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Discussion Starter · #1 ·
Hello everyone,
I am searching everywhere for information reagrding the tissue culture of aquatic plants...it was great to see many posts in APC reagrding the tissue culture...
I have some doubts specificly reagrding the tissue culture of aquatic plants :
1. How to sterilze the ex-plant ?
2. Which part is most suitable ex-plant ?
3. Multiplication medium & hormones per optimum rate ?
4. Rooting medium ?
5. Subculture period ?

Also it is my request to all members that if anyone has any sucessful protocols regarding any aquatic plants ....plz do share with me as it will guide me to the next level
 

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What you are asking would be probably covered by Plant Propagation in Vitro - probably a 4 hrs. university course....

Look for articles on the subject on Internet - that would be my best advise.
 

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1. How to sterilze the ex-plant ?
You want to use emersed grown plants, since they will be easier to sterilize. How you sterilize them exactly depends on the type of plant, the part of the plant you're trying to sterilize and the size of the part of the plant you're trying to sterilize. There is no black/white answer to this - but there are a few general guidelines.
1-2 seconds in alcohol followed by
10 minutes in 10% bleach
followed by 2 x rinses of 5 minutes each in sterile water.


2. Which part is most suitable ex-plant ?
Depends on the plant. Stem fragments, leaf fragments, the growing tip (meristem) are common parts used.

3. Multiplication medium & hormones per optimum rate ?
Depends on the type of plant. If you cannot find the protocol online, you must use trial and error to determine this.

4. Rooting medium ?
See above.

5. Subculture period ?
See above.
 

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Discussion Starter · #4 ·
@edwardn:
yup...a deep course. But wanted to know frm the experience from the hobbyists.

@hazanfar Ghori :
Thanks for the information .
For the exact thing , i want to propogate Staurogyne sp.
Another thing , can you give me any links where i can find some protocols online, this shall really help me a lot bro.
How many plants have you tried propogating via tissue culture ??
 

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I've cultured Staurogyne sp. though I must say, TC is only worth the effort for slow growing / rare / hard to propagate otherwise type plants.

In a nutshell,

Staurogyne sp.
Sterilize stem fragments that have atleast 1 node, minus the leaves for 10 mins in 10% bleach, followed by a 5 minute rinse in distilled water.

Initiate invitro using
Standard MS at standard concentration, with
30g of sucrose / liter
1 ml/l PPM
1 ml of 1 mg/l BAP
pH adjusted to 5.7
6.5-7g of Agar / l
 

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Discussion Starter · #6 ·
"TC is only worth the effort for slow growing / rare / hard to propagate otherwise type plants"
I completely agree with you .....

Coming on to the topic, have you obtained good results with this ??
I mean did you get good growth ?

My main aim is to start TC for the following plants and propogate them in numbers :rolleyes: :
Staurogyne sp.
Anubias rare species.
Echinodorus Rubin
Pogostemon Helferi

First i want to TC these then the more rare varities later....

Thanks again Ghazanfar Ghori for your response :p
 

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There is a lot of tissue culture done for the garden plants, hostas, daylilies and iris. Try looking up those.

There is also a lot of questions on whether it makes a true to form clone. You hear a lot of complaints about the plant not being quite right. Most I suspect are mislabling problems from the giant production houses that do it though rather than sports created by the tissue culturing.

A product called BAP is most commonly used.
 

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I'm taking a micropropagation class like edwardn suggested. I'm lucky enough to have Dr Kane as my professor. He gave a speech on the subject at last year's AGA convention. Dr Kane hooked me up with a micropropagation scholarship and is giving me free reign in the lab to experiment with any plants I choose. I'm currently working on Erio cinereum. I'm looking to use this method for hard to find or hard/slow propagating plants also.
It really is about trial and error with most plants. I'm using 4 different Stage 1 mediums with various cytokinin levels to get the Erio to grow. I'm using 2 different types of tissues as well. The buds should develop into new plants as long as they don't get contaminated. I'm also trying to get pieces of leaves to propagate through non-zygotic embryogenesis. This process will take much longer as the cells have to de-differentiate, then form a meristemoid for plant development.
One problem with tissue culture is possible callus development usually formed due to high auxin:cytokinin levels in the medium. With callus comes undetermined genetic variability. All plants appear to develop correctly through the various stages, but the problems are not apparent until the plants begin to grow after transplant. We were told a story in class today about a banana farm in Jamaica. This farm ordered several thousand plants from a lab. Calluses formed in culture, but the lab did not handle the callus correctly. Everything was fine until the plants were planted in the ground. Some plants were distorted or stunted, others did not fruit, some just died from unknown causes.
 

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Discussion Starter · #9 ·
@SueNH:
Had a look with the garden plants , including sugarcane, actually my friend is more into this he understands it more perfectly but both of us are now looking for some solution :(

@cah925:
"I'm taking a micropropagation class" :p Even i should join you because i need some knowledge regarding TC .
Good to hear that you are working with Dr.Kane , please do keep us updated on Erio cinerum . By chance if you need any cinerum's do let me know i will send them :D


Once the protocol is established ,i bet everything will go out perfectly.
 

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"I'm taking a micropropagation class" :p Even i should join you because i need some knowledge regarding TC .
Good to hear that you are working with Dr.Kane , please do keep us updated on Erio cinerum . By chance if you need any cinerum's do let me know i will send them :D
pm sent

Once the protocol is established ,i bet everything will go out perfectly.
Developing protocol on something that hasn't been documented (at least publicly) is very challenging. Part of this process with Dr Kane is doing research on plants in the same genus to help develop a plan using various hormones. Finding info like this is also challenging and I'm having to resort to looking at how other non-related aquatics are propagated. The Erio is for a class project, but I now have an ongoing relationship with Dr Kane to use the lab at my disposal to work on other plants as well. We are both hoping to get a research article published on my work with aquatic micropropagation.
 

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Discussion Starter · #12 ·
PM replied.I will send them for you [smilie=n: [ no $ required :D]

"Developing protocol on something that hasn't been documented (at least publicly) " - PM me :D
Very nice to see you working hard ....I hope to learn something and looking forward to see your published article.......
 

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Are you guys using 1mg of PPM in 50mg of water to disenfect explants and using PPM in the media. Or are you guys just using a Bleach or Alcahol to disinfect the plants. The last time I made my batches without PPM and had almost 80% contamination. The person I learned from doesnt use PPM but he told me I should have more luck with it.
 

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Hi All,

Ghazanfar is amazing! He visited us at GSAS for our January '09 meeting and did an absolutely excellent presentation on crypts with emphsis on emersed culture. I regularly visit his blog, he has become quite the TC expert it seems! Thank you Ghazanfar for sharing your experiences with us!
 

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Nice thread, when I get some free time I'd love to try some TC. Any recommendations on where I can go or books I can buy to get started? too bad I just graduated, I would have loved to take a class on TC.
 

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Nice thread, when I get some free time I'd love to try some TC. Any recommendations on where I can go or books I can buy to get started? too bad I just graduated, I would have loved to take a class on TC.
Here is a place to buy pretty much everything you need to get started http://www.kitchenculturekit.com/Catalog.htm there are other places also but I think your better off buying from here. I didnt and payed rediculious shipping cost. Go on you tube and do a search for tissue culture there is a guy that has a series of videos on the subject. Some advice take your tme learn as much as you can before you jump right in. This is all about trial and error dont get discouraged if you dont get results the first time and rememmber contamination is the enemy. Im no expert but if you have questions PM me and Ill gladly help if I can. Good luck!
 
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