I am working on Okra tissue culture but i am facing problem about okra seed germination , for seed treatment i am using HgCl2 0.1% for 3 minutes and after washing with dd water use 75 % Ethanol for 15 seconds and washed with dd water for 5 times and soaked in water for one night and next day put in 1/2 MS media ( 15 g sucrose, 8 g agar and 2.37 g MS powder media for 1 litter ) and place in dark for 1 to 2 days until the plumes come out and then placed in florescent light . i have tried many times, i have changed seed from different companies but some time good germination and next time 0%. can anybody help me about this problem ?
The mercury chloride may be killing the seeds since it is highly toxic (and also to humans). You could use bleach instead. Other reasons could be could be extremes of watering (although, pre-soaking is necessary for germination) or poor seed. You could do a germination trial in soil before using the seed in sterile media. The temperature should also be at least 21 degrees C.
Dear Hafiz Rizwan,
Mercury chloride is lethal effects on seed germination, if u want to use, it should be 0.01% for 1 or 0.5 minutes but it better to avoid for environmental issues. There is other option that without Mercury chloride u can use 10% (v/v) sodium hypochlorite solution with 2-3 drops of Tween-20 for 30 min. followed by washing with sterile distilled water 3-4 times, after washing with sterile dd water use 50% to 75 % Ethanol for 15 seconds and washed with sterile dd water for 3 times after all your seed would be ready for inoculation in different 1/2 to 1/3 basal MS media or inoculate in autoclave seed germination paper under in vitro test tube contain seed germination paper moister with only 1/2 or 1/3 MS basal nutrients media or only autoclave DD water without use agar or any carbohydrate source. By this protocol it will give positive result.
Thank u
why don't you use sodium hypocrite for surface sterilization? in my opinion sodium hypocrite should be the first priority for surface sterilization, try various concentrations of this disinfectant with different time period, then in second priority you can try hydrogen peroxide &.....
good luck
Yes Aida Shomali, first step can be use sodium hypocrite for surface sterilization then follow next step, that i did mention in my ans....
Thank u
Dear Rizwan
Acid scarification of okra seed for 3 min in 80% H2SO4 is the most effective treatment combination of breaking dormancy and enhancing germination since it had the highest germination rate of 96.6%. Soaking okra seed in water for 12 h at 30°C and dry heating for 5 min at 70°C also promotes high germination rates but germination is delayed to a period of 12 days.
http://www.academicjournals.org/journal/AJAR/article-full-text/27EB5A852548
Best wishes
Dear Hafiz
In addition to the toxicity of mercury chloride, okra shows a particular kind of seed dormancy, called delayed permeability, caused by the structure of the seed coat and particularly by the chalazal plug. The combination of temperature and time (40°C for 4 days and 100% RH) were pointed out to solve the dormancy.
Nada, E., Lotito, S. and Quagliotti, L. (1994). SEED TREATMENTS AGAINST DORMANCY IN OKRA (ABELMOSCHUS ESCULENTUS (L.) MOENCH).. Acta Hortic. 362, 133-140
- Badges
- Science topic
- Similar topics
- Social Science
- Media