Cryopreservation of cocoa (Theobroma cacao L.) somatic embryos for long-term germplasm storage

Fang, Jong-Yi; Wetten, A.; Hadley, P.

doi:10.1016/j.plantsci.2003.11.002

Cited by 60 publications

(36 citation statements)

References 17 publications

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“…These risks, together with the recalcitrant nature of cocoa seed with regard to storage, makes the establishment of a cryopreserved collection of key cocoa germplasm a logical safeguard. To this end over 600 accessions of cocoa are being cryopreserved at Reading University through the encapsulation-dehydration of floral-derived somatic embryos (SEs) (Fang et al 2004). This procedure involves the rapid cooling of the prolific secondary SEs obtained from cultured cotyledonary explants of primary SEs.…”

Section: Introductionmentioning

confidence: 99%

Use of secondary somatic embryos promotes genetic fidelity in cryopreservation of cocoa (Theobroma cacao L.)

Fang¹,

Wetten

Adu‐Gyamfi³

et al. 2008

AFSci

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The inability to conserve cocoa (Theobroma cacao L.) germplasm via seed storage and the vulnerability of field collections make the establishment of cryopreserved genebanks for the crop a priority. An effective encapsulation-dehydration based cryopreservation system has been developed for cocoa but because the somatic embryos used for freezing arise after a protracted period of callus culture there is concern about maintenance of genetic fidelity during the process. Microsatellite markers for seven of the 10 cocoa linkage groups were used to screen a population of 189 primary somatic embryo-derived emblings and the 43 secondary somatic embryos they gave rise to. Of the primary somatic embryos, 38.1% exhibited polymorphic microsatellite profiles while for secondary somatic embryos the frequency was 23.3%. The same microsatellite markers used to screen another population of 44 secondary somatic embryos cryopreserved through encapsulation-dehydration revealed no polymorphisms. Scanning electron microscopy showed the secondary somatic embryos were derived from cotyledonary epidermal cells rather than callus. The influence of embryo ontogeny on somaclonal variation is discussed.

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Section: Introductionmentioning

confidence: 99%

Use of secondary somatic embryos promotes genetic fidelity in cryopreservation of cocoa (Theobroma cacao L.)

Fang¹,

Wetten

Adu‐Gyamfi³

et al. 2008

AFSci

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“…During cultivations control explants developed axillary buds on bulb scales (Table 1). Literature reports on preservation of somatic embryos in liquid nitrogen are scarce, but indicate that improved tolerance to freezing and dehydration is achieved by using exogenous ABA and elevated sugar contents (Fang et al 2004).…”

Section: Resultsmentioning

confidence: 99%

Employment of encapsulation-dehydration method for liquid nitrogen cryopreservation of ornamental plant explants propagated in vitro

Pawłowska

2008

Folia Horticulturae

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In the present studies, an attempt has been made to develop a method of liquid nitrogen preservation of plant explants propagated in vitro in the laboratory of the Department of Ornamental Plants, of Agricultural University in Kraków: shoot apical and axillary meristems of Rosa 'New Dawn', somatic embryos of snowdrops Galanthus nivalis L. and G. elwesii Hook, and gametophyte of Phlebodium aureum (L.) J. Sm. (golden polypody). After encapsulation of plant material, it was dehydrated by quick method (capsules were placed in liquid media containing 0.75 M sucrose for 18 h) or by gradual method (capsules were transferred to liquid solutions of media with increasing sucrose concentrations from 0.3 M to 1 M for consecutive 7 days). Moreover, some explants for cryopreservation were treated with the medium containing elevated sucrose level (0.25 M) for 8 weeks.

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“…The cryopreservation procedure of Fang et al (2004), based on optimal somatic embryo survival of cocoa (plating encapsulated somatic embryos on ED medium enriched with 0.75M sucrose for seven days and dehydration on 30 g of silica gel for 4 h) was used. Somatic embryos (4-5 mm) were isolated from callus tissues and suspended for 30 min.…”

Section: Methodsmentioning

confidence: 99%

“…Florin et al (2002) also designed a cryopreservation protocol for embryogenic callus of cocoa which were prone to somaclonal variation (Hao and Deng, 2002). Fang et al (2004) established a cocoa-specific cryopreservation protocol for a few hours in which embryo survival was monitored under a series of encapsulation, pre-culture and dehydration regimes.…”

Section: Introductionmentioning

confidence: 99%

Effect of liquid nitrogen storage time on the survival and regeneration of somatic embryos of cocoa

Quainoo¹

2010

African Crop Science Journal

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Investigations were undertaken on the effect of liquid nitrogen (LN) storage time on survival and regeneration of somatic embryos of cocoa (Theobroma cacao l.). Somatic embryos from different cocoa genotypes (AMAZ 3-2, AMAZ 10-1, AMAZ 12, SIAL 93, and IMC 14) at 15.45% moisture content were cryopreserved in LN for one h, four and eight weeks. Somatic embryos of the genotypes emerged from the alginate beads at different periods 4 to 12 weeks post-cryopreservation. Individual genotypes subjected to low temperature storage time did not show significant differences in post-cryopreservation survival, although different genotypes responded differently with AMAZ 12 and IMC 14 recording the highest and lowest mean survival rates of 58% and 35%, respectively. Plantlets originating from five genotypes had been weaned and developed normally comparable to non-cryopreserved somatic embryo-derived plantlets in the glasshouse. The effects of low temperature storage time on survival and genetic stability of somatic embryos using genotypes from the same parental and distinct lines may contribute to long-term storage of cocoa germplasm.Key Words: Cryopreservation, genotypes, Theobroma cacao RÉSUMÉUne enquêtes relative à l'effet du temps de stockage de l'azote liquide (LN) sur la survie et la régénération des embryons somatiques de cacao (Theobroma cacao l.) avait été menées. Les embryons somatiques de différents génotypes du cacao (AMAZ 3-2, AMAZ 10-1, AMAZ 12, SIAL 93 et IMC 14) d'une tenneur en humidité de 15,45 % avaient été preservés dans LN pour une heure, quatre semaines et huit semaines. Embryons somatiques des génotypes émergés de perles d'alginate à différentes périodes de 4 à 12 semaines en post cryoconservation. Les génotypes individuels soumis à la basse température au temps de stockage n'ont pas démontré des différences significatives après la cryoconservation, bien que différents génotypes avaient réagi différemment. Les génotypes AMAZ 12 et IMC 14 avaient enregistré des taux moyens de survie de l'embryon somatique, respectivement 58 % et 35 %. Les plantules issue de cinq génotypes avaient été sevrées et développées dans une serre par voie normale en comparativement à l'embryon somatique des plantules non cryoconservées. Les effets du temps de conservation à basse température sur la survie et la stabilité génétique des embryons somatiques en utilisant de génotypes issue de meme lignées parentales et de lignée parentales distinctes peuvent contribuer au stockage à long terme de matériel génétique du cacao.

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Cryopreservation of cocoa (Theobroma cacao L.) somatic embryos for long-term germplasm storage

Cited by 60 publications

References 17 publications

Use of secondary somatic embryos promotes genetic fidelity in cryopreservation of cocoa (Theobroma cacao L.)

Use of secondary somatic embryos promotes genetic fidelity in cryopreservation of cocoa (Theobroma cacao L.)

Employment of encapsulation-dehydration method for liquid nitrogen cryopreservation of ornamental plant explants propagated in vitro

Effect of liquid nitrogen storage time on the survival and regeneration of somatic embryos of cocoa

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