Respected All,
I am facing a problem in an experiment. A necrotic leaf tissue have more chlorophyll content than a healthy one while quantifying with same amount of leaf (same weight). I tried to find out the reason by scanning electron microscopy (SEM) which showed that the cells become smaller and more compressed in tissues having necrosis. Is there any technique that can support my theory?
Can you upload some photographs of the leaves you are talking of? Cell shrinking should not have much to do with increasing chl concentration because you are using weight as criteria and not area.
First, you need to specify what is meant by "necrotic". In a sense you are describing tissues containing dead cells. If that is the case, usually there is no active chlorophyll to speak about. If you mention instead some mutants tended to form necrotic patches and interested in chlorophyll concentration of living cells, then you can calculate that by spectrophotometrical, SPAD or fluorescence ratio methods
Dear Dr Levinsh,
The dead cells was detected by the microscopy using dyes specific for detection of dead cells in plants. Yes, I am interested in measuring the chlorophyll content of living cells. I followed the spectrophotometrical techniques for the same.
In your photograph - the infected leaf appears green - an indication that we associate with living tissue - you have indicated detection of dead cells by microscopy - was it following TTC treatment - sometimes dry leaves also appear green for sometime - shrinking of leaves means loss of turgor - less moisture - that explains your observation of small cells in microscopy.
- the spectrophotometric detection of chlorophyll has issues with accuracy - due to artifacts etc - you can have a look at the several posts for the same in research gate and optimise - best is to go for fluorescence ratio -that will help you differentiate functional and non-functional chl - I am sure someone at NBRI has a mini PAM or dual PAM.
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