With Agrobacterium we have transformed potato tuber disc peeled and unpeeled ones and within one week the unpeeled potato tuber disc with an eye has some shoots! we are wondering if we should consider them transformed
Hello! I'm sorry, but the shoots that you got are not transgenic. These are just dormant buds that have grow up. for stable transformation, 1 week is not enough. You had to first obtain a callus (you must use phitogormones), and then regenerate it to shoots. it usually takes 3-4 months to get potato regenerants.
Thank you. I guess i will have to wait for the callus then. by phitogormones you mean phytohormones right?
Try partially peeled potatoes and unpeeled (pierce and make pin holes) and send me the result.
Yes, phytohormones. You can use 0.1 mg/L BAP and 1 mg/L NAA in standart MS-medium (1.6% glucose + 1 mg/L biotine + 5 mg/L Ca-pantothenate) for callusses genesis (about 1 month). Then you have to use 40 mg/L adenine, 2 mg/L trans-zeatine, 0.5mg/L gibberellic acid (GA3), 100 mg/L kanamycine and 500 mg/L claforan or cefotaxim in standart MS (like discribed before).
Dear Leonce, I believe If you use sliced microtuber with sharp pricks and treat with agro suspension, you can get your transgenics with approperiate harmonal combinations. You may need vigorous screening though.
Why do you use tubers?
For transgenic exp. I always use leaves and prick into the veins. Hairy roots appear sometimes already after 11 days.
And as you got recipees for getting shoots from callus you can use for getting shoots from the roots. Or even grow the roots under light on hormone free medium. This does not work for every plant spec. But potatoe is so weedy...
S. Wenkart the main reason i wanted to use tubers is that they seem to handle infections.can i get the protocol you are using it will be really helpful since I'm running out of time
You got shoots from the tuber pieces and they seem to be sterile and with leaves. As cut them off and use for transformation. One can insert the Agro. in the stem pieces too.