Looking for a resource on using gametophytes to seed lines
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Hi everyone. I have been developing a small seaweed hatchery and need a bit of support.
So far I have established a "clean room", and set up the entire environment to be welcoming and prepared for operations.
At this stage, we are early. We are looking to define our techniques.
One piece of this is obtaining gametophytes from a friendly source - which we have done! We are going to receive male and female gametophytes in separate containers.
Since our grow tank is configured (seed string lined PVC pipes with plenty of sea water ready for the tank), we want to use these gametophytes to attach to our seed strings and watch them grow.
??? But.... how ??!
We are struggling to find a good resource that helps us to understand this process. If you have a reference that you can point me to I would very much appreciate it over simple advice offered directly in this forum.
The best we've come up with is the following - which we lack confidence in - shown by lots of ???. We are continuing to research, but could use help being pointed to the right resource(s) to study.- ???? Combine the male/female gametophytes ????
- ???? Expose gametophytes to blue light for 24 hours ?????
- Use a caligraphy brush (after letting it sit in distilled water and dry off) to transfer the gametophytes to the seed string
- ???? Let sit in water tank without bubbler and without chiller on for 24 hours ????
- ???? Change to low white light and start the growth process ???
@adam_ornstein31 I followed this manual after collecting sorus tissue from a spot near me and successful few sugar kelp from spores to maturity…
https://maineaqua.org/wp-content/uploads/2020/06/OceanApproved_KelpManualLowRez.pdf
@mike_spranger this I'm familiar with, thanks so much for sharing it here.
It's a process that I'm looking forward to exploring in about 2 months time
But I don't see instructions in this document for how to use gametophytes to seed twine in this document (there is a minor discussion on the topic of gametophytes, but no for dives). - unless I missed it!? Can you tell me if you see that in this document?
@mike_spranger this I'm familiar with, thanks so much for sharing it here.
It's a process that I'm looking forward to exploring in about 2 months time
But I don't see instructions in this document for how to use gametophytes to seed twine in this document (there is a minor discussion on the topic of gametophytes, but no for dives). - unless I missed it!? Can you tell me if you see that in this document?
@adam_ornstein31 The process you outline at the bottom of your post looks accurate to me. I would suggest adding a grinding step to break up the gametophytes before you mix them and put them under blue light; we use cell tissue grinder to do this.
GreenWave is currently working on publishing a gametophyte seeding course to our Ocean Farming Hub. Until then, continuing to ask your questions here is a great way to get advice.
@adam_ornstein31 we combine either 2:1 or 1:1 F:M (optima vary with species and even strain…), blend to homogenize gametophyte culture for 10-15 seconds (looking for less small filaments under the microscope, not chunky colonies.) I found in my previous work, and other folks have also recently identified, that blending is the trigger for stimulating adhesion of the gametophytes to surfaces.
Spray or paint the blended culture onto the lines and store like you said. Exact densities aren’t published anywhere though I know folks are working on this, but a little bit goes a very long way with gametophytes. They need a bit of time, generally for a few hours to overnight, to adhere. Grow the spools in blue or full spectrum white light at the same intensities you would use for spore-seeded spools to induce and begin growing the sporophytes. Make sure to turn the aeration back on post-adhesion!
Unfortunately there isn’t a ton of published work on exact protocols for this, but this is what nurseries working with gametophytes are generally doing. Looking forward to updated public protocols for gametophytes once everything has been optimized!
@maisie_musor1057
First of all, thank you for sharing your process. It's insightful, useful and motivational to get this level of information.
Here I'm trying to process the information and I'm hopefully analyzing your experiences correctly.
Question - is your recommendation to (1) grind the gametophytes to size
(2) Mix m/f species
(3) Immediately paint the gametophytes onto the seed string, and leave seed string in the water in the dark.
(4) Let sit in still, dark water for up to 24 hours
(5) After 24 hours, expose to white (or blue) light, allow the bubbler and chiller to run fully in the tank
Again, thank you for sharing your insights from your own experience. It's greatly appreciated
@adam_ornstein31 Apologies for the delay, things have been hectic seeding our nurseries and I didn't see your response! Overall, yes, but let me clarify a few points.
1+2) Doesn't matter if you grind before or after mixing these. I only do it before because I find it easier to take representative measurements of gametophyte density when the are ground so I mix in the right ratios.
3-4) Yes, immediately paint on and leave in still water for 24 hours. However, you can and should start the white lights while they are adhering to the seed string. You don't need to put them in the dark, I just wrote "turn on the lights" too far along in my last post.
5) Correct- Water motion/aeration back on with the white lights to induce and grow and induce blades.
Best of luck this season! Hope things go well!!
-Maisie
@maisie_musor1057
First of all, thank you for sharing your process. It's insightful, useful and motivational to get this level of information.
Here I'm trying to process the information and I'm hopefully analyzing your experiences correctly.
Question - is your recommendation to (1) grind the gametophytes to size
(2) Mix m/f species
(3) Immediately paint the gametophytes onto the seed string, and leave seed string in the water in the dark.
(4) Let sit in still, dark water for up to 24 hours
(5) After 24 hours, expose to white (or blue) light, allow the bubbler and chiller to run fully in the tank
Again, thank you for sharing your insights from your own experience. It's greatly appreciated
@adam_ornstein31 Apologies for the delay, things have been hectic seeding our nurseries and I didn't see your response! Overall, yes, but let me clarify a few points.
1+2) Doesn't matter if you grind before or after mixing these. I only do it before because I find it easier to take representative measurements of gametophyte density when the are ground so I mix in the right ratios.
3-4) Yes, immediately paint on and leave in still water for 24 hours. However, you can and should start the white lights while they are adhering to the seed string. You don't need to put them in the dark, I just wrote "turn on the lights" too far along in my last post.
5) Correct- Water motion/aeration back on with the white lights to induce and grow and induce blades.
Best of luck this season! Hope things go well!!
-Maisie