Sophrolaelia Dry Seed Flasking - first attempt

[czayta]

Originally Posted by epiphyte78

Really excellent documentation! I'm quite impressed with how professional your first attempt was.

I'm also impressed that you went ahead and flasked the seeds even though you established that they didn't appear to be viable.

But I'm wondering whether the seeds really weren't viable because they were too old. What would the "viability over time" curve look like?

Day 1: 90% viability
Day 2: ...
Day 3: ...
Day 30: 0% viability

100% viability loss after 30 days strikes me as a bit off. You'd figure that most Cattleya alliance would have been naturally selected for seeds that had a greater margin of error. Because...what if the pod ripened and released its seeds 2 or 3 weeks before the start of the rainy season? Only the seeds with the greatest longevity would survive. Or perhaps orchids that failed to accurately predict the onset of the rainy season took themselves out of the gene pool long ago?

Personally I'm trying to germinate orchid seeds symbiotically. This year I've been pretty good at consistently pollinating my orchids. Right now I probably have 30+ pods in various stages of development.

The next time you plan on flasking...feel free to PM me if you're interested in additional seeds. Also, I could send you some fresh Cattleya alliance seed if you'd like to graph its viability over time. My Cattleya Penny Kuroda has 6 fat seed pods on it. Most of the pollen came from Mexican Laelias. I'd be interested to know if the seeds are viable and if so, what their longevity is.

Wow! That Cattleya Penny Kuroda is absolutely beautiful. I would love to try flasking some seeds for you. I prefer the green pod method (sterliziation is simpler and quicker) but I just bought 5 different types of dry seeds off ebay to try flasking as well. When the seeds from ebay arrive I plan on giving flasking a third attempt. I just bought some more P669 media from the Orchid Seed Bank so I will get to pressure cooking this weekend to prepare the mother flasks.

I also have 3 seed pods in progress, eta 5 months until they are ready for Green Seed Pod flasking. But they are just NoID Phalaenopsis so I am not very excited about them to be honest. :P

For your seeds, would you send them dry seed? I could look at them under a microscope every 3-5 days and record a viability estimate if you want for a month or two. I think in my case what happened was: I went to the greenhouse and he noticed his Sophrolaelia 'Cheerio' pod had split open. So he collected the pod that day, stuck it in an envelope and put it in a drawer. Then I returned 38 days later to his greenhouse again and he said, "Would you like to try flasking these? It's too late for me to send them to the lab." (i.e. he sorta new the viability might be low since they weren't stored in 2-8 degrees C). Since I had never flasked anything before I jumped on the opportunity.

You're using Mycorrhizal fungus? I think its really cool that you're trying symbiotic germination. Good luck! Post a thread with pictures and let us know how it goes. I'm very interested in that.

[epiphyte78]

Glad to hear that you would be interested in Penny Kuroda. I could send you a green pod if you prefer. You could flask some of the seed and store the rest at room temperature and test it every 3-5 days for viability. I store my seed at room temperature so would be very interested to know the rate of viability loss.

The drawback of sending one entire pod is that maybe that particular cross wouldn't be as vigorous as some of the others. Like I said, most of the pollen was from Mexican Laelias (~ 5 month old pollen)...but I'm pretty sure that one flower was pollinated with fresh pollen from one of the other flowering orchids in the picture...Lc Clayton Waglay var. alba. Clayton Waglay is Laelia anceps x (C. intermedia x C. loddigesii). But I didn't mark which flower was the recipient of this pollen. And within the Mexican Laelia pollen I mixed in the fresh pollen from one Psychilis krugii flower. So one Penny Kuroda flower ended up with pollen from one or more Mexican Laelias and Psychilis krugii. Of course I have no idea whose pollen won the race.

Rather than send you one entire pod...I could harvest all the pods when they are ripe, mix the seeds and send you a pod's worth of seeds. This way the genetic pool will be larger...which would increase your chances of flasking the most vigorous combination of traits.

Either way is fine with me...just wanted to share some information to help you make an informed decision. Not sure when the pods will be ripe enough to harvest...could be within a month or two.

Which orchid seeds did you buy on ebay? If you want some guaranteed fresh seeds...here are a couple options...

1. Papilionanthe vandarum (pod) x Vanda tricolor/suavis (pollen). I harvested the pod a couple days ago and could send you 1/4 of the seed.

2. Renanthera imschootiana (pod) x Rhynchostylis gigantea? (pollen). I can harvest the pod today and send you 1/4 of the seed.

Here's the thread where I've documented some orchid seeds that germinated on my tree. The success rate is far lower for symbiotic germination...but for conservation purposes it could be pretty important to try and cultivate the widest possible variety of fungus that orchid seeds need to germinate on trees in nature.

[Cntry]

Originally Posted by epiphyte78

Glad to hear that you would be interested in Penny Kuroda. I could send you a green pod if you prefer. You could flask some of the seed and store the rest at room temperature and test it every 3-5 days for viability. I store my seed at room temperature so would be very interested to know the rate of viability loss.

The drawback of sending one entire pod is that maybe that particular cross wouldn't be as vigorous as some of the others. Like I said, most of the pollen was from Mexican Laelias (~ 5 month old pollen)...but I'm pretty sure that one flower was pollinated with fresh pollen from one of the other flowering orchids in the picture...Lc Clayton Waglay var. alba. Clayton Waglay is Laelia anceps x (C. intermedia x C. loddigesii). But I didn't mark which flower was the recipient of this pollen. And within the Mexican Laelia pollen I mixed in the fresh pollen from one Psychilis krugii flower. So one Penny Kuroda flower ended up with pollen from one or more Mexican Laelias and Psychilis krugii. Of course I have no idea whose pollen won the race.

Rather than send you one entire pod...I could harvest all the pods when they are ripe, mix the seeds and send you a pod's worth of seeds. This way the genetic pool will be larger...which would increase your chances of flasking the most vigorous combination of traits.

Either way is fine with me...just wanted to share some information to help you make an informed decision. Not sure when the pods will be ripe enough to harvest...could be within a month or two.

Which orchid seeds did you buy on ebay? If you want some guaranteed fresh seeds...here are a couple options...

1. Papilionanthe vandarum (pod) x Vanda tricolor/suavis (pollen). I harvested the pod a couple days ago and could send you 1/4 of the seed.

2. Renanthera imschootiana (pod) x Rhynchostylis gigantea? (pollen). I can harvest the pod today and send you 1/4 of the seed.

Here's the thread where I've documented some orchid seeds that germinated on my tree. The success rate is far lower for symbiotic germination...but for conservation purposes it could be pretty important to try and cultivate the widest possible variety of fungus that orchid seeds need to germinate on trees in nature.

Epip how did the tree with the orchids do over the winter and what do they look like now. I just read your post and its so cool to think they could grow and be just fine.

[czayta]

Originally Posted by epiphyte78

Glad to hear that you would be interested in Penny Kuroda. I could send you a green pod if you prefer. You could flask some of the seed and store the rest at room temperature and test it every 3-5 days for viability. I store my seed at room temperature so would be very interested to know the rate of viability loss.

The drawback of sending one entire pod is that maybe that particular cross wouldn't be as vigorous as some of the others. Like I said, most of the pollen was from Mexican Laelias (~ 5 month old pollen)...but I'm pretty sure that one flower was pollinated with fresh pollen from one of the other flowering orchids in the picture...Lc Clayton Waglay var. alba. Clayton Waglay is Laelia anceps x (C. intermedia x C. loddigesii). But I didn't mark which flower was the recipient of this pollen. And within the Mexican Laelia pollen I mixed in the fresh pollen from one Psychilis krugii flower. So one Penny Kuroda flower ended up with pollen from one or more Mexican Laelias and Psychilis krugii. Of course I have no idea whose pollen won the race.

Rather than send you one entire pod...I could harvest all the pods when they are ripe, mix the seeds and send you a pod's worth of seeds. This way the genetic pool will be larger...which would increase your chances of flasking the most vigorous combination of traits.

Either way is fine with me...just wanted to share some information to help you make an informed decision. Not sure when the pods will be ripe enough to harvest...could be within a month or two.

Which orchid seeds did you buy on ebay? If you want some guaranteed fresh seeds...here are a couple options...

1. Papilionanthe vandarum (pod) x Vanda tricolor/suavis (pollen). I harvested the pod a couple days ago and could send you 1/4 of the seed.

2. Renanthera imschootiana (pod) x Rhynchostylis gigantea? (pollen). I can harvest the pod today and send you 1/4 of the seed.

Here's the thread where I've documented some orchid seeds that germinated on my tree. The success rate is far lower for symbiotic germination...but for conservation purposes it could be pretty important to try and cultivate the widest possible variety of fungus that orchid seeds need to germinate on trees in nature.

Your pictures are just amazing! It is so cool to see all of the orchids thriving out on the trees. I love the Vanda tricolor/suavis.

I'm not sure about the idea of mixing a bunch of seeds together to see what germinates/is the strongest in the genetic pool because it seems difficult to track/name the lineage that way. I'm very methodical and particular when it comes to breeding/pollinating/setting up crosses. You put two different types of pollen in one plant? :O How will you ever know what the true parentage is? I like crossing A x B. Not crossing A x B or C, but that's just me. :P

As for the seeds from ebay: I bought 5 different type of seeds from kumpol_p on ebay. Kumpol is a Thailand orchid seed seller Orchid Specie Seeds Vanda Denisoniana Year 2014 | eBay

I don't have any experience with this vendor, but I thought I'd give it a shot and see if anything germinates (and I was desperate for seeds because my green pods wont be ready for 5 months lol).

Tonight I prepared more P669 media for mother flasks. I would love to try some of the Papilionanthe vandarum (pod) x Vanda tricolor/suavis (pollen).

Also, as a side note: I recall seeing your symbiotic germination thread when I was just getting interested in orchids a while ago, and I couldn't believe how cool it was that your seeds had germinated on the tree!

[epiphyte78]

Today I cut the V. tricolor x P. vandarum pod open...it doesn't look like an obvious dud. There seems to be a decent amount of seeds and they were just starting to dry. So it's not too green. I'll give them a few days to dry out and then mail around 1/2 to you.

Regarding A + B or C...well...sometimes I even throw D into the mix! This increases the chances that I'll end up with a seed pod.

My overall goal is to select for tolerance. I think that more people will grow orchids on trees if they don't have to water them every day during summer and wrap a blanket around them when it gets cold.

Making progress in this direction is a numbers game. My guess is that you have to throw a lot of different combinations at nature to find individuals that are marginally more tolerant. If an individual survives the selection process...and you ask me what it is...my response would be..."it's a survivor!".

There's always room for improvement. This is true whether we're talking about orchid tolerance or my method for improving orchid tolerance! With your approach, if I only crossed A + B...and it didn't work...then I would avoid making that same cross again. Except, then I'd have to store all my pollen separately. I have 100s of species...and just as many hybrids...so I'd have little storage containers everywhere! If I kept them in the fridge I'd lose a lotta weight. I'd have one drawer in the fridge for food. With my haphazard method I have one drawer in the garage for orchid pollen.

What are your orchid breeding goals? To develop an orchid that smells like a peanut butter and jelly sandwich?

---------- Post added at 11:46 PM ---------- Previous post was at 11:37 PM ----------

Originally Posted by Cntry

Epip how did the tree with the orchids do over the winter and what do they look like now. I just read your post and its so cool to think they could grow and be just fine.

It didn't really get that cold this last winter. I don't think it dropped much lower than 40F.

The largest seedling should bloom in a couple of years. A somewhat smaller seedling put out two new growths for some reason.

[Cntry]

EPip thanks for the update on how the orchids did over the winter. Last few year we got hit hard here with snow or ice and sometimes both here in Maryland. So I am very glad your orchids in the tree is doing very well.

[czayta]

These are the six different types of dry seeds I flasked. 4 were from epiphyte78, thanks!


getting the dry seeds ready for sterlization


After I flasked all 6 types of seeds:


1. Papilionanthe vandarum (pod) x Vanda tricolor/suavis (pollen) - 2 flasks, clean so far


2. Renanthera imschootiana (pod) x Rhynchostylis gigantea? (pollen) - contaminated =(

3. Mixed Orchid seeds - both flasks contaminated, but I expected that much because I flasked way too many seeds and had a difficult time sterilizing them. I have 2/3rd of the original bag left, so I will try flasking them 2 more different times.

4. S. brevi - 2 flasks clean, no contamination yet


5. V. Denisoniana - 3 flasks clean so far


6. A. multiflora - 2 flasks clean so far


I have a really difficult time with properly sterlizing the dry seeds. I have 0% contamination (9/9 flasks clean after 6 weeks) when I flask green pod, but 50% contamination (3 out of 6 flasks contaminated after 6 weeks) when I flask dry seed. :/ Any suggestions on strength of bleach and length of sterilization time would be appreciated. I think I'm going to try Roby's sterilization technique next time I dry seed flask.

[epiphyte78]

Thanks for flasking the seeds and sharing the photos/process!

Regarding seed sterilization...this 2006 article by Fred Bergman...An Easy Approach to New Rewards...looks useful. You can compare it to this slice from an article he wrote a decade earlier...

*********************

The information in this article came from a four-year study that included an extensive literature search and an experimental investigation. The literature search covered 148 books and 165 papers. A total of 95 articles that described the use of calcium hypochlorite were located. Besides Wilson's original report, there were only four papers that described the concentration as AC (available chlorine). Hegarty (1995) reported using a 7.5 percent suspension of chloride of lime (calcium hypochlorite) containing 24 percent AC, for a final strength of 1.8 percent, and Castle and Nickell (1942), and Fennell (1956) used a 3.6 percent solution of calcium hypochlorite (70 percent AC) for a final strength of 2.5 percent AC. In addition, Liddel (1946) provided the information necessary to calculate that his solution contained 0.56 percent AC. All of the other articles described concentration only in weight percent so that the actual concentration used is unknown.

Apparently everyone assumes that the optimum concentration for calcium hypochlorite has been established. Not one paper was found that determined the minimum effective calcium hypochlorite concentration. The acceptance of 10g/140ml as optimum has continued in spite of the fact that literature contains numerous references to seed that appeared viable but failed to germinate. To obtain information on concentration effects, a serious of tests was run that indicated the concentration could be lowered to between 0.1 and 0.2 percent AC and remain contamination free. The tests were conducted with a contact time of 15 minutes with the seed recovered using filtration and two water rinses. It was found during these tests that as the concentration decreased, the germination improved.

Many orchid enthusiasts do not have access to a well-equipped laboratory. To assist in the preparation of a suitable disinfectant, I weighed several samples of calcium hypochlorite and found that 1/4 level teaspoon weighed close to 0.85g. To prepare a 0.15 percent AC solution, multiply 0.85 by the percent AC in the calcium hypochlorite (for example 70 percent) to obtain 0.595. Next, divide 0.595 by (0.15 x 100) to determine that 395 ml of water will be required to prepare the 0.15 percent AC solution. To use fluid oz instead of ml, divide 396 by 30 to find that 13.2 ounces of water will be required to prepare a 0.15 percent AC solution. The 13.2 ounces may be rounded to 13 ounces, producing an error of only 0.005 percent. - Fred J. Bergman, Disinfecting Orchids Seeds

*********************

That's quite a range of concentrations...from 2.5% to 0.1%. With the mixed orchid seeds a good hedge will probably err on the side of stronger concentrations.

[czayta]

Originally Posted by epiphyte78

Thanks for flasking the seeds and sharing the photos/process!

Regarding seed sterilization...this 2006 article by Fred Bergman...An Easy Approach to New Rewards...looks useful. You can compare it to this slice from an article he wrote a decade earlier...

*********************

Many orchid enthusiasts do not have access to a well-equipped laboratory. To assist in the preparation of a suitable disinfectant, I weighed several samples of calcium hypochlorite and found that 1/4 level teaspoon weighed close to 0.85g. To prepare a 0.15 percent AC solution, multiply 0.85 by the percent AC in the calcium hypochlorite (for example 70 percent) to obtain 0.595. Next, divide 0.595 by (0.15 x 100) to determine that 395 ml of water will be required to prepare the 0.15 percent AC solution. To use fluid oz instead of ml, divide 396 by 30 to find that 13.2 ounces of water will be required to prepare a 0.15 percent AC solution. The 13.2 ounces may be rounded to 13 ounces, producing an error of only 0.005 percent. - Fred J. Bergman, Disinfecting Orchids Seeds

*********************

That's quite a range of concentrations...from 2.5% to 0.1%. With the mixed orchid seeds a good hedge will probably err on the side of stronger concentrations.

Thanks for the link to the article. 0.1% to 2.5% seems very weak. =o I was only sterilizing for 10 minutes as well. Maybe the extra 5 minutes really count. I will try to prepare a 0.15% AC solution next time. And I am also planning on trying Roby's sterilization technique of just exposing the flask to an AC soaked gauze pad for 3-4 hrs after innoculation. The fumes should kill any bacteria, but not the seeds if done properly.

[epiphyte78]

What was the AC of the solution you used to sterilize the seeds? From what I've read online...most bleach contains 5.25% AC. So...

20% solution = 1.05% AC
10% solution = 0.525% AC
5% solution = 0.2625% AC

I wonder if there's much difference between sodium hypochlorite (the liquid chlorine in household bleach) and calcium hypochlorite (the powder chlorine used for cleaning pools).

The fume technique sounds promising. Found this short paper on the topic.