Cryopreservation of avocado embryogenic cultures using the droplet-vitrification method

Guzmán-García, E.; Bradaï, Fatiha; Sánchez-Romero, Carolina

doi:10.1007/s11738-012-1062-3

Cited by 12 publications

(19 citation statements)

References 21 publications

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“…Good results were obtained, for example, for somatic embryos of Quercus robur (Chmielarz et al, 2005), Picea sitchensis (Gale et al, 2008), and Alnus glutinosa (San-José et al, 2015) and also for embryogenic tissues of Picea omorika and Persea americana Guzman-Garcia et al, 2013), after application of new cryopreservation techniques, based on vitrification. Using a pregrowth dehydration method, almost a 100% recovery rate was obtained for Picea omorika embryogenic tissue .…”

Section: Cryopreservation Of Embryogenic Culturesmentioning

confidence: 99%

“…Using a pregrowth dehydration method, almost a 100% recovery rate was obtained for Picea omorika embryogenic tissue . For Persea americana, recovery rates from 77.78 to 100% were obtained (Guzman-Garcia et al, 2013) when applying the droplet-vitrification method. These results suggests that new methods may equal or in many cases exceed the results obtained for the traditional controlled-rate cooling method.…”

Section: Cryopreservation Of Embryogenic Culturesmentioning

confidence: 99%

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Tree somatic embryogenesis in science and forestry

Hazubska-Przybył

Bojarczuk

2016

Dendrobiology

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Somatic embryogenesis is the latest, and potentially the most efficient, method for the vegetative micropropagation of plants. Over the past three decades, numerous laboratory studies have investigated somatic embryogenesis of forest trees, yielding positive results for a number of economically important tree species. The first test trials were run and plantations were planted with interior spruce in the 90s by CellFor Inc. (Canada). However, at the beginning of the XXI century, the program to produce spruce and Douglas fir somatic seedlings was stopped for economic reasons. Thus, currently no operational program is ongoing except on a small scale in New Brunswick. In order to integrate somatic embryogenesis technology into operational reforestation programs, the production costs of forest tree somatic seedlings needs to be reduced, and the awareness of foresters and forest landowners that the material obtained through somatic embryogenesis is valuable needs to be increased. This awareness would enable implementation of this technology on a large scale for production and forest management throughout Europe including Poland. In this review, the importance of somatic embryogenesis in scientific research and in global and European forestry is presented. Our main aims are to provide basic information on the challenges in researching somatic embryogenesis of forest trees and to raise interest in this tree propagation technique in both scientists and foresters.

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Section: Cryopreservation Of Embryogenic Culturesmentioning

confidence: 99%

Section: Cryopreservation Of Embryogenic Culturesmentioning

confidence: 99%

Tree somatic embryogenesis in science and forestry

Hazubska-Przybył

Bojarczuk

2016

Dendrobiology

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“…The exact incubation time, however, has to be adjusted for each plant species because chemical toxicity may inhibit meristem growth. The vitrification method used in this study has already proven its efficacy for different species, resulting in high recovery rates [19,5]. Shoot meristems are often chosen as starting explants for cryopreservation because (i) they are highly regenerable and regenerate into a normal plant and (ii) meristematic cells contain relatively small vacuoles (with small amounts of water).…”

Section: Determination Of Pvs2 Toxicity and The Cryopreservation Suitmentioning

confidence: 99%

“…This method combines classical vitrification [29] and the droplet freezing method first described for potato cryopreservation [31]. Droplet vitrification is now successfully used for a wide variety of plant species like yam [8], banana [19], olive [30], pelargonium [4], thyme [9], and avocado [5].…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of Bituminaria bituminosa varieties and hybrids

et al. 2015

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25cryopreservation of Bituminaria species has been reported. We applied the ultra-fast cooling method, 26 using droplet vitrification on aluminum foil strips. First, we investigated the PVS2 toxicity and 27 cryopreservation damage in two genotypes, comparing three PVS2 treatments and two culture media. An 28 incubation of 30 min in PVS2 resulted in regeneration rates after cryopreservation higher than 80%. The 29MS medium was selected for optimal meristem outgrowth, in order to avoid the prominent callus 30formation that was observed in the presence of BAP. These conditions were subsequently used to 31 cryopreserve eight other genotypes. The results were highly variable; 45 days after cryopreservation, 32 survival ranged between 22 and 98% while regeneration ranged between 0 and 96%, depending on the 33 accession. A significant and positive correlation was observed between survival and regeneration. At 90 34 days post culture plantlets could be recovered from cryopreserved explants of all genotypes. This study 35shows that the droplet vitrification method is promising for the cryopreservation of eight of the 10 36 genotypes assayed and the method can thus be applied to develop a cryobank of B. bituminosa. 98Cryopreservation method and solutions 99We applied the ultra-fast cooling method using droplet vitrification on aluminum foil strips, as 122were used as controls. Two repetitions per treatment were performed, each consisting of two Petri dishes. 123All cultures were kept in darkness at 25 ± 1 ºC for a total of 7 days (from the beginning of the 124 experiment). Thereafter, the meristems were transferred to low-light conditions (1500 lx) and after 8 days 145The data were subjected to factorial analysis of variance (ANOVA). The percentage data were 146 subjected to an x 2 transformation prior to analysis. For means separation, the Duncan multiple-range test 147 was used. 149 Results and Discussion 164In Fig 173incubation lengthened (Fig. 1C and D). For cryopreserved meristems, the optimal time of incubation in 174PVS2 was 30 min, for both accessions and both media ( Fig. 1C and D). We postulate that, after a too- 180In conclusion, an incubation of 30 min in PVS2 resulted in more than 80% regeneration in both 181 genotypes after cryopreservation. Such percentages are promising for the establishment of a cryobank. 182The difference obtained between survival and regeneration shows the importance of observing 183 regeneration instead of survival; otherwise, the wrong conclusions are drawn. 184The growth aspects of the treated meristems are visualized in Fig. 2. A high amount of callus 185was already visible at 45 days of culture and the most prominent callus formation was observed for 186 8 explants cultured on MS BA, compared to MS (Fig. 2 A-B 201The ANOVA analyses of the survival and regeneration revealed highly significant effects of 202 genotype, treatment, and the interaction genotype × treatment ( after cryopreservation, the average regeneration rate over all the accessions at 45 dpc was 25%. 216Sign...

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“…This smaller increase is probably due to the fact that the regeneration of tips, after immersion in LN, had been delayed about seven days following that of the explants of the control treatments (-LN). Guzmán-García et al (2013) found that the cryoprotectant solutions caused some delay in reactivating the growth of embryogenic cultures of Persea americana Mill. after cryopreservation by the droplet method.…”

Section: Sobrevivência De áPices Caulinares De Cana-de-açúcar Após Crmentioning

confidence: 99%

Survival of sugarcane shoot tips after cryopreservation by droplet-vitrification

Nogueira

Pasqual

Scherwinski‐Pereira

2013

Pesq. agropec. bras.

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-The objective of this work was to evaluate the phytotoxicity of a plant vitrification solution (PVS2), and the survival of shoot tips of the sugarcane variety SP716949, after cryopreservation by droplet-vitrification. Shoot tips were precultured for 24 hours in MS medium containing 0.3 mol L -1 sucrose, and exposed to PVS2 for 0, 20 or 30 min. Shoot tips were then immersed in liquid nitrogen. Thawing was fast in concentrated sucrose solution (1.2 mol L -1 ). PVS2 is a nontoxic to shoot tips, which in turn are sensitive to liquid nitrogen. The best results occurred when shoot tips were maintained for up to 20 min in PVS2 solution, before freezing, with 20% survival.Index terms: Saccharum, in vitro conservation, PVS2, regeneration. Sobrevivência de ápices caulinares de cana-de-açúcar após criopreservação por "droplet-vitrification"Resumo -O objetivo deste trabalho foi avaliar a fitotoxicidade da solução de vitrificação de plantas (PVS2) e a sobrevivência de ápices caulinares de cana-de-açúcar, variedade SP716949, após criopreservação por "droplet-vitrification". Ápices caulinares foram pré-cultivados por 24 horas em meio MS com sacarose a 0,3 mol L -1 e expostos por 0, 20 ou 30 min a PVS2. Em seguida, os ápices foram mergulhados em nitrogênio líquido. O descongelamento em solução concentrada de sacarose (1,2 mol L -1 ) foi rápido. A PVS2 não tem efeito tóxico aos ápices que, no entanto, são sensíveis ao nitrogênio líquido. O melhor resultado ocorreu, quando os ápices foram mantidos por 20 min na solução PVS2, antes do congelamento, com 20% de sobrevivência.Termos para indexação: Saccharum, conservação in vitro, PVS2, regeneração.Preservation in liquid nitrogen at ultralow temperature (-196°C) is a potential technique for long-term conservation of plant germplasm. Recently, a new technique called droplet-vitrification cryopreservation has been developed, which combines the application of highly concentrated vitrification solutions with an ultrafast freezing and thawing rate (Panis et al., 2005). In this technique the duration for which explants are treated with cryoprotectant solutions (PVS2) is of extreme importance, since it determines the extent of cell dehydration and the number of components that will permeate the cells (Chen et al., 2011).According to Panis et al. (2011), depending on the target species, the development of a technique involving proper protocol may take years of study, since it depends on previous information available in the literature and on behavioral characteristics of the species, as for example, if it is tropical or temperate, woody or herbaceous, mono-or dicotyledonous.Sugarcane is a species of great economic importance. It propagates vegetatively and needs germplasm banks to meet breeding programs; however, few reports on the cryopreservation of this species using shoot tips and dropled vitrification technique are found in the literature. Barraco et al. (2011a) observed that the apices of two varieties have shown a relatively high tolerance to dehydration by the vitrificati...

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Cryopreservation of avocado embryogenic cultures using the droplet-vitrification method

Cited by 12 publications

References 21 publications

Tree somatic embryogenesis in science and forestry

Tree somatic embryogenesis in science and forestry

Cryopreservation of Bituminaria bituminosa varieties and hybrids

Survival of sugarcane shoot tips after cryopreservation by droplet-vitrification

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