were tested. The data were subjected to analysis of variance (ANOVA) and means compared by Tukey’s test at p < 0.05. We observe that explants inoculated in the Murashige and Skoog (MS) media with 0.05 mg L-1 of 6-benzilaminopurine (BAP) and cultivated under red-blue LED induced the highest number of shoots (3.67), number of buds (3.13), multiplication rate (15.67) and shoots length (22.03 mm) when compared with other treatments. MS and B5 media supplemented with 2.21 and 4.42 mg L-1 of 2,4-D induced 100% formation of friable callus cultivated under red-blue LED, demonstrating that the light quality significantly influenced callogenesis. Obtained results confirmed that in vitro regeneration and callogenesis is a useful strategy in the protection of endangered species. In this way, a new renewable source of biomass with high quality plant material is presented aiming at the bioprospecting of seedling extracts and friable callus to obtain secondary metabolites of this medicinal plant.

scholarly journals Optimized cell culture conditions promote ex-vivo manipulation and expansion of primitive hematopoietic stem cells for therapeutic gene editing.

2022 ◽  
Author(s):  
Rajeev Rai ◽  
Winston Vetharoy ◽  
Asma Naseem ◽  
Zohar Steinberg ◽  
Adrian James Thrasher ◽  
...  
Keyword(s):  
Gene Editing ◽  
Culture Media ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Fine Tuning ◽  
Great Promise ◽  
Hematopoietic Stem ◽  
Monogenic Diseases

During the last few years, gene editing has emerged as a powerful tool for the therapeutic correction of monogenic diseases. CRISPR/Cas9 applied to hematopoietic stem and progenitor cells (HSPCs) has shown great promise in proof-of-principle preclinical studies to treat haematological disorders, and clinical trials using these tools are now underway. Nonetheless, there remain important challenges that need to be addressed, such as the efficiency of targeting primitive, long-term repopulating HSPCs and expand them in vitro for clinical purposes. Here we have tested the effect exerted by different culture media compositions on the ability of HSPCs to proliferate and undergo homology directed repair-mediated knock-in of a reporter gene, while preserving their stemness features during ex-vivo culture. We tested different combinations of compounds and demonstrated that by supplementing the culture media with inhibitors of histone deacetylases, and/or by fine-tuning its cytokine composition it is possible to achieve high levels of gene targeting in long-term repopulating HSPCs both in vitro and in vivo, with a beneficial balance between preservation of stemness and cell expansion, thus allowing to obtain a significant amount of edited, primitive HSPCs compared to established, state-of-the-art culture conditions. Overall, the implantation of this optimized ex vivo HSPC culture protocol will improve the efficacy, feasibility and applicability of gene editing and will likely provide one step further to unlock the full therapeutic potential of such powerful technology.

scholarly journals Kultur in vitro pisang Kepok Merah (Musa paradisiaca) untuk mikropropagasi cepat

2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Efah FITRAMALA ◽  
Eva KHAERUNNISA ◽  
Nina Ratna Djuita Ratna DJUITA ◽  
Hadi SUNARSO ◽  
Diah RATNADEWI
Keyword(s):  
Woody Plant ◽  
Shoot Multiplication ◽  
Ms Medium ◽  
Musa Paradisiaca ◽  
In Vitro Micropropagation ◽  
The Media ◽  
B Complex Vitamins ◽  
Initiation Stage ◽  
Planting Materials

AbstractBanana (Musa paradisiaca L) cv. Kepok Merah has a high commercial value as it is used in food industries such as banana chip. Besides, Kepok Merah contains high B-complex vitamins that serve in energy metabolism and are important in the development of infant brain. The establishment of industrial plantations of this plant has been restricted by the lack of planting materials. This research aimed at ameliorating the capacity of plantlets multiplication up to rooting of this banana in a rapid way through in vitro multiplication techniques. Murashige and Skoog (MS) and Woody Plant (WP) were used as the basic media. For the initiation stage, the media was fortified with 0.2 mg/L IAA and two levels of BA at 3 and 5 mg/L.  For shoot multiplication, the concentrations of IAA as well as BA were increased. For rooting, 1 mg/L NAA or IBA was applied. The observations demonstrated that for shoots initiation, both basic media performed good results when enriched with 0.2 mg/L IAA and 5 mg/L BA. The highest rate of shoots multiplication at 6 – 17 shoots per explant, was obtained on MS medium added with 0.5 mg/L IAA and 5 mg/L BA.  NAA at 1 mg/L in MS medium produced more rooted plantlets, 3 – 16 roots per plantlet, than those of other treatments. Keywords: Musa paradisiaca cv. Kepok Merah, in vitro micropropagation, scalps.AbstrakPisang (Musa paradisiaca L.) kultivar Kepok Merah memiliki nilai komersial yang cukup tinggi yaitu sebagai bahan dalam industri pembuatan keripik pisang. Selain itu, pisang Kepok Merah memiliki kandungan vitamin B kompleks cukup tinggi untuk membantu produksi energi dan pembentukan sel-sel otak pada bayi. Pertanaman pisang ini dalam skala industri terkendala oleh kurangnya ketersediaan sumber benih. Teknik kultur jaringan diharapkan dapat menghasilkan benih secara massal dalam waktu yang relatif singkat. Tujuan dari penelitian ini adalah meningkatkan keberhasilan multiplikasi tunas in vitro hingga pengakaran tanaman pisang Kepok Merah secara cepat. Pada tahap inisiasi tunas digunakan media dasar Murashige and Skoog (MS) dan media Woody Plant (WP); ke dalam media dasar tersebut ditambahkan IAA 0,2 mg/Ldan 2 taraf BA yaitu 3 dan 5 mg/L. Multiplikasi tunas dilakukan pada media dasar yang sama namun dengan taraf konsentrasi IAA serta BA yang ditingkatkan. Tahap perakaran menggunakan media dasar MS dan WP dengan auksin NAA 1 mg/L atau IBA 1 mg/L. Hasil penelitian menunjukkan bahwa untuk inisiasi tunas, media MS dan WP yang diperkaya dengan IAA 0,2 mg/L dan BA 5 mg/L   sama baiknya. Untuk multiplikasi tunas, media MS dengan IAA 0.5 mg/L   yang dikombinasikan dengan BA 5 mg/L   memberikan jumlah tunas paling banyak, yaitu 6 – 17 tunas per eksplan, dan pertumbuhannyapun lebih baik. Pemberian NAA 1 mg/L   pada media MS dapat memberikan lebih banyak tunas yang berakar, dengan jumlah akar 3 – 16 per planlet.  Kata kunci: Musa paradisiaca cv. Kepok Merah, mikropropagasi in vitro, nodul meristematik

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