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Vol. 15(28), pp. 1503-1510, 13 July, 2016
DOI: 10.5897/AJB2016.15390
Article Number: 4B5FA1759443
ISSN 1684-5315
Copyright © 2016
Author(s) retain the copyright of this article
http://www.academicjournals.org/AJB
African Journal of Biotechnology
Full Length Research Paper
Micropropagation of Guadua angustifolia Kunth
(Poaceae) using a temporary immersion system RITA®
Luís Gonzaga Gutiérrez1*, Rodolfo López-Franco2 and Tito Morales-Pinzón3
1Grupo de Investigación en Biodiversidad y Biotecnología, Facultad de Ciencias Ambientales, Universidad Tecnológica
de Pereira, Carrera 27 #10-02 Barrio Alamos, Pereira, Colombia.
2Programa de Biología, Laboratorio de Biotecnología, Centro de Estudios e Investigaciones en Biodiversidad y
Biotecnología (CIBUQ), Universidad de Quindío, Carrera 15 Calle 12 Norte, Armenia, Colombia.
3Grupo de Investigación en Gestión Ambiental Territorial, Facultad de Ciencias Ambientales, Universidad Tecnológica
de Pereira, Carrera 27 #10-02 Barrio Alamos, Pereira, Colombia.
Received 9 April, 2016; Accepted 27 June, 2016
The micropropagation of Guadua angustifolia Kunth, commonly known as giant bamboo, using semi-
solid culture medium, is reported to have low multiplication rates. This study evaluated the
multiplication index of G. angustifolia in a temporary immersion system (RITA®), comparing results with
those obtained using a semi-solid culture medium. The treatments consisted of either three or four 2-
min immersions per day and use of semi-solid culture medium, which consisted of MS supplemented
with 3.0 mg L-1 of the cytokinin benzylaminopurine (BAP). Equipment consisted of 20 vessels for
automated RITA®, each containing 200 ml of culture medium. Immersions were performed for 2 min at
two different frequency intervals (6 and 8 h). Large clumps of G. angustifolia with 1, 2 or 3 stems were
inoculated depending on the treatment. Best results were obtained with four immersion cycles per day
(every 6 h), with a multiplication index of 2.7 shoots per original explant (axillary buds) and greater
rhizome growth. Overall, the temporary immersion system performed better than the semi-solid medium
in terms of shoot multiplication rates and rhizome growth. Further studies should be conducted to
develop an application for RITA® for use in the commercial production of G. angustifolia.
Key words: Giant bamboo, temporary immersion system RITA, rhizome.
INTRODUCTION
Guadua angustifolia, also known as giant bamboo,
belongs to the Poaceae family, one of the four largest
families of the plant kingdom, harboring from 600 to 700
genera and nearly 10,000 species (Soderstrom et al.,
1988). In Colombia, most populations of G. angustifolia
grow between 0 and 1800 m altitude, occupying diverse
habitats of tropical moist forests (Bh-T) and premontane
wet forests (Bmh-P), where they form small patches
along rivers or streams (Londoño, 1990).
Because of its widespread distribution, versatility and
*Corresponding author. E-mail: luisgon@utp.edu.co
Author(s) agree that this article remains permanently open access under the terms of the Creative Commons Attribution
License 4.0 International License
1504 Afr. J. Biotechnol.
physical and mechanical characteristics, G. angustifolia
has played an important environmental, cultural and
economic role in Colombia. However, for these same
reasons, it has also suffered indiscriminate exploitation.
Enormous pressure has been exerted on natural
populations of this species, reducing the areas planted
and endangering not only the species but also other
associated organisms (Cruz, 2009). Furthermore, G.
angustifolia produces seeds with low germination
capacity, which makes sexual reproduction difficult. The
deterioration of stands of G. angustifolia during vegetative
multiplication is further justification for exploring
micropropagation approaches to develop efficient mass
multiplication methods for this important species using
explants (Cruz, 2009).
Propagation studies are appropriate in the case of G.
angustifolia because in vitro multiplication techniques are
an alternative for species conservation, providing
abundant high-quality planting material. Manzur (1988)
reported that G. angustifolia plants perform similarly in in
vitro and in situ conditions. As a result, when micro-
cuttings from lateral branches are cultured in enriched
culture medium, their axillary buds activate the
development of intercalary meristems with roots emerging
from their basal nodes. The rhizome is immediately
formed, which then becomes the fundamental structure in
the micropropagation of G. angustifolia.
Only a few studies have been reported on the
micropropagation of G. angustifolia (Manzur, 1988;
Marulanda et al., 2005; Jiménez et al., 2006; Daquinta et
al., 2007). Most studies conducted on micropropagation
of bamboo have focused on Asian species (Muñoz et al.,
1989; Saxena, 1990; Gielis and Oprins, 2000; Sood et
al., 2002; García et al., 2007; Lin et al., 2007; Ogita et al.,
2008; Venkatachalam et al., 2015; Gantait el al., 2016).
In Colombia, several studies have been conducted on
the micropropagation of G. angustifolia, achieving partial
success at different stages of solid culture medium. The
first reports by Manzur (1988) showed low multiplication
rates (less than 2 shoots/explant); however, Marulanda et
al. (2005) subsequently reported a survival rate of 30%
during the establishment phase and Acosta and Guzmán
(1993) reported a better multiplication rate when 5.0 mg
L-1 6-benzylaminopurine (BAP) was added to the culture
medium, supplemented with kinetin 1.0 mg L-1 and
indoleacetic acid (IAA) 1.0 mg L-1.
Subsequently, Marulanda et al. (2005) and Jiménez et
al. (2006) used BAP (2.5 to 5.0 mg L-1) as growth
regulator, reporting low multiplication rates. Large losses
have also been recorded during the establishment phase
of G. angustifolia because of bacteria-induced
contamination (Cruz et al., 2007; Ramírez et al., 2009).
Depending on the consistency of the culture medium
(liquid, semi-solid and solid) used in plant micro-
propagation, several systems have been developed to
increase the multiplication index and reduce costs. Taking
into account the liquid condition of the medium, a system
was initially designed that consisted of a large, elevated
culture chamber that could be drained and then refilled
with fresh medium (Tisserat and Vandercook, 1985).
Subsequently a semi-automated system was developed
in which plants were cultured in a large container on a
medium that contained no gelling agent, with automatic
addition and removal of liquid medium at fixed intervals
so the plant material entered into contact with the
medium several times throughout the day (Aitken-Christie
and Davies, 1988).
Simonton et al. (1991) developed a programmable
apparatus, which intermittently applied culture medium to
the plants according to a pre-defined schedule, serving
as basis for the temporary immersion system (TIS) for
plant propagation (Alvard et al., 1993). This system,
whose commercial name is RITA for recipient for
automated TIS, has been successfully used with many
plant species, achieving significant increases in
multiplication rates and allowing the semi-automation of
processes (Lorenzo et al., 1998). Furthermore, the
automation of one or more phases of micropropagation
can help reduce not only handling costs but also costs
related to laboratory space, while increasing production
volume (Castro and González, 2000; Capote et al.,
2009).
The frequency and time conditions required to achieve
efficiency in TIS processes are determinants for system
optimization (Etienne and Berthouly, 2002). Albarrán et
al. (2002) confirmed that the massive regeneration of
somatic embryos of coffee (Coffea arabica) improved
when exposure in terms of frequency and duration was
optimized. Increases in daily frequency (1×1’, 2×1’, 6×1’)
stimulated embryo production without affecting embryo
quality.
The TIS (Figure 1A and 1B) offers multiple advantages
as compared with propagation using semi-solid culture
media (Figure 1C) in terms of shoot multiplication in
different plant species (González et al., 2005; Roels et
al., 2006; Barberini et al., 2011). In the case of banana,
increases have been achieved in high average
multiplication rates, with improved plant quality (Albany et
al., 2005; Berthouly and Etienne, 2005; Watt, 2012;
Pérez et al., 2013). Only a few bamboo species such as
Dendrocalamus latiflorus (Mongkolsook et al., 2005 cited
by García-Ramirez et al., 2014), Bambusa ventricosa and
Dracaena deremensis (Chaille, 2011) and Bambusa
vulgaris (García-Ramírez et al., 2014) have been
propagated by TIS. This study aimed to optimize shoot
production for micropropagation of G. angustifolia for
commercial purposes.
MATERIALS AND METHODS
Plant material
The study was carried out at the Plant Biotechnology Laboratory of
the Universidad Tecnológica de Pereira (UTP), where test plants
Gutiérrez et al. 1505
Figure 1. Guadua angustifolia shoot multiplication in RITA® (A and B). Shoots of G. angustifolia in
semi-solid medium (C). Plantlets of G. angustifolia obtained from semi-solid medium: non-segmented
(presence of rhizome) (D) and segmented (absence of rhizome) (E) and ex-vitro acclimatization of
plant produced (F). Scale bar, 5 cm.
had been previously established in vitro. The study compared the
multiplication indexes of G. angustifolia plants submitted to TIS with
the indexes reached when plants were cultured in semi-solid
medium (SSM). Explants, measuring 5 to 10 cm, were inoculated
according to the protocol proposed by Marulanda et al. (2005).
Culture conditions
The culture medium, containing MS salts and vitamins, was
supplemented with 100 mg L-1 myo-inositol and 30 g L-1 sucrose.
Gelrite® was used as gelling agent in the solid medium at 2.5 g L-1,
A
CD
E
B
F
1506 Afr. J. Biotechnol.
Figure 2. Rhizome size in Guadua angustifolia plants depending on culture media and condition of
plantlet during four weeks of culture, where RITA® A = 3 cycles, segmented; RITA® B = 3 cycles,
non-segmented; RITA® C = 4 cycles, segmented; RITA® D = 4 cycles, non-segmented; SSM A =
semi-solid medium, segmented; SSM B = semi-solid medium, non-segmented. Different letters
indicate statistically significant differences between groups (mean ± standard error, n = 10, Tukey
test, p<0.05).
and BAP was used as growth regulator at a concentration of 3 mg
L-1.
A total of 200 ml MS basal medium was added to each culture
vessel containing liquid medium (TIS) (Figure 1A and 1B), whereas
50 ml were added to each vessel (200 ml) containing SSM. Vessels
of both treatments were autoclaved at 121°C at 15 psi, using RITA®
vessels developed by CIRAD (Teisson and Alvard, 1995) (Figure
1A and B), being left for 30 min in the case of TIS and 20 min in the
case of SSM.
Five explants were inoculated per TIS vessel, whereas only one
explant was inoculated per SSM vessel. All plants were placed in a
growth chamber with an average temperature of 24±2°C and a 12-h
artificial photoperiod (lux).
The multiplication index of G. angustifolia plants using the TIS,
which consisted of three or four 2-min immersions per day (A = 3
cycles, segmented; B = 3 cycles, non-segmented; C = 4 cycles,
segmented; D = 4 cycles, non-segmented), was evaluated and
compared with the multiplication index of plants cultured in SSM (A
= segmented; B = non-segmented). Large 5 to 10 cm clumps of
segmented (absences of rhizomes) and non-segmented (presence
of rhizomes) of G. angustifolia were used in the inoculation. The
number of stems was used as basis to evaluate rhizome
generation, shoot multiplication index, and increase in plant height
over a 4-week period. Rhizome division was based on the following
variables: large clumps, either segmented (Figure 1D) or non-
segmented (Figure 1E), and number of stems (1, 2 and 3) per
inoculation unit.
Statistical analysis
A completely randomized block design (10 blocks) was used, with
each block corresponding to one of the established plants
(approximately 40 mm long) and a factorial arrangement of two
controlled factors: culture medium and conditions of plants at
planting.
Data were analyzed using a random block design with 20 replicates
(explants). Each replicate corresponded to one explant and each
variable depended on the study factors (rhizome size, shoot
multiplication index and increase in plant height after 4 weeks of
culture). The data obtained were submitted to the Tukey test to
compare the means of the applied treatments.
RESULTS AND DISCUSSION
Effect of culture system on rhizome size for
segmented and non-segmented G. angustifolia
Analysis of variance indicated that highly significant
differences exist in rhizome growth for large clumps of
segmented and non-segmented G. angustifolia,
depending on culture medium consistency (TIS or SSM).
The Tukey test, designed to make pairwise comparisons
among means, indicated that rhizome growth was
greater, but non-significant (p>0.05) in plants cultured in
the TIS with four 2-min immersions per day as compared
with plants cultured in the TIS with three 2-min
immersions per day or those cultured in SSM. For RITA®,
average rhizome growth was greater in non-segmented
large clumps than in segmented large clumps, presenting
significant differences (p<0.05) when 3 immersion cycles
was used (Figure 2).
Manzur (1989) confirmed that when the rhizome
originates in in vitro conditions, the established G.
angustifolia plant is capable of generating a large group
of successive plants. Furthermore, Londoño (1991)
considers bamboo rhizomes to be segmented axes,
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
RITA® A
RITA® B
RITA® C
RITA® D
SSM A
SSM B
Rhizome length (mm)
Treatments
a
ab ab
ab
b
b
segmented
non-segmented
Gutiérrez et al. 1507
Figure 3. Multiplication of Guadua angustifolia shoots depending on culture media and conditions of plantlets
during four weeks of culture, where RITA® A = 3 cycles, segmented; RITA® B = 3 cycles, non-segmented;
RITA® C= 4 cycles, segmented; RITA® D= 4 cycles, non-segmented; SSM A = semi-solid medium,
segmented; SSM B = semi-solid medium , non-segmented. Different letters indicate statistically significant
differences between groups (mean ± standard error, n = 10, Tukey test, p<0.05).
characterized by having a diameter larger than that of the
stem that it will produce. Mostly single rhizome buds
appear on nodes from which new plant structures will
emerge. Rhizome size was accordingly larger on bamboo
plantlets cultured in the TIS with four 2-min immersions
per day than when cultured on SSM (Figure 2).
Effect of culture system on shoot multiplication for
segmented and non-segmented G. angustifolia
The analysis of variance indicated that there are highly
significant differences (p=0.0023) in the number of shoots
produced per explant depending on culture medium
consistency. Significant differences also occurred
between segmented and non-segmented explants. For
the Tukey test, plantlets cultured in the TIS (Figure 3),
with four 2-min immersion cycles, showed a higher, but
non-significant (p>0.05) average number of shoots than
plants cultured in the TIS with three 2-min immersion
cycles and in SSM. Furthermore, non-segmented plants
presented a higher number of shoots than segmented
plants (Figure 3). These results agree with those of
studies carried out in sugarcane (Saccharum officinarum)
by Lorenzo et al. (1998) and in Eucalyptus grandis by
Castro and González (2000), two species that presented
a higher increase of biomass in the TIS than in solid and
semi-solid culture media.
Effect of culture system on growth of segmented and
non-segmented G. angustifolia
The analysis of variance revealed that the culture
medium consistency (TIS or SSM) significantly affected
growth of G. angustifolia. Highly significant differences in
plant height were also observed when segmented and
non-segmented plants were compared in RITA® (Figure
4). Plants cultured in SSM presented greater growth than
those segmented and cultured in the TIS (four and three
2-min cycles per day). Overall, non-segmented plants
presented higher average growth than segmented plants
(Figure 4).
Plant elongation during the multiplication phase in the
solid culture medium is similar to that reported by
Marulanda et al. (2005) and Jiménez et al. (2006). Plant
elongation in the TIS (four 2-min immersions per day) is
similar to that obtained using the solid culture medium
with non-segmented explants.
In contrast, Castro and González (2000) reported that
plant size of E. grandis increased in the TIS as compared
with plants cultured in solid medium. Lorenzo et al.
(1998) also found that sugarcane plants grew taller when
cultured in TIS (10.29 cm on average) as compared to
solid media (6.22 cm on average).
Fifty seedlings cultured in solid media and 50 cultured
in TIS were transferred to the nursery using the hardening
protocol developed by Marulanda et al. (2005) (Figure
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
RITA® A
RITA® B
RITA® C
RITA® D
SSM A
SSM B
Number of shoots
Treatments
a
ab ab
ab
b
b
segmented
non-segmented
1508 Afr. J. Biotechnol.
Figure 4. Increase in plant height of G. angustifolia plantlets depending on culture media and
conditions of plantlets during four weeks of culture, where RITA® A = 3 cycles, segmented; RITA® B =
3 cycles, non-segmented; RITA® C= 4 cycles, segmented; RITA® D= 4 cycles, non-segmented; SSM A
= semi-solid medium, segmented; SSM B = semi-solid medium , non-segmented. Different letters
indicate statistically significant differences between groups (mean ± standard error, n = 10, Tukey test,
p<0.05).
1F). No significant differences in plant acclimatization
were observed between groups, with a 95% survival rate
of plants (data not shown). Plants of G. angustifolia from
both treatments were then transferred to the field, where
plant growth and development are currently being
evaluated. Survival rates for other Poaceae in the
hardening phase have been reported to be higher than
70% (Gielis and Oprins, 2000; Marulanda et al., 2005;
Jiménez et al., 2006).
Effect of culture system on shoot multiplication for
segmented and non-segmented G. angustifolia
In this study, the average number of shoots produced by
plants cultured in the TIS (four 2-min cycles per day) was
higher than that of plants cultured in the SSM (Figure 5).
The TIS (3 stems) with four 2-min immersions per day
produced an average multiplication rate of 3.0 shoots per
original explant for non-segmented plants (Figure 5).
Marulanda et al. (2005) found an average multiplication
rate of 2 shoots per original explant for G. angustifolia
cultured on solid media, whereas Jiménez et al. (2006)
reported an average of 2.5 new individuals (referred to as
tillers) per each original plant established. When the
averages of all three studies were compared, the higher
multiplication efficiency in the TIS reached in this study is
clearly demonstrated. In addition, the multiplication rate in
solid culture medium yielded results similar to the
aforementioned results, with an average multiplication
rate of 2.7 shoots per explant.
Using micropropagation in the liquid system with
species of the subfamily Bambusoideae, Das and Pal
(2005) with Bambusa balcooa and Shirin and Rana
(2007) with Bambusa glaucescens reported successful
work in the in vitro multiplication phase. In addition,
bamboo species such as Dendrocalamus latiflorus
(Mongkolsook et al., 2005), Bambusa ventricosa and
Dracaena deremensis (Chaille, 2011) and Bambusa
vulgaris (Garcia-Ramirez, 2014) have been propagated
by TIS, with results similar to those found for G.
angustifolia in terms of rate of multiplication (3 to 5 shoots
per explant).
There are similar reports for other species. According
to Lorenzo et al. (1998), the in vitro multiplication of
sugarcane shoots per plant reached higher levels
(average of 8.13 shoots) in the TIS than when a solid
culture medium was used (average of 3.96 shoots). Yan
et al. (2010) also achieved better results with Siraitia
grosvenorii when RITA® was used for plant regeneration
than with solid culture media.
In this study, the number of shoots emerging from the
rhizome was higher for both SSM and RITA when plants
were cultured with three stems than when the culture was
performed with two and one stems per plant. When
cultured in the TIS (four cycles per day), G. angustifolia
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
RITA® A
RITA® B
RITA® C
RITA® D
SSM A
SSM B
Plant height (cm)
Treatments
aaa
b
ab
b
segmented
non-segmented
Gutiérrez et al. 1509
Figure 5. Multiplication rate depending on the number of non-segmented shoots per initial explant
inoculated in semi-solid medium and RITA® (four 2-min cycles per day). Different letters indicate
statistically significant differences between groups (mean ± standard error, n = 10, Tukey test,
p<0.05).
plants with three stems produced the highest number of
shoots (Figure 5).
For Ravikumar et al. (1998), the number of shoots
generated by bamboo plants differs depending on the
species and the inoculated explants. Kane (2000) has
also found that the type of explants used in micro-
propagation processes, whether nodal segments or
shoots, stimulate the generation of axillary buds
depending on each species.
Conclusion
Multiplying G. angustifolia using TIS highlights the
importance of the presence of the rhizome (non-
segmented explants) for optimal production of shoots.
This study also presented good results with a frequency
of three 2-min immersions per day, producing 2.7 shoots
per original explant. Further studies should be conducted
to develop an application for RITA® for use in the
commercial production of G. angustifolia.
Conflict of interests
The authors have not declared any conflict of interests.
ACKNOWLEDGEMENTS
The authors thank the Laboratorio de Biotecnología
Vegetal of the Universidad Tecnológica de Pereira and its
staff for their valuable collaboration in the development of
the present research project.
Abbreviations
BAP, 6-Benzylaminopurine; 2,4-D, 2,4-
dichlorophenoxyacetic acid; GA, gibberellic acid; IAA,
indoleacetic acid; MS, Murashige and Skoog (1962);
NAA, naphthalene acetic acid.
REFERENCES
Acosta V, Guzmán CC (1993). Propagación Vegetativa de la Guadua
(Guadua angustifolia K.et B. forma “macana”) mediante Cultivo de
Tejidos. Trabajo de Grado, Ingeniería Forestal, Universidad del
Tolima, Colombia.
Aitken-Christie J, Davies HE (1988). Development of a semi-automated
propagation system. Acta Hortic. 230:81-87.
Albany N, Jiménez E, Vilchez J, García L, De Feria M, Pérez N, Sarría
Z, Pérez B, Clavelo J (2005). Use of growth retardants for banana
(Musa AAA cv. Grand Naine) shoot multiplication in temporary
immersion systems. In. Hvoslef AK, Preil W (eds) Liquid Culture
Systems for In Vitro Plant Propagation, Springer, Dordrecht,
Netherlands, pp. 213-224.
Albarrán J, Bertrand B, Vásquez N, Etienne H (2002). La frecuencia y
duración de la inmersión afecta la cantidad y calidad de los
embriones somáticos de café regenerados en un biorreactor de
inmersión temporal. Instituto Nacional de Investigaciones Agrícolas,
Centro Nacional de Investigaciones Agropecuarias, Maracay,
Venezuela.
Alvard D, Cote F, Teisson C (1993). Comparison of methods of liquid
medium culture for banana micropropagation. Plant Cell Tissue
Organ Cult. 32:55-60.
Barberini S, Savona M, Ruffoni B (2011). Temporary immersion culture
of Lilium bulbiferum. Acta Hortic. 900:377-383.
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
1 stem
2 stems
3 stems
1 stem
2 stems
3 stems
SSM
RITA®
Number of shoots
Treatments
bc
ab
a
a
bc
c
1510 Afr. J. Biotechnol.
Berthouly M, Etienne H (2005). Temporary immersion system: A new
concept for use liquid medium in mass propagation; In: Hvoslef AK,
Preil W (eds) Liquid Culture Systems for In Vitro Plant Propagation,
Springer, Dordrecht, Netherlands, pp. 165-195.
Capote I, Escalona M, Daquinta M, Pina D, González J, Aragon C
(2009). Propagación in vitro de Vriesea en sistemas de inmersión
temporal. Biotecnol. Veg. 9:161-167.
Castro D, González J (2000). Eucalyptus (Eucalyptus grandis Hill)
micropropagación de eucalilto (Eucalyptus grandis Hill ex Maiden)
en el sistema de inmersión temporal. Agric. Tec. 62:68-78.
Chaille L (2011). Optimization of Tissue Culture Protocols for Cost-
Effective Production of Dracaena, Bamboo, and Succulent Plants.
M.Sc. Thesis, Department of Tropical Plant and Soil Science, Hawaii.
Cruz H (2009). Guadua angustifolia Kunth. Bosques Naturales de
Colombia. Plantaciones Comerciales en México. Editorial Gráficas
Olímpica S.A., Pereira. Colombia.
Cruz M, García Y, Sánchez C, Alvarado Y, Acosta M, Roque B, Leiva
M, Freire M (2007). Identificación y control de Bacillus sp.,
contaminante del establecimiento in vitro de Guadua angustifolia
Kunth. Biotecnol. Veg. 7:9-13.
Daquinta M, Gregori A, Cid M, Lezcano Y, Sagarra F (2007). Formación
de callos e inducción de brotes a partir de tejido intercalar de ramas
de plantas adultas de Guadua angustifolia Kunth. Biotecnol. Veg.
7:119-122.
Das M, Pal A (2005). In vitro regeneration of Bambusa balcooa Roxb.:
Factors affecting changes of morphogenetic competence in the
axillary buds. Plant Cell Tissue Organ Cult. 81:109-112.
Etienne H, Berthouly M (2002). Temporary immersion systems in plant
micropropagation. Plant Cell Tissue Organ Cult. 69:215-231.
Gantait S, Pramanik B, Banerjee M (2016). Optimization on planting
materials for large scale plantation of Bambusa balcooa Roxb.:
Influence of propagation methods. J. Saudi Soc. Agric. Sci. In Press.
García Y, Freire M, Fajardo L, Tejada M, Reyes M (2007).
Establecimiento in vitro de yemas axilares de Bambusa vulgaris var.
Vittata. Biotecnol. Veg. 7:155-159.
García-Ramírez Y, Gonzáles MG, Mendoza EQ, Freire-Seijo M,
Cárdenas MLO, Moreno-Bermúdez LJ, Ribalta OH (2014). Effect of
BA treatments on morphology and physiology of proliferated shoots
of Bambusa vulgaris Schrad. Ex Wendl in temporary immersion. Am.
J. Plant Sci. 5:205-211.
Gielis, J, Oprins J (2000). Micropropagation of temperate and tropical
woody bamboos from biotechnological dream to commercial reality.
In: Bamboo for sustainable development. Proceedings of the 5th
International Bamboo Congress and the 6th International Bamboo
Workshop, San José, Costa Rica, Pp. 333-344.
González JL, Fundora Z, Molina LA, Abdulnour J, Desjardins Y,
Escalona A (2005). New contributions to propagation of pineapple
(Ananas comosus L. Merr) in temporary immersion bioreactors. In
vitro Cell Dev. Biol. Plant 41:87-90.
Jiménez V, Castillo J, Tavares E, Guevara E, Montiel M (2006). In vitro
propagation of the neotropical giant bamboo, Guadua angustifolia
Kunth, through axillary shoot proliferation. Plant Cell Tissue Organ
Cult. 86:389-395.
Kane M (2000). Propagation from preexisting meristems. In: Trigano R,
Gray D (eds) Plant Tissue Culture Concepts and Laboratory
Exercises, 2nd ed. CRC Press, Boca Raton, USA, Pp. 75-97.
Lin ChS, Liang CJ, Hsaio HW, Lin MJ, Chang WC (2007). In vitro
flowering of green and albino Dendrocalamus latiflorus. New For.
34:177-186.
Londoño X (1990). Aspectos sobre la distribución y la ecología de los
bambúes de Colombia (Poaceae: bambusoideae). Caldasia 16:139-
153.
Londoño X (1991). Estudio botánico, ecológico, silvicultural y
económico-industrial de las Bambusoideae de Colombia. Cespedesia
16/17:51-78.
Lorenzo JC, González BL, Escalona M, Teisson C, Espinosa P, Borroto
C (1998). Sugarcane shoot formation in an improved temporary
immersion system. Plant Cell Tissue Organ Cult. 54:197-200.
Manzur D (1988). Propagación vegetativa de Guadua angustifolia
Kunth. Rev. Agron. 2:14-19.
Manzur D (1989). Bosques de guadua (Bambusa) en el laboratorio.
Rev. Agric. Trop. (Bogotá), pp. 53-55.
Marulanda MM, Gutierrez LG, Márquez M (2005). Micropropagación de
Guadua angustifolia Kunth. Actual. Biol. 27:5-15.
Mongkolsook Y, Tanasombut M, Sumkaew R, Likitthammanit P,
Wongwean P (2005). Temporary Immersion System (TIS) for
Micropropagation of Dendrocalamus latiflorus in Commercial
Production. Kasetsart Agricultural and Agro-Industrial Product
Improvement Institute, Kasetsart University, Bangkok.
Muñoz M, Guevara E, Montiel M (1989). Regeneración in vitro del
bambú gigante Dendrocalamus giganteus (Poaceae). Rev. Biol.
Trop. 4:50-56.
Murashige T, Skoog F (1962). A revised medium for rapid growth and
bioassays with tobacco tissue culture. Physiol. Plant. 15:473-479.
Ogita S, Kashiwagi H, Kato Y (2008). In-vitro node culture of seedlings
in bamboo plant, Phyllostachys meyeri McClure. Plant Biotechnol.
25:381-385.
Pérez, MB, Vega VM, Delgado MT, Torres JL, Pino AS, Cabrera
AR,Toledo MB, García YB, Ortiz AO (2013). Nueva alternativa para
la micropropagación en inmersión temporal del cultivar de plátano
vianda “INIVITPV -2011” (AAB) /New alternative for micropropagation
in temporary immersion system of plantain cultivar “INIVITPV -2011
(AAB ). Rev. Colomb. Biotecnol. 15(1):98-107.
Ramírez LA, Castaño SM, López R (2009). Identificación de bacterias
que afectan el establecimiento in vitro de segmentos nodales de
Guadua angustifolia Kunth. Rev. Investig. Univ. Quindio 19:151-158.
Ravikumar R, Ananthakrishnan G, Kathiravan K, Ganapathi A (1998).
In vitro shoot propagation of Dendrocalamus strictus nees. Plant Cell
Tissue Organ Cult. 52:189-192.
Roels S, Nocedal C, Escalona M, Sandoval J, Canal MJ, Rodríguez R,
Debergh P (2006). The effect of headspace renewal in a temporary
immersion bioreactor on plantain (Musa AAB) shoot proliferation and
quality. Plant Cell Tissue Organ Cult. 84:155-163.
Saxena S (1990). In vitro propagation of bamboo (Bambusa tulda
Roxb.) through shoot proliferation. Plant Cell Rep. 9:431-434.
Shirin F, Rana PK (2007). In vitro plantlet regeneration from nodal
explants of field-grown culms in Bambusa glaucescens Willd. Plant
Biotechnol. Rep. 1:141-147.
Simonton W, Robacker C, Krueger S (1991). A programmable
micropropagation apparatus using cycled liquid medium. Plant Cell
Tissue Organ Cult. 27:211-218.
Soderstrom TR, Judziewicz EJ, Clarck LG (1988). Distribution patterns
of neotropical bamboos. In: Heyer WR, Vanzolini PE (eds)
Proceedings of the Neotropical Biotic Distribution Pattern Workshop,
Academia Brasileira de Ciencias, Rio de Janeiro, Pp. 121-157.
Sood A, Ahuja PS, Sharma M, Sharma OP, Godbole S (2002). In vitro
protocols and field performance of elites of an important bamboo
Dendrocalamus hamiltonii Nees et Arn. Ex Munro. Plant Cell Tissue
Organ Cult. 71:55-63.
Teisson C, Alvard D (1995). A new concept of plant in vitro cultivation
liquid medium: Temporary immersion. In: Terzi M, Celia R, Falavigna
A (eds) Current Issues in Plant Molecular and Cellular Biology,
Kluwer, Dordrecht, Pp. 105-110.
Tisserat B, Vandercook CE (1985). Development of an automated plant
culture system. Plant Cell Tissue Organ Cult. 5:107-117.
Venkatachalam A, Kalaiarasi K, Sreeramanan S (2015). Influence of
plant growth regulators (PGRs) and various additives on in vitro plant
propagation of Bambusa arundinacea (Retz.) Wild: A recalcitrant
bamboo species. J. Genet. Eng. Biotechnol. 13(2):193-200.
Watt MP (2012). The status of temporary immersion system (TIS)
technology for plant micropropagation. Afr. J. Biotechnol. 11:14025-
14035.
Yan H, Liang Ch, Li Y (2010). Improved growth and quality of Siraitia
grosvenorii plantlets using a temporary immersion system. Plant Cell
Tissue Organ Cult. 103:131-135.
