As. Pac. J. Mol. Biol. & Biotech., Jan 2014 Vol. 1, 145-151
Organogenesis from chrysanthemum Dendranthema x grandiflora (Ramat.) Kitamura petals (disc and ray florets) induced by plant growth regulators
Jaime A. Teixeira da Silva1,2
1Faculty of Agriculture and Graduate School of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan.
2Current address: Miki-cho post office, Ikenobe 3011-2, P. O. Box 7, Kagawa-ken, 761-0799, Japan.
* Author for correspondence: Jaime A. Teixeira da Silva
Miki-cho post office, Ikenobe 3011-2, P. O. Box 7, Kagawa-ken, 761-0799, Japan.
The chrysanthemum (Dendranthema x grandiflora (Ramat.) Kitamura) is an important ornamental plant. The tissue culture of chrysanthemum is well explored, but the use of petal tissue in regeneration is less undertaken, most likely due to the risk of chimerism. This study provides a way to induce organogenesis (of callus, shoots, and roots) from the petals (disc and ray flowers) of a Japanese cultivar, ‘Shuhou-no-chikara’. The response to a wide range of plant growth regulators (PGRs), under light and dark conditions, and on a Murashige and Skoog basal medium, indicates that trial and error is necessary for the optimisation of a flower-based regeneration protocol. In this study, three trends were observed: 1) There was no difference between the response in the light and the dark, except for colour, or between disc and ray florets; 2) Twelve out of 30 PGRs, when tested alone, provided some organogenic response while the remaining 18 PGRs resulted in no or <5% explant response; and 3) Organogenesis is reported for the first time in chrysanthemum flower tissue for 5 PGRs: adenine sulphate, 2iP, picloram, coconut water (CW), and phloroglucinol (PG). Ads, 6-benzyladenine (BA) and Kin resulted in shoot formation, 2iP, picloram, 2,4-D and TDZ induced callus formation, CW formed shoots and calluses, while PG, IAA, IBA and NAA formed roots, each effective at different concentrations. In the presence of 2 mg L-1 TDZ, 621 mg of callus (fresh weight-basis) formed relative to 76 mg in the control (PGR-free basal medium) while 100% explant survival was observed in the presence of 2 or 4 mg L-1 BA (relative to 5% in the control). Use of petals allows a wider range of explants to be used when other parts of the plant may be recalcitrant, although material is limited to the flowering period unless in vitro flowering can be induced. This data set serves as a preliminary assessment of the response of floral tissue to a wider range of PGRs, applied alone, and serves as a spring-board for further studies that should examine a wider range of cultivars and PGR combinations. The ploidy, genetic stability and success of acclimatisation of such material should also be examined.