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Micropropagation: a general method for commercial bamboo production

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Micropropagation: a general method for commercial
bamboo production

Jan Oprins, Wim Grunewald1, Koen Gillis, Patricia Delaere, Hilde Peeters, Johan Gielis

Oprins Plant NV, Sint-Lenaartsesteenweg 91, B-2310 Rijkevorsel

1
Department Molecular Biotechnology, Ghent University, Coupure links 653, 9000 Ghent

Introduction
Bamboos are among the economically most important plants worldwide and interest in bamboo
as a source of biomass in forestry, agroforestry and sustainable agriculture systems has increased
rapidly in recent years. Bamboo is often considered as an ideal renewable resource for biomass,
useful for wood and paper industry. The feasibility of bamboo as a source of biomass has already
been shown convincingly in China and India, in a variety of applications, ranging from handicraft
(local industries), to industrial scale paper mills. Bamboo could play an important role as poverty
alleviator in many countries worldwide, in Asia, Africa and Tropical America. It has indeed been
stated that over 1 billion people live in houses made with bamboo, and over 750 million people,
mostly poor people, generate basic income cultivating, harvesting or transforming bamboo into
products.

Several international agencies including INBAR, FAO and UNIDO, national and regional
authorities, as well as private companies, are advocating and actively supporting the
establishment of agroforestry systems and or plantations. In NE India there is a lot of interest in
bamboo plantations with Cane and Bamboo Technology Centre CBTC, UNIDO and the NE
Council. Around the world large scale bamboo planting has been planned/ carried out, and,
several studies, pilot plantations or larger scale plantings are planned or carried out throughout
the world. Such approaches aim at sustainable supply of raw material for local industries. Natural
bamboo forests are not very productive and silvicultural management practices could partly
increase this productivity.

The whole downstream process of bamboo production and transformation consists of many
different steps. Each of these steps is important, and as in any successful industrial enterprise or
chain of processes, optimization in each step as well as integration of steps and feedback is
important. At the production side propagation and sustainable production, is crucial for mass
scale utilization of bamboo. At the very beginning, large scale propagation of bamboo with high
quality and low cost, is the pivotal upstream technology in the whole process. Estimates regarding
future use of bamboo all indicate that there will be a huge shortage for bamboo planting material
in medium and long term (Subramanlam, 1994; Nadgauda, 1997). To cope with this forecasted
shortage for the last two decades intensive research has been done on improved propagation
systems.

In Europe and North America, temperate bamboos originating from China and Japan, are used as
ornamentals for gardens, but there is increasing interest for uses in ecological applications and as

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energy crops. Traditionally, temperate bamboos are produced by division, but also here a number
of bamboo companies have tried to develop alternatives for this tedious and expensive process,
ranging from improved production techniques, to improved logistics.

For bamboo different propagation techniques are available, such as seed propagation, clump
division, rhizome and culm cuttings (Banik, 1994; 1995). In some cases the above propagation
methods have been applied successfully. Advantages of such methods are low cost and low
skilled labor. These methods are not general. Seeds are only available for some species, and
using seeds the genetic background and phenotypes are unknown. For temperate species seedling
propagation simply is not an option, nor is this method available for most tropical species.
Flowering and seed setting are very irregular in bamboos. The method of cuttings works
successfully in some cases, but it is impossible for most bamboos, especially temperate bamboos.
In many cases division is the only option.

Bamboo Tissue Culture and Micropropagation

While a variety of methods can be used for bamboo propagation, most of these methods are
mostly restricted to specific species and local conditions (such as availability of seeds). They are
not generally applicable methods. In addition, for mass scale propagation (> 500 000 plants per
year) classical techniques are largely insufficient and inefficient. Most of the classical techniques
for clonal propagation are only useful for the production on smaller scale (up to 10 000 plants per
year), although optimization or techniques such as macroproliferation (Adaresh Kumar, 1991), or
the ‘guaduines’ method in Guadua angustifolia can have higher production yields. Much will
depend of course on the availability of mother plants. Large scale propagation (between 10 000
and 500 000 plants per year) requires highly efficient techniques and lots of motherplants. For
mass scale propagation (> 500 000 plants per year) tissue culture is the only viable method.
Indeed, the order of magnitude of the demand for bamboo planting materials indicates that
micropropagation will inevitably be necessary for mass scale propagation (Rao et al., 1991;
Subramanlam, 1994; Gielis, 1998).

Since four decades many researchers and companies around the world have performed research to
develop efficient micropropagation technology for tropical and temperate bamboos. The first
paper on bamboo tissue culture was the germination of seed in test tubes (Alexander and Rao,
1969). Following the classical paper of Vasil and Vasil on the use of 2,4-D for the induction of
somatic embryogenesis, by the early eighties of the 20th century successful protocols had been
worked out based on somatic embryogenesis of tropical bamboos from seeds of Dendrocalamus
strictus and Bambusa bambos (Metha et al., 1982). For these same species also propagation by
axillary branching was successful (Nadgir et al., 1984). In that same decade groups succeeded in
producing plants through axillary branching, organogenesis or somatic embryos, from genera
such as Bambusa, Phyllostachys and Sasa in the classical series of five papers by LC Huang and
coworkers, Guadua angustifolia (Manzur, 1988), Bambusa beecheyana and Bambusa oldhamii
(Yeh and Chang, 1986). Many researches have focussed on somatic embryogenesis of seedlings
of tropical bamboos (INBAR, 1991). An inventory (BIC, 1994) shows that at least 21 labs in
South-east Asia were involved in bamboo tissue culture, mainly in India. Also in other countries
such as Singapore (Dekkers and Rao, 1987), and Thailand (Prutpongse and Gavinlertvatana,
1992), bamboo tissue culture research was very active. Also in Europe and the US several
research and commercial labs tried to develop micropropagation protocols for ornamental
bamboos.

2
Inevitably this short historical account is incomplete. In this busy period tissue culture was not
only used for propagation of bamboos, but research also included protoplast and cell cultures, salt
tolerant callus selection, and the induction of flowering. Especially, tissue culture flowering of
bamboos caused great excitement because it opened the way, at least theoretically, for
hybridization of bamboo (Nadgauda et al, 1990; Gielis, 1999). By now at least 130 papers on
micropropagation of bamboos have been published, original papers as well as reviews and some
in which tissue culture is described in a more general aspect (see Compendium of Research on
Bamboo, 1970-2003, Fernandez and Gielis, 2003). Other reviews on tissue culture of bamboo
can be found in INBAR, 1991; Saxena and Dhawan, 1994; Zamora, 1994; 2001; Nadgauda et al.,
1997; and Gielis, 1999).

At least 50 of these papers contain very specific details on the method developed. In annex to this
paper a table is given of selected research results, arranged per species, with details on the explant
types used, medium and culture conditions, and most important results. The references can be
found in the reference list, which is most complete list on bamboo tissue culture to this date.

When reviewing these data a number of trends are observed, with regard to commercial
propagation, the subject of this paper.

1. The main focus has been on tropical bamboos of the genera Bambusa and
Dendrocalamus. The main species covered are Bambusa bambos (in most papers as B.
arundinaceae), B. balcooa, B. oldhamii, B. ventricosa, B. vulgaris, B. tulda, Dendrocalamus
asper, D. giganteus, D. hamiltonii, D. latiflorus, D. longispathus, D. membranaceus and D.
strictus, with Bambusa bambos and Dendrocalamus strictus as absolute favorites.
2. Other species like Guadua angustifolia (Manzur, 1989) have been tested, but other
important bamboo genera like Gigantochloa, Schizostachyum, Melocanna, or African bamboos
have not been subject of successful research. Only three groups have focused on temperate or
semi-hardy bamboos, with Phyllostachys aurea (Huang and Huang, 1989) and Sasa pygmaea
(Pleioblastus pygmaeus; Huang and Huang, 1989; Watanabe et al., 2000), and Thamnocalamus
spathiflorus (Bag et al., 2000).
3. In addition, research has focused on one or two species only. Very few groups have
concentrated on more than one species. Only one paper mentions a large range of species tested
(Prutpongse and Gavinlertvatana, 1992).
4. Few protocols have been fully successful enough for conversion into commercial
production schemes, especially for axillary branching. The major problem always seems to be
rooting (see below). Only in the case of Dendrocalamus asper propagation through axillary
branching has been very successful.
5. Somatic embryogenesis SE has been successful for a limited number of species only,
mainly starting from seeds. Much less success has been achieved in inducing SE in adult
bamboos. However, in some well-detailed cases, somatic embryogenesis is also associated with
the induction of flowering (Rao and Raghav, 2002, Lin and Chang, 2003), an unwanted
phenomenon in production.
When reviewing all these data, the conclusion is that at the research level, much work will need
to be done further to develop good propagation methods, using tissue culture. In the last five
years an increase is observed in the number of papers published in international journals. This is
definitely a positive trend, and improvements of protocols may be expected in the following
decade. In publishing research results, focus must be placed, so some of these protocols may have

3
been further developed in these groups as a more general protocol of commercial propagation for
other species.
For tissue culture of bamboo the use of starting material (seeds or adult plants) and the choice of
the propagation method are crucial (Gielis, 1999; Gielis and Oprins, 2002). The two major
advantages of using seedlings are that seedlings establish a new generation, and that the
technology is easier. But the disadvantages are considerable: (1) insufficient or no knowledge of
genetic background, (2) restricted availability of seeds for most species and rapid loss of
germination capacity, and (3) comparison of in vitro to in vivo performance has not been
thoroughly evaluated. In addition there is a huge variability in responsiveness in tissue culture
(Saxena and Dhawan, 1994).

When using adult bamboos main problems are: (1) endogenous contamination, (2) seasonal
influences and instability of multiplication rates, and (3) many problems with rooting (also in
bamboos that root fairly well in classical propagation). Rooting percentages for adult bamboos
ranged from very low percentages of 10% for Bambusa vulgaris to 73% for adult Dendrocalamus
longispathus (Saxena and Dhawan, 1994). 55,9% rooting was found in Bambusa edulis (Lin and
Chang, 1998), and 68% for Dendrocalamus hamiltonii (Saxena and Bhojwani, 1991). The
combination of photomixotrophic in vitro multiplication and photoautotrophic in vitro rooting
stages may result in improved transplanting success (Watanabe et al., 2000). A rooting
percentage of 77% was obtained for adult Dendrocalamus giganteus in 3 or 4 weeks
(Ramanayake and Yakandawala, 1997). But, while 77% of success is good on 500 plants in a
laboratory experiment it still represents a loss of 33000 when you transplant 100 000 plants. Low
rooting frequencies still remain the major bottleneck to developing commercially viable protocols
(Saxena, 1993).

Large scale production of bamboo based on micropropagation

Micropropagation via tissue culture thus attracted a lot of attention but the translation and
transformation of these expectations into commercially viable propagation systems has been beset
with a number of problems that were either technological, or related to marketing. At present
only a very limited number of laboratories specialize in the production of bamboos at a
commercial scale (Gielis et al., 2001). One of the main problem is that micropropagation of
bamboos also has to deal with the development of new markets and market opportunities.
Logistics and supply chains such as in ornamental horticulture where tissue culture labs are
separate entities seem not to work. Instead the laboratories have to develop their own markets,
marketing and sales for bamboo plants (Gielis and Oprins, 1998).

This strategy of forward integration needs to go beyond the classical scheme of tissue culture.
The tissue culture phase terminates at Stage III, but in regard to marketing of plants one can also
distinguish subsequent stages: (1) Stage IV, the transplantation stage with the end product being
rooted plantlets in trays, (2) Stage V, the production of liners, either for production of saleable
plants or for use as micromotherplant, and (3) Stage VI, the production of saleable plants. This
distinction is important if the complete chain of production is integrated in a single company,
since this determines the added values. In our case the laboratory produces ornamental plants
exclusively for Oprins Plant’s own nurseries.

The lab currently has over 90 genotypes in culture initiated from adult genotypes of which about
40 are produced on a commercial scale (Gielis and Oprins, 1998, 2002). Species and varieties of
Arundinaria, Chimonobambusa, Fargesia, Phyllostachys, Pleioblastus, Pseudosasa, Sasa,

4
Sasaella, Semiarundinaria, Shibataea, Thamnocalamus and Yushania (temperate bamboos) and
Bambusa, Dendrocalamus, Dinochloa, Oxythenanthera and Thyrsostachys (tropicals) are
produced commercially. Rooting percentages of species are between 85 and 100%, depending on
species and season of transplanting (unpublished results).

These bamboos are produced either as ornamental, or for tropical forestry. Our main focus of the
production is in Europe with bamboo for ornamental purposes, and we produce between 750 000
and 1 million plants annually. Micropropagations is now fully integrated in the complete plant
production chain at Oprins Plants. Bamboos produced in tissue culture and hardened and rooted,
are shipped to production units of Oprins Plant in Belgium, France and Spain. There the plants
are grown further to the stage of saleable plants for the ornamental plant market. With
micropropagation we have been able to rapidly expand our production and enter into new
markets. Indeed, for horticulture, micropropagation of bamboos has allowed to develop a new
type of ornamental bamboo that can be produced year round with a high quality / price ratio and
distributed far more widely than classically propagated ornamental bamboos. Additional
advantages are vigorous growth and healthy leaves.

A balanced production and logistic chain, where bamboos are produced in Belgium and grown in
the nurseries in Spain and France, has allowed to produce high quality ornamental bamboos year
round at prices which are well below the current market prices for bamboo. These plants are
currently being marketed and sold under the brand name BambooSelectTM, our own brand.
Micropropagation has allowed for fast propagation of both tropical and temperate species, not
only ornamental bamboos. In an agricultural perspective it is now possible to produce the same
genotypes for large scale plantations in Europe at prices comparable to other energy crops (Gielis,
2000). Trial plantations with micropropagated plants have been set up in various parts of the
world.

Plant quality and genetic stability in micropropagation

In our propagation system, major emphasis is placed on quality and true-to-type production. In
tissue culture 3 basic methods exist for the propagation of plants (i.e. micropropagation): (1)
axillary branching, which mimicks the natural process of branching, (2) organogenesis, the
neoformation of organs or shoots in calli, and (3) somatic embryogenesis, the regeneration of
embryos from somatic cells. In bamboo all these three methods have been investigated, but in
bamboo both organogenesis and SE involve an intervening callus phase.

Among these methods axillary branching is definitely the most reliable. In our own production
we have tested this method for over 50 species. We have always strongly advocated using axillary
branching as the method of choice for commercial propagation of bamboo (Gielis and Oprins,
1998, 2002; Gielis, 1999), since somatic embryogenesis may cause several problems. Indeed, it
has been recently been shown that somatic embryogenesis produces the unwanted phenomenon of
flowering (Rao and Raghav, 2002; Lin and Chang, 2003). In addition, callus may cause problems
with genetic stability. In bamboo somatic embryogenesis has been successfully applied in some
tropical species initiated from seeds, but long term studies of the behavior of plants in the field
are not known.

However, even when using axillary branching, caution must be taken. The bulbous internodes of
Bambusa ventricosa were lost after tissue culture (Huang and Huang, 1995) and variegated leaves
appear very often during tissue culture in many species. For large or mass scale production,

5
efficiency of the propagation methods is important, but equally important is the genetic stability.
The evaluation of clonal stability is very difficult and time consuming in bamboos and since
feedback times are very long in bamboos to monitor the genetic stability, ensuring genetic
stability is very important. The molecular tools to test for genetic stability are in general very
rough approximations.

In most plants produced on large scale, axillary branching is used since this methodology has a
lesser risk of aberration. In our technology we have developed a Best Practice method to ensure
the best possible quality, which is also reflected in the BambooSelect brand. Some of these
practices are:

1. Micropropagation cultures are only initiated from selected mother plants, which are kept in
special section of our greenhouses. Only adult plants are used, with known phenotypes.
2. Cultures for large scale production are renewed annually, so that high production genotypes
are multiplied maximum of 16 subcultures using axillary branching.
3. Labelling of plants and cultures is fully computerized and a very specific management method
(HACPP) is being implemented.
4. Propagation rates (multiplication rates) are kept below five. Overproducing cultures are not
used in production.
5. People are trained in Positive Mass Selection PMS. During transfer of cultures only the
qualitatively best cultures are retained, the others are simply discarded. PMS is applied from the
very stage of initiation up to the final stage of the tissue culture process.
6. For ornamental bamboos we only sell large saleable plants, not liners or tissue culture plants.
This allows quality quality checks to the final moment of sales.

In addition we are using molecular methods to test if tissue culture have effects on the genome,
using general methods such as AFLP and methylation sensitive AFLP (Gielis et al., 2002: Gielis
et al., this conference). Our tests so far have shown that no differences in AFLP patterns can be
found (Gielis et al. 2002) when Phyllostachys-genotypes sampled in the laboratory were tested.
These included P. aurea, P. aureosulcata ‘Spectabilis’, P. aureosulcata ‘Aureocaulis’, P. bissetii,
P. humilis, P. nigra, P. nigra ‘Henonis’ and P. vivax. Of all these types, except P. nigra, samples
were also taken in the greenhouse of hardened tissue cultured plants. They included cultures
which had been kept for 1, 2 or 3 years. No differences could be shown using AFLP.
Additionally, we use AFLP to double check our accessions to well known bamboo collections for
clonal fidelity.

Conclusion
In the past 15 years we have developed not only a very general tissue culture methodology for
bamboo, but we have also been able to fully integrate micropropagation in the complete plant
production system and the logistics operations at Oprins Plant. The BambooSelect brand has
become a well-known trademark in horticulture and several new introductions have won prices in
exhibitions. In our technology the major focus is on quality production, with emphasis on true-to-
type production. Currently, research is ongoing to further improve propagation of bamboo for
more efficient production.

6
The main production is towards ornamental horticulture, but our technology is also suitable for
the large-scale production of tropical bamboos. The interest in bamboo plantations and
agroforestry systems, exerts a strong demand for quality planting material. Various studies are
underway to study the feasibility of technology transfer to bamboo countries.

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11
Annexe: Overview of published research from 1982 until 2004. References can be found in Compendium of Research on Bamboo 1970-2003.

Abbreviations used

Abbreviations: MS Murashige & Skoog (1962)

N6 Chu (1978)
B5 Gamborg et al.(1968)
2,4,5-T 2,4,5-trichlorophenoxyacetic acid
2,4-D 2,4-dichlorophenoxyacetic acid
ABA Absiscic acid
AC activated charcoal
AdS adenine sulphate
6
BA N -benzyladenine
6
BAP N -benzylaminopurine
BSA bovine serum albumin
CW coconutwater
GA3 gibberellic acid
IAA indole-acetic acid
IBA indole-3-butyric acid
Kn Kinetin
MES 2-(N-morpholino) ethanesulfonic acid
MMS MS with half normal concentration of
micronutrients
NAA 1-naphtaleneacetic acid
PEG polyethylene glycol
PVP polyvinylpyrrolidone
TDZ thidiazuron
- not mentioned

1
Species explant medium conditions results reference

Bambusa arundinacea surface-sterilized N6 basal (modified)+ meso-inositol (100 16 h daily light normal seedling Mehta et al. (1982)
grains mg/l) + sucrose (5%) + agar (1%); regime (1500
(pH=6) lux) at 27°C
-5
Bambusa arundinacea surface-sterilized + 2,4-D (3.10 M) callus obtained from the embryonal axis Mehta et al. (1982)
grains

-5 -6
Bambusa arundinacea surface-sterilized + 2,4-D (3.10 M) + BAP (5.10 M) + greater proliferation and somatic embryogenesis Mehta et al. (1982)
grains PVP (250 mg/l)
Bambusa arundinacea seedling explants MS + 2% sucrose + 5% CW + 2,2µM BA 24 h daylight at flowering cultures Nadgauda et al. (1997)
28°C
Bambusa arundinacea, node MS + CW(10%) + BAP(0,5) + Kn (0,2) multiple shoots Nadgir et al. (1984)
Bambusa vulgaris,
Dendrocalamus strictus
Bambusa arundinacea, shoot MS + IBA formation of roots Nadgir et al. (1984)
Bambusa vulgaris,
Dendrocalamus strictus
Bambusa arundinacea, shoot 1/2 MS + AC(0,25%) rooting Nadgir et al. (1984)
Bambusa vulgaris,
Dendrocalamus strictus
Bambusa arundinacea, seedling MS + CW(5%) + BAP(0,2) multiple shoots Nadgir et al. (1984)
Bambusa vulgaris,
Dendrocalamus strictus
Bambusa arundinacea, shoot MS + IBA(0,1) root induction Nadgir et al. (1984)
Bambusa vulgaris,
Dendrocalamus strictus
Bambusa arundinacea, shoot MS + NAA(1) + IBA(1) + 2,4-D(0,5) + Nadgir et al. (1984)
Bambusa vulgaris, phloroglucinol
Dendrocalamus strictus
Bambusa balcooa nodal cuttings 1/2 MS + sucrose (6%) + BAP (4.10-6 - sprouting of the axillaries after 3-4 d, multiple Roohi et al. (1991)
obtained from M) + Kn (1.10-5 M) shoots from 4 to 15 (average of 6)
mature culms

2
Bambusa balcooa shoots 1/2 MS + sucrose (6%) + BAP (4.10-6 - shoot clusters (continous growth and multiplication) Roohi et al. (1991)
M) + Kn (1.10-5 M)

Bambusa balcooa shoots 1/2 MS + sucrose (6%) + BAP (4.10-6 - 16% rooting and 75% survival of the rooted plants Roohi et al. (1991)
M) + Kn (1.10-5 M) + NAA (5.10-6 M) after acclimatization
-6 -5
Bambusa bambos surface-sterilized MS medium + BAP (8.10 M to 10 M); continuous seeds germinated within 1 week and produced up Rao & Raghav (2002)
seeds pH = 5,5 illumination to 30 shoots within 8-10 weeks of culture
(provided by
cool white
fluorescent
lamps) at 25°C
-6
Bambusa bambos multiple shoots in MS medium + BAP (5.10 M) identical; multiplication Rao & Raghav (2002)
clusters of 4-5 subculture
shoots period of 3
weeks
-7
Bambusa bambos multiple shoots in MS medium + BAP (5.10 M) - flowering was observed in nearly 40% of the culture Rao & Raghav (2002)
clusters of 4-5
shoots
-5 -5
Bambusa bambos multiple shoots MS medium + 2,4-D (10 M, 2.10 M) - compact callus at the base of the multiple shoots Rao & Raghav (2002)
which were raised within 2 weeks of culture
on MS + BAP for
10-12 weeks

Bambusa bambos green embryoids same medium - about 70% of the embryoids germinated and Rao & Raghav (2002)
in the compact flowered within 4 weeks of culture; 6-8 florets (1-1,2
callus cm) per spike, florets showed anthesis, pollen was
viable but no seed set

Bambusa beechayana young MS + myo-inositol + nicotinic acid + darkness or darkness leads to brown callus, further cultures Yeh & Chang (1986)
Munro var. Beecheyana inflorescences pyridoxine HCl + thiamine HCl + glycine 16/8 hours gave nodular structures who turned into clusters of
+ casein hydrolysate + sucrose + agar + daylight at 26 +- embryoids
2,4-D or Kn 1° C

3
Bambusa beecheyana sterile roots of MS (=MS basal + myo-inositol (100 darkness or a white and pale-yellow-colored callus (12 weeks) Chang & Lan (1995)
Munro var. Beecheyana regenerated mg/l) + nicotinic acid (0,5 mg/l) + dark/light cycle that became globular, cluster-like, translucent (28
bamboo pyridoxine HCl (0,5 mg/l) + thiamine HCl of 16/8 h of 25- weeks) and eventually embryogenic
-2 -1
(0,1 mg/l) + glycine (2 mg/l) + casein 40 µmol.m .s
hydrolysate (1 g/l)) + sucrose (30-60 g/l) at 20°C;
+ Kn (2 mg/l) + 2,4-D (3 mg/l) + Sigma subculture
agar (7 g/l); pH = 5,7 every 8 weeks

Bambusa beecheyana embryogenic identical medium but changing somatic embryos Chang & Lan (1995)
Munro var. Beecheyana callus osmolarity (2-8% PEG or mannitol) or
adding of abscisic acid (0,1-2 mg/l),
coconut milk (0,1-1%), polyamines (1-
100 mg/l), derivatives of salicylic acid
(10-200 mg/l) improved somatic embryo
formation and addition of activated
charcoal (0,2%) reduced browning and
senescence
Bambusa beecheyana somatic embryos MS (=MS basal + myo-inositol (100 dark/light cycle germination of embryoids and subsequent Chang & Lan (1995)
Munro var. Beecheyana mg/l) + nicotinic acid (0,5 mg/l) + of 16/8 h of 25- development of plantlets
-2 -1
pyridoxine HCl (0,5 mg/l) + thiamine HCl 40 µmol.m .s
(0,1 mg/l) + glycine (2 mg/l) + casein (PPF, day-light
hydrolysate (1 g/l)) + sucrose (30-60 g/l) fluorescent
+ Kn (2 mg/l) + 2,4-D (3 mg/l) + Sigma tubes, 40 W)(at
agar (7 g/l); pH = 5,7 20°C)
dramatically
enhanced
germination of
embryoids

Bambusa edulis nodal explants MS + sucrose (3%) + gelrite (2 g/l) + artificial light of healthy and vigorous shoots (average of 3,5 shoots Lin & Chang (1998)
-2 -1
from 10-year-old TDZ (0,1 mg/l); pH = 5,7 54 µmol.m .s from 1 shoot every 3 weeks)
field-grown culms with a light/dark
cycle of 16/8 h
at 26°C;
subcultured
every 3 weeks

Bambusa edulis regenerated MS + sucrose (3%) + gelrite (2 g/l) + 55,9% rooting and 100% survival in the field of Lin & Chang (1998)
shoots TDZ (0,01 mg/l) + 2,4-D (0,5 mg/l); pH = rooted plantlets without hardening
5,7

4
Bambusa edulis regenerated MS + sucrose (3%) + gelrite (2 g/l) + 13,5% flowering cultures Lin & Chang (1998)
shoots TDZ (0,01 mg/l); pH = 5,7
Bambusa edulis segment (ca. 2 MS medium + TDZ (0,1 mg/l) + 2,4-D (3 dark at 26°C callus Lin & Chang (2003)
mm in length) of in mg/l) + Gelrite (0,22%); pH = 5,7
vitro spikelets
Bambusa edulis somatic embryos MS medium + TDZ (0,1 mg/l); pH = 5,7 artificial lighting multiple shoots (4,19 culms; 0,856 spikelets; 25,9% Lin & Chang (2003)
with daylight spikelets/culms); 47,5% flowering
fluorescent
tubes (16/8 h
light/dark cycle);
subcultured
each 21 days

Bambusa edulis multiple shoots MS medium + 2,4-D (1 mg/l) - 3 types of rooted plantlets (spikelets only, Lin & Chang (2003)
vegetative culms only, spikelets and vegetative
culms); 67,7% rooting and 10% flowering after 1
month; 82,8% rooting and 23,3% flowering after 2
months

Bambusa edulis multiple shoots MS medium + sucrose (30 g/l) + TDZ 26°C at 16 h formation of de novo inflorescences Lin et al. (2003)
and (0,1 mg/l) photoperiode of
-2 -1
inflorescences 54 µmol.m .s

Bambusa edulis inflorescence MS medium + sucrose (30 g/l) + NAA (5 26°C at 16 h flower development, adventious root formation and Lin et al. (2003)
mg/l) photoperiode of de novo vegetative shoot formation
-2 -2
54 µmol.m .s

Bambusa edulis inflorescence MS medium + sucrose (30 g/l) + G A3 26°C at 16 h no development of normal flowers Lin et al. (2003)
photoperiode of
-2 -3
54 µmol.m .s

5
Bambusa flexuosa young leaf MS + 2,4-D (3 to 6 mg/l) - callus Vasana (1985)
Thyrsostachys siamensis
Bambusa arundinacea

Bambusa flexuosa young leaf MS + 2,4-D (1 to 3 mg/l) - callus Vasana (1985)

Thyrsostachys siamensis
Bambusa arundinacea

Bambusa flexuosa callus MS + BAP (2 mg/l) + NAA (0,5 mg/l) - plantlets Vasana (1985)
Thyrsostachys siamensis
Bambusa arundinacea

Bambusa glaucescens culm bud MS + BAP(1) + AC(3g/l) shoot formation Banik (1987)
Bambusa glaucescens shoot MS + BAP(5) + AC(3g/l) + NAA(1) rooting Banik (1987)
Bambusa glaucescens internodes MS + 100 mg/l myo-inositol + 0,5 mg/l elongated shoots Jullien & Van (1994)
'Golden goddess' nicotinic acid + 0,5 mg/l pyridoxin-HCl +
1 mg/l thiamine-HCl + 2 mg/l glycine +
15g/l + 20µM NAA

Bambusa glaucescens elongated shoots MS + 30 g/l sucrose + 250 mg/lcasein callus formation Jullien & Van (1994)
'Golden goddess' hydrolysate + 40 ml coconut milk + 8 g/l
agar + 500 mg/l PVP + 2,4-D + IAA +
BA + ABA
Bambusa multiplex plant regenerating medium + NAA (1 mg/l) + BA (1 mg/l) 16h/d to 45 adventious shoots Huang & Huang (1991)
-2 -1
Sasa pygmaea callus µmol.m .s
illumination at
25°C
Bambusa multiplex shoots medium + NAA 16h/d to 45 shoots readily rooted Huang & Huang (1991)
-2 -1
Sasa pygmaea µmol.m .s
Bambusa oldhamii illumination at
Phyllostachys aurea 25°C

6
Bambusa multiplex shoot apices MS salts + sucrose (30 g/l) + myo- 16h daily initiation of nodular callus Huang et al. (1989)
(Loureiro) Raeuschell inositol (100 mg/l) + thiamine HCl (1 exposure to 4,5
-2 -1
mg/l) + nicotinic acid (0,5 mg/l) + nEcm .s at
pyridoxine HCl (0,5 mg/l) + glycine (2 27°C
mg/l) + TC agar (8 g/l) (pH=5,7) + NAA
(1 mg/l) + BA (1 mg/l)

Bambusa multiplex shoot apices medium + NAA (1 mg/l) + BA (1-3 mg/l) adventitious shoots and organogenic calli Huang et al. (1989)
(Loureiro) Raeuschell
Bambusa multiplex protoplasts basal medium (Huang et al., 1988) + low intensity more than 30% of protoplasts had ondergone Huang et al. (1990)
(Loureiro) Raeuschell BSA (0,01%) + malt extract (0,05%) + illumination division in 3 d
Bambusa oldhamii Munro MES (10 mM) + arginine HCl (50 mM) + (16h/d with 22,5
-2 -1
mannitol (0,6 M) + FMC Sea Plaque µmol.m .s ) at
agarose (1,2%) 25°C
Bambusa multiplex liquid medium 0% division Huang et al. (1990)
(Loureiro) Raeuschell
Bambusa oldhamii Munro
Bambusa multiplex feeder cells more than 50% of protoplasts had divided after 42d Huang et al. (1990)
(Loureiro) Raeuschell
Bambusa oldhamii Munro

Bambusa multiplex lowering mannitol conc. 2 weeks 31,7% of protoplasts had divided and 30% had Huang et al. (1990)
(Loureiro) Raeuschell 0,6 M 4 weeks 0,4 M attained the 26-200 cells/cluster stadium
Bambusa oldhamii Munro 1 week 0,2 M
Bambusa oldhamii inflorescence MS + 2,4-D(3) + Kn(2) callus formation + embyoids Yeh & Chang (1986)
Bambusa oldhamii embryoid MS + Kn(2) embryoid germination Yeh & Chang (1986)
Bambusa oldhamii root MS + 2,4-D(3) + Kn(2) albino plantlets Yeh & Chang (1986)
Bambusa oldhamii shoot tips MS + sucrose (3%) + vitamins + glycine darkness at callus could be initiated and grew vigorously Huang & Murashige
Bambusa multiplex + myo-inositol (100 mg/l) + 2,4-D (3 27°C (1983)
Sasa pygmaea mg/l) + agar (0,8%)
Phyllostachys aurea
Bambusa oldhamii young leaf tissue - callus Huang & Murashige
Bambusa multiplex near the meristem (1983)
Sasa pygmaea
Phyllostachys aurea

7
Bambusa oldhamii Munro inflorescence MS basal + MS Vit + 2,4-D (3 mg/l) + darkness or 3 types of calli: 2 non-embryogenic lines and 1 Ho & Chang (1998)
tissue sucrose (6%) + agar (7 g/l) (pH=5,7) under embryogenic
fluorescent light
-
of 15-40 µmol.s
1 -2
.m on 16/8 h
day/night
regime at 26°C

Bambusa oldhamii Munro embryogenic MS basal + MS Vit + 2,4-D (3 mg/l) + darkness at long term maintenance of embryogenic callus (11 Ho & Chang (1998)
callus sucrose (6%) + Kn (2 mg/l) + Gelrite 26°C, explants year)
(2,2 g/l) were
subcultured
every 2 months

Bambusa oldhamii Munro subcultured MS basal + MS Vit + BA (1 mg/l) + NAA fluorescent light after 16 months of subculturing: 85% produced Ho & Chang (1998)
-
embryogenic (2 mg/l) + sucrose (6%) + Gelrite (2,2 of 15-40 µmol.s embryoids and subsequent plantlets
1 -2
callus g/l) .m on 16/8 h
day/night
regime at 26°C

Bambusa oldhamii Munro plant regenerating medium + NAA (1 mg/l) + BA (3 mg/l) adventious shoots Huang & Huang (1991)
callus

Bambusa oldhamii Munro medium + NAA (1 mg/l) + BA (1-3 mg/l) adventitious shoots and organogenic calli Huang et al. (1989)

Bambusa oldhamii Munro shoot apices (0,8- MS salts + sucrose (30 g/l) + myo- constant truly unorganized, highly friable, creamy white Huang & Huang (1991)
Bambusa multiplex 1,2 mm tall) inositol (100 mg/l) + thiamine HCl (1 darkness at callus
(Loureiro) Raeuschell mg/l) + nicotinic acid (0,5 mg/l) + 25°C
Phyllostachys aurea A. and pyridoxine HCl (0,5 mg/l) + glycine (2
C. Riviere mg/l) + 2,4-D (3 mg/l) + Gelrite (0,2%)
Sasa pygmaea (Miquel) E.
G. Camus
Bambusa oldhamii Munro shoot apices (0,8- identical medium + NAA + BA constant granular or nodular callus Huang & Huang (1991)
Bambusa multiplex 1,2 mm tall) darkness at
(Loureiro) Raeuschell 25°C
Phyllostachys aurea A. and
C. Riviere
Sasa pygmaea (Miquel) E.
G. Camus

8
Bambusa oldhamii Munro creamy, white liquid medium + 2,4-D medium on liquid suspension culture Huang & Huang (1991)
Bambusa multiplex callus shaker (50
(Loureiro) Raeuschell rpm),
Phyllostachys aurea A. and subculturing
C. Riviere every 3 weeks,
Sasa pygmaea (Miquel) E. 16h/d to 22,5
-2 -1
G. Camus µmol.m .s
illumination at
25°C

Bambusa oldhamii Munro protoplasts medium + BSA (0,1%) + arginine HCl 16h/d to 22,5 callus Huang & Huang (1991)
-2 -1
Bambusa multiplex (50 mM) + MES (10 mM) µmol.m .s
(Loureiro) Raeuschell illumination at
Phyllostachys aurea A. and 25°C
C. Riviere
Sasa pygmaea (Miquel) E.
G. Camus
Bambusa oldhammii shoots initiated MS medium + sucrose (3%) +BA (13 25 +/- 2°C 16 h propagation with culture browning to several Huang et al. (2002)
Munro, Phyllostachys nigra from 5-10 mm µM) for B. oldhammii + TDZ (0,0045µM) photoperiode of extends dependent on the medium
-2 -2
(Lodd.) Munro shoot tip of new for P. nigra + ascorbic acid, cysteïn, 45 µmol.m .s
laterals that ferulic acid, kojic acid, or thiourea (0-30
emerged on mM) + PVP (0-10 mM, pH 4,5-8
elongating culms
Bambusa tulda shoots obtained liquid 1/2 MS + BAP (8.10-6 M) + Kn - axillary shoot proliferation at a rate of 4-5x every 3 Saxena & Bhojwani
from 3-weeks-old (4.10-6 M) + sucrose (2%) weeks (1991)
aseptic seedlings

Bambusa tulda shoots liquid MS + BAP + Kn shoot multiplication Saxena (1990)

-5
Bambusa tulda shoots MS (NH4NO3 1/2x) + IBA (1.10 M) + - 92% rooting; 80% survival after transfer to the field Saxena & Bhojwani
-5
coumarin (6,8.10 M) (1991)
Bambusa tulda single node MS + BAP (1,2.10-5 M) + Kn (3.10-6 M) - 90% exhibited bud-break within 5-6 days Saxena & Bhojwani
segments from (1991)
lateral branches of
mature clumps

9
 -5 -6
Bambusa tulda excised shoots MS + BAP (1,5.10 M) + IBA (3.10 M) - 4x multiplication in 3 weeks Saxena & Bhojwani
(1991)

Bambusa tulda MS + IAA + coumarin rooting Saxena (1990)

Bambusa ventricosa node MS + BAP(3-40); BAP (5) + NAA(0,1- shoots Dekkers (1989)
10) +- AC(0,3%)
Bambusa ventricosa culm sheath base MS + 2,4-D(0,5 - 25); NAA(3-10); callus Dekkers (1989)
IBA(10); IBA + NAA(5+5)
Bambusa ventricosa internode MS + 2,4-D(5) callus Dekkers (1989)
Bambusa ventricosa culms sheath MS + sucrose (30 g/l) + 2,4-D (0,5-25 12 h callus induction Dekkers & Rao (1987)
Schizostachyum base explants mg/l) +NAA (3 and 5 mg/l) + agar (8 g/l) photoperiod at
brachycladum 27°C
Thyrsostachys siamensis

Bambusa ventricosa nodal explant from MS + sucrose (30 g/l) + BAP (5 mg/l) + 12 h 95% showed bud breaking Dekkers & Rao (1987)
McClure young leaves agar (8 g/l) photoperiod at
27°C
Bambusa ventricosa shoot apices (1 MS salts + MS Vit +myo-inositol (100 16h daily axillary shoot differentiation Huang & Huang (1995)
McClure mm) excised from mg/l) + sucrose (3%) + BA (4,44 µM) + exposure to 45
-2 -1
newly emerged TC agar (0,8%) or Gelrite (0,2%) µM.cm .s
laterals of young illumination at
culms 25°C;
transferred to
fresh medium
after 4 weeks
Bambusa ventricosa shoots MS salts + MS Vit + myo--inositol (100 rooted shoots Huang & Huang (1995)
McClure mg/l) + sucrose (3%) + BA (0,44 µM) +
NAA (5,4 µM) + TC agar (0,8%) or
Gelrite (0,2%)
Bambusa vulgaris single node MS + BAP (1,2.10-5 M) + Kn (3.10-6 M) - 70% bud-break Saxena & Bhojwani
segments (1991)

10
Bambusa vulgaris nodal segments MS + 555 µM myo-inositol + 4,06µM 16 h daylight at sprouted shoots after 10-12 days Rout & Das (1997)
with axillary buds nicotinic acid + 2,43 µM pyridoxine-HCl 25°C
+ 0,296 µM thiamine-HCl + 3%
saccharose + 0,8% agar
Bambusa vulgaris shoots MS + BAP (2,5.10-5 M) + Kn (5.10-6 M) - shoots have been multiplied for over 16 passages Saxena & Bhojwani
at a rate of 2,3x every 6 weeks (1991)
Bambusa vulgaris shoots (0-2 d) MS + IBA (100 mg/l) + coumarin - 10% rooting Saxena & Bhojwani
(100 mg/l); (2d-…) MS basal (1991)
Bambusa vulgaris sprouted shoots MS + BAP + Kn + IBA + 2,4-D dark or light (16 friable calli Rout & Das (1997)
h photoperiod)
at 25°C

Bambusa vulgaris friable calli MS + 13,33 µM BAP + 2,46 µM IBA 16 h daylight at regeneration Rout & Das (1997)
25°C
Bambusa vulgaris excised shoots 1/2 MS + 0,49 µM IBA 16 h daylight at root formation Rout & Das (1997)
25°C
Bambusa vulgaris excised zygotic MS basal + Kn (0,25 mg/l) + 2,4-D (3 dark at 25°C creamy-white, nodular and friable callus Rout & Das (1994)
Dendrocalamus giganteus embryos & nodal mg/l); pH = 5,7 with 55%
Dendrocalamus strictus explants of in vitro relative humidity
grown seedlings

Bambusa vulgaris callus 1/2 MS + Kn (0,5 mg/l) + 2,4-D (2 mg/l) 16h photoperiod greenish-white embryogenic calli and subsequent Rout & Das (1994)
Dendrocalamus giganteus + adenine sulphate (10 mg/l); pH = 5,7 with cool white somatic embryos and normal plants
Dendrocalamus strictus fluorescent light
-2 -1
(50 µE.m .s )
at 25°C and
55% RH

Bambusa vulgaris shoots from nodal liquid 1/2 MS + sucrose (3%) + adenine 60-70% in vitro flowering after 12 weeks of Rout & Das (1994)
Dendrocalamus giganteus explants taken sulphate (0,5 mg/l) + IBA (0,25 mg/l) + subculture
Dendrocalamus strictus from somatic GA3 (0,5 mg/l)
embryo
regenerated
plants
Bambuseae (54 species) seeds MS + BA (0,44 - 8,8µM) + NAA (2,7 or 16 h daylight at Prutpongse &
5,4 µM) + 2,4-D (0,45 - 27 µM) 25°C Gavinlertvatana (1992)

11
Bambuseae (54 species) inflorescences MS + 2,4-D (11,3 - 45,0 µM) + NAA (5,4 16 h daylight at callus (2,4-D 13,5 to 27,0 µM) Prutpongse &
µM) 25°C Gavinlertvatana (1992)

Bambuseae (54 species) stem-node MS + 88µM sucrose + 0,6% agar + 16 h daylight at multiple shoots (BA 22µM) Prutpongse &
sections NAA (2,7 - 10,8 µM) + BA (2,2 - 44,0µM) 25°C Gavinlertvatana (1992)

Bambuseae (54 species) meristem domes MS + 2,4-D (0,45 - 9,0 µM) + NAA (0,54 16 h daylight at callus (2,4-D and coconut water) Prutpongse &
- 5,4 µM) + BA (2,2 or 4,4 µM) + CW 25°C Gavinlertvatana (1992)
(10% or 20%)
Bambuseae (54 species) leavesections MS + 2,4-D (4,5 - 81,0µM) 16 h daylight at callus (2,4-D 13,5 to 27,0 µM) Prutpongse &
from underground 25°C Gavinlertvatana (1992)
shoots

Dendrocalamus asper axillary buds from liquid or semi-solid MS + sucrose (3%) + 16h illumination axillary bud break in all healthy cultures within 2 Arya & Arya (1996)
young juvenile myoinositol (100 mg/l) + different cycle of 3000 weeks
shoots concentrations of BA and Kn + agar- lux light
agar (0,7%) intensity from
cool white
fluorescent
tubes, at 26°C

Dendrocalamus asper surface sterilized MS + sucrose (3%) + myo-inositol (100 multiple shoots (1-20) were formed immediately Arya & Arya (1996)
seeds mg/l) + BA (1-10 mg/l) + agar-agar after seed germination
(0,7%)

Dendrocalamus asper shoots MS + sucrose (3%) + myo-inositol (100 very high shoot multiplication (15-20) rate in every Arya & Arya (1996)
mg/l) + BA (3-5 mg/l) + agar-agar (0,7%) 3-4 week cycle of subculture

Dendrocalamus asper shoots MS + sucrose (3%) + myo-inositol (100 95-98% rooting was achieved within 15 days Arya & Arya (1996)
mg/l) + NAA (3 mg/l) + IBA (10 mg/l) +
agar-agar (0,7%)
Dendrocalamus asper seeds MS + 100 mg/l myo-inositol + 30 g/l 16 h daylight at shoot initiation Arya et al. (1999)
sucrose + 7g/l agar + 1,0-10,0 mg/l BA 25°C

12
Dendrocalamus asper propagules of 3 MS + 1,0 - 5,0 mg/l NAA or 5,0 - 10,0 rooting Arya et al. (1999)
shoots each mg/l IBA

Dendrocalamus asper viable seeds MS medium + sucrose (3%) + BAP (20 12 h 32% germination 4,05 shoots/clump (average Semsuntud et al. (2000)
Backer µM), pH = 5,8 photoperiod at number)
25°C
Dendrocalamus asper young fresh nodes MS medium + sucrose (3%) + BAP (10 12 h 2,0 shoots/clump Semsuntud et al. (2000)
Backer µM), pH = 5,8 photoperiod at
25°C
Dendrocalamus asper shoots MS medium + sucrose (3%) + agar 12 h 90,5% of the shootlets rooted Semsuntud et al. (2000)
Backer (0,8%) + IBA (40 µM), pH = 5,8 photoperiod at
25°C

Dendrocalamus brandisii, individual shoots MS + 2% sucrose + 0,5ppm BAP + 5% 24 h daylight at shoot multiplication Nadgauda et al.(1990)
Bambusa arundinacea CW 28°C
Dendrocalamus giganteus apex and young MS basal medium (= myo-inositol (100 darkness at 70% of the explants formed calli; 20% of the calli Muñoz-Fonseca et al.
leaves from buds, mg/l) + thiamine-HCl (1 mg/l) + nicotinic 25°C; 4 weeks were organogenic (2002)
excised from acid (0,5 mg/l) + pyridoxine-HCl (0,5 on CIM
lateral branches mg/l) + glycine (2 mg/l)) + sucrose (30
g/l) + Gelrite (2 g/l) + 2,4-D (3 mg/l) +
zeatin (1 mg/l); pH = 5,5
Dendrocalamus giganteus organogenic calli MS medium + zeatin (1 mg/l) photoperiod each callus gave rise to 2-6 shoot primordia (a Muñoz-Fonseca et al.
(obtained in regime of 16/8 h cluster with a dominant shoot and axillary inhibited (2002)
darkness with 2,4- light/dark cycle smaller shoots)
D and zeatin for 8 (4 W/m_)
weeks and with
zeatin for another
8 weeks)

Dendrocalamus giganteus 70-year old field MS medium + 6mg/l BA shoots and spikelets Ramanayake &
clump Wanniarachchi (2003)
Dendrocalamus giganteus shoots, spikelets, MS medium + sucrose (4%) + 2,4-D (0 - 24+/-3 °C dark callus formation Ramanayake &
roots 67,8 µM) + NAA (0-40,3 µM) + CW (0 or Wanniarachchi (2003)
10%)

Dendrocalamus giganteus callus MS medium + sucrose (4%) + 2,4-D proliferation and formation of creamy white masses Ramanayake &
(33,9 µM) + NAA (0 or 5,4 or 16,1 µM) with early stage embryo and root formation Wanniarachchi (2003)

13
Dendrocalamus giganteus nodular callus MS medium + sucrose (4%) + 2,4-D light (450 lux) rooted shoots Ramanayake &
(33,9 µM) + NAA (0 or 5,4 or 16,1 µM) 12 h Wanniarachchi (2003)
photoperiod
Dendrocalamus giganteus single node MS medium + BA (26,6 µM) + Kn (0,5 - shoot proliferation Ramanayake et al.
segments µM) + sucrose (2 %) (2001)
Dendrocalamus giganteus in vitro shoots MS medium + BA (26,6 µM) + Kn (0,5 prolonged flowering Ramanayake et al.
µM) + + sucrose (2 %) culturing (29 (2001)
months)
Dendrocalamus giganteus nodal segments - (max.) 366 shoots/node Ramanayake et
from 4 to 5-year- al.(1995)
old juvenile plants

Dendrocalamus giganteus nodal segments of semi-solid MS + BAP (2 mg/l) + - (max.) 73 shoots/node Ramanayake et
Bambusa vulgaris var secondary Benlate/Benomyl (1 g/l) + Kn (0,1 mg/l) al.(1995)
striata branches from
mature culms
Dendrocalamus giganteus single node MS + sucrose (3%) + Sigma agar - mean bud-break of 19,8% (0-95%) Ramanayaka
Munro segments (0,35%) + BAP (2 mg/l) + Kn (0,1 mg/l) &Yakandawala (1997)
+ Benlate (1 g/l)
Dendrocalamus giganteus shoots (= nodal liquid MS + sucrose (3%) + BAP (6 mg/l) 16h photoperiod shoot proliferation (1,8x increase every 13 days Ramanayaka
Munro segments with in + Kn (0,1 mg/l) + coconut water (8%) at 24°C; shaker after a lag phase of 65 days) &Yakandawala (1997)
vitro formed at 55 strokes
axillary shoots) per min

Dendrocalamus giganteus shoots from the liquid 1/2 MS + sucrose (2%) + IBA (3 16h photoperiod 77,5% rooting and 70% propagules with new Ramanayaka
Munro shoot proliferation mg/l) + coumarin (10 mg/l) + biotin (2 at 24°C; shaker shoots and roots &Yakandawala (1997)
medium mg/l) + calcium panthothenate (2 mg/l) at 55 strokes
containing IBA (3 per min;
mg/l) in the last 2 transfered to
passages fresh medium at
2-3 weeks
interval

Dendrocalamus hamiltonii dehusked seeds MS + 3% sucrose + 0,7% agar germinated in hypocotyls Chambers et al. (1991)
Munro dark at 25°C
Dendrocalamus hamiltonii epicotyl tissue MS + 3% sucrose + 0,7% agar + BA 16h daylight at calli Chambers et al. (1991)
Munro (4,4µM) 30°C
Dendrocalamus hamiltonii hypocotyls MS + 3% sucrose + 0,7% agar + BA 16h daylight at calli Chambers et al. (1991)
Munro (4,4µM) 30°C

14
Dendrocalamus hamiltonii MS + 2% sucrose + BA (2,2µM) + 5% 16h daylight at multiple shoots Chambers et al. (1991)
Munro CW 30°C

Dendrocalamus hamiltonii single node 1/2 MS + sucrose (3%) + agar (0,8%) fluorescent light buds sprouted within 10 days Sood et al. (1994)
Munro cuttings taken (12h light/dark
from elite seedling cycle) at 25°C
plants
Dendrocalamus hamiltonii small segments MS + BAP ( 1 mg/l) + 2,4-D (1 mg/l); fluorescent light direct shoot bud regeneration Sood et al. (1994)
Munro (3-4 mm) from liquid cultures on shaker (50 rpm); (12h light/dark
newly sprouted subculturing at 15 day interval cycle) at 25°C
buds
Dendrocalamus hamiltonii buds 1/2 MS liquid + sucrose (3%) + IBA (0,5 rooting and subsequent rhizome formation within 6 Sood et al. (1994)
Munro or 1 mg/l) or NAA (0,5 mg/l) weeks
Dendrocalamus hamiltonii small segments MS + BAP (1 mg/l) + 2,4-D (1 mg/l) slightly compact, nodular, pale yellow and slow Sood et al. (1994)
Munro (3-4 mm) from growing callus (occasionally)
newly sprouted
buds
Dendrocalamus hamiltonii callus MS + BAP (1 mg/l) + 2,4-D (1 mg/l) + shoot bud differentiation Sood et al. (1994)
Munro GA3 (0,5 mg/l); repeated subculture
Dendrocalamus hamiltonii nodular and MS + BAP (1 mg/l) + 2,4-D (1 mg/l) + large numbers of thick, pale yellow and generally Sood et al. (1994)
Munro compact callus NAA (0,5 or 1 mg/l) unbrached roots within 3-4 weeks
lumps
Dendrocalamus hamiltonii shoots MS + sucrose (3%) + NAA (1 mg/l) + slightly brownish, thick finger like roots with root Sood et al. (1994)
Munro IBA (1 mg/l) + 2,4-D (0,5 mg/l) + hairs
phloroglucinol (0,1 mM)
Dendrocalamus hamiltonii MS + sucrose (3%) + choline chloride elongated, less hairy, white roots with secondary Sood et al. (1994)
Munro (3,9 mg/l) + IBA (0,5 mg/l) + IAA or NAA and tertiary roots
(0,1 mg/l) or 1/2 MS + 0,3% activated
charcoal
Dendrocalamus hamiltonii MS + sucrose (3%) + coumarin (9 mg/l) generally unbranched, profuse roots with negligible Sood et al. (1994)
Munro + IBA (0,5 mg/l) + IAA or NAA (0,1 mg/l) root hairs

Dendrocalamus hamiltonii stem segments (3- MS + BAP (1 mg/l) + 2,4-D (1 mg/l) or friable callus Sood et al. (1994)
Munro 4 mm) obtained MS + 2,4-D (1-10 mg/l)
from in vitro raised
shoots

Dendrocalamus hamiltonii callus continued subculture on MS + BAP (1 shoot bud differentiation in 8-10 weeks Sood et al. (1994)
Munro mg/l) + 2,4-D (1 mg/l)

15
Dendrocalamus hamiltonii nodal segments 1/2 MS medium + sucrose (3%) + agar 25 +/- 2°C 14-h bud sprouting Godbole et al. (2002)
Nees et Arn. Ex Munro (0,8%) photoperiod
(70+/- 5 µmol
m-2 s-1
Dendrocalamus hamiltonii small segments of 1/2 MS medium + sucrose (3%) + agar callus formation Godbole et al. (2002)
Nees et Arn. Ex Munro sprouted buds (0,8%) + BA (2 mg/l) + 2,4-D (2 mg/l) +
NAA (2 mg/l)
Dendrocalamus hamiltonii callus 1/2 MS medium + sucrose (3%) + agar embryoid differentiation Godbole et al. (2002)
Nees et Arn. Ex Munro (0,8%) + BA (2,5 mg/l)

Dendrocalamus hamiltonii callus 1/2 MS medium + sucrose (3%) + agar diffuse light 5 greenish embryoids that can be regenated Godbole et al. (2002)
-2 -1
Nees et Arn. Ex Munro (0,8%) + BA (2,5 mg/l) µmol m s

Dendrocalamus hamiltonii regenerants 1/2 MS medium + sucrose (8%) + agar vigorously growing green plantlets with high Godbole et al. (2002)
Nees et Arn. Ex Munro (0,8%) survival rate after hardening

Dendrocalamus hamiltonii single node 1/2 MS medium + sucrose (3%) + agar initiation aseptic culture and axillar bud break Sood et al. (2002)
Nees et Arn. Ex Munro segments (0,8%)

Dendrocalamus hamiltonii sprouted buds 1/2 MS medium + sucrose (3%) + agar shoot proliferation Sood et al. (2002)
Nees et Arn. Ex Munro from single node (0,8%) + BA (2,5 mg/l)
segments
Dendrocalamus hamiltonii shoots 1) MS medium + NAA/IBA (1 mg/l each), rooting Sood et al. (2002)
Nees et Arn. Ex Munro 2,4-D (0,5 mg/l) and phloroglucinol
(0,1mM), or 2) 1/2 MS with activated
charcoal (0,3%) or choline chloride (3.0 ,
9.0 mg/l) along with IBA (0,5 mg/l), IAA
or NAA (1mg/l), or 3)MS + coumarin (9
mg/l) + IBA (0, 0,5mg/l), IAA or NAA (0 -
0,1 mg/l)

Dendrocalamus hamiltonii sprouted buds MS medium + BA (1mg/l) + 2,4-D (1 callus formation on cutting edge Sood et al. (2002)
Nees et Arn. Ex Munro from single node mg/l)
segments
Dendrocalamus hamiltonii callus MS medium + BA (1mg/l) + 2,4-D (1 embryoids Sood et al. (2002)
Nees et Arn. Ex Munro mg/l) + GA3 (0,5mg/l)

16
Dendrocalamus hamiltonii embryoids MS medium rooted plantlets Sood et al. (2002)
Nees et Arn. Ex Munro

Dendrocalamus latiflorus
Dendrocalamus latiflorus
Dendrocalamus latiflorus
Dendrocalamus latiflorus cv
Machiku
shoot-tip MS + NAA + BAP callus, shoots and roots Huang (1988)
internode MS + 2,4-D(1-25) callus Zamora et al. (1989)
callus MS + BAP(1) + 2,4-D(1) shoots and plantlets Zamora et al. (1989)
internode tissue MS + sucrose (40 g/l) + 2,4-D (1 to 8 h photoperiod 60 to 70% of the cultures formed callus and 16 to Zamora et al.(1987)
25 ppm) with white 24% was compact callus
daylight
fluorescent
tubes at 25°C

Dendrocalamus latiflorus cv MS + sucrose (40 g/l) + BA (1 ppm) 2 to 16% compact callus Zamora et al.(1987)
Machiku
Dendrocalamus latiflorus cv callus MS + sucrose (40 g/l) + BA (1 ppm) + 75,9% callus regrowth; 51,8% loose callus; 24,1% Zamora et al.(1987)
Machiku 2,4-D (1 ppm) compact callus; 4,3% cultures with shootlike
structures and plant regeneration
Dendrocalamus latiflorus cv MS + sucrose (40 g/l) + BA (1 ppm) 82,6% callus regrowth; 65,2% loose callus; 17,4% Zamora et al.(1987)
Machiku compact callus; 3,4% cultures with shootlike
structures and plant regeneration
Dendrocalamus single node MS + BAP (1,2.10-5 M) + Kn (3.10-6 M) - 70% bud-break; 2-5 shoots/explant after 10-12 Saxena & Bhojwani
longispathus segments from days (1991)
actively growing
lateral branches of
3-year-old clumps
-5 -6
Dendrocalamus 3-4 shoots MS + BAP (1,2.10 M) + IBA (1.10 M) - shoots have been multiplied for over 15 passages Saxena & Bhojwani
longispathus clusters + coconut milk (10%) at an average rate of 3x every 4 weeks (1991)
-5 -5
Dendrocalamus shoots MS + IAA (1.10 M) + IBA (1.10 M) + - 68% of the shoots rooted and 85% survival after Saxena & Bhojwani
-5
longispathus coumarin (6,8.10 M) transferring to soil (1991)
Dendrocalamus 2-3 mm thick 1/2 MS + 2,4,5-T (5 mg/l) - white, smooth, globular embryo-like structures Saxena & Bhojwani
longispathus sections derived without a callus phase (1991)
from the
internodes of
newly emerging
culms

17
Dendrocalamus somatic embryos liquid 1/2 MS + 2,4,5-T (5 mg/l) dark for 14 multiplication Saxena & Bhojwani
longispathus passages (1991)

Dendrocalamus single node MS + sucrose (3%) + agar (0,8%) + 12h photoperiod 75% bud-break + 6,33 shoots/explant Saxena & Bhojwani
longispathus Kurz segments excised BAP (12 µM) + Kn (3 µM); pH = 5,8 with light (1993)
from the middle of intensity of 4000
young lateral lux provided by
branches cool white
fluorescent
tubes (40 W);
26°C

Dendrocalamus clusters of axillary liquid MS + sucrose (3%) + BA (15 µM) 12h photoperiod shoot multiplication rate of 3,2x every 4 weeks Saxena & Bhojwani
longispathus Kurz shoots + coconut water (10%); pH = 5,0 with light (1993)
intensity of 4000
lux provided by
cool white
fluorescent
tubes (40 W);
26°C; medium
on shaker

Dendrocalamus shoots 1/2 MS + sucrose (3%) + IAA (10 µM) + 12h photoperiod 73,3% rooting rate (2-6 roots within 2 weeks), Saxena & Bhojwani
longispathus Kurz IBA (10 µM) + coumarin (68 µM) + with light shoots remained green and healthy (1993)
gelrite (0,2%) intensity of 4000
lux provided by
cool white
fluorescent
tubes (40 W);
26°C

Dendrocalamus strictus caryopses B5 medium + 2,4-D + 0,8% agar + 2% 27 +- 2°C, 24 h callus formation, chlorophyllous embryoids Narang et al. (1985)
sucrose photoperiod
Dendrocalamus strictus embryoids B5 liquid + 2% sucrose + IBA + NAA plantlets Narang et al. (1985)
Dendrocalamus strictus node MS + NAA(1) + 1BA(1) + 2,4-D(0,5) + shoot growth and rooting Chaturvedi & Sharma
phloroglucinol (1988)

18
Dendrocalamus strictus scutellum of MS + sucrose (30 g/l) + 2,4-D (1 and 10 darkness at embryogenic callus Dekkers & Rao (1987)
zygotic embryos mg/l) + coconut water (10%) + agar (8 27°C
g/l)

Dendrocalamus strictus embryo MS + 2,4-D(1 and 10) + CW(10%) callus and embyoids Dekkers (1989)
-5
Dendrocalamus strictus seeds medium containing 2,4-D (3.10 M) - callus formed at the embryo end and formed Narang et al.(1985)
embryoids
Dendrocalamus strictus embryoids medium containing 2,4-D (3.10-5 M) - plantlets within 4 weeks Narang et al.(1985)
-
Dendrocalamus strictus plantlets 1/2 B5 liquid + sucrose(1%) + IBA(5x10 further development Rao et al. (1985)
7 -7
M) + NAA(10 M)
Dendrocalamus strictus caryopses B5 salts and vitamins + sucrose (20 g/l) - embryos germinated after 3-4 days and callusing Rao et al. (2002)
-5
+ agar-agar (8 g/l) + 2,4-D (3.10 M) started soon after germination; embryoid initiation
by the sixth weeks
Dendrocalamus strictus callus B5 salts and vitamins + sucrose (20 g/l) - secondary somatic embryogenesis Rao et al. (2002)
+ agar-agar (8 g/l) + 2,4-D (3.10-5 M) +
-6
BAP (10 M); pH = 5,8
Dendrocalamus strictus surface-sterilized semi-solid MS (0,8% agar) or rested on - 60-80% germination Saxena & Dhawan
seeds filter paper in liquid MS (1995)
Dendrocalamus strictus shoots from 3- liquid MS (shoots on filter paper strips) + - recurrent shoot multiplication (3,5x every 3 weeks) Saxena & Dhawan
weeks-old BAP (9.10-6 M) + IBA (1.10-6 M) + for more than 14 passages (1995)
seedlings coconut water (5%)
Dendrocalamus strictus shoots liquid MS (shoots on filter paper strips) + - 70% of the shoots rooted and 80% succes after Saxena & Dhawan
IAA (6.10-6 M) + IBA (5.10-6 M) + transferring to the greenhouse (1995)
coumarin (7.10-6 M)

Dendrocalamus strictus surface sterilized agarified MS + 2,4-D (3.10-5 M) - calli at the embryonic end (after 6 weeks) Saxena & Dhawan
seeds (1995)
Dendrocalamus strictus embryoids semi-solid MS + 2,4-D (1.10-5 M) + Kn - rapid multiplication at a rate of 2-5 fold/4 weeks Saxena & Dhawan
(5.10-6 M) + soluble PVP (250 mg/l) (1995)
Dendrocalamus strictus chlorophyllous MS + NAA (5.10-5 M) + Kn (5.10-6 M) + - 90% of the embryoids formed both shoot(s) and Saxena & Dhawan
embryoids PVP (250 mg/l) roots (1995)
Dendrocalamus strictus generated shoots 1/2 MS + NAA (3.10-6 M) + IBA (2,5.10- - rooted plants
6 M)
Dendrocalamus strictus seeds B5 medium + 2,4-D at 10 to 100 µM - callus after germination Rao et al. (1987)
Bambusa arundinacea

19
Dendrocalamus strictus callus B5 medium + 2,4-D at 10 to 100 µM - after 30 d the callus formed a modular structure, Rao et al. (1987)
Bambusa arundinacea cream in colour, some friable but some compacted

Dendrocalamus strictus embryogenic medium without 2,4-D - multiplication and some embryoids develloped into Rao et al. (1987)
Bambusa arundinacea callus plantlets

Dendrocalamus strictus seedlings MS + 2,4-D (3 mg/l) - callus that develloped into embryoids Chanthanuraksa (1991)
Dendrocalamus
membranaceus
Thyrsostachys siamensis

Dendrocalamus strictus juvenile seedling MS basal + BAP (3 mg/l) + sucrose 16 h light at 25-30 shoots Chatterjee et al. (1991)
Oxytenanthera stoksii explants (3%) {18 à 20 subculture cycles} 25°C
Dendrocalamus strictus clusters with 3 to 5 MS basal + IBA (1 mg/l) + activated 60% of the rooted plantlets were acclimatised in the Chatterjee et al. (1991)
Oxytenanthera stoksii shoots charcoal (0,25%) greenhouse
Dendrocalamus strictus matured explants MS basal + BAP (3 mg/l) + Kn (0,2 mg/l) 8-10 shoots per node Chatterjee et al. (1991)
Oxytenanthera stoksii + sucrose (4,5%) {10 à 12 subculture
cycles}
Dendrocalamus strictus single-node stem MS ( modified: NH4NO3 1500; KNO3 14 h daylight at axillary bud grew and proliferated Chaturvedi et al. (1993)
Nees segments 1500; thiamine HCl 0,2; sucrose 20000 ) 27°C
+ 0,5 mg/l IAA + 15 mg/lAdS
Dendrocalamus strictus nodal stem MS ( modified: NH4NO3 1500; KNO3 14 h daylight at root formation Chaturvedi et al. (1993)
Nees segments 1500; thiamine HCl 0,2; sucrose 20000 ) 27°C
+ 10mg/l l-argenine-HCl + 0,1 mg/l d-
biotin and 15 mg/l AdS + 1mg/l IBA +
1mg/l NAA + 1mg/l phloroglucinol + 0,5
mg/l 2,4-D
Dendrocalamus strictus seeds MS + agar + 30 µM 2,4-D embryogenic callus from the proliferation of the Saxena & Dhawan
Nees embryo (1999)
Dendrocalamus strictus somatic embryos 1)semi-solid MS + 10 µM 2,4-D + 5 µM rapidly multiplying of somatic embryos Saxena & Dhawan
Nees Kn + 2µM IBA + 250 mg/l PVP 2)MS (1999)
+ 50 µM2,4-D + 10 µM BAP + 250 mg/m
PVP
Dendrocalamus strictus somatic embryos MS + 5µM NAA + 5µM Kn + 250 mg/l healthy plantlets Saxena & Dhawan
Nees PVP (1999)

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Dendrocalamus strictus shoots 1/2 MS major salts + 3µM NAA + 2,5 µM rooting Saxena & Dhawan
Nees IBA (1999)
Dendrocalamus strictus embryogenic MS medium + sucrose (3%) + 2,4-D (3 - NaCl-tolerant callus Singh et al. (2003)
Nees callus from seeds mg/l) + agar (0,8%) + 13,6 µM 2,4-D + 0
to 200 µM NaCl)
Dendrocalamus strictus NaCl-tolerant MS medium + sucrose (3%) + 2,4-D (2 - somatic embryo's Singh et al. (2003)
Nees callus mg/l) + Kn (0,5 mg/l) + agar (0,8%) + 0
to 200 µM NaCl)
Dendrocalamus strictus somatic embryo's 1/2 MS medium + sucrose (2%) + NAA - NaCl-tolerant plantlets Singh et al. (2003)
Nees (0,02 mg/l) + IBA (0,1 mg/l) + agar
(0,8%) + 100 µM NaCl)
Dendrocalamus strictus embryo MS + 10µM 2,4-D embryogenic calli Zamora & Gruezo
Nees (1990)
Dendrocalamus strictus embryogenic calli MS + 10µM 2,4-D multiplication of embryogenic callid Zamora & Gruezo
Nees (1990)
Dendrocalamus strictus seed B5 + 2,4-D(10 or 30µM) callus formation Zamora & Gruezo
Nees (1990)
Dendrocalamus strictus excised embryos MS+ 10µM 2,4-D very good callus formation, embryogenic calli Zamora & Gruezo
Nees (1990)
Dendrocalamus strictus embryogenic calli MMS + CW + myo-inositol plantlets Zamora & Gruezo
Nees (1990)
Dendrocalamus stritus seeds 1/2 MS medium + sucrose (2%) + agar shoots Singh et al .(2000)
Nees (0,8%)
Dendrocalamus stritus excised shoots 1/2 MS medium + sucrose (2%) + TDZ 25 +/- 2°C, 16 h shoot bud formation from the basal node and Singh et al .(2000)
Nees (0-1 mg/l) (liquid) photoperiode shoots
-2 1
(50-70 µE S ,
21 days
Dendrocalamus stritus shoots 1/2 MS medium + sucrose (2%) (liquid) 2 months spikelet formation (from shoots regenerated with Singh et al .(2000)
Nees 0,5 and 1 mg/l TDZ

Otatea acuminata vegetative MS + ascorbic acid (100 mg/l) + 2,4-D - somatic embryos Woods et al. (1991)
aztecorum explants (leaf (3 mg/l) + BA (0,5 mg/l) + casein
sheath, internode, hydrolysate (250 mg/l) + sucrose (2%)
node, root and
rhizome explants)

Otatea acuminata zygotic embryo MS + ascorbic acid (100 mg/l) + 2,4-D darkness somatic embryos Woods et al. (1992)
aztecorum explant tissue (3 mg/l) + BA (0,5 mg/l) + sucrose (2%)

21
Otatea acuminata Woods et al. (1992)
aztecorum
Otatea acuminata somatic embryos B5 basal medium grown in the plants develloped and rooted efficiently (95%) + Woods et al. (1992)
aztecorum light 85% survival after 6 months in the greenhouse

Phyllostachys aurea plant regenerating medium + NAA (1mg/l) + BA (10 mg/l) adventious shoots Huang & Huang (1989)
callus

Phyllostachys aurea shoot-tip MS + 2,4-D(3) callus/cell suspension Huang (1988)

Phyllostachys aurea shoot-tip MS + NAA + BAP callus, shoots and roots Huang (1988)
Phyllostachys aurea A. & C. shoot-tip medium + NAA (1 mg/l) + BA (10 mg/l) initiation of nodular callus Huang et al. (1989)
Riviere
Phyllostachys aurea A. & C. shoot-tip medium + NAA (1 mg/l) + BA (10 mg/l) adventitious shoots and organogenic calli Huang et al. (1989)
Riviere
Pleioblastus pygmaeus propagules MS medium + sucrose (20 g/l) + BA (2 30 d; 100 multiple shoots Watanabe et al. (2000)
-2 -1
Nakai consisting of 2 mg/l) µmol.m .s
shoots photosynthetic
photon flux;
370-400
µmol/mol CO2;
1,9 air
exchanges/h
Pleioblastus pygmaeus multiple shoots MS medium (pH = 5,8) {magenta-type 40 d; 16 h NPR = 83 (+- 23) µmol CO2/gDW. h Watanabe et al. (2000)
Nakai vessels (370 ml) containing 40 ml MS photoperiod shoot DW = 230 (+- 110) mg/plantlet root
medium; autoclaved vermiculite (10 (25°C), 8 h dark DW = 75 (+- 60) mg/plantlet
g/vessel) as supporting material} (21°C); 2000
µmol/mol CO2
;(0-3d) 100
-2 -1
µmol.m .s
PPF, 1,9 air
exchanges/h;

22
Pleioblastus pygmaeus plantlets commercial soil mixture in trays + daily 25 d 100% survival; RGR = 0,031 (+- 0,003)/d; shoot Watanabe et al. (2000)
Nakai 40 ml water/tray DW = 1020 (+- 120) mg/plant; root DW = 610 (+-
110) mg/plant
Sasa pygmaea shoot-tip MS + 2,4-D(3) callus/cell suspension Huang (1988)
Sasa pygmaea shoot-tip MS + NAA + BAP callus, shoots and roots Huang (1988)
Sasa pygmaea (Miquel) E. medium + NAA (1 mg/l) + BA (1 mg/l) granular callus and adventitious shoots Huang et al. (1989)
G. Camus
Schizostachyum culm sheath base MS + 2,4-D(1,5,10); NAA(3,5) callus Dekkers (1989)
brachycladum
Schizostachyum internode MS + 2,4-D(10) callus Dekkers (1989)
brachycladum
Schizostachyum root MS + 2,4-D(1) callus Dekkers (1989)
brachycladum
Sinocalamus latiflora anthers N6 + 2,4-D (1 mg/l) + BA (1 mg/l) + 1 month at 25°C 6,7% of the anthers produced embryos and callus Tsay et al. (1996)
sucrose (9%) + charcoal (2 g/l) + agar followed by a
(0,8%); pH = 5,7 temperature of
30°C

Sinocalamus latiflora embryogenic same anther culture medium or an same conditions embryo germination and rooted plantlets (haploid) Tsay et al. (1996)
callus auxin-free medium for about 1
month

Sinocalamus latiflora mature zygotic MS + myo-inositol + NAA + pyridoxine darkness or darkness + 6mg/l 2,4-D + 3 mg/l kinetin + 5% Yeh & Chang (1987)
(Munro) McClure embryos HCl + thiamine HCl + glycine + sucrose 16/8 h daylight sucrose + 250 mg/l PVP gives callus formation
+ agar + Kn or 2,4-D or PVP at 26 +- 1°C kinetin (3mg/l) and sucrose (5%) and 2,4-D (6mg/l)
necessary for callus formation and somatic
embryogenesis
Sinocalamus latiflora anthers N6 + 1mg/l 2,4-D + 1 mg/l BA + 2 g/l darkness at embryogenic calli Yeh & Chang (1987)
(Munro) McClure charcoal + 0,8% agar + 9% sucrose 25°C
Thamnocalamus seeds MS medium + sucrose (2%) + 25 +/- 1°C, dark germination Bag et al. (2000)
spathiflorus (Trin.) Munro

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Thamnocalamus germinated seeds MS medium + sucrose (2%) 25 +/- 1°C, cool shoots Bag et al. (2000)
spathiflorus (Trin.) Munro white
fluorescent light
(16 hours
photoperiode,
-2 -1
40 µmol.m .s
Thamnocalamus germinated MS medium + sucrose (2%) + BAP (5 multiple shoot formation Bag et al. (2000)
spathiflorus (Trin.) Munro embryo's µM) + (IBA) (0,1 µM)
Thamnocalamus single-node stem MS + BAP (5 µm) + IBA (0,1 µM) Bag et al. (2000)
spathiflorus (Trin.) Munro segments with
healthy axillary
buds
Thamnocalamus clumps of 1/2 MS medium + IBA (150 µM) and 1/2 transfer of rooting Bag et al. (2000)
spathiflorus (Trin.) Munro microshoots MS tissue to IBA-
free medium

Thyrsostachys siamensis culm sheath base MS + 2,4-D(0,5; 1; 20) callus Dekkers (1989)

Thyrsostachys siamensis internode 1/2 MS + 2,4-D(5) callus Dekkers (1989)

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