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Flasking Setup

This is a discussion on Flasking Setup within the Flasking Equipment & Technique forums, part of the Orchid Propagation category; Hey Ross, Question for you, as I have just done my forst replate ever, a ...

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  1. #11
    ATester's Avatar
    ATester is offline Minster of Silly Flasking
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    Default Flasking process

    Hey Ross,

    Question for you, as I have just done my forst replate ever, a little nerve wracking I have to say...making sure nothing goes wrong), how do you replate? Do you have a glove box or do you have access to a hood?

    Having neither, I created a tent from wire and plastic sheeting with multiple layers of plastic in the opening where I put my hands through to keep as much outside air out as possible, I also wiped everything down with rubbing alcohol (91%) using a new package of sterile gauze everytime to keep contamination to a minimum. Though REALLY makeshift, I have not seen any fungal mats or mildew forming on the insides of the flasks yet. I am going to keep an eye on things though and hope that by a week or two without contamination, I should be in the clear (for interests of observation, after reflasking, I kept the mother without a lid and it took three full days for fungal mats to grow).

  2. #12
    montanum is offline Junior Member
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    Hi ATester,
    I replate inside a 10 gallon fish tank (fish not included). They cost $10. It's not as much room as I'd like, but I've managed ok.

    I used to be very cautious with replating: I would wipe down the flasks and fish tank with a bleach solution for every replate and I used bleach and H2O2 as my sterilizing agents for my tools. It took a long time.

    However, recently, I am trying to find the minimal requirements for sterility for time's sake. I still wipe down the inside of the fishtank, but usually just once beforehand. I should wipe the sides of the flasks with an alcohol soaked paper towel, but sometimes I am too impatient. I use bleach, H2O2, alcohol then fire on my tools, in that order. I use a camping stove (I know it says "Do not use indoors", but I only turn it on for 15 seconds at a time) for the flame. I, of course use latex gloves, for my skin more than anything else. I should use a dust mask to keep "wind" down, but I don't. I've been meaning to get some at the store, but always forget.

    More than anything, I rely on speed and accuracy. The time from flask opening to flask closing is all that really matters aside from the other necessary precautions. If I'm concentrating, I have less than 1 in 20 go bad (often 100% success). If I'm just replating because I have to and I have 30 more to do and I'm tired & hungry, then the results are worse.

    Best,
    Ross

  3. #13
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    Great info, Ross!

    Keep us abreast of your progress with your little critters!

    Julie

  4. #14
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    Ross, just want to say thanks for this... most informative...

    I'm actually learning heaps from what you guys are talking about in terms of hygiene when it comes to flasking.

    and as much as i hate to have to give up acaule... oh sweet lip of venus... errr... yes, kentuckiense, i'm studying over the net... start with one step and then the stars...

    again, thanks!

  5. #15
    montanum is offline Junior Member
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    Hi bench,
    Glad our banter is helping. Another important thing is to wear low lint clothes and move your body "slowly" to prevent drafts. Wind is the killer of flasks which is why the Laminar Flow Hoods are so important in commercial stuff. It's easy to sterilize tools and jars, but it's impossible to sterilize the air around us ;-)

    You should have a look at Cyp formosanum and japonicum since they both resemble acaule somewhat.

    And as I mentioned in the other post, Cypripedium, from Gk "Kypris pedilon" translates to "Shoe of Cyprus", a reference to the *slippers* of Aphrodite (Roman: Venus). Cyprus was her legendary birthplace. Hence the common name for the genus, Lady Slipper, is not just for any lady: It's the shoe of a God!

    I used to think that the word greek word for orchid, "orchis" (Gk: testicle) was a direct relation to Cyp acaule; however, I soon learned that it was actually a reference to the European genus Orchis. This genus has a healthy bulb which it grows from, and during the late part of the growing season, it still has the original from the current year as well as the new growth bulb which it spent it's time making: hence testicles.

    Best,
    Ross

  6. #16
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    Default replate

    well, the good news is, so far no contamination!!! I am still giving myself a couple weeks before I consider it "in the clear". I like the fish tank as a flasking set-up. I have been contemplating putting the expense into plexi and building a flasking case out of that, cutting round holes in the front face so I can attach gloves. the tent I set up was intended to be disposable (already went into the trash). My only problem was the fumes, I was retarded for about three hours afterwards (mostly from diluted bleach and 91% isopropyl alcohol), I was also probably holding my breath which didn't help, but I didn't dare open a window in case the air stirred up nasties. After flasking it was an emergency evacuation of the cats and myself until the smell disappated. I should have worn my mask, but I didn't think of it until after I was halfway through the process and I didn't have another pair of gloves to use as back up!

    Oh well, it was a learning experience. I jot down everything in my journal that I keep just for my orchid obsession

  7. #17
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    Default Replate

    So, so far the replate was a sucess! No contamination and I am finally able to discern some sort of structure to the first little green, for the lack of a better term, leaves.

    Ok, so Montanum...I have a question for you...say I was a glutton for punishment and am interested in flasking a particular Cyp species. What medium would you choose? And the temperature regimine?

    I have been doing a little light reading on the subject of Cyps and from what I understand there is one of many hurdles to Cyp flasking, and that is that there is a natural inhibiter to seed germination (some form of coating from what I can gather). What do you do (if anything) to overcome this barrier? From what I understand as well, as much as the temperature regimine is important, so is pH. At what pH do you make your medium?

    Since you are our expert on Cyps, I consider myself fortunate to pick your brain.

  8. #18
    montanum is offline Junior Member
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    Hi ATester,
    I am certainly not an expert, but I do have some relevant information :-)

    Your second question is easiest. The temperature should be about 68-70F degrees for best germination (Rassmussen: Terrestrial Orchids..., & Steele: Correspondence). However, in some cases a 3-5 month cold period directly after sowing is necessary to initiate germination. I always use cold treatment on acaule and pubescens just because I get better germination this way. Other people will have fine germination without it. So far, it hasn't ever hurt to give the seeds cold treatment. A rule for me is if it doesn't germinate after 3 months at 68F, into the fridge it goes. Often, this helps. Orchis mascula also requires 3 months in the fridge.

    Next, for destruction of the seed coat, most Cyps require a lengthy (compared to their sissy tropical cousins) bleaching in a 0.6% NaOCl sol'n (aka a 1:9 solution of Clorox ultra). Some people use Ca(OCl)2. Which Cyp do you want to do? The optimal scarification time varies (sometimes greatly) from species to species (And even plant to plant, unfortunately). For example, pubescens and acaule are good at about 1:30 to 2hr, kentuckiense and formosanum are good at about 0:35, and arietinum at 2:30. Let me know if you have a species in mind and I'll try to recommend something that's worked for me.

    Of course, the greenpod method is the only way to germinate some species such as calceolus, montanum and plectrochilum, among others.

    In general, germination can be anywhere from 0% like dry pod montanum, to 99% like the 4 blasted passerinum pods I sowed... On average, you should be happy with 40%, and very happy to see 70%.

    For medium: There are medium specifications per species. Below, I've included my list of media formulations for each species. Many are guesses (indicated by a ?). This list was compiled with help from several people in the biz, but mostly from Bill Steele at Spangle Creek Labs. You will notice that it is certainly not complete for the genus! I'm missing 30+ species, including all of the section Trigonopedia (spot leaved Asian species) and many others.

    Feel free to contribute your successful media for a given species if you have any. The pH for all of them is somewhere around 6.

    Compiled by Ross Kouzes Created 2005-07-08 Last Modified 2005-09-14
    macranthos@yahoo.com

    Sowing: reginae, acaule, fasciolatum?, tibeticum, macranthos?, calceolus
    T839 + 0.2g/L ammonium nitrate + 40cm3/L potato

    Sowing: flavum?
    T839 + 0.2g/L calcium nitrate + 40cm3/L potato

    Sowing: californicum, formosanum
    T839 + 0.2g/L ammonium nitrate + 20cm3/L potato

    Sowing: fasciculatum
    T839 + 0.2g/L ammonium nitrate + 80cm3/L potato

    Sowing: arietinum, montanum?
    T839 + 0.2g/L ammonium nitrate + 1mg/L kinetin + 40cm3/L potato
    (no kinetin for greenpods)

    Sowing: parviflorum, pubescens
    T839 + 0.2g/L ammonium nitrate + 120cm3/L potato

    Sowing: kentuckiense
    T842 + 40cm3/L potato


    Replate: reginae, flavum?
    T842 + 0.2g/L ammonium nitrate + 0.7g/L calcium nitrate + 40cm3/L potato

    Replate: acaule, californicum, formosanum, parviflorum, pubescens, kentuckiense, fasciculatum?, arietinum?, montanum?, fasciolatum?, macranthos?, calceolus
    T842 + 0.2g/L ammonium nitrate + 40cm3/L potato

    Replate: tibeticum?
    3/4 strength T839 + .1g/L calcium nitrate + 40cm3/L potato

    Hope this helps.
    Best,
    Ross

  9. #19
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    What a wonderful thread, thanks for sharing all of your knowledge.
    C

  10. #20
    yellow treasure is offline Junior Member
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    hi everyone,
    I'd like to know what you have to do to let cyp seedlings go into dormancy, and let them mature at the right time, according to your season,
    and what you have to do when it goes wrong.

    With regards

    Nick

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