About Agrobacterium mediated transformation in potato. Why explants turn white on media containing hygromycin?

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Rajesh Dev Sarkar

did you optimize the critical concentration of Hygromycin in potato culture?

Francesco Desiderio

Hi,

could you please attach some photo? Would be helpful in order to answer correctly to your question.

Best regards

S. S. Datir

Thanks Rajesh...I tried 10 and 25 ug per ml of hygro and I found them white. Currently, I am trying 5 ug/ml of hygro.

Hello Francesco...I will upload some photos.

First of all you can apply different concentrations of Hygro suppose 5- 50 ug in untransformed explants, to know at which minimum concentration of Hygro, cell growth is inhibited. Now if u get that at 10 ug, plant cell growth is well inhibited, then you can apply 10 ug in your transformed explants. So apply the minimum possible concentration.

Thanks Rajesh, I think I will do that and see what happens.

I would suggest you to check this article:

Article Effective hygromycin concentration for selection of Agrobact...

It appears to me (thanks for the picture) that there is some inhibition going on.

Agrreing with dr. Rajesh Dev Sarkar I would suggest you to try with lower concentration of antibiotic and then scale up to better understand the limits.

I think that those articles would be useful for a better understanding:

https://www.researchgate.net/post/Why_do_most_of_the_people_select_pCambia_vectors_for_cloning_in_transgenic_plants

Article Relationship between expression level of hygromycin B resist...

https://www.researchgate.net/post/Removing_agrobacterium_from_plant_tissue_culture_media2

If you have other antibiotics I would suggest you to try with others.

https://www.agriculturejournals.cz/publicFiles/73144.pdf

http://horticultureresearch.net/journal_pdf/20013-12.pdf

Thanks, I will.

Yuan-Yeu Yau

I agree with Rajesh Dev Sarkar.

In the attached paper, 25 mg/L of hygromycin is used to select transformed potato calli. Different potato cultivar/variety will response to selection agent concentration differently. It can depend on which variety of potato you are using for transformation. The authors used potato cultivar Iwa (see attachment).

Thanks Yuan