Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminium cryo-plates

2011 ◽  
Vol 10 (1) ◽  
pp. 14-19 ◽  
Author(s):  
Shin-ichi Yamamoto ◽  
Kuniaki Fukui ◽  
Tariq Rafique ◽  
Nayyar Iqbal Khan ◽  
Carlos Roman Castillo Martinez ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sucrose Solution ◽  
Shoot Tips ◽  
New Method ◽  
Fragaria Ananassa ◽  
Alginate Gel ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Strawberry Cultivars

Cryopreservation using an aluminium cryo-plate was successfully applied to in vitro-grown strawberry (Fragaria × ananassa Duch.) shoot tips. The shoots were cold-hardened at 5°C for 3 weeks with an 8-h photoperiod. The shoot tips (1.5–2.0 mm × 0.5–1.0 mm) were dissected from the shoot and pre-cultured at 5°C for 2 d on Murashige and Skoog medium containing 2 M glycerol and 0.3 M sucrose. The pre-cultured shoot tips were placed on the aluminium cryo-plate containing ten wells embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates in a loading solution (2 M glycerol and 0.8 M sucrose) for 30 min at 25°C. Dehydration was performed by immersing the cryo-plates in plant vitrification solution 2 for 50 min at 25°C. Then, the cryo-plate with shoot tips was transferred into an uncapped cryotube that was held on a cryo-cane and directly immersed into liquid nitrogen (LN). After storage in LN, shoot tips attached to the cryo-plate were directly immersed into 2 ml of a 1 M sucrose solution for regeneration. Using this procedure, the average regrowth level of vitrified shoot tips of 15 strawberry cultivars reached 81%. This new method has many advantages and will facilitate the cryostorage of strawberry germplasm.

scholarly journals Critical vitrification steps towards survival of Garcinia hombroniana (Clusiaceae) shoot tips after cryopreservation

2018 ◽  
Vol 66 (3) ◽  
pp. 1314
Author(s):  
Norafarain Sulong ◽  
Nurul Farhana Shahabudin ◽  
Normah Mohd Noor
Keyword(s):  
Water Content ◽  
Liquid Nitrogen ◽  
Shoot Tips ◽  
The Other ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Vitrification Solution ◽  
Cryopreservation Protocol ◽  
Recovery Medium

A cryopreservation protocol was developed for in vitro shoot tips of Garcinia hombroniana using the vitrification technique. Four critical steps in the technique were investigated, namely preculture, loading, dehydration with Plant Vitrification Solution 2 (PVS2), and unloading. Shoot tips precultured for 48 hr gave significantly higher survival (75 %) compared to 24 hr preculture (50 %) after cryopreservation. Treatment with 1 M glycerol plus 0.4 M sucrose as a loading solution gave higher survival (45.83 %) compared to the other treatments (0.4 M sucrose + 2 M glycerol; 0.4 M sucrose). Shoot tips dehydrated with PVS2 for 25 min gave the highest survival after immersion in liquid nitrogen. Stepwise PVS2 treatment for 15 min with 50 % PVS2 followed by 10 min with 100 % PVS2 solution improved survival of the shoot tips after cryopreservation (41.67 %). Murashige and Skoog medium with 0.4 M sucrose gave significantly higher survival (66.67 %) than MS with 1.2 M sucrose (25 %) as an unloading solution. Water content was shown to decrease throughout the whole vitrification steps from 6.83 ± 1.66 g g-1 dw for fresh shoot tips down to 2.93 ± 0.28 g g-1 dw after PVS2 treatment. Further study on each step including recovery medium is required to improve the survival. Nevertheless, the present study showed the potential of using the vitrification technique for cryopreservation of G. hombroniana.

MICROPROPAGATION OF Helianthus maximiliani (Schrader) BY SHOOT APEX CULTURE / MICROPROPAGACION DE Helianthus maximiliani (Schrader) POR EL CULTIVO DE VAINAS DE LOS VASTAGOS / MICROPROPAGATION DE L’Helianthus maximiliani (Schrader) AU MOYEN DE LA CULTURE DE L’APEX DE LA POUSSE

Helia ◽  
2001 ◽  
Vol 24 (34) ◽  
pp. 63-68 ◽  
Author(s):  
Vasić Dragana ◽  
Škorić Dragan ◽  
Alibert Gilbert ◽  
Miklič Vladimir
Keyword(s):  
Silver Nitrate ◽  
Shoot Apex ◽  
Casein Hydrolysate ◽  
Shoot Tips ◽  
Nodal Segments ◽  
Shoot Apices ◽  
Skoog Medium ◽  
Leaf Mesophyll ◽  
Murashige And Skoog Medium

SUMMARYH.maximiliani was micropropagated using culture of shoot apices on modified Murashige and Skoog medium (DV). Further propagation of in vitro grown plants was done by culture of their nodal segments and shoot tips on the same medium supplemented with phloridzin, silver nitrate and casein hydrolysate (DV'). Rooting was induced by dipping the explants into IBA solution prior culture. Viable protoplasts (90%) were isolated from leaf mesophyll. These protoplasts divided (18%) in culture in agarose droplets.

scholarly journals Shoot Proliferation and Rooting in Vitro of Pulmonaria

HortScience ◽  
1998 ◽  
Vol 33 (2) ◽  
pp. 339-341 ◽  
Author(s):  
Dennis P. Stimart ◽  
John C. Mather ◽  
Kenneth R. Schroeder
Keyword(s):  
Shoot Proliferation ◽  
Shoot Tips ◽  
Optimum Level ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Ba Genotype

Expanding shoot tips of Pulmonaria `Roy Davidson' and Pulmonaria saccharata `Margery Fish' were cultured in vitro on a modified Murashige and Skoog medium containing BA to establish proliferating cultures for use in comparing BA concentrations on shoot proliferation and rooting. The optimum level for shoot proliferation was 8.8 μm BA. Greatest rooting was on medium without BA. Genotype and time in culture influenced shoot and root counts. Chemical names used: N6-benzyladenine (BA)

scholarly journals REGENERATION OF CITRUS VIA SHOOT APECIES

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1112a-1112
Author(s):  
Suzanne M.D. Rogers ◽  
Kalyani Dias ◽  
David Byrne
Keyword(s):  
Growth Regulators ◽  
Shoot Tips ◽  
Sour Orange ◽  
Soil Medium ◽  
Skoog Medium ◽  
Murashige And Skoog Medium

Viral damage is a major problem in citrus. As most citrus are asexually propagated, it is necessary to have an alternative way of regenerating virus-free plants from infected plants. Shoot apicies are the most suitable explant material for this purpose because that part of the plant is virus-free. Fifty sour orange shoot tips and 22 Swingle shoot tips, 1 mm - 1.5 mm long, were excised from in vitro germinated seedlings and cultured on semisolid Murashige and Skoog medium, without growth regulators, containing 0.2 % Gelrite. After 8-10 weeks, shoots and leaves developed in 68'% of the sour orange explants, and in 77% of the Swingle explants. Some explants produced roots, after 11-12 weeks, and could be removed from culture and established in soil medium.

scholarly journals Cryopreservation of shoot tips of endangered Hayachine-usuyukiso ( Leontopodium hayachinense (Takeda) Hara et Kitam.) using a vitrification protocol

2008 ◽  
Vol 6 (02) ◽  
pp. 164-166 ◽  
Author(s):  
Daisuke Tanaka ◽  
Takao Niino ◽  
Yoshiko Tsuchiya ◽  
Kazuto Shirata ◽  
Matsuo Uemura
Keyword(s):  
Survival Rate ◽  
Liquid Nitrogen ◽  
Shoot Tips ◽  
Endangered Plant ◽  
Alpine Plant ◽  
Promising Technique ◽  
Skoog Medium ◽  
Murashige Skoog ◽  
Iwate Prefecture

Hayachine-usuyukiso (Leontopodium hayachinense) is an alpine plant native to Mt Hayachine. This unique chrysanthemum is listed as an endangered plant by the Department of Conservation, Iwate Prefecture, and as a threatened plant by the Ministry of the Environment, Japan. We successfully cryopreserved the shoot tips fromin vitro-grownL. hayachinenseshoots using a vitrification protocol. Cold-hardened shoot tips were excised and pre-cultured on a solidified Murashige–Skoog medium containing 0.3 M sucrose for 1 d at 5°C. The shoot tips were then treated with loading solution for 20 min at 25°C, dehydrated in plant vitrification solution 2 for 120 min at 25°C and immersed in liquid nitrogen. The survival rate of the vitrified shoot tips was 63.3% after 30 d of regrowth. This protocol appears to be a promising technique for the cryopreservation ofin vitro-grown shoots of this endangered plant.

scholarly journals In vitro Conservation and Cryopreservation of Nandina domestica, an Outdoor Ornamental Shrub

2013 ◽  
Vol 41 (2) ◽  
pp. 638 ◽  
Author(s):  
Aylin OZUDOGRU ◽  
Diogo Pedrosa Corrêa Da SILVA ◽  
Ergun KAYA ◽  
Giuliano DRADI ◽  
Renato PAIVA ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Storage Temperature ◽  
Sucrose Concentration ◽  
Aluminum Foil ◽  
Shoot Tips ◽  
In Vitro Conservation ◽  
Shoot Cultures ◽  
Storage Medium ◽  
Droplet Vitrification

The study focused on an economically-important ornamental outdoor shrub, Nandina domestica, with the aims to (i) optimize an effective in vitro conservation method, and (ii) develop a cryopreservation protocol for shoot tips by the PVS2 vitrification and droplet-vitrification techniques. For in vitro conservation of shoot cultures, the tested parameters were sucrose content in the storage medium (30, 45, 60 g/L) and storage temperature (4 °C or 8 °C). Cryopreservation was performed by applying the PVS2 vitrification solution, in 2-ml cryovials or in drops over aluminum foil strips, for 15, 30, 60 or 90 min at 0 °C, followed by the direct immersion in liquid nitrogen of shoot tips. Results show that N. domestica shoots can be conserved successfully for 6 months at both the temperatures tested, especially when 60 g/L sucrose is used in the storage medium. However, conservation at 4 °C showed to be more appropriate, as hyperhydricity was observed in post-conservation of shoots coming from storage at 8 °C. As for cryopreservation, a daily gradual increase of sucrose concentration (from 0.25 to 1.0 M) produced better protection to the samples that were stored in liquid nitrogen. Indeed, with this sucrose treatment method, a 30-min PVS2 incubation time was enough to produce, 60 days after thawing, the best recovery (47% and 50%) of shoot tips, cryopreserved with PVS2 vitrification and droplet-vitrification, respectively.

In vitro plant regeneration from leaf expiants of Lycopersicon esculentum (tomato)

1976 ◽  
Vol 54 (21) ◽  
pp. 2409-2414 ◽  
Author(s):  
R. M. Behki ◽  
S. M. Lesley
Keyword(s):  
Plant Regeneration ◽  
Lycopersicon Esculentum ◽  
Growth Regulators ◽  
In Vitro Plant Regeneration ◽  
Leaf Discs ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Napthaleneacetic Acid ◽  
In Vitro Plant

Leaf discs from 15 mutant clones of tomato were tested for their morphogenetic response in Murashige and Skoog medium supplemented with 12 combinations of the growth regulators napthaleneacetic acid (NAA) and benzylaminopurine (BA) and 4 combinations of NAA and zeatin. The results show that either callus, shoots, roots, or shoots and roots can be produced depending upon the hormone concentrations and ratios. Plants were regenerated from 12 of the 15 varieties tested.

scholarly journals Effects of Cytokinin on in vitro Propagation of Bauhinia variegata L.

2020 ◽  
Vol 1 (6) ◽  
Author(s):  
Belai Meeta Suwal Singh
Keyword(s):  
In Vitro Propagation ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Nodal Cuttings ◽  
Half Strength ◽  
Bauhinia Variegata ◽  
Mature Seeds

Mature seeds of Bauhinia variegata L were cultured on half strength Murashige and Skoog medium. For experimentation, nodal cuttings were used as explants from in vitro growing plants. Cytokinin, N-benzyl-9-(2-tetrahydropyranyl) (BPA), kinetin(6-furfurylaminopurine), zeatin, 6-(4-hydroxy-3-methyl-trans -2-butenyl amino purine), 2- isopentenyl amino purine (2-ip), and benzylaminopurine (BAP) were tested for best propagation. Well grown plants were achieved in medium supplemented with 5 µM BPA and 0.5 µM BAP. The propagated plants were acclimatized very well after transferred to the field.

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