The best basal medium and its strength from the previous experiment were prepared and supplemented with different cytokinin; 6-benzylaminopurine (BAP), kinetin (Kn), Zeatin (Zn), and 2-isopentenyl adenine (2iP) (Duchefa Biochemie, Netherlands) at different concentrations of 2.5, 5, 7.5, 10.0 µM. The control treatment was devoid of any plant growth regulators. The cultures were maintained for four weeks of incubation. The parameters taken for this experiment were the number of shoots, length of the shoot (cm) and the number of leaves. All the data collection was carried out after four weeks of culturing.
Zeatin
Manufactured by Duchefa Biochemie
Sourced in Netherlands
Zeatin is a type of plant growth regulator that belongs to the class of cytokinins. It plays a crucial role in plant cell division and differentiation, promoting cell growth and development. Zeatin is commonly used in plant tissue culture and biotechnology applications.
Automatically generated - may contain errors
4 protocols using zeatin
1
Optimization of Shoot Regeneration via Cytokinin Supplementation
2
Generating Transgenic Tobacco Plants
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Transgenic tobacco plants were generated through the agrobacterium-mediated transformation method as previously described (Horsch et al. 1989 ). Fully expanded unstressed leaves from a sterile cultured Nicotiana tabacum cv.Xanthi plant were cut into squares with diameter of 0.5–1.0 cm, and the leaf disks were incubated with LBA4404 agrobacterium harboring a plant expression vector for 10 min. After the inoculation, the disks were placed with their adaxial surfaces facing downwards on co-cultivation medium [3% sucrose, 4.4 g/L MS salt (Duchefa, Netherlands), 2 mg/L 2,4-D (Duchefa, Netherlands), and 200 μM acetosyringone (MBcell, Korea), pH 5.8]. After 2 days of co-cultivation, the disks were transferred into shoot induction medium [3% sucrose, 4.4 g/L MS salt, 50 mg/L kanamycin (Biosesang, Korea), 0.01 mg/L NAA (Duchefa, Netherlands), 0.1 mg/L GA3 (Duchefa, Netherlands), 1 mg/L zeatin (Duchefa, Netherlands), and 250 mg/L cefotaxime (Biosesang, Korea), pH 5.8]; the explants were transferred into fresh shoot induction medium every 2–3 weeks. The regenerated shoots were transferred to root induction medium (3% sucrose, 4.4 g/L MS salt, 50 mg/L kanamycin, and 300 mg/L cefotaxime, pH 5.8). The regenerated plantlets were transferred to soils.
3
Ginger Rhizome Propagation Protocol
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Fresh mature rhizomes of ‘Bentong’ ginger were collected from Bentong, a western part of Pahang in Malaysia. All the chemicals required for basal media were analytical grade purchased from R & M Chemicals, Essex, UK. Zeatin, TDZ, and Gelrite were purchased from Duchefa, Haarlem, The Netherlands. Kinetin, BAP, NAA, IAA, and IBA were purchased from Sigma Chemical Company, Saint Louis, Missouri, USA.
4
Callus Induction and Suspension Culture of Medicago truncatula
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Calluses were produced from leaf explants of Medicago truncatula cv. Jemalong, cultivated in a growth room at 24 ± 1 °C under a 16 h photoperiod of 70 µmol m−2 s−1 GreenLED (Philips), according to Orłowska et al. (2017 (link)). For callus induction, initial explants were placed on Petri dishes (ø 55 mm) with the SH medium (Schenk and Hildebrandt 1972 (link)) containing 0.5 µM 2,4-d (Duchefa Biochemie, Haarlem, The Netherlands) and 1.0 µM zeatin (Duchefa Biochemie) with 30 g l−1 sucrose solidified with 2.5 g l−1 gerlit and adjusted to pH 5.7. The cultures were kept at 28 ± 1 °C in the dark for 21 days. Suspension cultures were initiated from 0.5 g proliferated calluses by sub-culturing in 100 ml Erlenmeyer flasks in 25 ml liquid B5 medium (Gamborg et al. 1968 (link)) supplemented with 2.5 µM 2,4-d and 4.5 µM kinetin (Duchefa Biochemie) at 18 °C for 14 days.
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