It only takes one 
Here is the R.gigantea embryo which was densely waxy all over except for its cotyledon tips (which were pearly white)âŚthis pic has been cropped out of one of one of the original earlier embryo pix posted above, for ease of ID:
You cannot easily see the color difference in the above pic, you have to trust me about it, though LOL!!!
NowâŚ
Here is the same embryo having spent over 1 week in the WEC perlite+drinking glass germination set up:
(Scale: 1 gap = 1/16 inch)
Is there any correlation with dense clearly demarcated waxy embryo markings and subsequent embryo development??? I had wondered if this embryo might die off as its vital regions (radicle + âV junctionâ area between the cotyledons) were very densely waxyâŚ
Hmmmmmm⌠interesting!!!
Here again is the densely waxed embryo, as it appeared this afternoon:
Unfortunately, the greener of its two cotyledons was growing a visible mold on it, which I sprayed off it. I planted it, I canât see how it being in the perlite any longer is going to do any amount good for it.
One of the remaining embryos had not opened its cotyledons and was totally whiteâŚsince it had been in the wetted perlite for way over one week now, this was bad newsâŚI looked at it under 30X magnification and it had several cuts from the box cutter blade, and browning developing around those cutsâŚSo I eliminated it, as it was clearly dying off.
Here are the final two which remained in the perlite-drinking glass set up:
This one has hardly any greening:
This is the second of the tricots, also relatively lacking green (radilce pointing upwards):
These last two were also potted up today. I am hoping that trace amounts of âgermination stimulantâ chemicals are present in the perlite/seedling mix media (1:1), which might get some of these to finally convert to real plants. I donât think them sitting in a sterile media such as wetted perlite for many days more is going to benefit them, this is my geuss anyway.
Here is the heavily waxy one this morning:
I guess you can sort of appreciate that the tissues do not really appear to be normalâŚthese abnormal tissues correlate with the regions of heavy waxy change I noted on it as an embryo last week, at the time of its extraction.
If I were to describe the tissues today, to me they appear almost translucent and colorless.
The molding is now fairly global as you can just make out in this pic.
I cannot see this one progressing much further.
ERROR:
Further up, I said this one was injured with the box cutter blade leading to a flap type injury (original picture cropped for ease of ID):
I wish I had checked it under 30X magnification first, to verify what my eyes were seemingly telling me LOL!!!
In fact, it turns out it was one of the two tricotyledons of the batch. The mis-identified âflapâ seen in the above shot is in fact a smaller extra cotyledonâŚLOL!!!
It grew in the wetteed perlite/drinking glass to look like this:
\
Just to track the pix further, here is the same tricot yesterday:
and here it is now⌠still a pale lifeless ghostly lookinâ thing
Seedling update
Seedling deaths so far:
*Seedling #1: Failed to grow roots and did not green up sufficiently.
*Seedling #2: Decided after producing two true leaves it just wanted to wither away and it died despite showing lots of initial promise!
*Seedling #5: The â90% waxyâ seedlingâŚdied with fungus all over it.
Remaining live seedlings:
Seedling #3: Starting to develop a true leaf, and looks the best of all I have germinated so far.
Seedling #4: A tricoltyledonâŚtwo cotyledons faield to green up, one cotyledon is reasonably green and a new leaf is starting to show.
Seedling #6: Both cotyledons deficient in chlorophyl.
Seedling #7: A tricotyledon with all 3 cotyledons deficient in chlorophyl.
Lesson learned:
Very dried and waxy embryos + WEC---->horrendously low % germinations.
R.gigantea seedlings as seen today:
Seedling #3:
Starting to develop a true leaf, and looks the best of all I have germinated so far.
Seedling #4:
A tricoltyledonâŚtwo cotyledons faileld to green up, one cotyledon is reasonably green and a new leaf is starting to show.
Seedling #6:
Both cotyledons deficient in chlorophyl.
Seedling #7:
A tricotyledon with all 3 cotyledons showing worrying amounts of whitening (third cotyledon is mostly buried).
Here are the seedlings today:
Seedling #3:
Has made no additional progress compared to last pic:
Seedling #4 (tricot):
Appears to be making a second true leafâŚextremely sloooowwww progress!
The other two seedlings are not even worth showing pixâŚthey are not going to make it!
Hi George,
If you are unable to locate âfineâ perlite, and have access to a mortar & pestle, you may be able to produce small quantities.
Iâm enjoying these threads.
Rod
Hi Rod, it is good to hear from you again.
Thank you for your tip. Russ put me onto Gargen City Plastics in Melbourne for supply of fine (and all the other sized) perlite, amongst other thingsâŚapparently they are well priced bulk ordersâŚway to go!
Soon I am going to do a bulk order from MCP for perlite, and oddly I have actually come to love the ordinary grade perlite for the WEC runs (it turns out my initial worries about it were unfounded). So Iâll be ordering the usual grade perlite.
As for this WEC run, there is only one R.gigantea seedling left which has made it to this point, it is the tricotyledon. The two-cotyledon one which showed initially to be the most promising actually stagnated and died!!
Here is the successful tricot, pictured a few minutes ago:
This tricot was a big embryo.
This tricot was the embryo I wrote about further up here, which gave me lots of grief in attempting to remove a small cone of papery tissue which had firmly adhered to the entire radicle tip of the embryo! In the end, one extra desparate hard flick of the blade finally did the trick to dislodge it (usually such hard forcing near the radicle region causes the radicle to break away rendering the embryo useless).
In the case of this tricot, two of its three cotyledons failed to green up, but the one remaining greened up one seemed to be extra big long and curved. Maybe this provided for extra green surface area, which in turn might be able to provide extra fuel to get it to push out a good root system in this critical phase of establishment.
So far it is not showing any signs of decline, it is also taking full sun at the moment.
HmmmmmmâŚin case you are wondering where the third cotyledon is in the above picture, it is withering away, and hidden under the curved/green one (at that picture angle anyways).
This tricot seedling is still ok, and I think it is actually still growing out. I know if anything even very minor disturbs it now (like the pot getting knocked around by critters like pussy cats from next door), the root could become damaged and the plant will die.
I say this becasue I think minor root disturbance caused the other promising seedling which I had to decline and die so unexpectedly. There is no other plausible explanation. Pots have been displaced here and there, cats are likely out and about!
Must ge the dog to do his âworkâ outside, now that heâs betterâŚLOL.
Must treat this one like the best chinaware, at this critical stage!!!
Cockroaches have been disturbing the surface in pursuit of the snail baits as food (all eaten up as of last night), so I might have to just do without snail baits to keep those gross things away form the seedling, and also make sure I keep the pot indoors every night (plus if it rains), and not forget!!!
note to self
LOL
Howdy George, I read where you boiled your soil media to sterilize it, here is an other option, go and buy an el cheapo pressure cooker, place your soil media in containers and put the timer on for 20 min once it starts hissing, similar action to an autoclave.
Hope you have exprience with pressure cookers
cheers Wozza
Warren,
It is interesting you bring this up. I have obsessed over that dark chapter of WEC runs, of late LOL!
Something happened to the structure of the commercial seedling mix media after I treated it with boiling waterâŚdays / weeks later it started to have a slight odor (made me think it was prolly going a bit âacidâ)âŚ
If memory serves me right, I think there was also snail bait in it at the time I doust it with boiling water, so maybe that also broke down with the boiling water and may have released something toxic in the treated media. It was all a total disaster. Talk about losing control.
Slowly, every single seedling that had survived the fungus gnat attack (which led me to treat the media with boiling water in the first place) went on to show burning/blackening of the young leaves and all seedlings died, some even like many weeks later!
I lost all of David Zâs seedlings.
Murphyâs Law
Next time this sorta thing happens (eg. massive fungus gnat attack), Iâll throw out the media and get a fresh batch +/- different brand. So far that strategy has workedâŚOh⌠and I think throwing a blanket of perlite on top of the sowing media may prevent some of these critters creating a problem in the first place (that is just a guess tho).
BTW, your method of pasteurization is cute!
George, Pouring over boiling water want be enough to kill the pathogens, you need high temperature over a period of time to do it. you could use glass jars with the lids not tightened completely or Polycarbonate plastic containers which are heat tolerate. Plus you will need to make a metal mesh platform to keep them off the base of the cooker.
Warren,
Dousting the media with boiling water was only intended as an emergency quick measure to kill off a huge fungus gnat larval infestation. Pasteurization was not something I was aiming for, then.
For the initial germination of WEC-embryos, I am currently using sterile media (perlite, as per Don Holemanâs advice) and then transplanting to normal seedling mix after germination of the embryos has taken place.
Why?.. well because I would much prefer using media with balanced microbial populations for sprouted embryo-seedlings instead of well pasteurized (or even sterilized) media. I have a gut feeling that the microbe populations, when well balanced, are an essential component for getting seedlings to have a good healthy start in life.
I know, it sounds like some âprobioticâ commercial on TVâŚroflol.
George, you are likely correct. Thatâs why the Pro-Mix that I use comes with a Bacillus species added. The nurseries use it to reduce damping off and root rot. I use it because its what I can buy in bulk, but it is pricey ($25 USD/ 3 cu ft or so.)
Hi Larry, thx for sharing what you prefer to do, and why!
The R. gigantea seedling is not vigorous, but has lived longer than all its siblings before they all died.
Here it is as of a few minutes ago:
The second new leaf shrivelled and is hangning there dead in the picture at the 10 oâclock position.
The larger true leaf has started to develop a slight yellowing of its toothed margins.
There are signs of decline of the remaining (greened up) cotyledon.
There may be a hint of a tiny leaf emerging from the central growth region, but I am not really certain about this.
Looking at this situation, I will be rather surprised if this seedling progresses any further. Time will tell.