Try 2,4-D alone without kinetin. Also make us you germinate your seed in vitro under dark condition .

Hi,

I do agree with Lateefat Hassan, try 2,4-D alone.

I am working on tea which is a dicot. Callusing from leaves, finding a bit difficult. Please suggest a better medium.

Jameel sir, I'm working with Camellia sinensis, plant producing tea beverage.

Thanks Salamma. I'll definitely try this combination.

Hi,

I would also suggest 2,4-D- in range from 0.5-2 micromole will work, if not than you can increase the concentration.

Combining the Cytokinins with auxins may give you callus but it can be a morphogenic one and not friable or compact.

other things you can try is-

1) at the time of inoculation, wound the explant at the surface that touches to the medium, it would increase the chance and/or size of callus;

2) change the position of explant inoculation i.e. abaxial to adaxial or vice versa, as this can sometimes induce callus, i observed it in Bacopa monnieri leaf explant.

Hope it will work. 

Thanks Lateefat and Nagendra. Thanks for the papers Jameel sir. Ashutosh I have already tried wounding and surface changes. Now time to try the medium change. Hope it works. Once again thank you all for your valuable suggestions. Really appreciate them.

There is not general role to induce callus from any plant leaf. So, if the aim getting callus only, plant material, growth regulators and physical condition, it is enough, In addition all mention above, leaf Number on plant (leaf No. 3 or 4,...etc,) that was used as explant for callus induction, also as Ashutosh mention" position of explant inoculation", it sometimes considered critical factor, besides increase thiamine level. But if the aim for callus induction and regeneration, There are many factors affecting .Of them . auxin type and level ( 2,4-D singly or combined with BA or Kinetin) for callus induction,then reduce or omit 2,4-D or replace by NAA plus the different level of Kinetin or BA. Another way, using TDZ for callus induction, then replace it by BA in different level.............................hope it helps,

Thank you Metwally. At present my aim is just callus induction.

In addition to MS medium you can also try WP medium along with 2,4-D either alone or with BA or NAA. Can go through Brijwal et al.2015

In Vitro Cell.Dev.Biol.—Plant

DOI 10.1007/s11627-015-9716-7

Thank you Sushma.