I want to do some expression analysis experiment in wheat under drought stress, but a little confused in which method to use.may i go for hydroponics or soil pots or directly germinate the seeds on filter paper.
Dear Tinku Goutam
By growing plants in pots, you can investigate effects of drought stress on different growth stages and performance of wheat. Also in this condition, adjustment of stress treatments is easier compared to other planting substrate. While by cultivating on filter paper, only studying of a limited period of plant growth will be possible for you. Therefore, cultivation in soil pots is recommended.
Best wishes
Soil pot method will be better than others hydroponic will good for salinity stress or nutrition effect experiment.
Anyone can please suggest me that when to stop watering at the seedling stage for producing drought stress and when to harvest the leaf samples for RNA isolation. And how to prepare the pots if we have sandy soil type.
If you want to check effect of drought on seed germination, you may use PEG 6000, Petry dishes and filter paper. If you want to check effects of drought stress on seedlings or saplings, I suggest using pot soil.
If you want to study the effect on seedling stage you can use sub-stratum like blotting paper, cotton or soil. Water stress can be created by using polyethylene glycol (PEG-6000). You can also use slanting plate method for studying the effect during seedling stage and drought can be simulated by using PEG.
But to study the effect on growth and development, and also on yield, you can use soil pot method and for this water stress can be created by withdrawing water application at your desired stages of growth.
You can germinate wheat seeds in sterile water and subsequently transfer the germinated seedlings to osmotic stress which may be induced using PEG 6000 or 8000 grade for simulation of drought stress. For expression analysis through qRT-PCR, germination paper rolls of discs may serve the purpose and are easy to use compared to hydroponic set up and that also depends on number of genotypes you want to study.
I recommend soil pots for phenotypic and genotypic analysis. Hydroponic is also good but you have to make sure you are adding the exact same amount of PEG in each repetition...I think that soil pots could give more material though (leaf surface, root development)....
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