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I received a few inquires about how I germinated D.Regia seeds on compost. This was my first attempt at growing what I affectionately call the "uglies". They are by no means a plant I like, I am just an equal opportunity germinator. I read a couple of threads on other's experiences germinating and growing D.regia from seed. The take home message: 1) the seed had better be viable at the start and 2) feed the babies or they will suffer the consequences. Below, is a more detailed protocol of what I did. For those of you who I had previously responded to, please go by the notes below. After having sowed hundreds of containers in the last few months, I can't recall on the fly which babies got rolled in what. :-D

Soil
I used a 2:1:1 mix of peat:sand: pumice.
- Sand = HTH pool filter sand, 20 grit, 50 lb bag. If you use a lot of sand, this is very economical. I was buying 5lb bags of horticulural grade sand at OSH but this works just as well. I pour some in a 4L beaker and fill it up with milliQ water. I will mix the sand and change the water until is is clear, and no longer milky.
- Peat = Black gold peat. This has saponin as a wetting agent. I don't wash this - that is where I draw the line. ( FYI, saponin is a detergent (amphiphatic) and is used on various grains like quinoa and should be washed if one is going to consume whatever it is covering. It imparts a bitter taste.)
- Pumice = Black gold pumice (Yes, that is all they have at my local OSH). This stuff is VERY dirty (twigs/rocks/ probably my grandma if I looked hard enough..etc) and necessitates multiple washes/rinses.
The ingredients are mixed in the ratio above and should be moist, not soggy. It is placed in a 2L glass beaker, covered with foil and autoclaved for 20-25 minutes. Usually I make this ahead of time and have some on hand because it needs to cool to room temperature before use. Excess soil can be stored in the 22C.

Seed Pretreatment
In a small petri dish microwave some water to lukewarm temperature. It should be cool enough to touch but still warm. Add a "sprinkle" of trichoderma flowerable (Ampac Biotech, store 4C) - sorry that is what my notes say. Mix the trichoderma solution (color is light brown) and add the seeds. Allow the seeds to swim in the trichoderma mix at room temperature for 1 hour (at least).

Sowing Seeds
I sowed the pretreated seeds in a 4'' round plastic pot, the diameter which is perfect for making a dixie cup greenhouse. This is my emulation from RL7836 posts on the ICPS. I thought it would be a cheap way so I could watch them grow, as I am not a big fan of the ziplock baggies. (1) 2mm hole was made in the center top of the dixie cup with a surgical knife. In another clean glass beaker, I added the necessary volume of cool autoclaved soil mix and ~1/40th of a tsp of granular trichoderma (Ampac Biotech). The soil was not filled to the brim of the pot, ~1/2'' from the brim so the dixie cup could effectually "sit on top of the soil" snugly. Using a loop, I fished out each seed from the trichoderma brew and placed it onto the soil. I gave each seed a gentle "poke" to make sure it is snug in its new home. The seeds were evenly spaced from each other in the pot, all 1'' away from edge of the pot. I had already planned on dividing each little sprout into its own pot shortly after germination. This space between the seeds was to allow me to scoop each seedling + soil as a plug, and place it into a new pot without disturbing them (who likes earthquakes?). After all was snug, I placed the dixie cup over it, said a prayer, and into the incubator it went.

Germination Temperature, Lighting, and Watering
If you have read my other thread "Nepenthes Tissue Culture", you may remember that I have turned my planaria incubator into my germination chamber. (This is only for the interim until the planaria reach larger numbers - when I will have to move my plants elsewhere. Good thing one of my postdocs could not perform molarity calculations, or maybe not..).
Temperature -The incubator temperature is set to 21C, 24 hours a day, 365 days a year. The temperature inside the dixie cup is 76-78F high/69F low.
Light - Light cycle is 16hr ON/ 8 hr OFF. Light is provided by: 8watt, 320 lumens 13’’ F8T5CW 4200K + 24'' 20watt Phillips Daylight 6500K. The distance of the brim of the pot to the light bulb = 4''.
Watering - I mist with autoclaved milliQ h20 (I happened to have a lot on hand usually since we use a lot in the lab) every day or every other day. One sprout did not make it past sprouting and the verdict is still out if this was a result of desiccation, or if the seed was simply not up to snuff.

Seedling Temperature, Lighting, Feeding, Watering, and Transplanting
Temperature - The "uglies" have not seemed to fret over the intermediate conditions I have been providing them. Seedlings, with at least 2 true leaves, like to be closer, rather than farther, to the light provided that one can keep the temperature below 80F. There have been days when the incubator has been open for extended amounts of time and the temperature was at the low 80'sF.
Lighting -The 1'' "uglies" are separated onto 2 different shelves. One set is 9'' from the light and the other is ~3'' from light. Those 3'' from the light were 1.5X the size of those 9'' from the light at 5 weeks old. Yesterday, I moved those 9'' from the light closer to the 3'' by elevating them on empty pipet tip boxes.
Feeding - At 2 true leaves, the "uglies" are still tiny and I didn't expect much root formation so I placed the slow release oscomote pellets around the seedlings. The pellets were close, 1/2'' from base of each the "uglies". Basically, they were surrounded by at least 2 pellets each. This is critical. After having to starve oneself for many years to make weight on weigh in days, it makes one very unhappy and metabolic stress pathways turn on in full force. Feed them!
Watering - I HATE WATERING!! And even so, I must mist every 2 days at least. The dixie cups allow a lot of moisture to escape since the soil is not completely level, so they are checked on periodically. However, I don't water the plants. Instead, I give each oscomote pellet a good misting.
Transplanting - Each "ugly" was given its own 4'' pot + dixie cup when they started bumping into the sides of the dixie cup and I saw that the hairy real root started coming off its backside (4 true leaves at this point). I thought it prudent to move them before the root rooted. Using my trusty spoon (not the same one I eat with) which has been autoclaved, I spooned out each "ugly" + plug-of-soil into a new pot with same soil mix used originally. A little depression the size of the plug has been carved out to facilitate the process. Level the ground and surround the "ugly" with 5 or so, oscomote pellets. The pellets were placed a little farther this time, 1'' , to encourage the roots to grow out.

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"Uglies" got 2 true leaves so get their first feeding of oscomote pellets

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First week after oscomote pellets

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"Uglies" hairy backside -> time to transplant

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"Uglies" in their new homes with more pellets


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Incubator Growing Chamber - Second level is where D.Regia germinated

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Top level -> elevated the "Uglies" growing with empty pipet boxes

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Second level

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N. Attenboroughii sprout being dwarfed by its "ugly" neighbor

I sowed 11 seeds on compost. 7 germinated. Lost 1. 6 "Uglies" at the end of the day.

Hope this helps people in their endeavors.

Jen
 
Last edited:
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Details from seed indeed. Very informative Jen..
This should be a sticky!
 
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Yes, I have my own lab. 50ml centrifuge tubes are good. The ones we buy are usually the cloudy polypropylene type so not as good for light penetration. The clear polystyrene are much better for light penetration but they are brittle and are more susceptible to cracking. For smaller cultures I have been using 40ml glass scintillation vials with a rubber septum. On the positive side, they can be reused. On the downside, they need to be washed for reuse. We go through cases of plasticware for our tissue culture, I don't really want to add to that expense with my plant TC trifles:)
 
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HAHAHha, meth? Surely, there are better things to do in life.

No we do basic and applied research in the biological sciences so molecular biology, bioinformatics,..etc. This is similar to academia, with some translational projects.
 

jimscott

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We have something called Life Science Technologies in our area, with one building right next where I work, which is a pharmaceutical manufacturer. LST does a lot of cell culture work.

Back to the topic... I have a hard time getting D. regia past the cycle of one leaf postpeak; one leaf peaking; and one leaf emerging.
 
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You mean Life technologies, formerly Invitrogen? If so, they are one of the 2 big life sciences supplier companies. Thermo Fisher Scientific is the other. They have bought all the other larger-middle- and new startups in almost all fields (basic/applies/diagnostic/sequencing..etc) in the last decade. It is a dance between the companies, who can buy the companies faster and take the market share.

Really, the uglies have 6-8 leaves now. What do you mean postpeak, peaking.. Surely, it is different than my cute mental depiction that conjures.
 
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jimscott: Are you feeding your regias? I've tried feeding mine fish pellets and blood worms but the leaves always died off at the point of contact so I stopped. I find that adding 4-5 Osmocote pellets really helps with growth.

Mine have grown from 2" to over 8" single leaf span in a little over a year.
image_zps58748a88.jpg
 

jimscott

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You mean Life technologies, formerly Invitrogen? If so, they are one of the 2 big life sciences supplier companies. Thermo Fisher Scientific is the other. They have bought all the other larger-middle- and new startups in almost all fields (basic/applies/diagnostic/sequencing..etc) in the last decade. It is a dance between the companies, who can buy the companies faster and take the market share.

Really, the uglies have 6-8 leaves now. What do you mean postpeak, peaking.. Surely, it is different than my cute mental depiction that conjures.

This is about as good as it got for me:

Picture002-11.jpg


Post-peak means on the decline. Yes, Invitrogen. It's building next door from us. Our company, now Fresnius Kabi, in the 6 years I've been there, was called Abraxis Bioscience, American Pharmaceutical Partners, APP, and now the German owned Fresenius Kabi. It's like a corporate lava lamp.

Don: I feed my sundews and butterworts pulverized freezed dried bloodworms.
 
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Does the soil drain well? I see the cracks but the rest of the soil looks like my morning bowl of cooled congealed oatmeal (or polenta). Usually, if I try to add water to it, the water just slicks right off. You are using sand + milled sphagnum moss, correct? All the particles (sand,sphag) look the same size. In my experience sphagnum moss acts like a glue-sponge if processed well enough. The peat has particles of different sizes/geometries - twigs/round particles,..etc to help disrupts any compact matrix that might form. My uglie's compost is always dry (visually, at the top after) a day or two. You might try repotting it in different mix that has some thing of a different size thrown in or starting from seed on a compost that is not so uniform.

Eidetic memories of a college course on material sciences in the chemistry department.....
 

jimscott

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I actually have a better picture... somewhere... Anyhoo, that plant died. I have no D. regia. I thought I read that D. regia prefers wet conditions.

I had to look this up:

Eidetic memory
From Wikipedia, the free encyclopedia
This article is about the ability to perfectly recall images, sounds or objects. For the TV show, see Wogan's Perfect Recall. For the video game developer, see SCE Bend Studio. For the 2011 documentary film, see Photographic Memory (film).

This article needs additional citations for verification. Please help improve this article by adding citations to reliable sources. Unsourced material may be challenged and removed. (October 2012)
Eidetic memory (pron.: /aɪˈdɛtɪk/), commonly referred to as photographic memory, is a psychological or medical term, popularly defined as the ability to recall images, sounds or objects in memory with extreme precision. The word eidetic, referring to extraordinarily detailed and vivid recall not limited to, but especially of, visual images, comes from the Greek word εἶδος (pronounced [êːdos], eidos, "seen").[1]

It was new to me.
 
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It is one of my unfortunate attributes. There are people who can not forget a single day of their life which eventually pertubs them. Thankfully, I am not in that category. However, my cache is significantly large than average. I am dyslexic and my brain does not recognize correct spelling (thank god for spell check) but I can recall "pictures" of things up to my early childhood. I can tell you inane things like who wore what on a specific day or where a specific tube was on someone's bench years ago, or whole conversations...etc. In essence I am like a visual map for most people at the lab. It is like a somnabulistic function when I walk around, my brain just records everything pictures and sounds. Usually, I have a hard time focusing when I am around crowds since the brain is in overdrive memorizing every thing. I wanted to be a spy when I was little and thought it would be a good skill to have. Now, I am not so sure as it takes away some of brain's function for scientific creativity. It has proved invaluable for school and in the business world. But I did get into a lot of trouble as a result of it when I was younger :)
 
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Can you tell me which osmocote you use? Cause there are different kind of them in the store I last checked.
Thanks in advance
 
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