Biochemical analysis of proteins, peroxydasiques activities and polyphenols in different explants during micropropagation of Diospyros crassiflora Hiern

The objective of biochemical analysis was to determine the physiological status of uninodal stems in relation to bud break and axillary bud growth. This analysis will enable a biochemical study to be carried out during the process of callogenesis.
The biochemical analyses consisted of the extraction and determination of total soluble proteins, total polyphenols and peroxidases. The biochemical analyses were carried out on vitro plants taken from media suitable for the different in vitro culture techniques (stems, leaves, roots at different stages of growth as well as leaf callus, cotyledons), acclimatised vitro plants (leaves, stems, roots) and nursery vivoplants (leaves, stems and roots). The determination of total soluble protein was carried out according to the method of Bradford, (1976) using Coomassie Blue G250. The measurement of peroxidase activity is carried out in plant material according to the method of Thorpe (1978). The determination of phenolic compounds was carried out according to the method of Marigo et al. (1973). For each extract, three readings were made.
- The stems were taken during microcutting in the following months: 1st month; 2nd month; 3rd month.
- The leaves were removed during microcutting in the third month.
- Calls from leaves and cotyledons were taken during callogenesis in the following months: 1st month; 2nd month; 3rd month; 4th month.