coline
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Life's essence: patience
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Post by coline on May 20, 2013 12:45:11 GMT
And which concentration did you use?
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robert128
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http://www.plantculture.com.au/
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Post by robert128 on May 21, 2013 4:15:44 GMT
Hi coline sent you a message
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Post by won0507won on Jun 16, 2013 5:11:04 GMT
Hello, Robert!
First, for not good at English, please understand.
I am someone who is interested in tissue culture. I was surprised to see your nepenthes culture.
I tried to cultivate carnivorous plants that bybris and VFT, cephalotus, pinguicula. I am not still trying to incubation with nepenthes. For example, sterilization and hormone's ratio.
Please help for me the success of the culture of nepenthes. Thank you, and sorry for question.
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coline
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Life's essence: patience
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Post by coline on Jun 16, 2013 14:10:47 GMT
Well, good you are interested, but will you try nepenthes tissue introduction or seed introduction?
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Post by won0507won on Jun 17, 2013 0:27:20 GMT
Yes, I will try with nepenthes' tissue and seeds. When I see papers I know composition of media but I don't know sterilization process. Always too much or too little.
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coline
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Life's essence: patience
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Post by coline on Jun 17, 2013 3:18:18 GMT
Since I did not kill the seeds, and I see germination, I have used 1/3MS with 8%PPM for 7 hours in highland seeds as the only dissinfection process. And for tissue I followed Medina's protocol mentioned in this forum with 80% success.
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Post by won0507won on Jun 17, 2013 7:55:15 GMT
Oh thank you Coline! But in Korea there is not shop sells PPM. And i cannot find Medina's protocol.
I'll sterilize for seeds with bleach. In FTS(fly trap store) forum, I see your post and now I know that I must use Benomyl. Your TC also great..
Please help me coline!
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Post by won0507won on Jun 17, 2013 8:13:53 GMT
Oh, I find Medina's protocol.
But i wonder what 10% NaOCL is... It just mean 90ml plus 10g NaOCL or 10% bleach?
On your FTS post, you said that Benomyl effected like BAP. Then medium with Benomyl have anti fungi and BAP effect?
Very interesting! Thank you coline!!
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coline
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Post by coline on Jun 17, 2013 10:36:24 GMT
Well no, NaClO (as it is written correctly) is the common bleach, as it is a liquid, you dilute 10ml in 90 of water, try to see if they use 3,5% active ingredient or 5,25% (which are the most common concetrations sold) what I use is 5,25% a.i. Well, Benomyl indeed affects, but more sensitive species, as Sechium edule or maybe others, but with nepenthes, even with repeated dissinfections through it, has had no effect, or.. it may have helped shoot multiplication. PPM of course is not for sale in Korea, it is normally not sold anywhere, you would need to buy it from www.ppm4plant-tc.com/ (in some cases they may send a free sample) Or from kitchenculturekits.com
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Post by won0507won on Jun 17, 2013 11:48:21 GMT
Thank you very much..! It was a lot of help. I give permission to import chemicals is concerned.
Thank you very much once again.
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robert128
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Post by robert128 on Jul 18, 2013 10:35:41 GMT
here is the latest media formula (always trying to improve it)
NEPENTHES BASAL SALT MIXTURE and add full strength micronutrients 1 lt of distilled water
after you make this use 50 ml of above mix and add 2ml of PPM this will give you a good sterilizing treatment it works well Soak seed 4 to 24 hours in this don’t rinse just place seed strait on to media
the 1Lt of media you have made I add: 0.5 ml of PPM 25g sucrose (sugar) add 7.5 grams of growgel or about 5 grams of AGAR (test this out first with some plain water as some brands of agar require different ratios) adjust PH to 5.5 - 5.68 range (use white vinegar to lower PH and bi-carb soda (baking soda) to raise PH) Add media to your containers or jars and sterilize as normal. I have started to reduce the amount of PPM I use in my media as it is starting to show its limitations as an anti fungal treatment. plants with no signs of contamination suddenly start to become contaminated after a few months. Its seems to hide contamination rather than prevent it. For shoot multiplication use above media with 0.2mg/liter BAP and 0.2mg/liter Kinetin For callus use the same media with 0.2mg/liter BAP and 0.1mg/liter of TDZ. (TDZ is the way to go for callus production) for shoot formation use 0.5mg/liter of NAA
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robert128
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Post by robert128 on Jul 18, 2013 10:43:50 GMT
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coline
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Life's essence: patience
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Post by coline on Jul 18, 2013 13:42:35 GMT
The hoods you make look great!
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robert128
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http://www.plantculture.com.au/
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Post by robert128 on Oct 31, 2013 6:42:06 GMT
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robert128
Full Member
http://www.plantculture.com.au/
Posts: 57
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Post by robert128 on Nov 26, 2013 10:28:00 GMT
Hi all just a few more photos of the nepenthes This first one is of a callus i have been growing for about 18 months using TDZ and BAP this is a few sections of the callus that i removed from the main one and have been placed on media containing BAP and Kinetin they have been on this media for about three weeks the next two photos are the same pieces of callus a few months later
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