Articles
IN VITRO MANIPULATIONS OF ROSA BOURBONIANA L.
Article number
769_51
Pages
357 – 370
Language
English
Abstract
This study presents highlights of research centered on in vitro manipulations of Rosa bourboniana, an important scented rose species that yields rose oil.
The results describe optimization of micropropagation procedure using nodal explants, callus induction, and isolation of protoplasts for further manipulations.
Quicker response and multiple shoot induction were observed when nodal segments of R. bourboniana were cultured on Murashige and Skoog (MS) medium containing 4.44 µM BA and 0.27 µM NAA with 76% response and 7.5 ± 0.22 shoots per explant with an average length of 6.2 ± 0.41 cm after 45 d of cultivation.
These shootlets, when transferred to half strength MS basal medium supplemented with 2.46 µM IBA and 1.08 µM NAA, produced maximum number of roots (13.2 ± 0.32) with an average length of 6.5 ± 0.42 cm after 45 d.
The survival rate has been over 70% when the rooted plantlets were transferred for hardening.
Callus induction was studied from various explants such as juvenile leaves, shoot tips and nodes resulted from nodal explants on MS medium containing 4.52 µM 2,4-D and 2.22 μM BA. Optimum callus biomass production to the tune of 403.2 g L-1 fresh weight and 24.6 g L-1 dry weight was achieved in MS solid medium containing 2.26 µM 2,4-D, 2.22 µM BA and 4.65 µM kinetin after 24 d of culture.
When the above combination was used for suspension culture, optimum biomass production was achieved with 145.3 g L-1 fresh weight and 11.8 g L-1 dry weight after 21 d of culture.
Efficient protoplast isolation protocol has also been worked for suspension culture grown cells.
Maximum number of protoplasts was isolated using Cellulase (0.5%), Macerozyme R-10 (0.1%) and Pectinase (0.25%) and CPW13M was found suitable to stabilize osmotic pressure in washing and isolation solutions.
The highest protoplasts yield observed was 4.05 × 106 ml-1. Isolated protoplasts were purified and subjected for microcalli development.
The results describe optimization of micropropagation procedure using nodal explants, callus induction, and isolation of protoplasts for further manipulations.
Quicker response and multiple shoot induction were observed when nodal segments of R. bourboniana were cultured on Murashige and Skoog (MS) medium containing 4.44 µM BA and 0.27 µM NAA with 76% response and 7.5 ± 0.22 shoots per explant with an average length of 6.2 ± 0.41 cm after 45 d of cultivation.
These shootlets, when transferred to half strength MS basal medium supplemented with 2.46 µM IBA and 1.08 µM NAA, produced maximum number of roots (13.2 ± 0.32) with an average length of 6.5 ± 0.42 cm after 45 d.
The survival rate has been over 70% when the rooted plantlets were transferred for hardening.
Callus induction was studied from various explants such as juvenile leaves, shoot tips and nodes resulted from nodal explants on MS medium containing 4.52 µM 2,4-D and 2.22 μM BA. Optimum callus biomass production to the tune of 403.2 g L-1 fresh weight and 24.6 g L-1 dry weight was achieved in MS solid medium containing 2.26 µM 2,4-D, 2.22 µM BA and 4.65 µM kinetin after 24 d of culture.
When the above combination was used for suspension culture, optimum biomass production was achieved with 145.3 g L-1 fresh weight and 11.8 g L-1 dry weight after 21 d of culture.
Efficient protoplast isolation protocol has also been worked for suspension culture grown cells.
Maximum number of protoplasts was isolated using Cellulase (0.5%), Macerozyme R-10 (0.1%) and Pectinase (0.25%) and CPW13M was found suitable to stabilize osmotic pressure in washing and isolation solutions.
The highest protoplasts yield observed was 4.05 × 106 ml-1. Isolated protoplasts were purified and subjected for microcalli development.
Authors
B. Janarthanam, S. Seshadri
Keywords
Rosa bourboniana, in vitro culture, cell biomass, protoplast, microcalli
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