PRIMARY AND SECONDARY SOMATIC EMBRYOGENESIS OF CACAO: THE EFFECT OF EXPLANT TYPES AND PLANT GROWTH REGULATORS

<p class="abstrakinggris">The success of cacao somatic embryogenesis is affected by many factors, including the basal salt medium, the genotype, the explant type, and the concentration and composition of plant growth regulators (PGRs). The study aimed to evaluate the effects of PGRs composition on the primary somatic embryo (PSE) response and the effect of explant type and PGRs composition used in inducing PSE on the secondary somatic embryogenesis (SSE) response. PSEs were induced from basal petal and staminoid explants of MCC 01 and MCC 02 clones on DKW medium containing 2,4-D 2 mg l^-1+ kinetin 0.5 mg l^-1 or 2,4-D 2 mg l^-1 + kinetin 0.125 – 0.250 mg l^-1 + thidiazuron (TDZ) 2.5 – 5 µg l^-1 or 2,4-D 2 mg l^-1 + TDZ 10 µg l^-1. Genotype, explant type, and PGR composition dependently affected PSE response. The best PSE response was obtained from staminoid explant of MCC 02 clone on medium containing 2,4-D 2 mg l^-1 + kinetin 0.5 mg l^-1 (20%, 9 embryos). The explant type and PGR composition used in inducing PSEs affect the SSE response. The highest SSE response of MCC 01 clone was obtained from petal explant with medium containing 2,4-D 2 mg l^-1 + kinetin 0.5 mg l^-1. The formation of SSEs could increase the multiplication rate of MCC 01 clone by 7 times.</p>

Somatic Embryogenesis in Hedychium bousigonianum

An efficient primary somatic embryo (SE) and secondary somatic embryo (SSE) production system was developed for the ornamental ginger Hedychium bousigonianum Pierre ex Gagnepain. Addition of two ethylene inhibitors, salicylic acid (SA) and silver nitrate (AgNO3), to the culture media improved the system. Callus was initiated and proliferated on a medium containing Murashige and Skoog (MS) basal salts supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid and 4.6 μM kinetin. Friable callus was transferred to a liquid medium containing MS basal salts, B5 vitamins, 0.6 μM thidiazuron, and 8.9 μM 6-benzylaminopurine to induce somatic embryogenesis. The effects of various concentrations of SA (0, 25, 50, 75, 100, 125, 150 μM) and AgNO3 (0, 10, 20, 30, 40, 50, 60 μM) on callus growth, SE, and SSE development was further evaluated. The rate of callus growth decreased as the concentrations of SA or AgNO3 increased. AgNO3 and SA at all concentrations stimulated SE and SSE development better than the control although a decrease in embryo production was observed at higher concentrations of both SA and AgNO3. The best concentrations for SA were 75 and 100 μM, whereas for AgNO3, they were 30 to 50 μM for both SE and SSE production.

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

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.;