Current Issues in Plant Cryopreservation

Kaczmarczyk, Anja; Funnekotter, Bryn; Menon, Akshay; Phang, Pui Ye; Al-Hanbali, Arwa; Bunn, Eric; Mancera, Ricardo L.

doi:10.5772/32860

Cited by 60 publications

(77 citation statements)

References 102 publications

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“…Encapsulation-dehydration is a protocol developed by Fabre and Dereuddre (1990) that consists of seven (7) main stages: (i) preculture, (ii) formation of beads, (iii) osmoprotection, (iv) dehydration, (v) immersion in LN, (vi) thawing and (vii) culture in growth recovery (Kaczmarczyk et al, 2012). In this study, a simple and efficient method of encapsulation-dehydration has been established to cryopreserve IFEs for long term storage.…”

Section: Discussionmentioning

confidence: 99%

Establishment of encapsulation-dehydration technique for in vitro fragmented explants of Rosa hybrida L. cv. Helmut Schmidt

Mubbarakh¹,

Izhar²,

Rajasegar³

et al. 2014

Emir. J. Food Agric

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This study was carried out to evaluate the encapsulation-dehydration technique on IFEs of Rosa hybrida L. cv. Helmut Schmidt. The survival of IFEs was first assessed based on the effects of four sucrose concentrations (0, 0.25, 0.5 and 0.75 M) at different durations (0, 24 and 48 hours). The IFEs were then encapsulated and osmoprotected on shaker at 110 rpm with various sucrose concentrations (0, 0.1, 0.5 and 0.75 M). Subsequently, encapsulated IFEs were dehydrated under laminar air flow during three different time periods (0, 3 and 6 hours) on oven sterilized 50 g silica gel. The encapsulated IFEs were plunged into liquid nitrogen for a minimum duration of 24 hour. Encapsulated IFEs were thawed at 40ºC for 90 seconds and cultured on fullstrength MS medium supplemented with 3% sucrose for a week in dark, a week in semi-light and an additional 1 week under full light exposure. IFEs were then evaluated for survival by 2,3,5-Triphenyltetrazolium Chloride assay at 490 nm. The best conditions for the encapsulation-dehydration of IFEs of Rosa hybrida L. cv. Helmut Schmidt was obtained when IFEs were precultured in 0.25 M sucrose for 24 hours, osmoprotected in 0.75 M sucrose and dehydrated for 3 hours. Histological analysis showed cryopreserved IFEs with intensely stained nucleus but with severely damaged membranes.

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Section: Discussionmentioning

confidence: 99%

Establishment of encapsulation-dehydration technique for in vitro fragmented explants of Rosa hybrida L. cv. Helmut Schmidt

Mubbarakh¹,

Izhar²,

Rajasegar³

et al. 2014

Emir. J. Food Agric

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“…Penyimpanan secara kriopreservasi lebih menghemat area, tenaga, dan waktu untuk pemeliharaan (Kaczmarczyk et al, 2011). Dilaporkan bahwa aplikasi teknik kriopreservasi pada tanaman kentang dapat Tanaman buah-buahan merupakan salah satu kelompok tanaman yang sangat mendesak untuk disimpan secara kriopreservasi karena sebagian besar memiliki masa juvenil yang panjang dan dikembangbiakkan secara vegetatif.…”

Section: Pentingnya Aplikasi Teknik Kriopreservasi DI Indonesiaunclassified

Perkembangan Aplikasi Teknik Kriopreservasi untuk Konservasi dan Mendukung Program Pemuliaan Tanaman

Roostika

2016

J. AgroBiogen

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Development of the Application of CryopreservationTechniques for Conservation and Supporting Plant Breeding Program. Ika Roostika. In the beginning, cryopreservation technique was primarily used for germplasms long-term storage as passive collection because cell division and metabolism process can be stopped at super low temperature, commonly in liquid nitrogen. The technique is suitable for vegetative propagated plants and recalcitrant seeds. Recently, its application is extending for storing many species and orthodox seeds. In this paper, the development of cryopreservation application is discussed. In Indonesia, cryopreservation is being studied but the application of cryopreservation has been significantly developed abroad. The application of cryopreservation technique is not only for preserving passive collections but also for storing active collections, including to provide plant materials for hybridization, cellular engineering, genetic transformation, as well as pathogens eradication or cryotherapy. It is concluded that cryopreservation plays an important role in conventional and modern plant breeding program.

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“…The aim of effective cryopreservation is the long-term preservation of plant material, and it is important to know whether this induces permanent genetic changes. Changes in the level of DNA methylation were observed in papaya (Kaity et al 2008), chrysanthemum (Martín and González-Benito 2006;Martin et al 2011) currants (Johnston et al 2009), strawberries (Hao et al 2002a), citrus (Hao et al 2002b) and potatoes (Kaczmarczyk et al 2012). Changes in global DNA methylation may also be induced by cryovitrification agents and the conditions of in vitro culturing (Harding 2004).…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation changes the DNA methylation of embryonic axes of Quercus robur and Fagus sylvatica seeds during in vitro culture

Nuc

Marszałek

Pukacki

2016

Trees

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Key message Dehydration and cryo-stress (2196°C) caused a greater increase in the DNA methylation of embryonic axes (EAs) of beech seeds (tolerant to both stresses) than in EAs of oak (sensitive to both stresses). Simultaneously, in vitro survival of cryopreserved beech EAs was also significantly higher than in those of oak. Abstract DNA methylation plays a key role in the regulation of growth and differentiation in plants. The present study focused on global DNA methylation of embryonic axes (EAs) isolated from the seeds of pedunculate oak (Quercus robur L.) and European beech (Fagus sylvatica L.) after cryopreservation. Oak seeds can be labeled recalcitrant, i.e., sensitive to dehydration, while beech seeds fall under the suborthodox category, i.e., tolerant to dehydration. DNA methylation was determined using a specific antibody directed at the methylated cytosine in CpG dinucleotides. The analysis was carried out on DNA isolated from EA tissues and subjected to (1) dehydration (control), (2) a combination of dehydration and plant vitrification solution (PVS3) and (3) dehydration with vitrification PVS3, and rapid cooling in liquid nitrogen (LN 2 , -196°C) at a cooling rate of 32°C s -1 . Such treatments induced global DNA methylation primarily in tolerant EAs from F. sylvatica seeds. The level of global DNA methylation in this material was 30 % higher than in the control, i.e., dehydrated (not frozen) beech EAs. The same treatment in oak EAs did not evoke such changes. After 120 days of tissue growth, we observed substantially more DNA demethylation in the beech tissues after cryopreservation in LN 2 than in the controls or in tissues subjected to vitrification but not cryopreservation. Information on the global level of DNA methylation can be useful in monitoring procedures aimed at increasing the rate of recovery of EAs following cryopreservation.

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Current Issues in Plant Cryopreservation

Cited by 60 publications

References 102 publications

Establishment of encapsulation-dehydration technique for in vitro fragmented explants of Rosa hybrida L. cv. Helmut Schmidt

Establishment of encapsulation-dehydration technique for in vitro fragmented explants of Rosa hybrida L. cv. Helmut Schmidt

Perkembangan Aplikasi Teknik Kriopreservasi untuk Konservasi dan Mendukung Program Pemuliaan Tanaman

Cryopreservation changes the DNA methylation of embryonic axes of Quercus robur and Fagus sylvatica seeds during in vitro culture

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