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agar medium for stem propogation

Discussion in 'Introduce Yourself' started by April Anderson, Nov 29, 2018.

  1. April Anderson

    April Anderson New Member

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    Hello Orchid Forum,

    I am newly signed up!

    I need to buy or make some medium for stem propagating.

    My favourtie Phal and ALL her keikis became infected with a disease - a bacterium or virus - I don't know what it is, I tried every thing recommended to stop it to no avail.

    I have never ever had any disease in my orchids before (20+ years) and I've had this particular orchid for 10 years with nothing but robust and vibrant health.

    Now in 2 months all her keikis (which were huge vibrant plants as well) have died, and the original "mother" has now succumbed.

    This orchid has been so remarkable in her propensity for making keikis, so hoping to coax one more from her before she dies, I removed a healthy stem with 3 healthy nodes, and this is now soaking in the 10% bleach. This disease progresses so rapidly that I decided I must remove the stem now while part of the plant still lives ... so now here I am with the healthy nodes, on green stems, soaking in 10% bleach. I will remove them and put them in sterile water … how long can I keep the stems in water, waiting to "plant" them in medium?

    Please if someone can tell me how to make a medium for the stems, using agar and … what else? I found one recipe only online, and it calls for beef bouillon and sugar! Can this be right? I could buy medium somewhere if it can be shipped immediately.

    I am EXTREMELY fond if this orchid, losing her keikis was devastating and now her too.

    None of my other orchids (all Phal) have any sign at all of infection; could this particular orchid be genetically inclined to be susceptible to disease?

    Please send advice if you can, thank you!

    April Anderson
    Slocan Park Bc
    [email protected]
     
  2. RustyExotics

    RustyExotics Nicholas - It's a terrestrial thing

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    I'm sorry to say that I don't have much experience at all with sterile tissue culture... I buy the majority of my nutrient medium or prepared-medium from PhytoTechnology Laboratories. I would suggest trying some prepared (including agar) basal medium from them or buying a basal nutrient (salt) mixture and adding agar yourself (~8g agar for 1L water - adjustable if necessary).

    I wish I could tell you more, but I really haven't done any tissue culture. A heads up though: if you plan on using agar, sterilization is absolutely key to success. After preparing the agar, exposing it to any sort of contaminated environment (even by opening the jar for a second) can lead to contamination, so it isn't the easiest to do at home.

    I would suggest putting the stem into a sealed container with some moist sphagnum moss before going into anything with agar. I know that sometimes this can induce keikis to form. Sterilize the moist sphagnum by putting it in the microwave for a few minutes and allowing it to cool some. This will help minimize mold growth in the container. I would also treat the medium and the stem with a gentle fungicide. Thiomyl works brilliantly and is what I use when deflasking plants.

    Hopefully someone else who has more experience with this than me will respond soon, but I also hope this helps.

    Also, maybe you could post pictures of what you have to see what kind of stem cutting you're dealing with, how much, and what the mother plant looks like? That may help to identify the best way to go about saving the plant.
     
  3. April Anderson

    April Anderson New Member

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  4. April Anderson

    April Anderson New Member

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    Hello Rusty Exotics Nicholas,

    Thank you very much for the reply!

    Yes, I did find a utube where a woman simply "planted" a stem in sphagnum and a keiki did develop from a bud on the stem, and I have been thinking that's what I should try too, very low-tech but does not incur the dangers of contamination that agar does.

    I will take some pics of the stem with exposed buds; pics of the dying mother plant would just be too painful!

    Also thank you for the info re PhytoTechnology Laboratories, I hope they ship to Canada, I will google them.

    I am a total amateur orchid collector so I don't have fungicides etc on hand but I hope I can find Thiomyl locally; there are a few plant grow support stores here.

    What really amazes me the most about this situation is the rapid infection rate of my plants. I've been lucky so far I guess, but now there is some invisible undetectable horrible disease destroying my orchids, and I've done nothing different in my care routine. Creepy!

    To try to stop the disease, I sterilized scissors is alcohol, cut away the diseased leaves, sprayed entire plant with hydrogen peroxide (one plant I totally immersed in hydrogen peroxide (always using brand new unopened bottles of hp)) … repotted the plants in fresh medium.

    To no avail.

    It's very disturbing to not know what's going on.

    Anyway back to the subject of trying to get some new clones from the stem buds, I will try the sterilized moss in a sterilized jar, do I then close the jar up tight; how will the step breathe if I do this?

    Thanks very much for your reply!

    April Anderson
    Slocan Park BC Canada
     
  5. RustyExotics

    RustyExotics Nicholas - It's a terrestrial thing

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    Hmm... I didn't think about the possibility of them not shipping to Canada...

    Anyways, I'm wondering if you're dealing with black rot. Black rot is a very, very fast spreading fungus that appears as dark, black patches that extend into the interior of the plant's tissue, even healthy-looking tissue. That in mind, when you cut away the necrotic tissue, make sure to check the inside of the leaf where you made the cut and ensure that it's green and not dark brown inside, as that means that the infection has spread further than the cut you made. Black rot is bad because it spreads easily and quickly, but it likely not impossible to treat chemically if done quickly. The AOS suggests using StopRot (contains Bordeaux which is a Copper Sulfate mixture, CuSO4 being a very powerful fungicide and is also usually commercially available). I have never used this, so I'm not sure how common it is. A member of this forum named Ray tends to be the expert when it comes to chemicals, so hopefully he'll add onto this.

    This is just a guess since I haven't seen any pictures yet, but this is a good procedure whenever you're fighting some sort of fungal or bacterial infection.

    If there's any of the mother plant remaining, keep doing what you're doing (cut off infected parts at least 1 cm away from the dead/infected tissue) and then treat it with a fungicide. Thiomyl is pretty commonly available, but it might be something you have to order online because I don't know how common it is in stores. Also, I would put a small amount of cinnamon powder on the cuts. Cinnamon is a natural fungicide and will also help to seal the wound by drying it out.

    As for the stem cutting, I would actually suggest placing the stem on the surface of the sphagnum in a small pot. It may benefit to treat the fresh tissue of the stem with a rooting hormone powder to help induce root and keiki production. After that, I would cover the pot with some plastic wrap or put it in a sealed plastic bag to keep the moisture in. Oxygen isn't a large concern since the plant constantly produces and consumes Oxygen and CO2 (this is how sealed terrariums/vivariums work).
     
  6. April Anderson

    April Anderson New Member

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  7. April Anderson

    April Anderson New Member

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    Hi Rusty Exotics,

    When the disease first appeared I googled the symptoms and thought it must be Erwinia. The symptom was that big circular areas on the leaf very rapidly (in half a day) became wet and soft and if I squeezed the wet soft area I would go right the way through the leaf. Very quickly the whole leaf was in that state.

    Unfortunately I can't send photos because I have burned all the damaged material as soon as I cut it off the plant (and burned all the medium too (I have a woodstove!)). I didn't even know about this website or I would have consulted first! I discovered this website by googling "agar, medium for stem cuttings", etc.

    Regarding the "mother plant", when I saw that it too had succumbed I cut off two big leaves and then repotted the orchid (the roots looked to be in excellent condition) and the result is that seems to be in a state of collapse. The remaining leaves are soft and droopy, but not like the "disease", just like transplant shock … like it is not yet able to take up water. If any more of the "rot" develops I will certainly photograph it.

    I will take some pics of the stem buds and post them.

    I will see if I can find "StopRot" or Copper Sulfate in Canada. When I first googled "Erwinia", none of the recommended treatment products could be purchased in Canada! I tried the local plant grow supply store, and they said it's very difficult / impossible to import the chemical that were recommended for Erwinia (I can't remember what they were called).


    Thank you for your continued conversation!

    April Anderson
    Slocan Park BC Canada
     
    Last edited: Dec 1, 2018
  8. RustyExotics

    RustyExotics Nicholas - It's a terrestrial thing

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    Ohhh, okay. Yeah, Erwinia and Black Rot look absolutely nothing alike. I definitely recommend treating the open cuts with cinnamon. That may act as a barrier between healthy tissue and the infection. I would also suggest some sort of systemic fungicide, although it may be bacterial instead of fungal...

    I did a little bit of research and found that Copper based fungicides may help. As I mentioned, Copper Sulfate can usually be found in common fungicides. I don't know if you have them in Canada, but Lowe's, Home Depot, Ace Hardware, etc. likely have a large selection of chemicals for plants/yards. I would try to find a store with a large selection and browse through them, looking at the active ingredients of each one and trying to find one with Copper in it. Southern Ag and Bonide both make Copper Sulfate fungicides that might help.
     
  9. April Anderson

    April Anderson New Member

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    Well I spoke too soon, here is a picture of the original orchid, now showing signs of rot but now it's as you described above, it's at the center of the orchid.

    This is not what the other orchids had … the "rot" was on the leaves, not at the center of the plant, and it was not brown, it was mushy green soft tissue in big circles.

    I really cannot understand how this started, after all these 20+ years with never a rot in sight.

    I will try to attached photos … how do I do that here …
     

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  10. April Anderson

    April Anderson New Member

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  11. April Anderson

    April Anderson New Member

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    Hello RustyExotics,

    I seem to have to reply in the same box as your message now … hmmm

    Anyway yes we do have Home Depot and Ace Hardware here so I will gather all the copper sulfate fungicides and any other things they may have, asap.

    Cinnamon I do have!

    So I will try that on newly cut areas.

    as you can see from the pics, this orchid is really sick now. Terrible, very hard to understand why this is happening. phal rot.2.jpg phal rot.3.jpg
     
    Last edited: Dec 1, 2018
  12. April Anderson

    April Anderson New Member

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    I am going to proceed with "planting" the stems in sterilized sphagnum now as you instructed.

    Here is a pic of the stems with the healthy looking buds.

    I don't think anything can be done with the section of stem with the un-opened buds … can it?
     

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  13. April Anderson

    April Anderson New Member

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    Thank you very much for all the time you took to advise me, a total amateur, with a very insignificant collection!

    Looking at the photos on your site I am reminded of how amazing orchids are, I hope to have some other (than Phalaenopsis) orchids again one day, but not until I have a better controlled environment.

    I hope to post a success story re the stem propagation … soon!

    I'll go get out the cinnamon now,

    Best Regards,

    April Anderson
     
  14. RustyExotics

    RustyExotics Nicholas - It's a terrestrial thing

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    That's never a great sign; rot in the center of the plant may lead to crown rot which could be either the result of the infection or may be due to excess water sitting in the plant's crown. Luckily, it doesn't it totally out of control in my opinion, so that's good.

    I do hate to say this, but you might want to cut the flower spike off. Flowers take an immense amount of energy from the plant. When you have a sick plant with a flower spike, it very well may use the majority of its energy on the flower spike instead of fighting off whatever pathogen is causing damage. Because of that, I may suggest cutting the spike off. I've had to do this a few times (and it hurts every time), but it ends up being better for the plant in the long run. That decision is up to you; it is possible for the plant to survive with the spike, but it will just be an energy distraction for the plant.

    Also, make sure to monitor that closely. When it comes to infections in the crown of an orchid, it can be very hard to control. Brad from Brad's Greenhouse on Youtube (he actually lives in Canada, too, I believe) made a video on he he successfully controlled it. Fairly simple: keep the crown dry, apply cinnamon to the infected area, wait. I'll put the link here in case you'd like to watch it.



    As for the stem with the unopened buds: I don't think that has a high chance of creating keikis, and may actually just lead to the dieback and likely rotting of the unopened buds, so I would leave those out. If anything, you could put that part in a separate container and cross your fingers!

    Regarding the Copper fungicide: Just make sure to read the instructions carefully! Copper Sulfate is great, but it's also a harsh fungicide (which I guess is why it's so effective, though). As long as you mix the solution as required by the packaging or doing very slightly less, I wouldn't worry.

    Best of luck and make sure to keep us updated! I really do hope this turns out the best for you.
     
  15. April Anderson

    April Anderson New Member

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    RustyExotics likes this.