PJB-2020-344
In vitro shoot multiplication in gerbera (Gerbera jamesonii)
Dr. Monis Hussain Shah
Abstract
Gerbera (Gerbera jamsonii L.) is one of the most famous and beautiful cut-flower of tropical and subtropical climate. It is one of the most famous flowers known as Transvaal daisy or Barberton daisy. The efficacy of different explants (Clumps, callus and embryo rescue) for multiple shoot induction for further research in Pakistan. The clumps were used for callus induction while seeds were sterilized for embryo rescue. The variety subjected for both callus formation were cv. Submarine, Danallen, savanna and stanza, while for embryo rescue/embryo culture Mexican eye catcher Red Bicolor with White was used. Callus induction through clumps was achieved followed by washing with running tap water for an hour followed by the 30 minutes with 1g/L of Topsin M fungicide. The explant were further subjected to washing of explants for 5 minutes with 10% NaOCl (Bleach) followed by rinsing in 70% ethanol followed by washing with autoclaved distilled water. The explants were further rinsed in 0.1% of HgCl2. The seeds of cv. Mexican eye catcher were washed with 0.1% of HgCl2 followed by 99.9% Ethanol. The seeds were finally treated with 1g/L of Topsin M fungicide. The culture conditions for clumps and seeds were 16 hr. light and 8 hr. dark in the growth rooms of laboratory while seeds were placed in the dark. The explants were cultured in plane MS-Medium initially followed by the MS-Medium modified with Naphthalene acetic acid (0.2, 0.4, 0.6, 0.8 and 1.0 mg/L), 2, 4-Dichlorophenoxy acetic acid (1, 2, 3, 4 and 5mg/L), Naphthalene acetic acid+6-Benzyladenine (0.2+1, 0.4+2, 0.6+3, 0.8+4 and 1.0+5 mg/L) and inverse dose of Naphthalene acetic acid+6-Benzyladenine for callus induction from clumps. The MS-Medium was modified with 6-Benzyleadenine (1, 2, 3, 4 and 5mg/L), Indole acetic acid (0.5, 1.0, 1.5, 2.0 and 2.5mg/L), 6-Benzyleadenine+Indole acetic acid (1+0.5, 2+1.0, 3+1.5, 4+2.0 and 5+2.5mg/L) and 6-Benzyleadenine+Indole acetic acid (1+2.5, 2+2.0, 3+1.5, 4+1.0 and 5+0.5mg/L) were used for shoot mortification of plant lets form embryo rescue. The Sterilization protocol was successful for clumps and seeds. The culture sustainability rates in Clumps (95 %) and seeds (99%) were observed. The clumps were subjected in 2, 4-D for callus induction. The callus induction in plain MS-medium was also seen however it was non-regenerative. The mean best and friable callus were observed in various levels of 2, 4-D (4.39). The maximum wait of callus was (1.30g), with leafy green color of with small globular embryos. The callus was than subjected to various doses of BA and BA+IAA (3+1.5mg/L) for shoot induction however no significant shoot regeneration was observed through callus. To generate the population seeds were planted into the culture medium. The seed was vulnerable as viable when its coat is burst and it became open. The seeds were picked and coats were removed at the seed coat bursting stage followed by cultured again in vertical position from embryonic side. The plantlets were formed and subjected to the multiplications in various doses of BAP, Kinetin and combinations as well. The least days for start of shoot induction was observed in MS-Medium modified with 2.5mg/L 6-Benzyleaminopurine (BAP) in cv. Mexican eye catcher (5.67), while the largest number of shoots were observed at 2mg/L of (BAP) (13.67), while greater number of shoots were observed at 3mg/L (11.46).
Key Words: Gerbera, 6-Benzyl adenine, multiple shoot induction, Callus and clumps
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