landzt

hi all ,is it best to to keep media a little on the soft side than to firm ?also with initiation does it if you use less 1/2 strenght m/s in media?

Gabe15

I have found it doesn't matter too much if its soft or firm, some systems don't use any gelling agent at all.

I would use at least 1/2 strength for standard micropropagation, though you may find that some varieties will be slow and not grow well. These will sometimes like full strength MS better, but I have not ever needed less than 1/2 strength.

__________________
Growing bananas in Colorado, Washington, Hawaii since 2004. Commercial banana farmer, 200+ varieties.

Faisar Mamacotao

Hello everyone. I have tried tissue culturing bananas using MS + 5 ppm BAP but the results are not good. The plants showed blackening and contaminated.

What shall I do? Are there any modifications to the MS that work good for bananas?

Please help. Thank you.

Cheers,

Faisar

jmoore

There is a broad spectrum fungicide and biocide on the market called 'ppm' which stands for plant preservation mixture, I think. The concentration is 1ml per litre of media. This may help, I'm using it at the moment, bit early to tell how effective it is yet.

Did you sterilise the explant in bleach before putting it into the media?

How good is your autoclave? Are all of your intruments sterilised prior to use?

The media you're using sounds OK, I think.

novisyatria

I also use PPM and the results is efective. i add 0,5 ml PPm in to 1 liter MS SOlution. we can add PPM 0,5 -2 PPM (based on manual instruction) but based on my experience 0,5 PPM is enough
regards

__________________
Novi Syatria
Lab of Tissue Culture , Dept of Forestry, Faculty of Agriculture, Univercity of Bengkulu, Indonesia www.kuljar.com email: novi_syatria@yahoo.com

Faisar Mamacotao

Thank you sir for that reply. Though I haven't used the ppm yet, so far contamination has been minimal, using bleach to sterilize the explants. But still, the explants are not growing well.

Since MS is a generic formula, which components of the MS should I vary?

Thank you.

novisyatria

if explant not growing maybe bleach is too hard or you soak too long so explant not good.

__________________
Novi Syatria
Lab of Tissue Culture , Dept of Forestry, Faculty of Agriculture, Univercity of Bengkulu, Indonesia www.kuljar.com email: novi_syatria@yahoo.com

landzt

Hello ,and thank you for the replys to thread i have learned a little more and have a few things to try .Do many people us PPM ? and is it only used in initiation or if a problem a rises.

novisyatria

i use PPM for all culture i have, but for initiation aybe more efectice with 1 ml/liter. for subculture 0,5 ml/lt is enough

__________________
Novi Syatria
Lab of Tissue Culture , Dept of Forestry, Faculty of Agriculture, Univercity of Bengkulu, Indonesia www.kuljar.com email: novi_syatria@yahoo.com

landzt

I have just recieved the PPM ,i have read were it is possible to sterilize media with ppm in ,but also read you can mix ppm in liquid sterilize it and add it to as a cover -like another lay in laminar cabinet i hope i have described it right ,is it best in media and only use liquid on light contamination were ppm is not in media?thanks

novisyatria

landt
sorry i dont full undrstand what do you mean. i add PPM to media. and never add to laminar. do you mean spraing IT to laminar? based on manual from plant cell biotechnology, i dont fine some information about sterilize laminar with PPM. its only used as broad spectrum fungal and bacterial and added in media.
regards

__________________
Novi Syatria
Lab of Tissue Culture , Dept of Forestry, Faculty of Agriculture, Univercity of Bengkulu, Indonesia www.kuljar.com email: novi_syatria@yahoo.com

stevethelizard

Carol at the home tissue culture group uses a liquid overlay on top of agar media i have never tried but it seems a good idea

novisyatria

ohh i got what lantd mean now. maybe its good idea but i think we have to prepare the media 2 times. first wait until media/agar is cold. second we add PPM liquid. thats mean PPm solution better if in steril condition. so we have to filtering firts with syringe filter and when we add PPM must in laminar air flow. give it to media in bottle one by one. i think its needs more time. its oke if we just make a little media but not eficient if a lot media. i make 100-200 bottle/day and if i do like carol maybe not efficient. based on my experience add PPM to media directly when we make media is good enough.
regards

__________________
Novi Syatria
Lab of Tissue Culture , Dept of Forestry, Faculty of Agriculture, Univercity of Bengkulu, Indonesia www.kuljar.com email: novi_syatria@yahoo.com