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ISHS Acta Horticulturae 1143: VII International Symposium on Edible Alliaceae

The use of thin cell layer (TCL) explants in micropropagation of shallot (Allium ascalonicum L.)

Author:   I.M. Hidayat
Keywords:   hyperhydricity, bulblet, seed production, nuclear seed, cytokinins, auxins
DOI:   10.17660/ActaHortic.2016.1143.36
In shallot seed production system, a healthy nuclear seed by means pathogen tested plant material is a prerequisite for further multiplication in a control environment. In vitro techniques offer possibility to be applied for micropropagation of healthy materials. However, micropropagation of shallot has been hampered by hyperhydricity that is a physiological disorder which inhibits a normal planlet growth and development. Reports indicated that Thin Cell Layer (TCL) explants have been used successfully in micropropagation of many ornamental species. Study in using transversal TCL of shoot tip explants of shallot cv. Sumenep was conducted in 2014, using BDS (Dunstan and Shoort, 1977) basal media supplemented with 30 g L-1 sucrose, 2 g L-1 gelrite, pH 5.8, and autoclaved at 121C, 15 psi for 15 min. auxin (2,4-D, IAA and NAA) at 0.1 mg L-1 alone and in combination with cytokinins (benzylamino purine, 2-isopentenyladenine, and thidiazuron) at each 1.0 mg L-1. Subculture was conducted in four weeks interval. Results indicated that tTCL explants in media combination NAA and BAP gave the highest average 8.6 shoots explant-1, and 10.43 shoots explans-1 at 9 and 15 week of culture (WoC) respectively compared to shoot tip eksplant at average 2.16 and 4.65 shoots explant-1. Cultures in media with 2,4D in combination with cytokinins generally performed hyperhydricity symptom compared to other media. For shoot multiplication and bulblet formation Murashige and Skoog basal media was used. Bulblets were initiated after 24 WoC. Some of the shoots were transferred and growth to normal plants in the screen house without passing through in vitro bulblet formation. This means that shoots can be transferred directly to the screen house for mini bulb production. On the other hand, a sub culture into liquid media in 3 L bulb bioreactor with 15 tTCL cultures gave 240 planlets in 10 WoC.

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