How to increase the length of in vitro multiplying shoots of any explants. Do we should increase the concentrations of Cytokinin or add gibberellins? suggest me if you have experienced in your lab.
Cytokinin promotes shoot proliferation but may inhibit elongation if used in excess. Therefore, you can use gibberellins or combinations of ½ MS + 0.5 mg/L GA₃ + 0.1 mg/L NAA + 3% sucrose.
The general rule is that the more shoots you have, the smaller they are. An optimal cytokinin concentration can induce many shoots, but the trade-of is that the shoots will be smaller as they compete for nutrient and space. Therefore, you may want to use a cytokinin concentration that is a bit "sub-optimal" to induce just enough shoots and have them grow better. Or you can do an initial "kick" with cytokinin, then move the shoot cluster to a medium with no cytokinin for the shoots to develop and elongate.
Light can also be a factor. From my experience, monochromatic red light greatly induce elongation in plants. However, I've yet tried it on in vitro shoot clusters.
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