*Corresponding author (A. Jala). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail address: [email protected]. 2013. American Transactions on Engineering & Applied Sciences. Volume 2 No. 3 ISSN 2229-1652 eISSN 2229-1660 Online Available at http://TuEngr.com/ATEAS/V02/213-222.pdf 213 American Transactions on Engineering & Applied Sciences http://TuEngr.com/ATEAS , http://Get.to/Research Callogenesis and Organogenesis from Inflorescence Segments of Curcuma Alismatifolia and Curcuma Hybrid ‘Laddawan’ Anchalee Jala a* a Department of Biotechnology, Faculty of Science and Technology, Thammasat University, 12120 THAILAND A R T I C L E I N F O A B S T R A C T Article history: Received February 21, 2013 Received in revised form April 22, 2013 Accepted May 03, 2013 Available online May 09, 2011 Keywords: Curcuma sp.; Callogenesis; Shoot organogenesis; 2,4-Dichlorophenoxy acetic acid. Callogenesis of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ were proliferated from young inflorescences when cultured them on Murashige and Skoog (MS) medium supplemented with various concentration (8, 10, 12, and 14 mg/l) 2,4-dichlorophenoxyacetic acid (2,4-D) at dark period for 4 weeks to form somatic embryo. The highest percentage growth rate is 90 percent for somatic embryo proliferated on MS medium supplemented with 14 mg/l 2,4-D. When induced these somatic embryo to form new shoot organogenesis by culturing them on MS medium supplemented with various types ( glucose, sucrose and maltose ) of sugar, the highest percentage of new shoots were proliferated from Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ with 0.25 g/l maltose which were 50.00 and 46.67, respectively. 2013 Am. Trans. Eng. Appl. Sci. 1. Introduction The genus Curcuma (Zingiberaceae) comprises more than 80 species of rhizomatous perennial 2013 American Transactions on Engineering & Applied Sciences.
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Callogenesis and Organogenesis from Inflorescence Segments of Curcuma Alismatifolia and Curcuma Hybrid ‘Laddawan’
Callogenesis of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ were proliferated from young inflorescences when cultured them on Murashige and Skoog (MS) medium supplemented with various concentration (8, 10, 12, and 14 mg/l) 2,4-dichlorophenoxyacetic acid (2,4-D) at dark period for 4 weeks to form somatic embryo. The highest percentage growth rate is 90 percent for somatic embryo proliferated on MS medium supplemented with 14 mg/l 2,4-D. When induced these somatic embryo to form new shoot organogenesis by culturing them on MS medium supplemented with various types ( glucose, sucrose and maltose ) of sugar, the highest percentage of new shoots were proliferated from Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ with 25 g/l maltose which were 50.00 and 46.67, respectively.
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*Corresponding author (A. Jala). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail address: [email protected]. 2013. American Transactions on Engineering & Applied Sciences. Volume 2 No. 3 ISSN 2229-1652 eISSN 2229-1660 Online Available at http://TuEngr.com/ATEAS/V02/213-222.pdf
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American Transactions on Engineering & Applied Sciences
http://TuEngr.com/ATEAS, http://Get.to/Research
Callogenesis and Organogenesis from Inflorescence Segments of Curcuma Alismatifolia and Curcuma Hybrid ‘Laddawan’
Anchalee Jala a*
a Department of Biotechnology, Faculty of Science and Technology, Thammasat University, 12120 THAILAND A R T I C L E I N F O
A B S T R A C T
Article history: Received February 21, 2013 Received in revised form April 22, 2013 Accepted May 03, 2013 Available online May 09, 2011 Keywords: Curcuma sp.; Callogenesis; Shoot organogenesis; 2,4-Dichlorophenoxy acetic acid.
Callogenesis of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ were proliferated from young inflorescences when cultured them on Murashige and Skoog (MS) medium supplemented with various concentration (8, 10, 12, and 14 mg/l) 2,4-dichlorophenoxyacetic acid (2,4-D) at dark period for 4 weeks to form somatic embryo. The highest percentage growth rate is 90 percent for somatic embryo proliferated on MS medium supplemented with 14 mg/l 2,4-D. When induced these somatic embryo to form new shoot organogenesis by culturing them on MS medium supplemented with various types ( glucose, sucrose and maltose ) of sugar, the highest percentage of new shoots were proliferated from Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ with 0.25 g/l maltose which were 50.00 and 46.67, respectively.
2013 Am. Trans. Eng. Appl. Sci.
1. Introduction The genus Curcuma (Zingiberaceae) comprises more than 80 species of rhizomatous perennial
2013 American Transactions on Engineering & Applied Sciences.
214 Anchalee Jala
herbs and is widespread in tropical Asia, extending to Africa and Australia (Purseglove et al.
1981). Within the genus Curcuma, Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ is a
highly valued ornamental, which is indigenous to Burma and Thailand (Apavatjrut et al. 1996;
Schaffer et al. 2011). It is considered as a novel ornamental plant in the floricultural market and
grows well after having been introduced to Guangzhou, China. It is usually used in tropical and
subtropical landscape design as a potted, foliage, and flower plant. It has a long flowering period
(April to November) and vase cut flowers can last 20 days. For these reasons it has seen an
increasing demand in the world ornamental market. However, seeds obtained are limited and
can’t germinate during cold season due to their dormancy, as traditional multiplication involves
propagation by rhizome. Since it is difficult to keep pace with the demands created by market
exploitation, it is thus essential to establish an efficient propagation and regeneration system,
preferably in vitro, to ensure the clonal propagation of desired genotypes.
In this study, anthers were used as explants to induce somatic embryo, which was then used to
form shoots organogesis. This study reports on an efficient callus and plant regeneration for
Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’.
2. Materiasls and Method Young inflorescence of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ were used as
explants and cleaned surface with liquid detergent and washed with tap water for 2 min., immersed
in 70% alcohol for 5 min. and sterilized with 20% sodium hypochlorite for 20 min. then with 5%
sodium hypochlorite for 10 min and immersed in sterile distilled water 3 times for 2 min each.
Petal and Sepal and the end of inflorescence were removed from the sterilized inflorescence.
Sterilized inflorescence about 2.5–3.5 cm long sections were inoculated on Murashige and Skoog
(MS.1962) supplemented with 2mg/l BA. Explants were incubated on agarified MS medium.
All medium contained 30 g m/l sucrose and pH was adjusted to 5.8 with 1.0 N HCl or 1.0 N NaOH
before adding 2.5 gram Phytagel (Sigma, USA) and autoclaving at 121°C for 20 min and incubated
at culture room at 27 ± 2°C exposed to 60 µmol m-2 s-1 fluorescent light (TLD 36W/84 3350lm
Philips Thailand) in a 16-hour photoperiod. All cultures were subcultured every two weeks, for
three times to multiply callus.
*Corresponding author (A. Jala). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail address: [email protected]. 2013. American Transactions on Engineering & Applied Sciences. Volume 2 No. 3 ISSN 2229-1652 eISSN 2229-1660 Online Available at http://TuEngr.com/ATEAS/V02/213-222.pdf
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2.1 Somatic Embryo Induction Proliferated callus was transferred to MS medium supplemented with various concentration
(2, 4, 6, 8, 10, 12 and 14 mg/l ) 2,4-D , 3% sucrose and pH was adjusted to 5.8 with 1.0 N HCl or 1.0
N NaOH before adding 2.5 gram Phytagel (Sigma, USA) and autoclaving at 121°C for 20 min and
incubated at culture room at 27±2°C in dark period for four weeks to induce somatic
embryogenesis. Percentage number of somatic embryos is recorded.
2.2 Shoot Organogenesis Callus from Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ which cultured on MS
medium supplemented with 14 mg/l 2,4-D and 0.25 gram per liter of different types of sugar
(glucose, sucrose, maltose ) and pH was adjusted to 5.8 with 1.0 N HCl or 1.0 N NaOH, before
adding 2.5 gram Phytagel (Sigma, USA) and autoclaving at 121°C for 20 min and incubated at
culture room at 27 ± 2°C exposed to 60 µmol m-2 s-1 fluorescent light (TLD 36W/84 3350lm
Philips Thailand) in a 16-hour photoperiod. All cultures were subcultured every three weeks for
four times. Shoot proliferation was investigated after culturing for 45 days.
3. Data Analysis Somatic embryo experiment was used Randomized Complete Block Design with 8 treatments
three replicates and each replication was 10 explants. Shoot Induction was used Randomized
Complete Block Design with three treatments three replicates and each replication was 10 explants.
Means were analyzed by Analysis of Variance and significant differences between means were
compared by the Duncan’s New Multiple Range Test (DMRT) using SPSS v. 13.0. For all
comparisons, statistical significance was considered at p<0.01.
4. Result Somatic embryos from Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ were
proliferated after cultured on MS medium supplemented with different concentration of 2,4-D in
dark period for 4 weeks. It was found that number of somatic embryos proliferated on MS
medium supplemented with 8, 10, 12 and 14 mg/l 2,4 – D. When compared average percentage
216 Anchalee Jala
Table 1 Effect of 2,4-D which induced somatic embryo from young inflorescence of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ after cultured for 4 weeks. 2,4-D (mg/l) Percentage of somatic embryo proliferated from young
inflorescence of Curcuma (%) C. alismatifolia** C. hybrid ‘Laddawan’ .**
0 0.00 e 0.00 d 2 0.00 e 0.00 d 4 0.00 e 0.00 d 6 0.00 e 0.00 d 8 36.67 d 60.00 c
10 46.67 c 66.67 bc 12 80.00 b 76.67 b 14 90.00 a 90.00 a
** - highly Significant difference p ≤ 0.01 abc - in the same row was not significant difference when compared their mean with DMRT at p ≤ 0.01
Figure 1 Young inflorescence Curcuma alismatifolia and Curcuma hybrid ‘Laddawan cultured on
MS Medium supplemented with 12 and 14 mg/l 2,4-D after cultured for 4 weeks 1a.Compact callus ( C. alismatifolia) proliferated on MS medium supplemented with 12 mg/l 2,4-D 1b.Friable callus(C. alismatifolia) proliferated on MS medium supplemented with 14 mg/l 2,4-D 1c.Compact callus(C. hybrid ‘Laddawan’) proliferated on MS medium supplemented with 12 mg/l 2,4-D 1d. Friable callus (C. hybrid ‘Laddawan) proliferated on MS medium supplemented with 14 mg/l 2,4-D.
number of somatic embryos in statistic. They were highly significant difference (p<0.01) and in
MS medium supplemented with 14 mg/l 2,4-D gave the highest average percentage number of
somatic embryos in both Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ which are 90
as shown in Table 1. When compared type of callus in 12 and 14 mg/l 2,4-D in each Curcuma
alismatifolia and Curcuma hybrid ‘Laddawan,’ they showed that characteristic of callus which
proliferated in 12 mg/l 2,4-D their cells are smaller and more compact than from 14 mg/l 2,4-D as
shown in Figure 1.
*Corresponding author (A. Jala). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail address: [email protected]. 2013. American Transactions on Engineering & Applied Sciences. Volume 2 No. 3 ISSN 2229-1652 eISSN 2229-1660 Online Available at http://TuEngr.com/ATEAS/V02/213-222.pdf
217
The callus originated from the young inflorescence surface and not from inner tissue in
Curcuma alismatifolia (Figure 1a) and Curcuma hybrid ‘Laddawan (Figure 1c). The callus
which proliferated from MS medium supplemented with 14mg/l 2,4-D are friable , their cells are
bigger and uninucleated which ready to form somatic embryo in both Curcuma alismatifolia
(Figure 1.b) and Curcuma hybrid ‘Laddawan (Figure 1d).
4.1 Shoot Induction from Somatic Embryo The callus originated from the young inflorescence surface and not from inner tissue in
Curcuma alismatifolia and Curcuma hybrid ‘Laddawan and formed somatic embryo .When culture
these somatic embryo on MS medium supplemented with 0.25g/l different types of sugar (glucose
sucrose and maltose) for 12 weeks. It was found that somatic embryo responded to different types
of sugar. They proliferated to young shoot in MS medium supplemented with different type of
sugar. When compared the result in statistic, induction shoot from MS medium supplemented with
0.25g/l maltose was highly significant difference (p ≤ 0.01) from glucose and sucrose as shown in
Table 2. The highest average percentage of shoot induction in Curcuma alismatifolia and
Curcuma hybrid ‘Laddawan were 50.00 and 46.67, respectively.
Table 2 Shoot Induction of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan from somatic embryo which cultured on MS medium supplemented with different types of sugar after cultured
for 12 weeks. Types of sugar
(0.25 g/l) Percentage of shoot induction (%)
C. alismatifolia ** C. hybrid ‘Laddawan’ ** No sugar 0.00 c 0.00 c glucose 23.33 b 13.33 c sucrose 23.33 b 30.00 b maltose 50.00 a 46.67 a
** - highly Significant difference p ≤ 0.01 abc - in the same row was not significant difference when compared their mean with DMRT at p ≤0.01
218 Anchalee Jala
Figure 2 Shoot induction from somatic embryo on MS medium supplemented with 0.25g/l maltose
on Curcuma alismatifolia and Curcuma hybrid ‘Laddawan after culturing 12 weeks: (a) Somatic embryo proliferation in C. alismatifolia (b) Differentiation of Somatic embryo in C. alismatifolia (c) Shoot proliferation in C. alismatifolia (d) Somatic embryo proliferation. In
C. hybrid ‘Laddawan’ (e) Differentiation of Somatic embryo in C. hybrid ‘Laddawan’ (f) Shoot proliferation in C. hybrid ‘Laddawan’
5. Discussion Young inflorescence of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan cultured on
MS medium supplemented with different concentration of 2,4-D in dark period for 4 weeks
showed different responses. Callus proliferated on MS medium supplemented with 8, 10, 12 and
14 mg/l 2,4-Dand MS medium supplemented with 14mg/l 2,4-D gave the highest average
percentage of friable callus which were 90 in both curcuma. As worked of Yaping et al. (2013)
showed that 2,4-D in the induction medium could induce a high percentage of callus (56.2 %). This
callus was compact and friable. In C. alismatifolia, regeneration has been reported from young
inflorescences (Wannakrairoj, 1997). Also, Toppoonyanont et al. ( 2005) had reported that C.
alismatifolia inflorescences were used as explants and, when inoculated on MS basal medium
containing 44.0M BA and 0.57M NAA for 1 month they developed and reverted to vegetative
shoots directly from flower organs (or floral buds) and not via callus. Mohanty et al. (2008)
reported that plant regeneration from callus cultures of C. aromatica was possible with 2,4-D
(9.1M) and 2.3M KT. Callus could be induced from the base of adventitious shoots of C.
kwangsiensis on MS medium containing 1.4M TDZ, 4.4M BA and 2.3M 2,4-D (Zhang et al.
2011). Plant regeneration from the culture of immature of C. longa inflorescences was possible
by direct shoot development on MS basal medium supplemented with BA (22.0 or 44.0M) in
combination with 0.9M 2,4-D or 0.54M NAA and 4.5 or 9.0M TDZ in combination with 0.57M
*Corresponding author (A. Jala). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail address: [email protected]. 2013. American Transactions on Engineering & Applied Sciences. Volume 2 No. 3 ISSN 2229-1652 eISSN 2229-1660 Online Available at http://TuEngr.com/ATEAS/V02/213-222.pdf
219
IAA (Salvi et al. 2000). Moreover, they located at the same positions and were arranged spirally
within the bracteole, similarly to ex vitro shoots (Udomdee et al. 2003). All of these reports
showed that callus were induced from different explants by using of 2,4-D single or NAA in
combination with BA, KT or TDZ. The callus doubled in size in less than two weeks when
subcultured in a fresh MS medium supplemented with 1 mg/l 2,4-D.
In this treatment, the concentration was maintained at 1 mg/l due to its response in producing
favorable rapid results. In plant tissue culture, sucrose is the most commonly used carbohydrate
source because of the wide spread of this disaccharide as a transporter molecule, and its high
solubility in water. Many in vitro studies have proven that sucrose supports near optimum rates of
growth and also plays multiple roles in the provision of carbon and energy that promotes cell
growth and division (Balachandran et al. 1990). Viu et al. (2009) reported that Callus was
suspended in a liquid medium to enhance proliferation. For initiation, only friable callus was used.
Prior to any new treatment, 1 g of the friable callus was proliferated in a liquid medium with same
concentration as callus induction medium (MS medium supplemented with 1mg l-1 2,4-D and 30g
/l sucrose).
The highest average percentage of shoot proliferated from somatic embryo of Curcuma
alismatifolia and Curcuma hybrid ‘Laddawan, when cultured on MS medium supplemented with
0.25g per liter of maltose which are 50.00 and 46.67, respectively. But Wannakrairoj(1997) and
Jala, (2012) reported that plant regeneration from the culture of immature of C. longa
inflorescences was possible by direct shoot development on MS basal medium supplemented with
BA (22.0 or 440 M) in combination with 0.9 M 2,4-D in combination with 30g/l sucrose. And the
same as Zhang et al (2011) report that callus could be induced from the base of adventitious shoots
of C. kwangsiensis on MS medium containing 1.4M TDZ, 4.4 M BA and 2.3M 2,4-D. Normally
they used sucrose for carbon source. Sucrose is the most commonly used carbohydrate in plant
tissue or cell culture (Vu et al., 1993). Many factors affecting the growth of excised plant organs,
tissues and cells in vitro have been studied extensively over the last 30 years, including plant
growth regulators, light, temperature, medium pH, humidity, gas exchange and the presence of
harmful microorganisms (Leifert et al., 1995). On the other hand, plant carbohydrate
220 Anchalee Jala
requirements in vitro have rarely been studied in detail and still are poorly understood (Leifert et
al., 1995; Swedlund & Locy, 1993). It has also been well documented that certain plant tissues
may contain and/or utilize different carbohydrates at the same time. It is then not surprising that
carbon sources other than sucrose might be effective in promoting in vitro tissue specific growth
responses in a given species (Swedlund & Locy, 1993). Furthermore, several reports have pointed
out these carbon sources as inducers of cell differentiation in some plant species (Lemos & Blake,
1996; Karhu, 1997; Tamil et al. 2012). Galactose, glycerol and sorbitol are examples of
alternative carbon sources that have been tested in vitro for different purposes in a wide range of
plant species (Swedlund et al.1993).
6. Conclusion Young inflorescence of Curcuma alismatifolia and Curcuma hybrid ‘Laddawan could
proliferated to somatic embryo when cultured on MS medium supplemented with 8, 10, 12, and 14
mg/l 2,4-D in dark condition for 4 weeks and got the highest average percentage at 14mg/l 2,4-D
which are 90 percent. Somatic embryo proliferated to shoot organogenesis from friable callus that
cultured on MS medium supplemented with 0.25g/l maltose. The highest average percentage of
shoot proliferated in Curcuma alismatifolia and Curcuma hybrid ‘Laddawan’ were 50.00 and
46.67 , respectively.
7. References Apavatjrut P, T. Sirisawad, P. Sirirugsa, P. Voraurai,C. Suwanthada. (1996) Studies on
chromosome number of seventeen Thai Curcuma species. In: Proceedings of 2nd national conference on flower and ornamental plant. 2: 86–99.
Balachandran, S.M., S.R. Bhat, and K.P.S. Chandel. 1990. In vitro clonal multiplication of turmeric (Curcuma spp.) and ginger (Zingiber officinale Rosc). Plant Cell Reports. 8: 521-524.
Jala, A. (2011) Effects of NAA BA and Sucrose On Shoot Induction and Rapid Micropropagation by Trimming Shoot Of Curcuma Longa L. American Transactions on Engineering & Applied Sciences, 3: (2) : 101 -109.
Karhu, S.T. (1997) Sugar use in relation to shoot induction by sorbitol and cytokinin in apple. Journal of the American Society for Horticultural Science. 122: 476-480.
Leifert , C., K.P. Murphy, P.J. Lumsden. (1995) Mineral and carbohydrate nutrition of plant
*Corresponding author (A. Jala). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail address: [email protected]. 2013. American Transactions on Engineering & Applied Sciences. Volume 2 No. 3 ISSN 2229-1652 eISSN 2229-1660 Online Available at http://TuEngr.com/ATEAS/V02/213-222.pdf
221
cell and tissue cultures. Critical Reviews in Plant Science.14: 83-109.
Lemos, E.E.P., J. Blake. (1996) Micropropagation of juvenile and mature Annona muricata L. Journal of Horticultural Science.71: 395-403.
Mohanty S, M.K. Panda, E. Subudhi, S. Nayak. (2008) Plant regeneration from callus culture of Curcuma aromatica and in vitro detection of somaclonal variation through cytophotometric analysis. Biol Plant 52:783–786
Murashige, T. and F. Skoog. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15: 473-479.
Purseglove JW, E.G. Brown, C.L. Green, S.R. Robbins. (1981) Spices. Longman, London, New York.
Salvi N.D., L. Geoge, S. Eapen. (2000) Direct regeneration of shoots from immature inflorescence cultures of turmeric. Plant Cell Tissue Org Cult. 62:235–238.
Schaffer M, P.M. Schaffer, J. Zidan, G.B. Sela. (2011) Curcuma as a functional food in the control of cancer and inflammation. Curr Opin Clinical Nutrition Metabolic Care. 4:588–597
Swedlund B and R.D. Locy. (1993) Sorbitol as the primary carbon source for the growth of embryogenic callus of maize. Plant Physiol. 103: 1339-1346.
Tamil C.M. Sundram, M. Suffian, M. Annuar and Norzulaani Khali.( 2012) Optimization of culture condition for callus induction from shoot buds for establishment of rapid growing cell suspension cultures of Mango ginger (Curcuma mangga) Australian Journal of Crop Science. 6(7): 1139-1146.
Toppoonyanont N, Chongsang S, Chujan S, Somsueb S, Nuamjaroen P. (2005) Micropropagation scheme of Curcuma alismatifolia Gagnep. Acta Hort (ISHS) 673:705–712
Udomdee W, S. Fukai, L. Petpradap, J.A. Teixeira da Silva. (2003) Curcuma: studies on tissue culture, pollen germination and viability, histology and flow cytometry. Prop Ornamental Plants. 3:34–41.
Viu A.F.M., M.A.O. Viu, A.R. Tavares, F. Vianello, G.P.P. Lima. (2009) Endogenous and exogenous polyamines in the organogenesis in Curcuma longa L. Sci Hortic. 121: 501–504.
Vu, J.C.V.; R.P. Niedz; G. Yelenosky . ( 1993) Glycerol stimulation of chlorophyll synthesis, embryogenesis and carboxylation and sucrose metabolism enzymes in nucellar callus of "Hamlin" sweet orange. Plant Cell, Tissue and Organ Culture. 33:75-80.
Wannakrairoj S (1997) Clonal micropropagation of patumma (Curcuma alismatifolia gagnep).
222 Anchalee Jala
Witthayasan Kasetsart. 31:353–356.
Yaping Kou; Guohua Ma, A. Jaime, Teixeira da Silva, Nian Liu. (2013). Callus induction and shoot organogenesis from anther cultures of Curcuma attenuata Wall. Plant Cell, Tissue and Organ Culture (PCTOC).112 : 1-7.
Zhang S.J, A.W. Liu N, G.H. Sheng, Ma, G.J. Wu. (2011) In vitro plant regeneration from organogenic callus of Curcuma kwangsiensis Lindl. (Zingiberaceae). Plant Growth Regul. 64:141–145.
Dr.Anchalee JALA is an Associate Professor in Department of Biotechnology, Faculty of Science and Technology, Thammasat University, Rangsit Campus, Pathumtani , THAILAND. Her teaching is in the areas of botany and plant tissue culture. She is also very active in plant tissue culture research.
Peer Review: This article has been internationally peer-reviewed and accepted for
publication according to the guidelines given at the journal’s website.