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Thread: Heli pollen observations

  1. #9
    Av8tor1's Avatar
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    Mach,

    I dunno.... almost everything Ive read on the subject state anywhere from 2-20% with most refs saying 20%

    So that is why I started there... If still not results with 20% next time ill drop back to 10%

    But Im wayyyyyyyyyyy out of my field of expertise, I am open to any and all suggestions.

    I tried harvesting some pollen a few mins ago, only got a tiny amount... the anthers are still a little green,

    So I tried the onion epidermis and 20% sucrose, the leeks are going to go well with a steak Im about to grill LOL

    ---------- Post added at 01:46 PM ---------- Previous post was at 01:43 PM ----------

    example ref:

    Sucrose concentration: various sources list concentrations from 1 per cent to 20 per cent w/v. The SAPS protocols recommend a starting point of 1.2 M sucrose, which is about 40% w/v, diluted by half on mixing with mineral salts culture medium. The optimum sucrose concentration varies with species – so it may be necessary to use trial and error to find a reliable concentration. 10% - 20% final sucrose concentration seems to be ideal for most. This solution will keep for some months in a refrigerator.

    Im betting Ill end up needing the boric acid and calcium nitrate for the sucrose based media

    This is from the ref that seems the most "pollen tubes for dummies" friendly

    To Germinate the Pollen:
    GERMINATION MEDIA (Since germination occurs rapidly, there is no need to autoclave unless you are preparing them in advance.)
    NOTE: Media C and E are used for extensions, only.

    Medium A - 10% sucrose
    Medium B - 10% sucrose, 100mg/L boric acid, 300 mg/L calcium nitrate
    Medium C - 10% sucrose, 100mg/L boric acid, 300 mg/L calcium nitrate, 200 mg/L
    magnesium sulfate, 100 mg/L potassium nitrate
    Medium D - the same as Medium B with the addition of 1% agar. Heat to boiling and pour into
    petri dishes.
    Medium E - the same as Medium C with the addition of 1% agar. Heat to boiling and pour into
    petri dishes.
    Last edited by Av8tor1; 11-20-2011 at 11:55 AM.

  2. #10
    Hello, I must be going... Not a Number's Avatar
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    http://www.practicalbiology.org/area...s,127,EXP.html

    Says:
    1 Sucrose concentration: various sources list concentrations from 1 per cent to 20 per cent w/v. The SAPS protocols recommend a starting point of 1.2 M sucrose, which is about 40% w/v, diluted by half on mixing with mineral salts culture medium. The optimum sucrose concentration varies with species – so it may be necessary to use trial and error to find a reliable concentration. 10% - 20% final sucrose concentration seems to be ideal for most. This solution will keep for some months in a refrigerator.

    1.2 M of sucrose = 410.76 grams
    Grand Hotel... always the same. People come, people go. Nothing ever happens.

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    Av8tor1's Avatar
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    Im really hoping the onion epidermis works LOL

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    then someone is pickling sweet onions, chopped steak and onions, grilled onions, bacon and onions

  5. #13
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    Kula onion kimchi.
    Grand Hotel... always the same. People come, people go. Nothing ever happens.

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    with ogo

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    If you could see the T-bone that I am letting rest at the moment

    omg yum!
    T-bone, sauteed Leek, broiled asparagus with olive oil and sea salt.... and cold Guinness

    you two better get over here quick

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    Quote Originally Posted by Av8tor1 View Post
    Warren,

    From what all I've read on the subject I'm not sure using another type of pollen would tell me much.

    It seems that what media works for one pollen type may or may not work for another.

    I got a Spanish onion and some leeks today; I will try a simultaneous run with onion, leek and 20% sucrose with uber fresh pollen.

    (From flower to slide)
    Always the optimist! No result with pollen tubes = french onion soup on a cold Sunday night . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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