Application of cryoprotectants to improve low temperature storage survival of orchid seeds

Huehne, Pattana Srifah; Bhinija, Kisana

doi:10.1016/j.scienta.2011.11.026

Cited by 14 publications

(8 citation statements)

References 20 publications

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“…Similarly, the germination of seeds of Phaius tankervilleae was maintained at ∼70% during storage in liquid nitrogen for at least 12 mo when seeds were first precultured on a sucrose solution and then treated with PVS2; in the absence of PVS2, no survival was recorded (Hirano et al 2009). For seeds of two species of Dendrobium and the hybrid Vanda Miss Joaquim, which do not survive storage at Ϫ70ЊC, the cryoprotectants mannitol and meso-inositol enhanced survival; after 5 yr of storage, up to ∼30% germination was achieved for Dendrobium crystallinum seeds (Huehne and Bhinija 2011). These compounds, along with glucose, were also found to be effective at enhancing seed survival for a number of other Dendrobium spp.…”

Section: Vitrificationmentioning

confidence: 97%

“…Seed viability was maintained for a number of orchid species-including Dactylorhiza spp., D. anosmum, E. gonychila, and Paphiopedilum rothschildianum-during storage for up to 12 mo at Ϫ70Њ and Ϫ196ЊC (Pritchard et al 1999). In a study of 12 epiphytic and terrestrial orchid species from Thailand and Singapore, after 5 yr of storage at Ϫ70Њ, Ϫ20Њ, and 4ЊC, germination was found to be greatest following storage at Ϫ70ЊC (Huehne and Bhinija 2011). Storage at Ϫ80ЊC was also suitable for maintaining the viability of a number of Australian terrestrial species over 2 yr of storage .…”

Section: Low-temperature Storage and Cryopreservation Of Orchid Seedsmentioning

confidence: 99%

“…This is achieved by the addition of cryoprotective solutions, such as glycerol, dimethylsulfoxide, glycols, and plant vitrification solution 2 (PVS2), a mixture of 30% (w/v) glycerol, 15% (w/v) ethylene glycol, and 15% (w/v) dimeth-54 INTERNATIONAL JOURNAL OF PLANT SCIENCES ylsulfoxide (Sakai and Engelmann 2007). Vitrification techniques-developed principally to allow for the cryopreservation of desiccation-sensitive plant material, such as in vitro tissue culture-derived shoot tips, somatic embryos, and cell cultures-are becoming increasingly common for the cryostorage of orchid seeds and protocorms (Ishikawa et al 1997;Hirano et al 2005Hirano et al , 2011Nikishina et al 2007;Vendrame et al 2007;Huehne and Bhinija 2011;Galdiano et al 2012;Watanawikkit et al 2012). Vitrification appears to enhance survivability of orchid seeds.…”

Section: Vitrificationmentioning

confidence: 99%

“…These compounds, along with glucose, were also found to be effective at enhancing seed survival for a number of other Dendrobium spp. as well as Grammatophyllum speciosum and Pholidota articulata over a 6-mo storage period at Ϫ70ЊC (Huehne and Bhinija 2011). Vitrification techniques have also been used to good effect by allowing for the cryostorage of immature seeds of Bletilla striata, providing readily germinable seeds on retrieval from storage by avoiding the dormancy present once seeds mature (Hirano et al 2005).…”

Section: Vitrificationmentioning

confidence: 99%

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Ex situ Conservation and Cryopreservation of Orchid Germplasm

Merritt

Hay

Swarts

et al. 2014

International Journal of Plant Sciences

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Premise of research. Orchids are among the most enigmatic of plant species. Yet the Orchidaceae comprises more species at risk of extinction than any other plant family. The collection and storage of orchid germplasmprincipally seeds and associated mycorrhizal fungi but also protocorm-like bodies using encapsulation and vitrification techniques-allows for secure ex situ conservation. This article reviews the approaches and techniques used for the ex situ conservation of orchid germplasm, with a focus on seed banking and the use of cryopreservation techniques to improve the longevity of germplasm.Pivotal results. It is increasingly apparent that cryopreservation-the storage of germplasm at ultra-low temperatures (e.g., in liquid nitrogen)-is required for the long-term and low-maintenance conservation of all types of orchid germplasm. For orchid seeds, desiccation tolerance is common, but longevity in storage is poor. Cryopreservation of orchid seeds shows promise, but some complexities in low-temperature storage behavior still require explanation and resolution. The application of more advanced cryopreservation techniques, including encapsulation-dehydration and vitrification, is becoming increasingly common. These techniques provide for the simultaneous storage of orchid propagules with their compatible fungus, while for seeds, vitrification techniques show potential for improving tolerance to the stresses of cryopreservation. Conclusions.A renewed focus on describing the low-temperature storage physiology of orchid seeds to more precisely define the relationship between seed water content, storage temperature, and seed survival is required, as is perhaps the wider adoption of the use of cryoprotectants for seeds. This research, coupled with the development of improved methods of seed viability testing, will support the growing work of germplasm banks to protect orchid biodiversity in the face of habitat loss and potential species extinction.

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Section: Vitrificationmentioning

confidence: 97%

Section: Low-temperature Storage and Cryopreservation Of Orchid Seedsmentioning

confidence: 99%

Section: Vitrificationmentioning

confidence: 99%

Section: Vitrificationmentioning

confidence: 99%

See 2 more Smart Citations

Ex situ Conservation and Cryopreservation of Orchid Germplasm

Merritt

Hay

Swarts

et al. 2014

International Journal of Plant Sciences

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“…For the long-term storage of seeds, the survival after thawing depends on the quality, physiological status, and water content of seeds (HUEHNE and BHINIJA, 2012). The water content is a meaningful factor for the successful cryopreservation.…”

Section: Discussionmentioning

confidence: 99%

Germinação in vitro e criopreservação de sementes de Zinnia elegans

Prudente

Nery

Paiva

et al. 2015

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Zinnia elegans Jacquim is a species of big cutting and ornamental potential, but its seeds have low germination percentage. This study aimed to establish a protocol for in vitro germination and water content for the cryopreservation of seeds. The length, width, thickness, and weight of one thousand seeds were determined. MS and WPM culture mediums, as well as the concentrations of gibberellic acid (GA 3 ) were tested regarding in vitro culture. For cryopreservation, the seeds were subjected to drying on silica gel or laminar flow for different times. Then, the seeds were stored in liquid nitrogen (-196° C) for 24 hours. After this period, the seeds were thawed and inoculated into culture medium. The biometric results of seeds showed 8.6 mm average length, 4 mm width, and 0.9 mm thickness. The weight of one thousand seeds was 851 mg, characterizing them as small, lightweight, and easy to disperse. The use of MS medium with no addition of GA 3 enhanced germination (67%). The initial moisture content of Z. elegans seeds was 9%. Seeds subjected up to 2 hours of drying in both treatments obtained 23% germination in silica gel and 19% in laminar flow. Z. elegans seeds may be desiccated by 4% moisture content and cryopreserved with no loss of germination potential. Keywords: In vitro conservation, ornamental plant, tissue culture. RESUMO Germinação in vitro e criopreservação de sementes de Zinnia elegansZinnia elegans Jacquim é uma espécie de grande potencial para flor de corte e ornamentação, porém suas sementes apresentam baixa porcentagem de germinação. Objetivou-se estabelecer um protocolo para germinação in vitro e o teor de água para a criopreservação das sementes. O comprimento, largura e espessura das sementes e o peso de mil sementes foi determinado. Quanto ao cultivo in vitro testou-se os meios de cultura MS e WPM e cinco concentrações de ácido giberélico (GA 3 ). Para a criopreservação, as sementes foram submetidas à secagem em sílica gel ou fluxo laminar por diferentes tempos. Em seguida, as sementes foram armazenadas em nitrogênio líquido (-196º C) por 24 horas, após esse período, as sementes foram descongeladas e inoculadas em meio de cultura. Os resultados de biometria das sementes apresentaram comprimento médio (mm) de 8,6; largura de 4,0 e espessura de 0,9. O peso de mil sementes foi de 851 mg, caracterizando-as como pequenas, leves e de fácil dispersão. A utilização do meio de cultura MS sem adição de GA 3 favoreceu a germinação in vitro (67%). O teor de água inicial das sementes de Z. elegans foi de 9%. As sementes submetidas até 2 horas de secagem em ambos os tratamentos obtiveram germinação de 23% na sílica gel e 19% no fluxo laminar. Sementes de Z. elegans podem ser dessecadas até graus de umidade em torno de 4%, e criopreservadas sem perda do poder germinativo. Palavras-chave: Conservação in vitro, planta ornamental, cultura de tecidos.(1)

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In vitro conservation of Dendrobium germplasm

et al. 2014

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Dendrobium is a large genus in the family Orchidaceae that exhibits vast diversity in floral characteristics, which is of considerable importance to orchid breeders, biotechnologists and collectors. Native species have high value as a result of their medicinal properties, while their hybrids are important as ornamental commodities, either as cut flowers or potted plants and are thus veritable industrial crops. Thus, preservation of Dendrobium germplasm is valuable for species conservation, breeding programs and the floriculture industry. Cryopreservation represents the only safe, efficient and cost-effective long-term storage option to facilitate the conservation of genetic resources of plant species. This review highlights 16 years of literature related to the preservation of Dendrobium germplasm and comprises the most comprehensive assessment of thorough studies performed to date, which shows reliable and reproducible results. Air-drying, encapsulation-dehydration, encapsulation-vitrification, vitrification and droplet-vitrification are the current cryopreservation methodologies that have been used to cryopreserve Dendrobium germplasm. Mature seeds, pollen, protoplasts, shoot primordia, protocorms and somatic embryos or protocorm-like bodies (PLBs) have been cryopreserved with different levels of success. Encapsulation-vitrification and encapsulation-dehydration are the most used protocol, while PLBs represent the main explant explored.

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Application of cryoprotectants to improve low temperature storage survival of orchid seeds

Cited by 14 publications

References 20 publications

Ex situ Conservation and Cryopreservation of Orchid Germplasm

Ex situ Conservation and Cryopreservation of Orchid Germplasm

Germinação in vitro e criopreservação de sementes de Zinnia elegans

In vitro conservation of Dendrobium germplasm

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