As part of my PhD project I am using sterile micro-propagated potato plants for experiments.
I do this by cutting nodes from a mother plant, growing in full strength MS media supplemented with 2% sucrose, adjusted to pH 6,4 and 0,8% noble agar.
The plants are then grown in the same media in a closed container with a breathable membrane in the lid.
The plants grow fine, but I get a lot of variation in the size of the new seedlings both within each container but especially between containers. Its seems like for some reason plants in some containers are just growing way faster and some are barely growing.
Anyone has experience with how to get homogeneously sized seedlings?
Thanks in advance!
The cuttings can vary in performance for several factors. Assuming the medium is uniform, two likely factors are microenvironment and the cuttings themselves. Often your growth chamber will have some microenvironment differences (by the door vs in the back, sides vs center. You can change the positions of the containers to try and rotate them through all the different zones. Another possibility is the cuttings themselves. If the nodes vary in thickness or in distance from the main plant you might get performance differences. It could be worth tracking cutting size/distance to see if that is indeed important.
In the meantime, you can set a specific size parameter for your experimental cuttings and try to use uniformly sized and aged plantlets. Just be sure to propagate more than you'll need so you can discard the ones that are too big or too small.
Jonas Agergaard I would support opinion of Amy Klocko : 1) you should set a specific size parameter for potato cuttings and use uniformly sized and aged parts of mother plants; 2) rotate containers through all the different zones. Plants are very sensitive to small differences in temperature and light.
Some more exotic ideas: "sterile" plants are not always sterile (endophytic fungi, bacteria, and plant viruses). Article RNA-Seq of potato plants reveals a complex of new latent bac...
At least, all potato/tomato plants carry para-retroviruses (similar to CaMV) integrated into plant genome, and they are sometimes expressed, sometimes - do not.
If you rotate the containers to ensure equal environmental conditions, then the variation will be taken into account by your statistical tests if you have the correct replication :)
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