Cryopreservation of shoot tips from in vitro plants of sweet potato [ Ipomoea batatas (L.) Lam.] by vitrification

Pennycooke, Joyce C.; Towill, Leigh E.

doi:10.1007/s002999900171

Cited by 73 publications

(63 citation statements)

References 12 publications

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“…Recently developed methods, such as DMSO droplet method, vitrification, encapsulation/dehydration, encapsulation/vitrification and droplet vitrification apply rapid cooling by plunging precultured and cryoprotected shoot tips directly in LN. The fastest cooling rate (24,000°C min −1 ; Pennycooke and Towill 2000) is implemented by using partially solidified nitrogen (LN slush, approximately −208°C (Pennycooke and Towill 2000)), which was also applied in potato by Kim et al (2006). The cooling rate of 360°C min −1 is reached using PVS2 solution within cryovials (Towill and Bonnart 2003) and can be further increased by using aluminium foil strips (7,800°C min −1 , Towill and Bonnart 2003).…”

Section: Coolingmentioning

confidence: 99%

Potato Shoot Tip Cryopreservation. A Review

2010

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Potato is one of the most important crops worldwide. Genetic resources of potato (Solanum tuberosum L. ssp. tuberosum) and related cultivated species are conserved through storage of tubers, in vitro plants and in cryopreservation. Cryopreservation, storage in or above liquid nitrogen, is the best option to maintain vegetatively propagated plants in the long term. The present review gives comprehensive information about various cryopreservation techniques for potato published from 1977 until the present. It discusses factors that affect the process and success of cryopreservation, such as donor culture conditions, preculture, cooling, warming and post-culture treatments. Studies are presented that analyse the histological and ultrastructural changes after different cryopreservation steps and the morphological pathways during regeneration of plants after rewarming. The maintenance of genetic stability in potato after cryopreservation has also been demonstrated by various phenotypic and molecular methods. The first thermal analyses on potato shoot tips are presented using differential scanning calorimetry to analyse the state of water during cooling and warming. Biochemical analyses of different compounds, such as soluble sugars and proteins, have been performed to understand and improve existing cryogenic methods. Potato is an example where successful virus elimination has been obtained via cryopreservation of shoot tips (cryotherapy). There are already cryopreserved collections of potato shoot tips in

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Section: Coolingmentioning

confidence: 99%

Potato Shoot Tip Cryopreservation. A Review

2010

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“…Notable changes include alterations in membrane composition (Steponkus, 1984) and accumulation of compatible solutes such as soluble sugars, Pro, and Gly betaine (Guy, 1990). Sugars have been shown to be effective cryoprotectants in vitro (Carpenter and Crowe, 1988;Pennycooke and Towill, 2000), and there is evidence indicating a role in freezing tolerance in cold-acclimated membranes (Sanitarius, 1973), cells (Sanitarius and Milde, 1977), and plants (Olien and Clark, 1993;Taji et al, 2002;Strand et al, 2003).Raffinose family oligosaccharides (RFOs), particularly raffinose, play a role in the acquisition of cold tolerance in many plant species, including herbaceous (Bachmann et al woody plant species (Stushnoff et al, 1993). Concentration of RFOs in Ajuga reptans has been shown to be the lowest in summer and highest in fall and winter (Bachmann et al, 1994).…”

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confidence: 99%

Down-Regulating α-Galactosidase Enhances Freezing Tolerance in Transgenic Petunia

Pennycooke

Jones²,

Stushnoff³

2003

Plant Physiology

157

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␣-Galactosidase (␣-Gal; EC 3.2.1.22) is involved in many aspects of plant metabolism, including hydrolysis of the ␣-1,6 linkage of raffinose oligosaccharides during deacclimation. To examine the relationship between endogenous sugars and freezing stress, the expression of ␣-Gal was modified in transgenic petunia (Petunia ϫ hybrida cv Mitchell). The tomato (Lycopersicon esculentum) Lea-Gal gene under the control of the Figwort Mosaic Virus promoter was introduced into petunia in the sense and antisense orientations using Agrobacterium tumefaciens-mediated transformation. RNA gel blots confirmed that ␣-Gal transcripts were reduced in antisense lines compared with wild type, whereas sense plants had increased accumulation of ␣-Gal mRNAs. ␣-Gal activity followed a similar trend, with reduced activity in antisense lines and increased activity in all sense lines evaluated. Raffinose content of nonacclimated antisense plants increased 12-to 22-fold compared with wild type, and 22-to 53-fold after cold acclimation. Based upon electrolyte leakage tests, freezing tolerance of the antisense lines increased from Ϫ4°C for cold-acclimated wild-type plants to Ϫ8°C for the most tolerant antisense line. Down-regulating ␣-Gal in petunia results in an increase in freezing tolerance at the whole-plant level in nonacclimated and cold-acclimated plants, whereas overexpression of the ␣-Gal gene caused a decrease in endogenous raffinose and impaired freezing tolerance. These results suggest that engineering raffinose metabolism by transformation with ␣-Gal provides an additional method for improving the freezing tolerance of plants.Many plants increase in low temperature and/or freezing tolerance in response to low nonfreezing temperatures, a phenomenon known as cold acclimation. For decades, the study of low temperature stress has had a primary goal of cataloging and understanding the biochemical and physiological changes occurring during cold acclimation (Levitt, 1980;Guy, 1990;Thomashow, 2001). It is known that the disruption of cellular membranes, particularly the plasma membrane, is the primary site of injury during a freeze-thaw cycle and that this injury usually results from dehydration associated with freezing (Steponkus, 1984). Other consequences of freezeinduced cellular dehydration include the generation of reactive oxygen species that are damaging to other cellular components (McKersie, 1991). As such, plants have developed mechanisms to deal with these multiple stresses.It has long been established that changes in gene expression occur upon exposure to cold acclimation (Guy et al., 1985). In the last decade, extensive research to identify and characterize cold-responsive (COR) genes has been undertaken. Hajela et al. (1990) isolated the COR genes from Arabidopsis that encode polypeptides thought to have protective roles against dehydration. Expression profile experiments in Arabidopsis demonstrated that extensive changes in gene expression occur during cold acclimation and that a substantial number of the genes that are up...

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“…Matsumoto et al (1994) reported that the absence of preculture in 0.3 M sucrose results in a reduced recovery rate in wasabi, which is similar to the results reported for cassava (Charoensub et al, 1999). Even in sweet potato, there was no survival observed without preculture (Pennycooke and Towill, 2000). Sucrose preculture provides some protection against dehydration from PVS2 exposure, even when no cryoprotectant preculture is used (Pennycooke and Towill, 2001 …”

Section: Resultsmentioning

confidence: 99%

“…According to Pennycooke and Towill (2001) (Benson, 2008;Pennycooke and Towill, 2000;Towill and Jarret, 1992). Direct exposure without osmoprotectant preculture did not yield viable shoot tips in sweet potato (Towill and Jarret, 1992).…”

Section: Effects Of Preculture With Osmoprotectants On Recoverymentioning

confidence: 99%

Cryopreservation of in Vitro Grown Shoot Tips of Sweet Potato (Ipomoea batatas L.) by the Encapsulation-Vitrification Method

Yi¹,

Lee²,

Lee³

et al. 2016

Korean Journal of Plant Resources

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-Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulationvitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at 25°C. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

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Cryopreservation of shoot tips from in vitro plants of sweet potato [ Ipomoea batatas (L.) Lam.] by vitrification

Cited by 73 publications

References 12 publications

Potato Shoot Tip Cryopreservation. A Review

Potato Shoot Tip Cryopreservation. A Review

Down-Regulating α-Galactosidase Enhances Freezing Tolerance in Transgenic Petunia

Cryopreservation of in Vitro Grown Shoot Tips of Sweet Potato (Ipomoea batatas L.) by the Encapsulation-Vitrification Method

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