A New Micropropagation Technology of Tilia amurensis:In VitroMicropropagation of Mature Zygotic Embryos and the Establishment of a PlantRegeneration System

Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos.

Micropropagation of Curcuma Sp., a Threatened Medicinal Plant

A rapid and improved micropropagation protocol was developed for Curcuma sp., a threatened and high value medicinal plant, using main bud sprout and top of rhizome sprout as explants. Stepwise optimization of different plant is to change the growth regulator, reduce the level of macro-nutrition and add humate. The present study has created multiple shoot and root induction and plantlet in vitro culture, transfers the plantlet to ex vitro. The M2 medium (MS’s macronutrition 1/4, MS’s micronutrition + Morel’s vitamine + coconut milk 10% + sucrose 25 g/l + humate 1.0 ml/l + agar 7.5 g/l + 2,4D 0.5 mg/l + BAP1.0 mg/l + TDZ 1.0 mg/l) is the highest ratio of callus induction. The TA3 medium (MS’s macronutrition 1/4 + MS’s micronutrition + Morel’s vitamin+ coconut milk 10% + sucrose 25g/l + humate 2.0 ml/l + 2,4 D 0.5 mg/l + Kinetin 2.0 mg/l + TDZ 1.0 mg/l + BAP 1.0 mg/l2 + NAA 2.0 mg/l + activated carbon 2.0 g/l) is able to create buds and regeneration multiple bud for Curcuma sp. TA3 medium adding IAA 2 mg/l and IBA 0.5 mg/l has resulted in the highest indices of quantity, healthy shoot and large diameter of roots. A large number of healthy plantlets are induced by the medium of MS’s macronutrients 1/2, MS’s micronutrients full, Morel’s vitamin, humate 3 g/l, coconut milk 150 ml/l, activated carbon 3 g/l, composition phytohormone: IAA 2 mg/l + BAP 2 mg/l + TDZ 0.5 mg/l. Further studies should focus on optimizing the humate concentration for these species of Zingiberaceae. The duration of the research is from 5/2015 to 4/2016 at Faculty of Agriculture and Forestry, Tay Nguyen University, Vietnam.