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avip04
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 Posted: Thu Aug 01, 2013 12:07 pm Post subject: Aquatic Plant Tissue Culture - Anubias Barteri |
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Plant Tissue Culture - Anubias Barteri
Anubias Barteri [Aquired from Aqua Star, Kolathur, Chennai, India]
Used MS medium with 1mg/ml of Benzlaminopurine
Washing Explants:
washing with Tween 20:
After washing:
MS agar medium in sterile bottle:
Sterilized Explants on MS agar:
Rhizomes on MS agar:
Anubias Leaves of agar:
Reference:
Kanchanapoom K. et a., Micropropagation of Anubias barteri var. Nana from Shoot Tip Culture and the Analysis of Ploidy Stability., Not Bot Horti Agrobo, 2012, 40(2):148-151
http://www.notulaebotanicae.ro/index.php/nbha/article/download/7520/7019 ., 31st July 2013
Its been around 48 hours since inoculation there seem be no signs of contamination.
Everything going good except for the agar solidification problem. |
Last edited by avip04 on Thu Aug 01, 2013 3:51 pm; edited 1 time in total |
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Nidhi
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Joined: Nov 23, 2011
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Location: Bangalore, Prakashnagar.
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| Posted: Thu Aug 01, 2013 12:26 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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Good project.. would love to see the results.
The image size is very large, please reduce the size so that people with slow net speed will not have problem. |
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Ryel
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| Posted: Fri Aug 02, 2013 4:10 am Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| Keep updating this thread. Would like to see how this turns out. |
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Sunny_2K_99
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| Posted: Fri Aug 02, 2013 11:53 am Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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 Keep it up... |
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avip04
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| Posted: Fri Aug 02, 2013 3:19 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| Bad news, air conditioning in the lab stopped working and the temperature is getting higher [arnd 40 degs, optimum would be around 28 degs]. . The rhizomes are beginning to turn dark. No problem with the leaves though. Mayb i'll put the agar bottle in some water to cool it down. . . will post pics soon. |
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superbsite
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| Posted: Fri Aug 02, 2013 4:16 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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+1 avip04
Glad to see that youve taken up this
Thanks for keeping us updated |
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coonooranand
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| Posted: Sat Aug 03, 2013 12:42 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| After using tween you directly transfered it to a MS media? What did you use for sterilization? |
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avip04
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| Posted: Sat Aug 03, 2013 7:15 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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This was followed :
Washing and sterilization:
The parts of the plant cut using a scalpel blade and were washed the with tap water for 5 minutes after which the water was discarded and fresh water was collected, to this 2-3 drops of tween 20 was added and then the plant was washed again for 5 minutes. This was followed by washing with double distilled water and then with 3-4 drops of 4% Sodium Hypo chloride solution in fresh distilled water for 5 minutes. Finally the plant was rinsed twice with distilled water and then with 1mg/ml of mercuric chloride solution and then rinsed again using sterile distilled water in aseptic conditions to remove the residual chemical substances. |
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avip04
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| Posted: Sat Aug 03, 2013 7:49 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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There were two tubes which got contaminated with some kind of bacteria. The partially solidified agar started bubbling up, so i had opened the one i brought home , there was smell of fermentation and the piece of plant was soft and dead, had to discard it. The other one in the lab had a large chunk of rhizome in it which showed similar conditions. The other rhizomes are turning dark and showing signs of rotting, but not really sure as of now.
Contaminated Tubes:
Rhizome with leaf:
Rhizome:
Rhizome in bottle turning Dark:
On the "UP" Side: 
There is no problem with the leaves they are green and bright.
Leaves: [Got some 5 tubes of these]
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Ryel
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| Posted: Thu Aug 08, 2013 9:41 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| @avip04: how is the tissue culture going about. Any success. |
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avip04
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| Posted: Fri Aug 09, 2013 9:52 am Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| The leaf parts are doing fine , no sign of cell differentiation, mayb cos its a slow growing plant. Will post pics soon. Given up on the rhizomes, they are dark and rotting, no contamination though. |
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avip04
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| Posted: Fri Aug 09, 2013 10:04 am Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| The leaf parts are doing fine , no sign of cell differentiation, mayb cos its a slow growing plant. Will post pics soon. Given up on the rhizomes, they are dark and rotting, no contamination though. |
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Ryel
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| Posted: Fri Aug 09, 2013 5:51 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| how many days have become since you started this process. |
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avip04
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| Posted: Fri Aug 09, 2013 10:19 pm Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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| Around 11 days. |
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avip04
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| Posted: Mon Aug 19, 2013 9:58 am Post subject: Re: Aquatic Plant Tissue Culture - Anubias Barteri |
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Project failed, even the leaves started dying  |
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