Hibiscus syriacus is a difficult species in micropropagation due to its endogenous contamination and recalcitrant shoot formation; therefore, studies on using explants other than shoot tip or axillary buds of growing shrubs were initiated. Three different seedling fragments (root, hypocotyl, and leaf petiole) from aseptically germinated seedlings of hibiscus (var. Aphrodite) were evaluated for adventitious bud formation, shoot and leaf development. The explants were cultured on McCown's woody plant basal salt medium supplemented with KNO3 (800 mg/L), adenine sulfate (80 mg/L) and MS vitamins containing BA or 2iP or TDZ at 0.5, 1.0, 2.2, 4.4 and 10 mM. Adventitious buds were present on all of the three different explants grown on medium containing TDZ; however, the most abundant bud formation, with many small leaves originating from callus was observed on hypocotyl explants cultured on medium with 1 mM of TDZ.
Petiole explants were the most frequent to develop short shoots (≈15 mm) and one to nine leaves without callus formation, where 70% of hypocotyl and the root explants formed leaves originating from callus. Callus was induced on all explant types regardless of the level or type of cytokinin used. However, the number of shoots produced by any explant type was low, petioles cultured on 0.5 and 1mM of TDZ were the most suitable material for non-callus shoot development in H. syriacus. Hypocotyl explants proved to be an excellent source for adventitious bud formation but their ability to develop shoots needs to be investigated.
Natural Variation in Plant Pluripotency and Regeneration