TITLE:
Embryo and Protoplast Isolation from Barlia robertiana Seeds (Orchidaceae)
AUTHORS:
Mehmet Aybeke
KEYWORDS:
Barlia; Orchid; Embryo; Protoplast Isolation
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.4 No.6A,
June
19,
2013
ABSTRACT: In the present study, it is aimed to investigate embryo and protoplast
isolation from orchid seeds, because of know-how deficiency. As material Barlia robertiana seeds were subjected enzymatic
process. Globular embryos with suspensor were fully removed by enzymatic
maceration for between 100-180 min of incubation. The
highest obtained embryo yields were found in Rapidase
EX Color and Rapidase Vino Super, with degree of 89.2-90.3 × 103·g-1 MF and 86.4-91.3 × 103·g-1 MF, respectively.
According to multicomponent maceration tests, a treatment comprising 150 min of
the enzymatic reaction with 0.18% (E/S) at 50℃ was the most
optimal for obtaining high embryo yields. Regarding protoplast isolation tests,
enzyme combinations C (1% Cellulase “Onozuca” R-10 + 0.2% Macerozyme R-10 and
0.1% Driselase) and F (1% Cellulase “Onozuka” + 1% Macerozyme + 0.5% Driselase)
after 15 hours produced the best results for protoplast isolation and were significantly
different compared to other enzyme combinations. Greater protoplast yields were
obtained using a rotatory system with protoplasts incubated in the dark. Also
the present findings were discussed from the point of view of in vitro plant manipulation and orchid
cultivation in cultural media and then their importance emphasized in molecular
biological/genetical studies.