Hi, i started working with UCB1 pistachio micro propagation. I made some media according to Parfitt, D.E. and A.A. Almehdi. 1994.(i've attached the link to the paper). The media is composed of DKW supplemented with 30g sugar, Garmborg's vitamins, 85.5mM Zn, 156mM B and 9.9mM KNO3. I also added 1mg/L BAP and 0.01 mg/L IBA. Although the plants managed to grow, they don't callus properly. The stem above the media become swollen and the stem below the media just turn black. Over time, the stem just become woody/hard and turn reddish. The leafs also looks vitrified to me. I've tried to culture the plants on MS and 1/2 MS with no success. Can anyone suggest any papers or protocols i can follow to micro propagate them? Thanks!
http://www.sciencedirect.com/science/article/pii/0304423894900507
http://ijagcs.com/wp-content/uploads/2013/02/280-284.pdf
http://link.springer.com/chapter/10.1007%2F978-94-010-0125-0_19
Hello, Iu Wang
From your description, it seems rather a physics problem than chemistry. Have you tried to reduce the water in the medium, decrease sugar content, and change to 8h photoperiod? That might reduce vitrification and oxidation of the explants while increasing the number of calli. Good luck!
Hi, Lu
I am very pleased to hear someones studies on pistachios. I have a couple of suggestions for you, but first I should know what type of material used for the initiation of in vitro culture. Were they juvenile or mature? The more juvenile the explant the easier to establish the in vitro culture.Secondly why do you want to establish callus culture? and Swollen tissues, turning coloring of the tissues and vitrification are directly related to environmental and chemical culture conditions. What I mean is that I can properly explain solutions in order to sort out the culture problems you faced if you explain the aim of your studies. I have loaded most of my papers related to the micropropagation of pistahios in to the Researchgate portal. However, currently the UCB-1 is commercilly produced in Paramount Farming Company.
If you let to know me your aim porperly I shall explain my suggestion.
Kind regards
Ahmet
Hi Wang
My Sugestion is:
Using WPM media (For Woody Plants)
and using TDZ and NAA for Callogenesis
Good Luck
Hi
one question
Is the UCB 1 tissue culture same other pistachio variety?
Dear Lu Wang
Hi,You must remove phenolic compounds from the callus with scalpel. Use absorbent and antioxidants compounds reduce the proliferation. You can improve proliferation: the decreased concentration of ammonium nitrate (20%) and increased potassium nitrate (20%) in MS Basal medium.
vitrification depends on the quality of agar, matherial (chemical culture conditions) especially ammonium nitrate and growth chamber conditions, Which are easily controlled.
STN (shoot tip necrosis) can be controlled by increasing the concentration of calcium but But it affects the type of calcium.
I am using nano-calcium compounds to improve the protocol previously reported by myself.
best regards
seyed reza Nezami
TMU, I.R. Iran
Good louk.
Dear Iu Wang, could you tell us sth about the results? I am so interested about this :)
sure :) sorry for the late update Seray. I sticked with DKW media (but i believe you can use MS media too), but i switched EDDTA-ferrous sulfate to EDDHA and incubated them in the dark to reduce the phenolic compounds..the other thing i guess is that i need to transfer them more often (2 week worked for me)
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