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Thread: B52 . . .

  1. #9
    kulamauiman's Avatar
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    Quote Originally Posted by mmlr38 View Post
    PPM is an excellent biocide though I've not had as much luck with it yet as I have with the traditional bleach method...most likely because of the amount of time spent working with each.
    I was going to try the peracetic acid again. Seemed like less hassles I did get nice well decontaminated capensis explants. Just over cleaned them....

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    Peatmoss's Avatar
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    So cool! I have gotta try TC, it boggles my mind how fast the plants grow.

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    Tropical Fish Enthusiast jimscott's Avatar
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    Quote Originally Posted by Peatmoss View Post
    So cool! I have gotta try TC, it boggles my mind how fast the plants grow.
    My thoughts exactly!

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    Quote Originally Posted by mmlr38 View Post
    PPM is an excellent biocide though I've not had as much luck with it yet as I have with the traditional bleach method...most likely because of the amount of time spent working with each.
    While I still use bleach solutions from time to time, especially with some "explants" grown outside, I generally reserve the PPM to sterilize sensitive seeds, such as those of Nepenthes; and, as a matter of fact, it was some of that leftover solution that I had used on the B52 flower stalk. I can generally re-use the PPM solution about six times (filtered after each use), before it declines or loses its efficacy . . .
    “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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    kulamauiman's Avatar
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    could one use dichloroisocyanuric acid in pace of bleach with similar times?

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    BigBella's Avatar
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    Quote Originally Posted by kulamauiman View Post
    could one use dichloroisocyanuric acid in pace of bleach with similar times?
    Dichlor is very similar in action to bleach, and I have known some orchid growers who have used it on seed to some success; though I do not know of the preferred concentration or duration . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

  7. #15
    Grow Pitcher Plants! DroseraBug's Avatar
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    Nice Work.

    I use traditional bleach methods with explants and seed of several different carnivorous plant families. Would like to here more about your ppm methodology. For me, it is cost/benefit. If I have to use ppm (more expensive when you rule in concentration) to sterilize something I'm interested in propagating, yet, I can get comparable results with something as cheap as bleach or peroxide, I'll stick with traditional kitchen micropropagation. I do use ppm but minimally.

    I'm definitely interested in hearing more about this because from my research, have not read defined methods (i.e. concentrations of ppm and methods used that work in tissue sterilizatinon with explants or tougher seed to sterilize such as Heliamphora/Nepenthes/Sarracenia).

    Could you give a recommended concentration of ppm to use when sterilizing vft or pitcher plant seed?
    Thanks for posting.
    Last edited by DroseraBug; 02-18-2012 at 11:05 PM.
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    BigBella's Avatar
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    Thanks . . .

    Quite to the contrary, I have found PPM (Plant Preservative Mixture) economical to use for a number of reasons -- chiefly its very predictable success in the establishment of TC cultures, with no issues (in my experience) with either vitrification or phenolic bleeding when used with cuttings: an all-too-common occurrence with bleaching, hydrogen peroxide, and peracetic acid sterilization practices. Also, the lion's share of failures I have seen with those pursuing Nepenthes culture -- especially for the first time -- have always stemmed from over-bleaching. I cannot tell you the number of photos and queries sent my way in the last couple of years -- shots of all-too sterile cultures with very dead translucent seed; and considering the work I have done with some expensive material for clients over the years, PPM has proved to be highly predictable and consistent. There had been some discussion -- more lip-flapping than anything else -- that its use (either in media or simply as a disinfectant) could inhibit germination; but I have not found that to be the case (http://icps.proboards.com/index.cgi?...ay&thread=5073).

    I currently use four and eight percent solutions of PPM (though I have experimented with others), mixed with thirty percent MS (regardless of what your final media will be) -- just the salts dissolved in distilled water -- no sucrose, PGRs (plant growth regulators), or pH adjustment. I generally make small batches, usually not exceeding 50 ml at a time (so only 2 ml of PPM is used for 4% and 4 ml for 8%). Both solutions can be reused upwards of six times (the mix filtered between use and kept refrigerated) For highland Nepenthes, four to six hours in 4% percent seems to be effective; for lowlanders, twelve hours or more in 8% percent (the seed typically being far more contaminated by algal and mold issues than their highland counterparts -- even if freshly-collected).

    For Dionaea, Sarracenia, or Heliamphora seed (or leaves or rhizomes), I would treat them as though highland Nepenthes.

    As an aside, my 2011 contamination rate with that methodology was 1.05% -- and there were hundreds of cultures . . .
    Last edited by BigBella; 02-19-2012 at 11:09 AM.
    “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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