Plant vitrification solution 2 lowers water content and alters freezing behavior in shoot tips during cryoprotection

Volk, Gayle M.; Walters, Christina

doi:10.1016/j.cryobiol.2005.09.004

Cited by 134 publications

(101 citation statements)

References 37 publications

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“…Prolonged PVS2 exposure (40 min) led to excessive dehydration, resulting in cell death. Therefore, balancing the water content is crucial to minimize desiccation damage and freezing injury (Volk and Walters, 2006). Regrowth of Dendrobium nobile improved to 80% following thawing at 38 ± 2 °C for 2 min (Mohanty et al, 2012), while regrowth of Anemarrhena asphodeloides Bunge was observed at 60% after thawing was conducted at 35 °C for 5 min (Hong and Yin, 2012).…”

Section: Discussionmentioning

confidence: 99%

An improved PVS2 cryopreservation technique for Ascocenda Wangsa Gold orchid using protocorm-like bodies

Rajasegar¹,

Mansor²,

Poobathy³

et al. 2015

Turk J Biol

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Ascocenda Wangsa Gold is a new and fascinating orchid hybrid in the Malaysian flower industry. An efficient plant vitrification solution 2 (PVS2) cryopreservation technique was developed for protocorm-like bodies (PLBs) of Ascocenda Wangsa Gold orchid. Parameters assessed included the effect of PVS2 exposure periods, thawing duration, temperature, and culture conditions based on 2,3,5-triphenyltetrazolium chloride absorbance readings and regrowth rates. A regrowth rate of 33.3% was obtained after 2 months when the PLBs were dehydrated in PVS2 for 30 min. The growth rate was improved to 47% when thawing was conducted at 45 °C for 85 s. The highest growth rate (53.3%) was obtained when the PLBs were subjected to a 7-day dark treatment before being transferred to a 16-h/8-h light/dark photoperiod. Histological analyses were conducted to study the morphology of cryopreserved and noncryopreserved PLBs of Ascocenda Wangsa Gold. The vitrification protocol developed in this study is a feasible and safe method for strengthening the germplasm conservation of this orchid for commercial purposes.

show abstract

Section: Discussionmentioning

confidence: 99%

An improved PVS2 cryopreservation technique for Ascocenda Wangsa Gold orchid using protocorm-like bodies

Rajasegar¹,

Mansor²,

Poobathy³

et al. 2015

Turk J Biol

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“…The lack of shoot regeneration after cryopreservation without preculture in sucrose shows that the PVS2 treatment alone was not able to protect shoot tips from the lethal effect of freezing in liquid nitrogen. Plant vitrification solutions induce a certain level of dehydration in cells, but the extent of dehydration is not known (Volk and Walters, 2006). When applying cryopreservation, it is important that the stored material is capable of producing plants that are true-to-type to the nontreated genotype.…”

Section: Discussionmentioning

confidence: 99%

Genetic integrity assessment of cryopreserved tomato(Lycopersicon esculentum Mill.) genotypes

Coste¹,

Şuteu²,

Băcilă³

et al. 2015

Turk J Biol

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Shoot tips of five tomato (Lycopersicon esculentum Mill.) genotypes were successfully cryopreserved by droplet vitrification. Recovered plants were studied for genetic stability by two different assays: amplified fragment length polymorphism (AFLP) using 4 primer combinations and sequencing of lycopene β-cyclase gene (LCY-B) from leaves. The highest shoot regeneration after cryopreservation was 70% (Pontica) following 24 h osmoprotection in 0.5 M sucrose and 30 min dehydration in plant vitrification solution 2 (PVS2). The data inferred from AFLP showed no genetic dissimilarities between in vitro regenerants derived from cryopreserved tissues compared with the noncryopreserved controls. Although a single nucleotide polymorphism, a G→T transversion, was identified in position 1139 in Capriciu and Darsirius, this was not due to the cryopreservation process itself, since it was detected in both cryopreserved and control samples. Thus, sequencing of LCY-B gene from leaves revealed no DNA damages after cryopreservation and subsequent in vitro regeneration. Our results indicate that cryostorage by droplet vitrification is an efficient and reliable technique to preserve the selected tomato genotypes and to regenerate true-to-type plants.

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Section: Vitrificationmentioning

confidence: 99%

“…Volk and Walters (2006) demonstrated that the extent of penetration of PVS2 into mint and garlic shoot tips was directly proportional to exposure time. The water content of the shoot tips also significantly decreased with an increase in exposure time to PVS2 (Volk & Walters, 2006). Greater penetration of CPAs can be useful as it helps to increase the internal solute concentration and may contribute to maintaining cell volume, thus preventing damage to the cells (Meryman, 1974).…”

Section: Vitrificationmentioning

confidence: 99%

Current Issues in Plant Cryopreservation

Kaczmarczyk¹,

Funnekotter²,

Menon³

et al. 2012

Current Frontiers in Cryobiology

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Plant vitrification solution 2 lowers water content and alters freezing behavior in shoot tips during cryoprotection

Cited by 134 publications

References 37 publications

An improved PVS2 cryopreservation technique for Ascocenda Wangsa Gold orchid using protocorm-like bodies

An improved PVS2 cryopreservation technique for Ascocenda Wangsa Gold orchid using protocorm-like bodies

Genetic integrity assessment of cryopreserved tomato(Lycopersicon esculentum Mill.) genotypes

Current Issues in Plant Cryopreservation

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