Tissue culture protocols have generally following stages

Please see details about Banana Tissue Culture on follwing links

http://www.slideshare.net/sathes32/tissue-culture-techniques-of-banana

http://www.agbiotek.com/Banana_Tissue.php

I think the first link (slide show) provided by Uttar is a pretty good one which has all the details and pictures of all stages of banana tissue culturing. I like it.

By the way, for different variety/genotype of banana the explants of each stage can be different. So, just use other people's publications as references, because the experimental results of your banana can be different from others'.

Thanyou Uttar and Yuan. Actually I have gone throught that already and I am not yet sure that how the culture looks when they start to respond. I mean like color changes, Peeling of outer layer of tissue or swelling like that. In most of the data they have said that changes occur  in 3 weeks. But I need to know the visible changes that occur in a week or two after inoculation. 

Dear Santosh

During banana micropropagation following stages occurs.

Initiation of aseptic culture

Culture establishment and swelling in shoot tips

Multiplication

Elongation of developed plantlets

Rooting

Hardening

The in vitro of cultures vary from genotype to genotype Ie. AAA, AAB, AB genomes responded differently.

During establishment phase shoot tip starts swelling after 2 weeks. U can't see any remarked changes within week.

For better understanding you focus on daily observations.

By the way, during initial phase, developing cultures seen creamish white, because of dark incubation. When these cultures transferred to light, converted to green.

For better understanding download some good research articles according your variety of interest or type of genome. Ie.AAA  genome- Grand naine.....

Dear Rajesh,

Thank you For the answer. 

I am trying to culture a local variety of banana. So I donot have the exact idea about the genotype of the cultivar. So I have been trying by varying the concentration of hormones from the papers published. 

And also can you please tell me the principle behind the Dark incubation during the culture initiation? I mean how the dark incubation prevents the phenolics production?

Regards,

Santosh Dahal

Dear Dahal

Initial incubation in darkness helps reduce the production of secondary metabolites including phenolics. In addition, darkness prevents the oxidation of such phenolic compounds.

Addition of antioxidant compounds such as ascorbic acid, citric acid to the medium is very helpful

Dear Omer,

Thankyou for your response. As per the papers I incubated the explants in the dark for a week and then transferred to light. The explant turned green and showed swelling. 

But there is no any bud or growth even after a month. So can you please provide me your expertise knowledge that what might be the reason for that. 

I have also included the pics of the explant. Please have a look on it.

Regards,

Santosh Dahal

Induction of somatic embryogenesis in explant culture should yield embryos regenerating into plant.

Santosh

After a certain period you need to cut off the explant to expose fresh tissue as the blackened surface is dead due to phenolisation. Generally within 2 weeks response is quite visible. Also check photo gallery by Novyastria on bananas.org she gives a practical insight than would be discernible from a collection of papers. Media composition  (hormones) and use of antioxidants are also critical to your success. Goodluck