In our study to improve faba bean plant to be tolerant to salt and drought stresses using genetic modification, we want to confirm the obtained results with some chemical and physiological studies.
you could germinate the seeds of faba bean in vitro, and then treat the seedlings with different concentrations of Na Cl starting with the threshold level of salinity tolerated by the plant. for drought tolerance you could use different levels of PEG. then you could determine vegetative growth characteristics of the plants as well as determination of proline, total soluble carbohydrates, and anti oxidant enzymes, such as peroxidase, as well as endogenous hormones. you could also determine the protein pattern of the genetically modified plants compared with control.
We assayed salt stress in vitro with citrus seedlings using test tubes. Germination was carried out in agar water and then for salt stress the dose was 40 mM NaCl in 4% agar modified hoagland nutrient media. Root growth can be measured weekly, leaf area damaged at the end, as well as Cl and Na content in plant parts. Whole plant growth is also important. Whole plant transpiration can be assayed in seting the plant in a separate isolated glass with water and measuring water loss by weighting the system along a light cycle.
First of all you have to study the PH of the soil where faba bean grows well.Then maintain that Ph of the culture media. Add different concentration of Na cl and prepare the media will give the salinity suitability.For drought study in tissue culture method i do not have any idea.
I confirmed with Muayed Fadhil Abbas in the method described.
Different concentrations of NaCl and PEG can be employed in tissue culture tubes itself for accounting survival percentage. In regenerated seedling stage after appropriate treatments the following physiological parameters may be analyzed for relative salt and drought tolerance:
1. Chlorophyll content
2. Leaf proline content
3. Leaf water potential
4. PS II activity and
5. A few ROS scavenging enzyme assays (peroxidase, SOD and catalase etc)
I agree with previous responses, but you could also assess the integrity of cell membranes and quantify the amount of ions released to the medium (deionized water)
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