Hi Guys,
Is there any specific way to sterilise Rice seed, I've got many protocols to do that. I appreciate if you can recommend the best way with the reference for that.
Regards
Dear Hyder
Sterilization protocol in the following article may help you.
Kind Regards
Shiwani
RIce has a hard seed coat, so you can use a harsh method to sterilise. Wash with soap and water, Rinse under running water, under tap. 1 min in 75 % EtOH followed by 30 % household bleach for 20 min on a shaker. Rinse with sterile water 3X. This should remove all external contaminants. If the seed coat is compromised during harvesting etc, there may be microbes underneath. In that case it may help to dehull and sterilise. Same procedure should work.
Rinse with sterile water for about 1 mts and 1-2 mts in 75% Ethyl Alcohol would be sufficient or repeat one more time will work
Try this; Full text available at Researchgate
Sodium Hypochlorite: Effect of Solution pH on Rice Seed Disinfestation and Its Direct Effect on Seedling Growth S.-C. Chun · R. W. Schneider · MA Cohn
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ABSTRACT: Bacteria were completely eliminated from rice seeds following immersion in household bleach solutions (50% bleach and 2.6% NaOCl) adjusted to pH 7.0 in 0.5 M potassium phosphate, while fungi were eliminated at pH 5.0 and below. Mercuric chloride was used to disinfest nondormant rice seeds in order to test the direct effect of sodium hypochlorite on rice seedling growth apart from an indirect effect related to elimination of microbial contaminants. Seeds were immersed for 2 h in each solution or water in each of the following treatments: (i) sterile water followed by sterile water, (ii) HgCl2 (1,000 mu g/ml) followed by sterile water, (iii) HgCl2 followed by KH2PO4 (0.3 M), (iv) HgCl2, followed by NaOCl, and (v) NaOCl followed by sterile water. Sodium hypochlorite solutions were prepared in 0.3 M potassium phosphate, which gave a final pH of 7.3. Seedling growth in treatments in which seeds were treated with HgCl2 followed by NaOCl, and NaOCl followed by sterile water, was significantly greater than those treatments in which seeds were treated with sterile water followed by sterile water, HgCl2 followed by sterile water, and HgCl2 followed by KH2PO4. Sodium hypochlorite stimulated rice seedling growth directly, as opposed to an indirect effect related to elimination of microbial contaminants or alleviation of dormancy.
Plant Disease 07/1997; 81(7):821-824. DOI:10.1094/PDIS.1997.81.7.821
Dear Hyder Shihab,
These article will be helpful to you.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC92979/pdf/am003046.pdf
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC92979/
It is more safe to apply the following steps:
1- Take about 20 rice seeds into 2 mL Eppendorf tube.
2- Add 5% Calcium hypochloride or 5% Sodium hypochloride to about 3/4 of Eppendorf tube with seeds
3- Mix well for 30 -60 min. using vortex with low strength.
4- Remove washing solution and change it with sterilized water for 5 or 6 times.
5- Completely remove all water from seeds
6- Place the seeds in the sterilized tissue towel, and directly put them media you decide to grow.
Good luck
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