Micropropagation of three Heliconia species of commercial interest in Mexico via direct organogenesis

E. Hernández-Meneses1; M. C. G. López-Peralta2*; A. A. Estrada-Luna3

1. Posgrado en Recursos Genéticos y Productividad (PREGEP)-Fisiología Vegetal, Colegio de Postgraduados., Colegio de Postgraduados, Posgrado en Recursos Genéticos y Productividad, (PREGEP)-Fisiología Vegetal, Colegio de Postgraduados, Mexico , 2. PREGEP-Genética, Colegio de Postgraduados, 56230, Montecillo, Texcoco, Estado de México., Colegio de Postgraduados, PREGEP, Genética, Colegio de Postgraduados,

<postal-code>56230</postal-code>
<city>Texcoco</city>
<state>Estado de México</state>
, Mexico , 3. Centro de Investigación y de Estudios Avanzados - Unidad Irapuato. Departamento de Ingeniería Genética., Centro de Investigación y de Estudios Avanzados, Unidad Irapuato, Departamento de Ingeniería Genética

Correspondence: *. Corresponding Author: López-Peralta, María Cristina Guadalupe. PREGEP-Genética, Colegio de Postgraduados, 56230, Montecillo, Texcoco, Estado de México, México. Phone: +52(595) 9520200 Ext. 1540. E-mail: E-mail:


Abstract

Heliconias are tropical ornamentals plants cultivated mainly as a cut flower. Its cultivation in Mexico has experienced limited growth due to some problems that include obtaining vegetative material of high phytosanitary quality. In this research, a protocol to obtain plants of Heliconia collinsiana, H. nickeriensis and cv. Golden Torch via direct organogenesis was developed. The lateral buds were disinfected with commercial detergent, sodium hypochlorite (30 % v/v), colloidal silver (3 % v/v) and Tween® 20. Benzylaminopurine concentrations (4.4-17.8 μM) were evaluated to induce and multiply shoots as well as auxins to promote rooting. Histological sections were performed to determine the anatomical origin of the shoots. In the acclimatization, three substrates and three doses of the nutrient solution Steiner (1961) were tested. In H. collinsiana 4.8 shoots per explant was induced in MS culture medium added with 13.3 μM of benzylaminopurine, while in H. nickeriensis and cv. Golden Torch the induction of 3.0 and 2.1 shoots per explant was reached with 11.1 μM of benzylaminopurine, respectively. The shoots originated at the base of the explant and in the axils of the leaves. The shoots multiplication was obtained with 11.1 μM of benzylaminopurine combined with 1 μM of indoleacetic acid in the three species; 6.9 shoots per explant in H. collinsiana, 4.1 shoots in cv. Golden Torch and 3.6 in H. nickeriensis. The shoots originated at the base of the explant from axillary buds and shoots primordia differentiated from procumium. The rooting was induced with 1 μM of naphthaleneacetic acid and the acclimatization of the plants was achieved in peat and perlite mixture with 50 % Steiner nutrient solution after 55 days of the transplant.

Received: 2018 April 10; Accepted: 2018 August 23

revbio. 2020 May 30; 5: e487
doi: 10.15741/revbio.05.e487

Keywords: Keywords: H. collinsiana, H. nickeriensis, Golden Torch, micropropagation, regeneration, native.

Introduction

Heliconias are perennial herbaceous plants mainly cultivated as cut flowers, though its cultivation is also important as pot plants destined to interiors and gardens (Criley, 2000; Loges et al., 2007). Flowers of Heliconias are highly appreciated in national and international markets due to the exotic morphology of their inflorescence and bright colors of their bracts. Their cultivation and commercialization in Hawaii and Colombia represents a prosperous industry (Santos et al., 2009).

Heliconias originate from the tropics of Central and South America and from a wide area in the Pacific Ocean, west of Indonesia. They include a family integrated by approximately 200 species. Mexico has the native genetic resources like Heliconia uxpanapensis C. Gutierrez Báez (Suárez-Montes et al., 2011), H. champneiana Griggs cv. Maya Gold, H. latispatha Bentham cv. Orange Gyro, H. vaginalis Bentham and H. wagneriana Petersen (Santos et al., 2009). In Costa Rica, Honduras, Colombia, Ecuador, Jamaica, Guyana, Barbados, Trinidad and Tobago, and Surinam, Heliconias have become the main ornamental cultivation for cut flower thanks to the diversity of commercial cultivars (Jerez, 2007). The most popular Heliconias are those that produce erected and pendant inflorescences with red and pink bracts like H. bihai (cv. Pink Peach), H. chartacea (cv. Sexy Pink, cv. Sexy Scarlet), H. rostrata, H. stricta (cv. Iris Red, cv. Tagami), among others (Baltazar et al., 2011).

Despite of the diversity of species of Heliconias in Mexico, being the main center of origin and having favorable environmental conditions, the growth of this crop at commercial scale has been limited by several factors. The most important factors are the lack of experience on care and handling of native species and commercial cultivars, the lack of efficient propagation systems of vegetative material of high phytosanitary quality and the diversification with genetically superior varieties (high winter and annual productivity and long postharvest life). This last aspect has propitiated the cultivation of native species for which no culture systems nor agronomic handling have been established (Murguía et al., 2007; Iracheta-Donjuan et al., 2013). Currently, species and commercial varieties are cultivated in southeastern Mexico (Chiapas, Tabasco, Veracruz) like H. bihai (cv. Pink Peach), H. caribaea (cv. Black Magic, cv. Green Thumb), H. latisphata, H. orthotricha (cv. Edge of Nite, cv. She), H. psittacorum (cv. Lady Di, cv. Sassy), H. rostrata, H. stricta (cv. Las Cruces, cv. Tagamy) and H. wagneriana.

The main method of propagation of Heliconia is by division of rhizomes because it is the most efficient way to maintain clonal populations. However, the risk of transmission of bacterial and fungus diseases is high, and the rate of reproduction of shoots is variable. There are species and cultivars that produce up to 50 new plants a year (H. psittacorum), while others only produce from three to five plants (H. caribaea and H. vellerigera). The sexual propagation is not commercially used because germination rates are low as a consequence of rigidity of the endosperm and latency, which also can slow it down from three months to three years (Simão & Scatena, 2003; Iracheta-Donjuan et al., 2013).

Plant tissue culture has played an important role in the propagation of many plant species, including ornamentals, since efficient protocols have been established to clone elite genotypes (Jain & Ochatt, 2010). In addition, histological methods allow understand in vitro culture systems and of anatomical and histochemical changes provide information on the cellular processes which lead to a determined morphogenic route (Yeung, 1999). In literature, in vitro propagation of some Heliconia have been reported (Nathan, et al., 1992, Nogueira et al., 2004; Sosa et al., 2008); nevertheless, the new improved cultivars and some native species with outstanding agronomic characteristics have not yet been micropropagated.

In this sense, the need for develop an efficient system to multiply species and varieties, which recently stand up as new and contributing to increase the diversity of produced flowers, was raised. The objective of this research was to develop a protocol for in vitro regeneration of three species of Heliconia via organogenesis from lateral buds and to determinate the anatomic origin of shoots produced.

Material and Methods

Vegetal material, explants and disinfection

Adult and healthy plants of H. collinsiana Griggs, H. nickeriensis Mass & Rooij and H. psittacorum L.f. x H. spathocircinata Aristeguieta cv. Golden Torch from commercial plantations were used. Lateral buds of 0.5 cm of length were dissected and then washed using commercial detergent (Roma®) and tap water for 5 minutes. After five rinses with distilled water, they were submerged for 15 min in a mixture of stable colloidal silver (3 % v/v), NaOCl (Cloralex®, 30 % v/v) and Tween® 20 (4 % v/v). They were rinsed five times with sterile distilled water and each explant was established in 10 mL of culture medium in 45 mL glass flasks.

Culture medium and incubation conditions

Mineral salts from MS culture medium (Murashige & Skoog (1962) were used, added with sucrose (30 g L-1), myo-inositol (200 mg L-1), thiamine (2 mg L-1), pyridoxine (0.5 mg L-1), glycine (2 mg L-1), nicotinic acid (0.5 mg L-1) and solidified with agar-agar (Merck®, 7 g L-1). The pH of the culture medium was adjusted to 5.7 with NaOH (1N) y HCl (1N) and it was sterilized in a vertical autoclave (AESA 300®) at 121 ºC and 1.5 kg cm-2 of pressure during 20 min. Cultures were maintained at 25 ± 2 ºC with a photoperiod of 16/8 h (light/darkness) (45 μmol m-2 s-1).

Shoots induction

The organogenic capacity of lateral buds was explored by incubation in MS medium supplemented with seven concentrations of benzylaminopurine (BAP; 4.4, 6.6, 8.8, 11.1, 13.3, 15.5 and 17.8 μM). The experiment had a completely randomized and factorial design 3X7=21 (three Heliconia species and seven BAP concentrations). Each treatment had 12 repetitions and the experimental unit was one explant per glass flask. At 12 weeks of culture, shooting (% of explants that generated shoots), number of shoots per explant and shoot length (cm) were quantified.

Histological analysis

Explants tissues were sampled at the first, fourth, and tenth weeks during the phase of induction of shoots. Tissues were fixed in formalin-acetic acid-alcohol (FAA 50:5:10 v/v) for 24 hours; they were dehydrated with isopropylic alcohol (30, 40, 50, 70, 90, 100 and 100 %), xylene-isopropylic 3:1, 1:1, 1:3 and two changes in xylene for 6 hours in each solution; they got soaked in liquid paraffin (60 °C) with two changes (12 hours). The samples were placed and cut in a manual rotatory microtome in sections of 10 μm of thickness, which were placed on a slide with chrome adhesive, they were dyed with safranin (0.05 % of time for 12 hours) and Fast Green FCF (0.12 % for 1 minute) (López et al., 2005). The sections were observed and photographed using an optical microscopy (Zeiss® modelo Axiostar Plus with Paxcam® digital camera).

Shoots multiplication

Shoots 2 cm in length of the three Heliconia species resulting from the induction were transferred to a MS culture medium added with BAP (11.1, 15.5, 20.0 and 24.4 μM); 1 μM of indoleacetic acid (IAA) was added for each dose of BAP. Subcultures to fresh medium were performed at four weeks; and after eight weeks of culture the same variables as in the phase of shoots induction were quantified. The experiment had a completely randomized design and factorial arrangement; 12 treatments were formed by the combination of four doses of BAP and three Heliconia species; each treatment had 12 repetitions and the experimental unit was of one shoot per glass flask.

Plants rooting

Shoots 3 to 4 cm in length of the three Heliconia species were cultivated in MS culture medium added with 1 μM of alpha-naphthalene acetic acid (ANA), 1 μM of indolebutyric acid (IBA) and in MS culture medium free of plant growth regulators (control). After four weeks, the number and length (cm) of roots were quantified, as well as plant height (cm). The experiment had a completely randomized design and factorial arrangement 3X3 (three auxin levels and three Heliconia species). Each treatment had 12 repetitions and the experimental unit was one shoot per glass flask.

Plants acclimatization

Rooted plants 7 cm in length were transplanted into 175 mL polystyrene pots with peat moss:perlite (1:1) as substrate and the nutritive Steiner (1961) solution (0, 50 and 100 %) was assessed. They were cultivated in a growth chamber (Environmental chamber, Lab-Line Instruments®) at 25 ± 2 °C with a photoperiod of 12/12 h (light/darkness) and 60% of relative humidity during 40 days. Then, they were transferred to greenhouse conditions and at two weeks, plants survival rate (%), height (cm) and the diameter of pseudo-stem base (cm) were quantified. The experiment was conducted in a completely randomized design and factorial arrangement 3x3=9 (three doses of nutritive solution and three Heliconia species). Each treatment had 20 repetitions and the experimental unit was one plant.

Statistical analysis

Data obtained from each experiment were submitted to analysis of variance using Statistical Analysis Software SAS (SAS Institute, 2003) and the Tukey test (p≤0.05) was used to compare means. Values in percentages were transformed using square root.

Results and Discussion

Shoots induction

The genotype, doses of BAP and its interaction had significant effects on shooting and on the number of shoots per explant. The shoot length was only affected by the dose of BAP and the interaction; in the three variables the highest effect was produced by the dose of BAP (p≤0.05). BAP caused the activation of the axillary meristems and the resulting shooting of lateral buds on the explants. In general, shooting increased as the dose of BAP increased and was of 100 % with 11.1 and 13.3 μM. This optimal dose produced the highest number of shoots per explant on each species; 4.8 in H. collinsiana, 3.0 in cv. Golden Torch and 2.1 in H. nickeriensis. It also favored the highest length of shoots; 1.3 cm in H. collinsiana, 1.4 cm in cv. Golden Torch and 1.5 cm in H. nickeriensis (Table 1; Figure 1a-c). Shoots induction occurred at 10 weeks of culture in H. collinsiana and cv. Golden Torch, while it appeared at week 12 in H. nickeriensis. BAP concentrations higher than the optimal level inhibited the induction and drastically reduced the number of differentiated shoots.

Table 1.

Effect of benzylaminopurine (BAP) on shoot induction from lateral bud explants of three Heliconia species after 12 weeks of culture.


Species BAP
(μM)
Shooting
(%)
Shoots per
explant (Num.)
Shoot length (cm)
H. collinsiana 4.4 0.0f 0.1gh 0.0e
6.6 50.0bcde 0.8efg 0.4cde
8.8 83.3abc 1.0def 0.8abcd
11.1 91.7ab 2.1c 0.9abcd
13.3 100.0a 4.8a 1.3a
15.5 75.0abcd 1.7cd 1.3ab
17.7 0.0f 0.0h 0.0e
cv.Golden Torch 4.4 0.0f 0.0h 0.0e
6.6 83.3abc 1.0def 1.0abc
8.8 100.0a 1.3de 1.0abc
11.1 100.0a 3.0b 1.4a
13.3 58.3abcd 1.7cd 1.0abc
15.5 41.7cdef 0.4fgh 0.9abcd
17.7 0.0f 0.0h 0.0e
H. nickeriensis 4.4 0.0f 0.0h 0.0e
6.6 16.7ef 0.2gh 0.2de
8.8 41.7cdef 0.4fgh 0.5bcde
11.1 100.0a 2.1c 1.5a
13.3 91.7ab 1.0def 1.5a
15.5 33.3def 0.3fgh 0.5bcde
17.7 0.0f 0.0h 0.0e
MSD 47.0 0.7 0.7

TFN1Means with equal letters per column are not statistically different (Tukey, 0.05). MSD= Minimum significant difference.



[Figure ID: f1] Figure 1.

Organogenesis in Heliconia spp. [a] Shoots induction in H. collinsiana with 13.3 μM of BAP; [b] Shoots induction in cv. Golden Torch and [c] H. nickeriensis with 11.1 μM of BAP. [d] Shoot primordia of H. collinsiana originated from procambium (pr) at the base of the explant. [e] Differentiation of axillary bud (ya) of cv. Golden Torch at the base of the explant. [f] Shoots multiplication in H. collinsiana with 13.3 μM of BAP and 1 μM of IAA; [g] Multiplication in cv. Golden Torch and [h] H. nickeriensis with 11.1 μM of BAP and 1 μM of IAA. [i] Rooting of H. nickeriensis plants with 1 μM ANA. [j] Plants acclimatization of cv. Golden torch. cm = meristematic cells; h = leaf.


Shoots promoted by BAP in this phase differ from the ones reported by Nathan et al., (1992) in H. psittacorum cv. Choconiana where shooting was only possible with 10 μM of BAP after 24 weeks of culture. In contrast, in H. stricta, the highest quantity of shoots (2.4 and 2.3) was in induced with 8.8 and 17.7 μM of BAP, respectively, after 8 weeks (Nogueira et al., 2004). Also, in H. curtisphata 8.9 μM of BAP was the optimal concentration to induce 5.6 shoots after three weeks from six-months-old in vitro plantlets (Alarcon et al., 2011). In explants of floral buds of H. bihahi cv. Lobster Salmon the highest results of shoots induction were also obtained with 8.9 μM of BAP but combined with 5.7 μM of IAA after six weeks, although the number of shoots was not specified (Marulanda-Ángel et al., 2011). In H. psittacorum cv. Choconiana BAP (4.4 μM) also resulted effective to induce shoots in explants of thin cell layer, but combined with 2,4-D (4.5 μM) after eight weeks (Meneses et al., 2009). In contrast, in Heliconia ortotricha cv. Total Eclipse the optimal dose to induce from 7.5 to 10.1 shoots was of 17.8 μM of BAP, twice that reported in other species and cultivars, although the response was modulated by the light source (Takeui et al., 2016).

The breaking of meristems latency and shoots induction is a response promoted by BAP because it stimulates cellular division and results effective in shoots initiation in many plant species. Its effects on organogenesis can be magnified when combined with auxins (Machakova et al., 2008; van Staden et al., 2008). Although BAP stimulated the production of new shoots, differences in morphogenic capacity among genotypes of Heliconia are appreciated by the number of shoots generated per explant, since some species are naturally more prolific than others. In this study, the quantity of shoots emitted by H. collinsiana was 2.2 times higher than H. nickeriensis and 1.6 times more than cv. Golden Torch.

Histological analysis

Produced shoots were formed in two specific sites of the explant. Buds primordia were observed to emerge from the axils of the fourth and fifth leaf, near the apical meristem of the shoot, after 4 weeks of culture in H. collinsiana. In H. collinsiana and cv. Golden Torch observed the formation of adventitious shoots originated at the base of the explant, from meristematic cells derived from procambium after ten weeks of culture (Figure 1d). In cv. Golden Torch the activation of lateral buds present at the axil of the leaves was observed (Figure 1e); while in H. nickeriensis only the elongation of the leaves surrounding the apical meristem of the shoot was appreciated. The action of BAP is due to its capacity to modify apical dominance and promote growth of lateral buds (Sakakibara, 2006).

Shoots multiplication

In this phase, the genotype, BAP concentration and the interaction between both factors only significantly affected the number of shoots per explant. Shooting and shoot length were only affected by the dose of BAP (p≤0.05). Similarly, to that obtained in shoots induction, the highest effect was produced by BAP. The shooting in the three species was 100 % with 11.1 μM of BAP, while the highest doses decreased it. This concentration also stimulated the highest number of shoots per explant (6.9 in H. collinsiana, 4.1 in cv. Golden Torch and 3.6 in H. nickeriensis) and the highest shoot length (2.1 cm in H. collinsiana and H. nickeriensis; 1.9 cm in cv. Golden Torch). Doses of BAP superior to 11.1 μM decreased the number and size of shoot per explant (Table 2; Figure 1f-h).

Table 2.

Effect of benzylaminopurine (BAP) and indoleacetic acid (IAA) in the in vitro shoots multiplication of three Heliconia species after eight weeks of culture.


Species BAP + IAA
(μM)
Shooting
(%)
Shoots per
explant (Num.)
Shoot length (cm)
H. collinsiana 11.1 + 1 100.0a 6.9a 2.1a
15.5 + 1 100.0a 3.0cd 2.0ab
20.0 + 1 100.0a 2.1def 1.9ab
24.4 + 1 50.0b 0.6h 1.1bc
cv.Golden Torch 11.1 + 1 100.0a 4.1b 1.9ab
15.5 + 1 91.6ab 2.5de 1.6abc
20.0 + 1 83.3ab 1.6efg 1.4abc
24.4 + 1 50.0b 0.5h 0.8c
H. nickeriensis 11.1 + 1 100.0a 3.6bc 2.1a
15.5 + 1 66.6ab 1.4fgh 1.6abc
20.0 + 1 58.3ab 1.1gh 1.3abc
24.4 + 1 58.3ab 0.5h 1.1bc
MSD 48.8 0.9 1.0

TFN2Means with equal letters per column are not statistically different (Tukey, 0.05). MSD = Minimum significant difference.


Shoots number produced in the multiplication phase in each species was superior to the one generated during the shoots induction phase and the shoots were generated in a shorter time (eight weeks of cultivation). These values indicate that the combination of cytokinins and auxins favored the proliferation of axillary shoots. The evaluated doses of BAP were combined with IAA (1 μM) because auxins attenuate the inhibitory effect of cytokinins on shoot length and increased the number of shoots (Gaba, 2005; Kane, 2005).

The effectiveness of BAP in shoots multiplication coincides with that reported by Ulisses et al., (2010) in H, bihai cv, Lobster Claw Two, where the best results of shoots multiplication (2.3 shoots per explant) were obtained with 11.1 μM of BAP but in MS culture medium with half concentration of salts after six weeks. In H. standley, the maximum quantity of shoots per explant (4.6) was obtained with a lower dose of BAP (8.8 μM) combined with 3.7 μM of IAA (Sosa et al., 2008). In contrast, shoots multiplication in H bihai cv. Lobster Salmon requires two subculture of eight and four weeks with 26.6 and 8.8 μM of BAP, respectively, to obtain a maximum of 3 shoots per explant (Marulanda-Ángel et al., 2011). Even when the highest effect on shoots multiplication of the three species was due to the action of the cytokinin, the genotype also influenced. H. collinsiana surpassed cv. Golden Torch and H. nickeriensis, both in shoots induction and multiplication, showing that the species of the genus exhibited different morphogenic capacities.

Shoots obtained in this multiplication phase were subcultured three times (with six weeks of interval) in the MS culture medium added with 11.1 μM of BAP and 1 μM of IAA. With these doses of plant growth regulators, the multiplication rate was maintained until obtaining 328.5 shoots per explant in H. collinsiana, 68.9 in cv. Golden Torch and 46.6 in H. nickeriensis, in a period of 18 weeks.

Plants rooting

The number and length (cm) of roots, as well as plants height were significantly affected by the genotype, by plant growth regulators and by the interaction between both factors (p≤0.05). Although rooting in the three species was produced in every treatment, auxin was the factor that most influenced the three variables with 97, 92, and 63 % of the total variation, respectively. Plants of the three species reached the highest values in the three variables related to roots when they were cultivated with 1 μM of ANA. This auxin allowed to duplicate the quantity of roots obtained in comparison with the IBA and the control (Table 3).

Table 3.

Effect of naphthalenacetic acid (ANA) and indolebutyric acid (IBA) in the in vitro rooting of three Heliconia species after eight weeks of culture.


Species Auxin
(μM)
Roots
(Num.)
Root length
(cm)
Plant height
(cm)
H. collinsiana 0 5.1d 2.9e 6.2f
1 ANA 9.0b 6.3c 9.1b
1 IBA 3.9f 3.0e 5.3h
cv. Golden Torch 0 5.1d 2.0f 5.9fg
1 ANA 8.1c 7.2b 7.8d
1 IBA 4.2ef 5.1d 5.2h
H. nickeriensis 0 4.6de 2.1f 8.5c
1 ANA 9.9a 8.8a 10.8a
1 IBA 4.7de 2.8e 7.1e
MSD 0.5 0.7 0.6

TFN3Means with equal letters per column are not statistically different (Tukey, 0.05). MSD= Minimum significant difference.


The emergency of roots originated in the fourth week of cultivation and the maximum values were registered in the eighth week. Heliconia nickeriensis was the species in which taller plants were obtained and with higher quantity and length of roots, followed by H. collisiana and cv. Golden Torch (Figure 1i).

Even though shoots of the three species of Heliconia in the multiplication phase formed roots in the MS culture medium with no growth regulators, the rooting was considerably improved with the use of ANA. The easiness to form roots without any presence of auxins was also reported in H. psittacorum cv. Choconiana and H. bihai cv. Lobster Clow Two (Nathan et al., 1992; Ulisses et al., 2010). On the other hand, the rooting in H. bihai cv. Lobster Salmon was induced with 7.4 μM de IAA (Marulanda-Ángel et al., 2011).

Plants acclimatization

The genotype, the dose of nutritive solution and the interaction between both factors had significant effects on plants height and pseudostem diameter; survival rate was only affected by the dose of nutritive solution (p≤0.05). However, the main effect was produced by the dose of nutritive solution in all measured variables. Survival rate was of 100 % in the three species when the nutritive solution was applied at half concentration. In contrast, plants that achieved a higher growth rate, and therefore a larger size, were obtained with the complete nutritive solution (Table 4, Figure 1i).

Table 4.

Survival and growth of micropropagated plants of three species of Heliconias after 55 days.


Species Steiner
Solution (%)
Survival
(%)
Plant height
(cm)
Pseudostem
diameter (cm)
H. collinsiana 0 30b 11.9de 0.5e
50 100a 22.2abc 1.1a
100 50b 28.2a 0.8c
H. nickeriensis 0 45b 9.0e 0.4e
50 100a 19.8bc 0.8c
100 50b 24.7ab 0.6d
cv. Golden Torch 0 35b 9.7e 0.4e
50 100a 17.9cd 0.8c
100 55b 23.3abc 0.6d
MSD 40.6 6.1 0.1

TFN4Means with equal letters per column are not statistically different (Tukey, 0.05). MSD= Minimum significant difference.


Results obtained with the nutritive solution applied at 50 % were due to the fact that the requirements of the plants were optimally covered, which produced a higher activation and accumulation of plant growth. Although plants height was higher with the solution at 100 % concentration of salts, pseudostem diameter in the three species was considerably reduced, which produced weaker plants. Plants treated with 50 % concentration of salts presented medium size but with thick pseudostems, which was reflected by vigorous plants, ideal for their transplant into pots and greenhouses. Fertilization requirements of plants during acclimatization were similar to the ones used in the establishment of new commercial plantations of Heliconias, where more nitrogen is provided (3N-1P-2K) for stimulating plant growth during the first year; then the proportion of potassium is increased in order to induce blooming (Jerez, 2007; Alarcón & Bernal, 2012).

Conclusions

A protocol for the in vitro propagation of Heliconia collinsiana, H. nickeriensis and H. psittacorum x, H. spathocircinata cv. Golden Torch by direct organogenesis from lateral buds culture was developed. Meristems activation and shoots induction was achieved with 11.1 and 13.3 μM of BAP in the three species. The histological analysis revealed that the shoots originated in the base of the explant and from the differentiation of adventitious shoots. The highest shoots multiplication rate was achieved with 11.1 and 13.3 μM of BAP combined with 1 μM of IAA in the three species. Rooting was induced in MS culture medium without plant growth regulators and was improved with 1 μM of ANA. Plants survival in acclimatization was 100 % in peat moss: perlite (1:1) and fertilized with 50 % of Steiner nutritive solution. This protocol constitutes a valuable contribution for the propagation of native species, new hybrids and commercial cultivations of Heliconia as well as the rescue of plantations devastated by pathogens.


fn1Cite this paper: Hernández-Meneses, E., López-Peralta, M. C. G., Estrada-Luna, A. A. (2018). Micropropagation of three Heliconia species of commercial interest in Mexico via direct organogenesis. Revista Bio Ciencias 5, e487.Doi: https://doi.org/10.15741/revbio.05.e487

Acknowledgements

To Colegio de Postgraduados Campus Montecillo, for the support provided in the development of this research and to Finca las Abejas de Tacotalpa, Tabasco, for the plant material provided.

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Revista Bio Ciencias, Año 12, vol. 8,  Enero 2021. Sistema de Publicación Continua editada por la Universidad Autónoma de Nayarit. Ciudad de la Cultura “Amado Nervo”,  Col. Centro,  C.P.: 63000, Tepic, Nayarit, México. Teléfono: (01) 311 211 8800, ext. 8922. E-mail: revistabiociencias@gmail.com, revistabiociencias@yahoo.com.mx, http://revistabiociencias.uan.mx. Editor responsable: Dr. Manuel Iván Girón Pérez. No. de Reserva de derechos al uso exclusivo 04-2010-101509412600-203, ISSN 2007-3380, ambos otorgados por el Instituto Nacional de Derechos de Autor. Responsable de la última actualización de este número Dr. Manuel Iván Girón Pérez. Secretaria de Investigación y Posgrado, edificio Centro Multidisciplinario de Investigación Científica (CEMIC) 03 de la Universidad Autónoma de Nayarit. La opinión expresada en los artículos firmados es responsabilidad del autor. Se autoriza la reproducción total o parcial de los contenidos e imágenes, siempre y cuando se cite la fuente y no sea con fines de lucro.

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Revista Bio Ciencias por Universidad Autónoma de Nayarit se encuentra bajo una licencia de Creative Commons Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional

Fecha de última actualización 03 de Noviembre de 2021

 

licencia de Creative Commons Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional