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.

Preliminary Study on Induction and Identification of Polyploidy of Shandong Salvia miltiorrhiza Buds Treated by Colchicine

[Objective] The aim was to study the feasibility of using colchicine to induce polyploidy from the buds of Shandong Salvia miltiorrhiza.[Method] The buds of Shandong Salvia miltiorrhiza were induced by colchicine.[Result] When the concentration of colchicine supplemented in culture medium was 12 mg/L,the induction rate of polyploidy was the highest（36.6%）,but its survival rate was only 16.7%.When the concentration of colchicine was 8 mg/L,the induction rate of polyploidy was 15.9%,and its survival rate was up to 40.0%.Polyploidy plant of Shandong Salvia miltiorrhiza was strong and had thick,large and dark leaves,and long stomata on the lower epidermis of leaves had small density.[Conclusion] Colchicine could induce buds effectively,and the polyploidy breeding material of Salvia miltiorrhiza was obtained,which laid a foundation for shortening breeding process.

Effects of Exogenous Hormones on Subculture Multiplication and Root Induction of Tissuecultured Seedlings of Mulberry

Objective The purpose of this experiment was to investigate the effects of exogenous hormone factors on subculture multiplication and root induction of tis-suecultured seedlings in mulberry. [Method] The aseptic seedlings of Guisangyou 12 were used as the materials and the comparative experiment on subculture multi-plication and root induction used different kinds,concentrations and combinations of exogenous hormones. [Result] The hormone combination of 0.1 mg/L IBA＋1.5 mg/L 6-BA＋0.03 mg/L TDZ showed the best effects to bud multiplication, and the multiple of bud multiplication was 5.28. The effects of NAA was better than IBA and IAA on root induction. The hormone combination of 2.0 mg/L NAA＋1.0 mg/L PP333 was most suitable to the root induction, in which the rooting rate arrived to 100%, the root number was 7.01 and the root length was 1.38 cm on average. [Conclusion] The results wil provide some technical reference for large-scale propagation of mul-berry seedlings in vitro.

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.

In vitro Axillary Bud Multiplication of Citrus Nobilis Lour. in Indonesia

This research aimed to get the best composition ofcytokinin （BAP） and auxin （NAA） concentration on in vitro axillary bud multiplication of Tawangmangu orange （Citrus nobilis Lour.）. The research was conducted in Plant Physiology and Biotechnology Laboratory of Agriculture Faculty, Sebelas Maret University Surakarta from February 2008 to September 2008, used factorial design with two factors based on Completely Random Design. The first factor was BAP concentration consisting of five levels： 0, 0.5, 1, 1.5, 2 ppm and the second factor was NAA concentration consisting of five levels： 0, 0.5, 1, 1.5, 2 ppm. So there were 25 combination treatments, each of them was repeated for three times. The result observation data of shoot initiation time and number of shoot were analyzed with analysis of variance and if significant continued with Duncan＇s Multiple Range Test at 5% level. Data of shoot color and amount of leaves were analyzed descriptively. The result showed that there was interaction between BAP and NAA on shoot initiation time. Combination of 0.5 ppm BAP and 1.5 ppm NAA was able to accelerate the shoot initiation time and produced the largest number of leaves that was eight leaves. BAP was able to increase the number of shoots. Average largest amount of shoot on 0.5 ppm BAP treatment was 3.93 shoots/explant. The dominant color of leaves was light green.