Dear All,
I have to mix mycelia of an endophytic fungi with the seeds of barley and incubate it in an optimum temperature until germination.
Question 1: Is there any method of mixing mycelia and barley seeds together until germination of barley seeds?
Question 2: The germination rate if very low, Is there any international manual for seed testing?
Thanks
My suggestion for the biopriming of barley is to use polyethethylene glycol to create an ability to swell the seed without having the radicle break the seed coat or testa. When the seeds are rinsed and dried under ambient temperature and the viability will be maintained. As for the priming this should be done in an aerated solution of the proper PEG osmotic potential.
If you interested in positive endophytic fungus I would suggest Pirisporoforma indica which would be grown vermiculite perlite infused with nutrient. The fungus grown on the solid media to produces its chlamydospores in the material without contamination and embedded in the carrier material. This material mixed with the bioprimed seeds will give the colonization of the root systems and the positive interaction.
Hope these thoughts are useful. Another potential methodology would be to use the PEG biopriming to colonize the seed. It is possible that acremonium fungi which are positive and have good ability to colonize seeds might be able to be proliferated in the priming stage.
Dear Prof. Dr. Paul Reed,
Could you elaborate further about the use of polyethylene glycol? How can I use it and what concentration for no of seeds. I want the plants roots to be infect by a fungus. I want to check GFP tagg fungi in the roots, what would be the ratio of vremiculite and perlite. Do I need to add the vermicultie and perlite with the nutrient soil for GFP tagg fungi? PEG is good idea, it swells the seeds. We have many plants, Rice barley and wheat could the procedure would be same for all these plants?
Thanks
Hepperly, P. R., and J. B. Sinclair. 1977. Aqueous polyethylene glycol solutions for treating soybean seeds. Seed Science and Technology 5:727-733.
PEG 6000 molecular weight is nontoxic and forms aqueous solutions when water is add. Adding the proper amount of PEG can produce a swelling of the seeds without the seed coats being rupture. If these aerated solutions are rinsed off and dried the dried seeds will germinated more uniformly and faster.
I believe the technique can be widely adapted to various grain seeds.
I would suggest the colonization of endophytic fungi be done by growing the inoculum on vermiculite perlite with nutrients and then applying the inoculum with the seeds at planting. The endophyte will enter the germinating seedling roots and colonization of plants will spread alone with the root systems.
For this type of inoculation Pirisporoforma indica and acremonium endophytes which produce plant stimulating alkaloids would be good candidates. The first fungus would be a good selection based on its good ability to grow on media and soils.
In relation to the inoculum one volume of perlite is mixed with one volume vermiculite then a potato carrot broth could be the nutrient source.
Grow the inoculum in stoppered Erlenmeyer flask for mason jars using aseptic techniques.
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