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Micropropagation of an exotic ornamental plant, Calathea crotalifera, for production of high quality plantlets.

Rozali SE, Rashid KA, Taha RM - ScientificWorldJournal (2014)

Bottom Line: A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera.Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem.This is the first report of rapid mass propagation for C. crotalifera.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia ; Center for Foundation Studies in Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera. The effects of different sterilization techniques, explant type, and the combination and concentration of plant growth regulators on shoots induction were studied. The axillary shoot buds explants sprouted from rhizomes in soil free conditions showed high induction rate of shoots with lowest contamination percentage when treated with combination of 30% (v/v) NaOCl, 70% (v/v) ethanol, and 0.3% (w/v) HgCl2. In the present study, the highest number of multiple shoots was obtained in MS basal medium supplemented with 3.5 mg/L 6-Benzylaminopurine (BAP), 1.0 mg/L 1-Naphthaleneacetic acid (NAA), 3% sucrose, and 6 g/L plant agar for both varieties and was used as multiplication medium. Microshoots were highly induced when the young shoot bud explants were incised longitudinally prior subculture. Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem. The maximum roots induction was recorded on MS medium supplemented with 1.0 mg/L 1-Naphthaleneacetic acid (NAA) compared to indolebutyric acid (IBA). The complete regenerated plantlets were successfully acclimatized in the soilless medium under greenhouse condition. This is the first report of rapid mass propagation for C. crotalifera.

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Related in: MedlinePlus

Development of high quality plantlet from shoot bud explants of Calathea crotalifera on multiplication media (MS medium with 3.5 mg/L BAP and 1.0 mg/L NAA) and rooting media (MS medium with 1.0 mg/L NAA). (a) Induction of microshoots from incised shoot bud explants. (b) High quality plantlets with dark green leaves in multiplication medium supplemented with 1 g/L activated charcoal. (c) Plantlets with light green leaves without necrosis problem on multiplication medium supplemented with 1 g/L L-glutamine. (d) Plantlets with high multiple shoots induction. (e) Induction of healthy root in rooting media supplemented with 1.0 mg/L NAA. (f) Roots induction on rooting media supplemented with 1.0 mg/L IBA. (g) and (h) Healthy plantlets ready to be acclimatized. (i) Development of successfully acclimatized plantlets.
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Related In: Results  -  Collection


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fig4: Development of high quality plantlet from shoot bud explants of Calathea crotalifera on multiplication media (MS medium with 3.5 mg/L BAP and 1.0 mg/L NAA) and rooting media (MS medium with 1.0 mg/L NAA). (a) Induction of microshoots from incised shoot bud explants. (b) High quality plantlets with dark green leaves in multiplication medium supplemented with 1 g/L activated charcoal. (c) Plantlets with light green leaves without necrosis problem on multiplication medium supplemented with 1 g/L L-glutamine. (d) Plantlets with high multiple shoots induction. (e) Induction of healthy root in rooting media supplemented with 1.0 mg/L NAA. (f) Roots induction on rooting media supplemented with 1.0 mg/L IBA. (g) and (h) Healthy plantlets ready to be acclimatized. (i) Development of successfully acclimatized plantlets.

Mentions: Complete plantlets with high quality shoots and roots (Figures 4(g) and 4(h)) were removed from the rooting medium and the roots were carefully cleaned under running tap water to remove the residual agar. Each plantlet was then transplanted into plastic pots containing mixture of black soil, river sand, coconut husk, and vermiculite (4 : 2 : 2 : 1). The plantlets were then allowed to grow for two months under greenhouse conditions at 25 ± 2°C with direct sunlight for 12 h daily. Each pot was watered with distilled water for everyday and was covered with transparent plastic which was gradually removed. The morphology of leaf stomata from two months acclimatized plantlets were examined under scanning electron microscope (SEM). The leaf disc with 0.25 cm2 area from the in vitro and in vivo grown plantlets was used to compare the morphological development of the leaf surface and leaf stomata from two different conditions. The samples were fixed in 2% aqueous osmium tetroxide, OsO4 for overnight at 4°C. The treated samples were rinsed with distilled water for two times at 15 minutes each prior to dehydration through ethanol series (10, 20, 30, 40, 50, 60, 70, 80, 95, and 100%). The samples were then infiltrated through ethanol, acetone mixture at 15 minutes each, and ended with pure acetone for 1 hour before dried using a Critical Point CO2 Dryer. The samples were finally viewed under SEM after being gold coated. The presence of wax and morphological development of stomata aperture on the abaxial and adaxial leaf surfaces were observed and recorded.


Micropropagation of an exotic ornamental plant, Calathea crotalifera, for production of high quality plantlets.

Rozali SE, Rashid KA, Taha RM - ScientificWorldJournal (2014)

Development of high quality plantlet from shoot bud explants of Calathea crotalifera on multiplication media (MS medium with 3.5 mg/L BAP and 1.0 mg/L NAA) and rooting media (MS medium with 1.0 mg/L NAA). (a) Induction of microshoots from incised shoot bud explants. (b) High quality plantlets with dark green leaves in multiplication medium supplemented with 1 g/L activated charcoal. (c) Plantlets with light green leaves without necrosis problem on multiplication medium supplemented with 1 g/L L-glutamine. (d) Plantlets with high multiple shoots induction. (e) Induction of healthy root in rooting media supplemented with 1.0 mg/L NAA. (f) Roots induction on rooting media supplemented with 1.0 mg/L IBA. (g) and (h) Healthy plantlets ready to be acclimatized. (i) Development of successfully acclimatized plantlets.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4127217&req=5

fig4: Development of high quality plantlet from shoot bud explants of Calathea crotalifera on multiplication media (MS medium with 3.5 mg/L BAP and 1.0 mg/L NAA) and rooting media (MS medium with 1.0 mg/L NAA). (a) Induction of microshoots from incised shoot bud explants. (b) High quality plantlets with dark green leaves in multiplication medium supplemented with 1 g/L activated charcoal. (c) Plantlets with light green leaves without necrosis problem on multiplication medium supplemented with 1 g/L L-glutamine. (d) Plantlets with high multiple shoots induction. (e) Induction of healthy root in rooting media supplemented with 1.0 mg/L NAA. (f) Roots induction on rooting media supplemented with 1.0 mg/L IBA. (g) and (h) Healthy plantlets ready to be acclimatized. (i) Development of successfully acclimatized plantlets.
Mentions: Complete plantlets with high quality shoots and roots (Figures 4(g) and 4(h)) were removed from the rooting medium and the roots were carefully cleaned under running tap water to remove the residual agar. Each plantlet was then transplanted into plastic pots containing mixture of black soil, river sand, coconut husk, and vermiculite (4 : 2 : 2 : 1). The plantlets were then allowed to grow for two months under greenhouse conditions at 25 ± 2°C with direct sunlight for 12 h daily. Each pot was watered with distilled water for everyday and was covered with transparent plastic which was gradually removed. The morphology of leaf stomata from two months acclimatized plantlets were examined under scanning electron microscope (SEM). The leaf disc with 0.25 cm2 area from the in vitro and in vivo grown plantlets was used to compare the morphological development of the leaf surface and leaf stomata from two different conditions. The samples were fixed in 2% aqueous osmium tetroxide, OsO4 for overnight at 4°C. The treated samples were rinsed with distilled water for two times at 15 minutes each prior to dehydration through ethanol series (10, 20, 30, 40, 50, 60, 70, 80, 95, and 100%). The samples were then infiltrated through ethanol, acetone mixture at 15 minutes each, and ended with pure acetone for 1 hour before dried using a Critical Point CO2 Dryer. The samples were finally viewed under SEM after being gold coated. The presence of wax and morphological development of stomata aperture on the abaxial and adaxial leaf surfaces were observed and recorded.

Bottom Line: A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera.Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem.This is the first report of rapid mass propagation for C. crotalifera.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia ; Center for Foundation Studies in Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera. The effects of different sterilization techniques, explant type, and the combination and concentration of plant growth regulators on shoots induction were studied. The axillary shoot buds explants sprouted from rhizomes in soil free conditions showed high induction rate of shoots with lowest contamination percentage when treated with combination of 30% (v/v) NaOCl, 70% (v/v) ethanol, and 0.3% (w/v) HgCl2. In the present study, the highest number of multiple shoots was obtained in MS basal medium supplemented with 3.5 mg/L 6-Benzylaminopurine (BAP), 1.0 mg/L 1-Naphthaleneacetic acid (NAA), 3% sucrose, and 6 g/L plant agar for both varieties and was used as multiplication medium. Microshoots were highly induced when the young shoot bud explants were incised longitudinally prior subculture. Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem. The maximum roots induction was recorded on MS medium supplemented with 1.0 mg/L 1-Naphthaleneacetic acid (NAA) compared to indolebutyric acid (IBA). The complete regenerated plantlets were successfully acclimatized in the soilless medium under greenhouse condition. This is the first report of rapid mass propagation for C. crotalifera.

Show MeSH
Related in: MedlinePlus