You selected nice study it is useful for Indian system of medicines. Literature available on google

You can visit the plants in the wild to find out

I think this attachment my be useful in your work..regards

The most important issue, is to determine the best method to get their seeds to sprout on top of moist sterile cotton--what temperature is needs, how much time, and if there are still dormant ungerminated seeds after 14-20 days.

Then, if you can get to where the plants grow wild, get a soil sample from around where each plant grows, to a depth of only 5 cm and do a soil test for both macro-nutrients like nitrogen, phosphorus, potassium, calcium and pH, and the micronutrients like iron, copper, zinc, manganese, magnesium, etc.

Once you know the needs and speed of sprouting, then copy your soil medium as your test indicate, and never sow your seeds more than 3x their diameters.

Thank you for sharing your knowledge

For those two species, you need to start with absolutely fresh seeds, and send 10 seeds to a commercial seed testing lab where they can do a tetrazolium staining test for you, which will tell you what the potential total germination would be--the process stains the living embryos.

If the tetrazolium staining show that you have a high percentage of living seeds, but you produce a lot of dormant seeds with your top of cotton tests, you have one or more dormancy issues--and there may be an "after-ripening" requirement of the embryo that needs a few months more to develop, before it will sprout.

Or a dormancy issue, that is resolved by the seeds going through a cold moist period in sand or soil, and that is very common for wild seeds from parts of the world that have cold winters for example. For temperate climate seeds it could be 1-3 months cold-moist treatment, to get to sprout.

Also it can be a dormancy to light, where the seed must be in total darkness in order to sprout.

Another kind of dormancy, is will not sprout until seeds surrounded by the right amount of nitrogen, that can be added by putting some potassium nitrate into distilled water to make up a 3% solution and use that to water the top of cotton tests.

Sometime, but probably not with these seeds, there is a hard seed coats, which keeps seeds from absorbing water.

But might be a problem with these seeds, is not enough heat during the day, and most of the time, the day and night temps. must fluctuate in order for the seeds to sprout. Check for studies on these seed for germination studies done in the past, like on GOOGLE SCHOLAR.

I forgot another dormancy--that there are seeds that MUST have light to germinate. So, if they sprout on top of your cotton tests, but are dormant when you plant them in your soil-mix, they may need to be planted on the surface and not covered with any soil, because they need light to sprout.

So as I said the first time, find out what your seeds need to sprout properly, then copy the natural soil conditions where they grow. And if you can get a sample of that soil to mix into your growing medium, all the better. That way, you will be adding the soil micro-organisms that they may also need to thrive.

Thanks a lot for the suggestions! Craig Carlton Dremann , Wisam Thamer Al-Mayah , Ajay Kumar Meena , Andrew Paul McKenzie Pegman

The tetrazolium (TZ) tests maybe expensive-- in the USA usually under $50-- but if they only test 10 seeds of each, they may give you a break on the price. Usually the seed labs test for legal labeling by using 400 seeds. But since you just need to know what is alive in your sample, then a lower price may be possible.

You can start with the top-of-cotton test first, but if you run into problems, then the TZ will tell you exactly what the potential is for that batch of seeds. Image is from a slide show by Ransom Seed labs at https://static1.squarespace.com/static/5cdf03346b058f0001162855/t/5ce05b2e306d590001c17ef6/1558207281734/TZtest.pdf

studying the agro-pdeological conditions of naturally grown endangered planst and later underatking in-vitro multiplication is quite task . These two exercises have heaven to hell difference. But , all the best to you....

Good work..lot of paper and literature available in medicinal plants related Journal. You can use different media,. Conditions for study

Loss of native plant habitat is the main reason for plants becoming endangered. Conservation of endangered plants is best done by in vitro methods such as tissue culture. There is an extreme pressure on natural environment to maintain rare plant species and tissue culture can be used to relieve the pressure and propagate those species. Plants need to have adequate variation in their genes if they are to adapt and survive in natural environment. Agriculture depends on the conservation of natural resources such as water, soil and climate and hence assessment of soil, irrigation and climatic requirements are necessary for better vigor.