In vitro Embryogenesis Derived from Shoot Tips in Mass Propagation of Two Selected-Clones of Phalaenopsis (original) (raw)
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Turkish Journal of Agriculture and Forestry, 2021
Phalaenopsis is one of the most remarkable commercial genera in Orchidaceae family. Using immature explants for tissue culture of orchid results in undesired characteristics such as genetic variation and long-lasting juvenility. In this study, we optimized a protocol for somatic embryogenesis of Phalaenopsis amabilis cv. Jinan, by using mature thin cell layer (TCL) explants to minimize immature explant limitations. We examined somatic embryogenesis from two types of mature explants, viz. leaf lTCL and flower stalk nodes tTCL at various concentrations of thidiazuron (TDZ) (0, 1, 2, and 3 mg L-1). The results demonstrated that lTCL explants cultured on medium supplemented with 3 mg L–1 TDZ produced the highest number of somatic embryos (SEs) (21.5 embryos per explant), the lowest yellowish SE (7%), the highest plantlet regeneration (20.5 plantlets per explant) and the highest survival rate (94%) in comparison with tTCL explants. This protocol can be used in commercial mass propagation of Phalaenopsis to overcome the limitations such as long-term juvenility and genetic variation of plantlets regenerated in vitro by using immature explants.
In Vitro Cellular & Developmental Biology - Plant, 2001
Embryogenic calluses were induced from 73% of Phalaenopsis shoot-tip explants excised from flower stalk buds by culturing for 7 mo. on New Dogashima Medium (NDM) containing 0.5 mM a-naphthaleneacetic acid (NAA), 4.4 mM 6-benzylaminopurine and 29.2 mM sucrose. The sucrose concentration was increased to 58.4 mM 4 mo. after initiation of the callus culture. These calluses were successfully subcultured as cell suspension cultures in liquid NDM supplemented with 5.4 mM NAA and 58.4 mM sucrose. By simply reducing the sucrose concentration to 29.2 mM, the cells grew into plantlets through a developmental process similar to that of Phalaenopsis seedlings. The occurrence of somaclonal variants was less than 10% in six out of eight genotypes examined. These results suggest that the embryogenic callus and cell suspension culture could be utilized as the materials for micropropagation and breeding of Phalaenopsis orchids.
Plant Tissue Culture and Biotechnology, 2011
High frequency direct shoot proliferation was induced from the shoot tip explants derived from the in vitro grown seedlings of a critically endangered and horticulturally important ground orchid Phaius tancarvilleae (L'Her) Blume. Shoot tip explants cultured on solidified MS with alone or combination of various concentrations of NAA and BAP produced shoots and multiple shoots. The maximum number of healthy shoots was observed on MS with BAP (1.0 mg/l) with an average of 13.3 shoots per culture in 20 weeks; where shoot multiplication was initiated after 4 weeks of culture. Regenerated shoots rooted on MS with various concentrations of NAA, IAA, IBA. MS with NAA (0.5 mg/l) was the most appropriate condition for rooting. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing cocopeat and sphagnum moss in the ratio of 2 : 1. Key words: Mass propagation, Phaius tancarvilleae, shoot multiplication D. O. I. 10.3329/ptcb.v21i2.10241...
In vitro Propagation of orchid (Dendrobium ovatum (L.) Kraenzl.) through Somatic Embryogenesis
Plant Tissue Culture and Biotechnology
An efficient in vitro regeneration protocol through somatic embryogenesis was established from longitudinally bisected protocorm (lTCL) of an endangered orchid Dendrobium ovatum. The efficiency of EC and SEs from the protocorms significantly relied on the concentration of PGRs. MS medium supplemented with TDZ (1.0 mg/l) induced optimum of EC (31.8%) and SEs (28.1/explant). Similarly, ZEA (0.5 mg/l) induced EC (27.6%) and SEs (17.1/explant). The combined treatment of TDZ (1.0 mg/l) and NAA (0.5 mg/l) resulted in the maximum induction of EC (58.6%) and SEs (39.8/explant) in an upright incubated explant. In another combined effect of ZEA (1.0 mg/l) and NAA (0.5 mg/l) induced EC (43.8%) and SEs (24.3/explant), whereas the combination of BAP (0.5 mg/l) and NAA (0.5 mg/l) produced EC (34.4%) and SEs (16.8/explants). The induced EC and SEs where healthier on medium containing TDZ + NAA than on the medium containing ZEA+NAA and BAP+NAA. The orientation of lTCL explants seemed to interact wi...
Efficient Method for Direct Embryogenesis in Phalaenopsis Orchid
Journal of Horticultural Science, 2021
Light spectrum is one of the environmental cues that influence plant growth and development. Light is a stimulating factor for induction of somatic embryos during tissue culture practices. To accelerate the direct embryogenesis, six different light spectra including: white (W), red (R), blue (B), green (G), red + blue (R+B) and red + far red (R+FR) together with dark condition (D), in combination with thidiazuron (TDZ) in four concentrations (0, 0.5, 1.5 and 3 mg L-1) were used. Inter-simple sequence repeat was used for identification and genetic stability analysis of somatic regenerated plantlets. Intact protocorm explants showed higher potential for direct somatic embryogenesis (DSE) than the other explants. The rate of DSE was highly dependent on the concentration of TDZ and its interaction with light spectra. R and R + FR spectra with 3 mg L-1 TDZ on intact protocorms and R+FR with 3 mg L-1 TDZ were efficient treatments to induce DSE without somaclonal variation. G light spectru...
IN VITRO CLONAL PROPAGATION OF PHALAENOPSIS THROUGH YOUNG LEAF
Phalaenopsis is an elegant orchid genus containing 62 species and mainly used for commercial production as cut flower and potted plant. Phalaenopsis orchid brings high profit in flower markets around the world. In Myanmar, Phalaenopsis is one of the famous and expensive genera among the orchids. Tissue culture became the important method for propagation of Phalaenopsis because of its growth habit of being difficult to propagate vegetatively. Explant types, plant growth regulators and culture system are mainly attributed to the success in the tissue culture propagation technique. In this experiment, in vitro leaf explants were studied for their PLBs induction potential in response to different kinds of plant growth regulator (BA, TDZ and NAA), different concentrations (2, 4 BA, 1, 2 TDZ and 0.5 NAA) and combinations of PGRs (BA + NAA, TDZ + NAA) and two culture systems (solid and cotton support liquid medium). Among the PGRs, highest PLBs induction was observed on ½ MS medium containing 2 mg.L-1BA. Interaction effect was observed between plant growth regulators and culture system. The effects of culture systems were not statistically different from each other. The PLBs obtained from TDZ supplemented media gave better result in solid culture than cotton culture. In contract, PLBs obtained from BA supplemented media showed better result in cotton culture than solid culture. Therefore, the use of PGRs should be selected depending on the culture system.
Acta Physiologiae Plantarum, 2010
Two Phalaenopsis orchids, Phalaenopsis amabilis and Phalaenopsis 'Nebula', were used to test the effects of induction period (30, 45 and 60 days), subculture period (30, 45 and 60 days), and explant length (1, 1.5 and 2 cm) on direct somatic embryogenesis from different regions (leaf tip, adaxial side, abaxial side and cut end) of leaf explants from in vitro grown seedlings. The results showed that the cut end had a highest competence to form embryos than the other regions of the leaf explants from both orchids. In addition, the suitable culture conditions were 60 days for induction period in darkness, 45 days for subculture period in light and 1 cm for explant length. Besides, the combinations of N 6 -benzyladenine (BA) and naphthaleneacetic acid were tested on their effects on plantlet conversion and further development of leafderived embryo. It was found that 0.5 mg/l of BA showed the highest response on plantlet conversion rate and the lowest browning rate of explants. In this communication, the embryo structures and development were proved by scanning electron microscopy.
Notulae Scientia Biologicae
Phalaenopsis ‘AMP 17’ is an important orchid commodity in Indonesia with high market demand; however, scaling up the orchid commercially is constrained by the availability and sustainability of qualified seedlings. To overcome the problem, a reliable in vitro propagation protocol, especially via secondary embryogenesis, was undertaken. In the present study, in vitro secondary embryogenesis derived from meta-topoline (mT) treatment on mass propagation of Phalaenopsis ‘AMP 17’ was successfully established. Embryos, as explant sources, were prepared by culturing meristem tips of flower stalk shoots on Murashige and Skoog (MS) medium containing 1.5 mg/L thidiazuron (TDZ) and 0.25 mg/L N6-benzylaminopurine (BAP) for ± 3 months. High secondary somatic embryo (SSE) formation up to 64.90% with 12.30 SSEs regenerated per embryo was determined on half-strength MS augmented with 0.5 mg/L BAP and 2.5 mg/L mT. The combination also stimulated the result of high multiplication rate of SSE formatio...
In vitro clonal propagation of pigeon orchid (Dendrobium crumenatum Sw.) through callus from bud culture was carried out. The suitable conditions for three main stages of plant regeneration, callus proliferation, PLBs formation and proliferation as well as regeneration of shoots were examined. The callus proliferation required the combination of 0 I mg l-r NAA, I mg ll BA, 2 g l-r AC and 2 g l-' peptone to promote growth. PLBs formation stage needed additive 10% CW while the stage of shoot regeneration had occasion to the condition from previous steps ofexperiment. Peptone could be added in the media for all three developmental stages but it must be used as a mlnor substance only. Histological observation suggested that plant regeneration was developed through both embryogenesis and organogenesis.
International Journal of Current Microbiology and Applied Sciences, 2017
Phaius tankervilliae (L’Her.) Blume is one of the most attractive and horticulturally valuable ground orchids of the world. Due to illegal collection for trade and consumption and loss of habitat, the population of this species has decreased rapidly at an alarming rate in natural habitats. In vitro seed germination and seedling development of Phaius tankervilliae technique was successfully established for rapid multiplication using 0.8% (w/v) agar solidified MS medium supplemented with different concentrations and combinations of Kinetin (Kin) and NAA. MS medium supplemented with 1.0 mg L-1 Kin + 1.0 mg L-1 NAA was the most ideal condition for early seed germination (2.87 weeks), fastest protocorm formation (5.07 weeks), tallest shoots (6.85 cm) and maximum number of roots per plant (6.20). However, maximum number of shoots (52.27) per culture and maximum number of shoots with roots (50.27) were produced when the plantlets were transferred to MS basal medium supplemented with 1.5 mg L-1 Kin + 1.0 mg L-1 NAA. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing only vermiculite (86.00 %) followed by vermiculite + vermicompost (71.00 %) at 1:1 ratio.