On the propagation studies of Embelia ribes seeds. To check the viability, which is the best method to adopt?
limitations are seeds are very small, some times seeds are empty. both the case X-ray, Tetrazolium it is very difficult to make a conclusion on Embelia ribe seeds.
Viability of the seeds in general is the per cent (%) of seeds that able to germinate and form seedlings. This depends on many enviromental conditions (such as temperatue, moisture, presence of some hormones, stratification ets.) The viability estimation in general is simply estimation the count of germinated seeds to all seeds (usually use 100 or more seeds) in the experiment in standard conditions.
Using of x-rays and another agents (for example tetazolinum, other chemicals or physical agents or simply terms and conditions of store etc.) can show only the resistance of seeds to such action. You can estimate seed resistance to any factor through their viability estimation. But in such case you must to choose factor and compare obtained results whith control group without any specific factors action. Also in such way you can estimate efficient doses of every used factor and/or their complex action.
Probably the in vitro germination test as is closely associated with micropropagation!
@ Andrii, Thanks for valuable suggestion. This seed normally shows very less germination. out of 100 very few individuals only succeed.
Embelia ribes seeds mostly shows the absence of embryo, to find out fully mature or well developed embryo for success.
X-ray, Tetrazolium both are well know test to check the viable. but in this case, seed is very small so not possible to identify or interrupter under X-ray, and in Tetrazolium test, seed not showing any considerable result. Please suggest some more test like fluorescein diacetate to test viable.
@ Tobias, thanks. that's true. it will give good result, but naturally whats the status of the seed. if out of 100 seed one seed leads to new plant, what about 99 seed. why this condition not only Embelia ribes so many seed are there in case.. out of 1000 only 10 seeds showing the next generation. let see the viability of seed and moving further in natural condition other than micro culturing.
Dear Sebastian. What is the size of the seeds? We have successfully performed Tetrazolium stainings with seeds as small as those from Arabidopsis (Tesnier et al. (2002) Seed Science and Technology 30 149-165).
Can you remove or count the empty seeds by fluid density sorting?
very good question. I will store seeds under ideal conditions and keep checking viability by germinating them after 30 day intervals. If the seeds are being released of dormancy than you will find out and if they do not germinate at all then they are most probably dead. However, there may be a number of exceptions but this will give you some information to go forward. Age of seed is very important and use of fresh seeds is recommended.
If TTZ test is not possible then go for Hydrogen peroxide test to see either embryo showing any sign of viability or not. For very low percentage of germination, please check the kind and level of dormancy. I think Myrsinaceae family possesses physiological dormancy and needs either warm or cold or combination of warm + cold stratification to over come dormancy.
Dear Dr. Steven, fruit size approximately 2-4 mm, I not tried fluid density sorting. Thanks. I try IDS.
@ Dr. Shyam, I hope Physical trigger will work. I will try this also.
Dear Sebastiaan,
Tetrazolium Is the most frequently used method for seed viability testing and used by many official seed testing labs. Also dormant seeds will stain red if viable. Difficulty is the penetration of the substrate. Many seeds have an impermeable layer surrounding the embryo. Therefore seed testing labs make a cut in the seed. Your seed size is on the edge of making incisions, especially since you need to analyse maybe thousand seeds. Arabidopsis seeds are small as sand grains and it is impossible to make incisions. In the paper I mentioned, we used diluted hypochlorite to make the Arabidopsis seeds permeable. That worked fine and we could see the red staining of the viable seeds through the testa. I don’t know about your seeds and if you succeed in penetration of the substrate , it is possible to discriminate between viable and non-viable seeds. My advice is to give it a try.
Cut test to examine embryo, tetrazolium staining, or flotation test.
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