Shoot tissue culture of Robinia pseudoacacia f. decaisneana

Robinia pseudoacacia f. decaisneana is a transfiguration of Robinia pseudoacacia. For enhancing propagation coefficient of the species, the experiment of shoot tissue culture of Robinia pseudoacacia f. decaisneana was conducted in Forestry College of Shenyang Agricultural University from July 1999 to July 2001. The experiment included medium selection of explant induction survival, initial culture, subculture as well as rooting culture, and forming seedling with callus. The results showed that shoot segment in vitro survive rate is larger in spring than in autumn, and green dense callus could form plantlet. The best medium for initial culture was SH+0.5mg/L BA+0.05 mg/L NAA, with a propagation coefficient of 4.1 (per micro-cutting in a month), and for subculture it was B5+0.5 mg/L BA+0.05 mg/L NAA+ 10 mg/L Glu., with a propagation coefficient of 4.7. The best rooting medium was 1/2MS+0.5 mg/L NAA+10 mg/L Glu., with a rooting rate of 84.4%. These results provide reference data for reproduction of superior individuals of Robinia pseudoacacia f. decaisneana.

Direct Regeneration of Plants Derived from in vitro Cultured Shoot Tips and Leaves of Poplar （Populus×euramericana ＇Neva＇）

The purpose of the present study was to establish a regeneration procedure for Populus × euramericana ＇Neva＇ by using in vitro shoots tips and leaves. For sterilization, 0.1% （w/v） mercuric chloride （HgCl2） solution for 8 to 10 min was the optimal treatment for this poplar cultivation. The effects of benzyladenine （BA） and α-naphthaleneacetic acid （NAA） added to Murashige and Skoog （MS） medium were tested on organogenesis. The highest regeneration rate and numbers of shoots/explant from shoot tips （96.7%, 9.8） and leaves （90.0%, 8.7） were obtained on the half-strength MS medium supplemented with 0.5 mg/L BA and 0.1 mg/L NAA. The optimal medium for in vitro rooting of shoots was on a half-strength MS medium containing 1 mg/L indolebutyric acid （IBA） with the highest rooting frequency （93.3%） and numbers of roots/explant （8.2）. For acclimatization, in vitro rooted plantlets were transferred to plastic cups containing vermiculite and peat （1： 1）. After acclimatization, transplanted plantlets grew well in a shade house. Therefore, we believe that this efficient plant regeneration protocol especially by leaf explants is very important for in vitro clonal propagation of Populus×euramericana ＇Neva＇.

Contributions of Chinese Botanists to Plant Tissue Culture in the 20th entury

This paper looks back to the development of plant tissue culture in China in the last century. Since 1934, tissue culture studies in China has kept up with the international development in the fields. Progress has been made by Chinese in nearly every branches of tissue culture, including in vitro organogenesis, shoot tip culture, anther culture, ovary culture, endosperm culture, protoplast culture as well as mass cell culture. On the basis of reviewing the articles written by Chinese on plant tissue culture, the internationally recognized contributions are specially mentioned. The applications of plant tissue culture to agriculture and industry in China are also introduced.

An efficient method for clonal propagation and in vitro establishment of softwood shoots from epicormic buds of teak(Tectona grandis L.)

Softwood shoots were produced from 40 cm long stem segments placed horizontally in flat trays containing sterilized sand under natural light or shade conditions for subsequent rooting and micropropagation studies in teak （Tectona grand＆ L.）. Higher number of shoots （6.17） per log was produced under natural light as compared to shade conditions. Forcing was also better in natural light as compared to shade in terms of shoot length, number of nodes or leaves. For rooting, 2-4 cm long softwood shoots were excised and treated with either indole-3-butyric acid （IBA） or α-naphthyl acetic acid （NAA） at 0, 1000, 2000 or 3000 μmol.L^-1 each or with combinations （1000 ＋ 1000, 2000 ＋ 2000 or 3000 ＋ 3000 μmol.L^-1） and then placed in flat trays containing autoclaved sand at 25 ± 2℃ in 16 h photoperiod at 35 μmol.m^-2.s^-1. After 28 days, softwood cuttings treated with IBA ＋ NAA （3000 ＋ 3000 μmol.L^-1） had highest rooting percentage （89.3%） with 5.5 mean roots. Shoot apex and nodal explants of softwood cuttings were pretreated with 0.1% （w/v） ascorbic acid, boric acid, activated charcoal, citric acid, glutamine or polyvinylpolypyrollidone （PVP） for 24 h to remove phenolic compounds before surface disinfestation. Glutamine （G1） and PVP were equally effective resulting in 60% establishment of shoot apices on MS medium supplemented with 10 μmol.L^-1 6-benzylaminopurine （BAP） ＋ 5 μmol.L^-1 NAA. Using shoot apices, highest （42.80） number of multiple shoots with 54.33 mm shoot length were obtained on MS ＋ BAP （8.8 p.mol.L 1） ＋ IBA （2 μmol.L^-1） after 45 days. Shoots were successfully rooted and acclimatized to greenhouse conditions.

Effect of 24-Epibrassinolide on Growth of <i>in Vitro</i>Shoot Tips of Different Yam (<i>Dioscorea</i>Spp.) Species

In this work we compared the effect of the growth regulator content of the culture medium on the growth of in vitro shoot tips of five yam accessions belonging to four yam species (one Dioscorea alata, one D. rotundata, one D. cayenensis and two D. trifida). Medium S contained 0.6 μM benzyl adenine, 1.07 μM naphthalene acetic acid and 0.23 μM gibberellic acid while medium EBR contained 0.23 μM gibberellic acid and 0.1 μM 24-epibrassinolide. After 2 months of culture, oxidation level was significantly reduced on medium EBR compared to medium S for four of the five accessions tested. By contrast, medium EBR did not have any positive effect on shoot length since length of shoots produced after 2 months of culture on medium S and EBR were similar, except with accession 3-45T, for which shoot length was shorter on medium S compared to medium EBR. These results underline the potential of 24-epibrassinolide to reduce oxidation phenomena during in vitro culture and call for its utilization for regeneration of cryopreserved yam shoot tips, which is often impeded by oxidation phenomena.

Morpho-Histological Analysis of Direct Shoot Organogenesis Induced in Flower Buds Cultures of <i>Allium altissimum</i>

The present work gives a detailed study of in vitro shoot organogenesis of the ornamental onion A. altissimum Regel from the buds of the middle layer of the inflorescences. The course of morphogenesis was examined by light and scanning electron microscopy. Histological observation revealed that during 3 - 5 days of culture on the BDS medium supplemented with 2.0 mg·L-1 of BA and 2.0 mg·L-1 of NAA the epidermal cells of the stamen filament in the area of its fusion with the tepal became competent and dedifferentiated. Originally the organogenesis involved several initial epidermal cells. The formation of meristematic centers was observed from day 3 to day 14. The apical shoot meristems and leaf primordia in a roller shape formed from day 14 to day 28 of culture on the same media. The further development of vegetative shoots and formation of the bulblets were observed when the explants were stimulated by triapenthenol (2.0 mg·L-1).

Study on Shoot Tip Culture of Illciaceae Ornamental Plants

[Objective] This study aimed to establish a technology system for tissue culture and rapid propagation of Illciaceae ornamental plants. [Method] Effects of medium components and anti-browning agents on the survival and growth of shoot tips were investigated by using apical buds of IItciaceae plant Haierlian as experiment material and MS as basic medium. [Result] The results showed that apical buds at the early germination period in spring were the most suitable explants for tissue culture of IIIciaceae plant Haierlian. Sterilization with 0.1% HgCI2 for 6 min achieved the best effect, while conventional surface-sterilization with ethanol would affect the survival of explants. The optimal medium for primary culture was MS-D （with modifications in major elements and organic components） ＋ anti-browning agents （equa~ volume） ＋ 2.0 mg/L of 6-BA ＋ 0.5 mg/L of NAA. The optimal subculture medi- um was MS-F （with modifications in inorganic and organic components） ＋ anti-brown- ing agents （equal volume） ＋ 2.0 mg/L of 6-BA ＋ 0.1 mg/L of NAA. [Conclusion] This study laid the foundation for establishment of tissue culture and rapid propagation technology system for Haierlian.

Tissue Culture and Rapid Multiplication Techniques of Apocynum L.

[Objective] This study aimed to investigate rapid multiplication of Apocynum by tissue culture so as to provide plantlet sources for its industrialized cultivation. [Method] The asepsis seedlings were obtained by dealing with Apocynum seeds. Its cotyledons, hypocotyls and shoot tips were cultured on the media containing different concentrations of hormones. Finally, the influence of different hormone combinations on differentiation of cotyledons and hypocotyls, rapid multiplication of shoot tips, rapid multiplication of regenerated shoots, and rooting of test-tube plantlets was com- pared. [Result] MS＋2.0 mg/L BA＋0.03 mg/L NAA and MS＋0.07 mg/L NAA were the optimum medium for inducing regenerated buds from cotyledons and hypocotyls re- spectively; MS＋2.0 mg/L BA＋0.02 mg/L NAA was the best medium for rapid multi- plication of shoot tips; MS＋1.9 mg/L BA＋I.7 mg/L NAA was the best medium for rapid multiplication of regenerated buds： and 1/2MS＋0.6 mg/L NAA was the best medium for inducing roots. [Conclusion] The optimum hormone combination was de- termined for Apocynum rapid multiplication by tissue culture, which provides technical support on Apocynum industrialized cultivation.

Study on the optical property and biocompatibility of a tissue engineering cornea

AIM: To study the optical property and biocompatibility of a tissue engineering cornea. METHODS: The cross-linker of N- (3-Dimethylaminoropyl)-N'ethylcarbodiimide hydrochloride (EDC)/ N-Hydroxysuccinimide (NHS) was mixed with Type I collagen at 10% (weight/volume). The final solution was molded to the shape of a corneal contact lens. The collagen concentrations of 10%, 12.5%, 15%, 17.5% and 20% artificial corneas were tested by UV/vis-spectroscopy for their transparency compared with normal rat cornea. 10-0 sutures were knotted on the edges of substitute to measure the corneal buttons's mechanical properties. Normal rat corneal tissue primary culture on the collagen scaffold was observed in 4 weeks. Histopathologic examinations were performed after 4 weeks of in vitro culturing. RESULTS: The collagen scaffold appearance was similar to that of soft contact lens. With the increase of collagen concentration, the transparency of artificial corneal buttons was diminished, but the toughness of the scaffold was enhanced. The scaffold transparency in the 10% concentration collagen group resembled normal rat cornea. To knot and embed the scaffold under the microscope, 20% concentration collagen group was more effective during implantation than lower concentrations of collagen group. In the first 3 weeks, corneal cell proliferation was highly active. The shapes of cells that grew on the substitute had no significant difference when compared with the cells before they were moved to the scaffold. However, on the fortieth day, most cells detached from the scaffold and died. Histopathologic examination of the primary culture scaffold revealed well grown corneal cells tightly attached to the scaffold in the former culturing. CONCLUSION: Collagen scaffold can be molded to the shape of soft contact corneal lens with NHS/EDC. The biological stability and biocompatibility of collagen from animal species may be used as material in preparing to engineer artificial corneal scaffold.

In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.

Tissue Culture of Betula microphylla Bunge var. paludosa

[Objectives]This study aimed to study the tissue culture technique of Betula microphylla Bunge var. paludosa. [Methods]Taking seeds as explant,seedlings of B. microphylla were cultured. Then,stem sections with leaves were cut off and subjected to induction of clustered shoots. Finally,the rooting and transplanting of adventitious shoots were completed. Thus,the tissue culture system of B. microphylla was established. [Results]The natural seed germination induction medium was Murashige and Skoog( MS),and the germination rate was31%. The most suitable shoot induction medium was WPM medium added with 1. 0 mg/L of 6-benzylaminopurine( BA),and the multiplication coefficient was 13. 7. The rooting medium was Woody Plant medium( WPM) added with 1. 0 mg/L of indole-3-butytric acid( IBA),and the rooting rate was 100%. The transplanting substrate for tissue-cultured seedlings was composed of humus soil,vermiculite and perlite( V∶ V∶ V =4∶3∶3),and the survival rate reached 88. 75%. [Conclusions]The experimental materials are easy to obtain and preserve,and the proliferation is rapid. This study provides technical support for the rapid acquisition of the tissue-cultured seedlings of B. microphylla.

<i>In Vitro</i>Organogenesis of <i>Quisqualis indica </i>Linn.

Shoot organogenesis and plant regeneration were achieved on callus derived from leaf section and stem base explants of Quisqualis indica (Combretaceae). In vitro cultures were established using nodal segments obtained from mature field-grown shrubby plants. For the development of optimized protocol, different types and concentrations of plant growth regulators were used to induce adventitious shoot regeneration via callus from leaf section and one-node stem base explants obtained from in vitro regenerated micro shoots and direct field-grown newly flush-off shoots. The TDZ was considered to be the best among the cytokinins (6-benzyladenine (BA), 6-(?-?, dimethylallyamino purine) (2-iP) and thidiazuron (TDZ) added to the Murashige and Skoog’s medium (MS) for adventitious shoot productions. A combination of 1.0 mg/L TDZ and 0.5 mg/L GA3 was most effective in stimulating callus induction and adventitious shoot regeneration from the leaf section derived calli with an average of 6 shoots per callus explant and an average of 8 shoots per callus explant originated from one-node stem base explants. In vitro raised shoots were sub-cultured on MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L GA3 for further shoot growth. Maximum rooting of in vitro regenerated shoots was obtained on MS medium supplemented with either 0.5 mg/L indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) individually or a combination of 0.5 mg/L IAA and 0.5 mg/L IBA. Plantlets raised in vitro were acclimatized and subsequently transferred to experimental field.

Effect of Vitamins on <i>In Vitro</i>Organogenesis of Plant

Vitamins are necessary compounds synthesized and utilized in plants. In tissue culture media, vitamin addition is not always common;since the amount needed by plants is relatively unknown and varies. Vitamins, in combination with other media constituents, have been shown to have direct and indirect effects on callus growth, somatic growth, rooting, and embryonic development. For example, different studies have shown that thiamine is associated with cytokinin and has a role in inducing callus growth and rooting. Moreover, thiamine was essential in facilitating the production of more secondary metabolites such as proteases in pineapple. Both biotin and riboflavin play a role in callus development as well. Specifically, riboflavin exerts different effects on plant rooting either positively and negatively. Vitamin D known to cause uptake of calcium in animal tissue, exerts a similar effect in plants. In addition, vitamin D causes cell elongation and meristematic cell division. Vitamin C, known for its anti-oxidative properties, has also enhanced shoot growth and rooting.

In Vitro-Propagation of Agave tequilana Weber cv.azul in a Temporary Immersion System

In Mexico,there is a need to produce large quantities of plantlets for the establishment and replanting of blue(cv.azul)agave production areas.Most of these plots are within the origin denomination area(DOT,Spanish acronym)of the distilled product of this plant,known as tequila.The objective of this study was to develop an in vitropropagation protocol for Agave tequilana Weber cv.azul using segmented stems in both:solid and liquid media.A disinfection and in vitro technique were developed to obtain shoots,through plantlets collected in commercial plots,which attained 100%surface-disinfection and budding rate.At the multiplication stage,the effects of 6-Benzylaminopurine(BA)(0.0,4.4 and 13.2μM)and kinetin(0.0,9.4,18.8 and 37.6μM)were evaluated on lateralshoot production of segmented sagittal stems.These were cultivated on Murashige&Skoog(MS)medium,with the addition of 3.0%sucrose and 8 g L−1 agar.It was observed that BA and kinetin increased the number of shoots per explant,obtaining up to 18 and 26,respectively.Furthermore,it was found that just the sagittal segmentation of explants increased axillary budding.On the other hand,segmented-stem bases were grown in MS liquid medium with 3.0%sucrose,inside a RITA
system,programmed by a 5 min immersion step with a frequency of every 4 h.The effect of Indole−3-Acetic acid(IAA)(0.57,2.9,5.7μM)was evaluated,while maintaining a concentration of BA(13.2μM).It was observed that the greatest concentration of IAA led to the formation of more than 20 buds per explant.These results offer a new methodology to increase the efficiency of A.tequilana Weber cv.azul-in vitro multiplication by sagittal segmentation of stems and the addition of BA and/or IAA.

Influence of Watering Solution and Phenotype on the Growth of <i>in Vitro</i>Propagated Pineapple (Smooth Cayenne Cultivar) Plantlets during Acclimatization

Pineapple is the first fruit crop cultivated in south Benin that greatly contributes for food and nutritional security and farmers’ income. But the lack of homogenous planting material constitutes the major constraint for improving pineapple yield. In vitro micropropagation is now used in the production of homogenous and free disease planting materials of pineapple. However, the acclimatization to natural condition of pineapple plantlets is an important step in planting material production of this crop. Here, we determined the intrinsic and extrinsic factors which influence the behavior of plantlets during the acclimatization process. For this purpose, plantlets from different categories were selected, trimmed and planted on a horticultural substrate made up of potting soil, white sawdust and compost previously sterilized. The plantlets were then incubated in under acclimatization greenhouse with average temperature of 29˚C and 70.2% of humidity. A batch of plantlets was subjected to two different watering solutions: Shive and Robbins solution and NPK 14-6-5 foliar fertilizer. The results obtained initially showed high rate (100%) of survival and growth of the plantlets watered with Shive and Robbins solution against 50% of the plantlets watered with the foliar fertilizer solution. In addition, the plantlets with spread pores exhibiting the characteristics of which the number of leaves varies between 9 and 11, the weight between 1.2 and 1.5 g, the size of 4.5 to 5.5 cm, and a good junction between the aerial part and the root system were those which were successfully grown in acclimatization phase under greenhouse, unlike plantlets with erected pores having lower success rate. This study goes a long way in providing good procedures of acclimatization of homogenous and free disease planting material of pineapple to the famers.

Efficient Micropropagation of Chestnut Hybrids(Castanea spp.)Using Modified Woody Plant Medium and Zeatin Riboside

The chestnuts genus(Castanea spp.)is comprised of economically important trees native to the Northern hemisphere that are used as food and hardwood timber.Here,a very efficient method for micropropagation of European×Japanese chestnut hybrids(Castanea sativa×C.crenata)is described.Woody Plant Medium was used as the basal medium.In vitro shoots of four rootstock cultivars were micropropagated without shoot-tip necrosis on multiplication medium containing 5.7 or 11.4μmol·L^(−1)zeatin riboside,and were rooted on rooting medium containing 2.46μmol·L^(−1)indolebutyric acid.Monthly shootmultiplication rates for each cultivarwere 2–5 folds.In vitro rooting percentages for four cultivars were 87%for‘Maraval’,67%for‘Marigoule’,93%for‘Marsol’,and 97%for‘Précoce Migoule’.Within a 5 week period,80%–95%of rooted shoots were successfully acclimated under high humidity conditions after they were planted in either soil or rockwool.

Micropropagation of yellow mangosteen:a valuable endemic tree of India

We developed a method for in vitro regenera- tion of Garcinia xanthochymus （yellow mangosteen） from matured seed segments. Multiple shoots were induced on woody plant （WP） medium supplemented with cytokinins. An average of 11 shoots per explant were regenerated from mature seed segments on WP medium containing 20 μM 6-benzylaminopurine. Histological analysis revealed that hypodermal cells of seed segments were initially involved in active division, which later developed into meriste- moids, subsequently leading to the formation of shoot buds. Shoot elongation was achieved by repeated subculturing of seed explants in shoot regeneration medium. Rooting of shoots was achieved on WP medium supplemented with indole-3-butyric acid or s-naphthalene acetic acid. Plant- lets were transplanted to pots containing soil： compost （1：1） and survival rate was 90 %.

High frequency wheat regeneration from leaf tissue explants of regenerated plantlets

The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from leaf tissue explants of regenerated plantlets, resulting in difficulty in obtaining homoplastic plants via multiple rounds of antibiotic selection of chloroplast transformants. Thus, a repeated in vitro regeneration system from leaf tissues was studied in this research. Our results showed that 2 mm leaf basal segments of the 4 cm high leaves from regenerated plantlets can give the best callus induction at present study. The best callus induction medium was Murashige and Skoog (MS) basal medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L naphthalenacetic acid, which gave a callus induction rate of up to 87.2%. The optimal differentiation medium was MS basal medium supplemented with 10 mg/L silver nitrate and 1 mg/L 2,3,5-triiodobenzoic acid, which gave a regeneration rate up to 33.7% for the wheat lines tested. This is the first report showing that leaf basal segments of in vitro regenerated plantlets can be used for regeneration of wheat. The establishment of a repetitive regeneration system should pave the way for the development of chloroplast transformation and the plant regeneration systems starting from leaf material of in vitro regenerated wheat and other cereal crops.

Photoperiod Affects <i>in Vitro</i>Flowering in Wild Peanut <i>Arachis paraguariensis</i>

Arachis paraguariensis, a wild peanut species, is a potential experimental system for studying the molecular mechanisms of flowering in the genus Arachis. The present study was carried out to investigate the effect of photoperiod on in vitro reproductive behavior of five genotypes of A. paraguariensis. Day-lengths of 12, 16 and 24 h were tested to monitor in vitro flowering using growth chambers kept at 26?C ± 1?C and 60% ± 5% relative humidity under an illumination of 40 μmol?m–2?s–1. Flowering percentage of plantlets ranged from 35% to 93%, 20% to 75%, and 5% to 53% for 12, 16 and 24 h day-lengths, respectively. Genotype PI 262842 displayed the highest frequency of flowering under all the day-length treatments but in vitro flower bud initiation was delayed. The highest mean flowering percentage of 65% across all the genotypes for plantlets exposed to 12 h photoperiod is indicative that flowering induction actually occurred. The results presented in this paper provide evidence for photoperiodic flowering response as well as the occurrence of short day-length-enhanced flowering in A. paraguariensis.

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.