I want to take data from tissue cultured plant but I am confused how and from where I should start.
Dear Farhan
In the question you have not mentioned the plant species. The date recording will depend on the objective of experiments and most importantly on the habit of plant; i.e whether it is herb, shrub or tree.
I suggest you may look in literature about similar work on the species you are working on or plant species belonging to the family in case no report exists on the species in question.
Ideally the data should be taken on important morphological, yield, biochemical and molecular parameters to assess if the tissue culture generated plants are true to type.
Best regards
Neelam
Actually i am working on Impatiens Balsamina plant that is ornamental plant..a herb belong to balsamenseae family...actually i tissue cultured the plant but bit confused how i can take the data from roots,shoot etc. the objective of the study is produce standard tissue culture procedure for the Impatiens species.
dear farhan, you can take the wet weight of callus, the optimum concentration of hormone, etc
If the purpose of developing this protocol is 'propagation of the species' then parameters like number of shoots per explant in 'x' no of days may be important. I assume once you obtain shoots you will seperate the shoots and transplant them to a rooting media. How many shoots germinate (% rooting) may be another important number to look at.
This will tell you your tissue culture efficiency.
Other parameters are shoot elongation ( every 'n' number of days), shoot length during transplantion, % rooting, and the total time of propagation (time for shooting, rooting etc are important to predict tissue culture efficiency. A short yet replicable and consistent (less variability) protocol is always desirable for commercial production.
Okay thank you so much i want to take morphological data. i got your point.
you start taking your reading from the meristematic region of the cultured plants, either from the shoot apical meristem or the root apical meristem. These areas are found in the shoot tip or root tip.
First of all you have to mention that what is you objective or what pathway of plant tissue culture is you are going to use? I am giving you example
First of all you have variable conditions for experiments along with this you have to set number of replicates for each treatment.
Now coming to the point..
To decide number of replicates you have to choose as much no. of replicates that SD value/ SE becomes low. Which gives uniform data and right conclusion of your results bcz many times less number of replicates resulted in more SD values and Slandered errors in the experiments
1, Number of shoot/s per explant and shoot length, Number of leaves
2. Percentage response
3.Shoot length
4.Number of roots and root length
5 type of callus i.e. embryogenic, compact, color of callus etc
6. Somatic embryo (SE): Number of embryo and percentage for each stage and conversion rate. Conversion rate is very important factor bcz 80% of soamatic embryogenesis related papers they reported different stages of SE but they are lack with information abt conversion rate form globular to torpido..Cotyledonary and finally acclimatization stage. and always keep a good quality of camera with you to record morphological developmental stages along with practical record book for data collection.
7. Keeping camera with record book is very important bcz day to day various small changes observed. Many times vitrification, contamination rate, effect of season, time of collection and number of repeated experiments etc are few more e.g.
Best wishesh
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