SLOW GROWTH STORAGE for the mid-term conservation of a large number of species, including tropical and temperates. In this case, reducing the in vitro growth through the application low temperatures and less hours of light.
Dear Mônica Bossardi Coelho there's not one light or one temperature. Although plants growth in vitro under general conditions (light and temperature), every species has its own requirements. Thus, you must to determine the suitable conditions for the species you’re working with. I’ve worker with species (walnut) that can't resist short periods of low temperature/dark; whereas other (garlic) can be storage at 4°C in the dark for several weeks without any observable damage.
Light intensity is a very complex term to define/ explain and measure as there are so many units involved, lux, foot-candle, micromol per second per metre, lumen per square metre etc...
The right answer for you will depend on your source of light as well, whether you are using fluorescent lamps or LEDs.
In most common scenarios we use fluorescent lamps and I shall suggest that you go by trial and error for the species you are working on. Each species has its own requirement.
As a rule of thumb, 5 mol per sq. m per day for plants loving low light conditions.
please use an online light intensity convertor to work out according to the light measuring device you are using.
Full sunlight is 108, 000 lux = 2000 µmol m-2 s-1
Supplemental lighting in a plant growth chamber would be 100 - 400 µmol m-2 s-1
most commonly 150 µmol m-2 s-1
So for germplasm conservation, I may suggest 50 -100 µmol m-2 s-1
5 mol per sq. m per day for plants loving low light conditions is equivalent to 58 µmol m-2 s-1
Play around with these figures and test them on your plant species.
Dear Mônica Bossardi Coelho . See the following useful link: https://www.atophort.com/news/led-grow-light-for-plant-tissue-culture.html
Kindly see also the following RG useful link : Chapter In Vitro Conservation Through Slow-Growth Storage
Also check please the following useful link: Article Modeling the Effects of Light and Sucrose on In Vitro Propag...
Aref Wazwaz shared a paper where it is stated:
The model suggests the best condition for obtaining higher quality acclimatized plantlets is the combination of 2.3% sucrose and photon flux of 122-130 µmol m(-2) s(-1).
Thank you Aref Wazwaz
The following link is also very useful: https://www.pacc.in/e-learning-portal/ec/admin/contents/2_MBO34A_2020113010045341.pdf
Visit kindly the following useful RG link: Article Effect of light conditions and ABA on cold storage and post-...
Ricardo Julian Licea-Moreno , Navindra Boodia and Aref Wazwaz thank you for your answers and clarifications. I also read the links you shared with me. They are very useful to me. Indeed, for several species stored in a same enviroment it is hard to find common conditions of luminosity. In my search I found a varied range that can be used, wich depends directly on the species being stored, as you mentioned. Well, it sounds challenging, since in my case I need to adapt this condition for hundreds of species/varieties stored in the same chamber.
Would it be possible to intersperse days of total darkness, followed by days of low light intensity, and so on, to keep the plants preserved for longer? I wonder if there is some study approaching this and if the plants in general would be able to tolerate this kind of situation.
It is very interesting that you want to apply for hundred plant species stored in the same chamber. Besides the low light intensity, you should consider low temperature and low sucrose concentrations. By the way, how long do you want to keep them in the slow growth condition?
Dear Mônica,
For grapes: we decrease light intensity to ~10 uM/m/s to promote more etiolation and we also use a deficient medium.
I am sharing a nice paper on medium-term in vitro storage attached.
Best Regards,
Jean
Dear Jean Carlos Bettoni thanks for your message and the paper you shared.
Please, do you use light intensity to ~10 uM/m/s for how many hours a day?
Dear Mônica Bossardi Coelho
The StarPac bags are sealed and held for 2 weeks in a growth room at 25 °C with a 16 h photoperiod provided by fluorescent lights (40 μM m-2·s-1). Then the StarPac bags are transferred at 4°C in low light. You can work with 12 to 16 h of photoperiod.
Best Regards,
Jean
Dear Jean Carlos Bettoni
Thanks for the details.
All the best,
Mônica
Dear Mônica Bossardi Coelho
It can be difficult to choose the right light intensity for different plant species (light can be a stress factor under slow-growth conditions).
I suggest You to do a little experiment on species that are evidently photophilic - do they tolerate culture in the darkness (even a commercial refrigerator can be used if you plan to lower the temperature of the culture).
Good luck,
Monika
First of all, I agree with Monika Kamińska, but in general, reducing growth factors (temperature, light intensity, carbon sources and so on) plant growth rate decrease.
Also, I woud like to recommend these articles:
1. Singh, S., Thangjam, R., Harish, G. D., Singh, H., Kumar, R., Meena, D. P. S., & Agrawal, A. (2021). Conservation protocols for Ensete glaucum, a crop wild relative of banana, using plant tissue culture and cryopreservation techniques on seeds and zygotic embryos. Plant Cell, Tissue and Organ Culture (PCTOC), 144(1), 195-209.
2. Ray, A., & Bhattacharya, S. (2008). Storage and plant regeneration from encapsulated shoot tips of Rauvolfia serpentina—an effective way of conservation and mass propagation. South African Journal of Botany, 74(4), 776-779.
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