Micropropagation of Rosa damascena Mill. through transverse thin cell layer culture

2012 ◽  
Vol 2 (4) ◽  
pp. 184-189
Author(s):  
Kavita Kshirsagar ◽  
V. J. Braganza

A basic factor underlying the success of large‐scale micropropagation and genetic transformation of any plant species is regeneration. In order to regenerate propagules of Rosa damascena Mill. on a large scale, an efficient and improved in vitro propagation system has been established using transverse thin cell layer culture (tTCL). By optimizing the position of the tissue and applying an improved selection procedure, in vitro shoots were elongated in 8 weeks of culture. Modified Murashige and Skoog (1962)(MS) medium fortified with 4.0 mg l‐1 6‐benzylaminopurine (BAP) and 0.4mg l‐1 anaphthalene acetic acid (NAA) gave optimal shoot regeneration. The explant was inoculated on this medium in the upright position and exhibited a high frequency of shoot regeneration (~96.66%), and it also gave the highest number of shoots (22.33/explant). The horizontally placed explant on an average 7.66 shoots/explant. Our experiments indicate that explant orientation strongly influences the organogenesis response. The frequency of shoot initiation and the number of multiple shoots produced from each explant were significantly dependent on the plant source, concentration of plant growth regulators and the orientation of the explants and contributed significantly to in vitro regeneration. Rooting of well developed shoots was achieved on hormone free ¼ strength MS medium with 4% sucrose.

2017 ◽  
Vol 14 (2) ◽  
pp. 24-31 ◽  
Author(s):  
S S Riva ◽  
A Islam ◽  
M E Hoque

An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration.  It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium.  For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.The Agriculturists 2016; 14(2) 24-31


2021 ◽  
Vol 19 (2) ◽  
pp. 337-347
Author(s):  
Hoang Thanh Tung ◽  
Hoang Thi Van ◽  
Huynh Gia Bao ◽  
Le The Bien ◽  
Hoang Dac Khai ◽  
...  

In vitro culture establishment is one of the most important stages in micropropagation. The disinfectant effectiveness depends on the type of surface disinfectant, concentration and the time treatment. In this initial study, silver nanoparticles (AgNPs) were used as a disinfectant for petioles, flower stalks and stems of Begonia tuberous. In addition, thin cell layer culture (TCL) technique has been applied for the purpose of somatic embryogenesis. The results showed that AgNPs were effective in eliminating infectious microorganisms on B. tuberous explants; which were identified included 4 species of fungi (Fusarium sp., Aspergillus aculeatus, Trichoderma sp. and Penicillium sp.) and 1 species of bacteria (Pseudomonas sp.). At concentrations of 200 ppm and 300 ppm, AgNPs were not only effective in disinfection but also increased the induction rate of somatic embryogenesis in flower stalk TCL explants (approximately 40.00%); a similar effect was observed in stem TCL explants at the same concentration. Meanwhile, for petiole TCL explants, the induction rate of somatic embryogenesis was optimal when using AgNPs at a concentration of 100 - 300 ppm to disinfected the explant. In contrast, at high (400 ppm) or low (50 ppm) concentrations of AgNPs did not play a disinfecting role and stimulated somatic embryogenesis. In addition, explants derived from AgNPs sterilization did not show any abnormalities in somatic embryogenesis with shapes such as globular, heart, torpedo, and cotyledon. AgNPs showed double efficacy in sterilization of explants and improved efficiency of somatic embryogenesis from TCL petioles, flower stalks and stems explants; thus increasing the efficiency micropropagation of B. tuberous.


2019 ◽  
Vol 8 (4) ◽  
pp. 6934-6938

The present work was carried out to investigate the effects of gamma radiation on regeneration of Capsicum annuum L. var Kulai via in vitro. Seeds of C. annuum were irradiated with various doses of gamma ray (0, 20, 40, 60, 80, 100, 200, 300, 400, 500, and 600 Gy) emitted from the Caesium-137 source at the rate of 4.31 Gy per minute. Irradiated seeds grown on MS medium without hormone for hypocotyl and cotyledon preparation as explant for in vitro regeneration. Seed germination rate revealed significant variation between treatments, and seeds started to germinate between 6 to 17 days. Irradiated seeds between 0-60 Gy were observed to germinate in less than 10 days. All explants including hypocotyl and cotyledon were cultured on MS medium with different concentrations of BAP in combination with AgNO3 to observe the response of these explants to different hormone concentrations. From the observation, calluses were induced in 90% of hypocotyl and cotyledon explants in all treatments. The characteristics of calluses were varied with greenish friable, greenish compact, yellowish watery, yellowish friable and yellowish compact. In other treatments, calluses were found in purple, bright yellow and yellowish orange. On the other hand, shoot regeneration was observed in treatment between 40-100 Gy. In conclusion, gamma radiation gave impact on seed germination, seedling growth performance, in vitro callus formation and shoot regeneration of Capsicum annuum var. Kulai


2014 ◽  
Vol 6 (1) ◽  
pp. 85-91
Author(s):  
Dikash Singh THINGBAIJAM ◽  
Devi Sunitibala HUIDROM

An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL) sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%). Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25). Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%). Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.


2012 ◽  
Vol 47 (2) ◽  
pp. 211-216
Author(s):  
KK Paul ◽  
MA Bari

An efficient in vitro regeneration protocol was developed in medicinal aroid, Ghetkachu (Typhonium trilobatum Schott) using field grown corm bud explant. Highest percentage (75 %) of direct multiple shoot regeneration obtained in MS media supplemented with 5.0 mgL-1BAP + 1.5mg L-1NAA. Callus formation occur (80 %) in MS media containing 0.5mgL-1BAP + 2.0mgL-1NAA. The appearance of calli was white, creamy white light green in colour and the texture of calli were soft, friable and semi hard and compact. Shoot regeneration (85 %) obtained from calli in MS medium having 5.0mgL-1BAP +1.0mgL-1NAA. The regenerated plantlets were successfully acclimatized with loamy fertile soil and survived cent percentage in natural condition.   DOI: http://dx.doi.org/10.3329/bjsir.v47i2.11454   Bangladesh J. Sci. Ind. Res. 47(2), 211-216, 2012  


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