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seed pod

#21 User is offline   Yeong 

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Posted 07 January 2016 - 12:56 PM

Gel Preparation:
Ingredients needed:
- Activated Charcoal ( It is not easy to buy it, so we can grind activated charcoal tablets for diarrhoea and use it)
- Agar Powder ( It is usually packed in a 50-kg bag for industrial use, difficult to buy small quantity, so we can use agar strip to
replace it; agar strip can be bought from provisional shop, about $1.20 per package.Do not use gelatine powder for
making cake)
- Vitamin B1 tablet ( Can buy from medical hall, if you happen to go to Thailand, can buy it there as it is much cheaper)
- Sugar
- Banana Pulp ( blend a small banana with 100ml of water)
- Coconut water ( can buy a small Thailand coconut from supermarket, about $1.20 per piece and get the juice)

Procedure:
1) Measure 2 g activated charcoal (grind 4 tablets), 7 g Agar strip (cut into
small pieces), 1.2 g vitamin B1 (grind 3 tablets , each contains 100mg Tiamine)and 20 g sugar in a boiling pot
2) Add 900 ml tap water and 60 ml banana pulp or 30 mi coconut water.
3) Bring the mixture to boil using small fire, keep stirring the mixture until all the agar strips are dissolved.
4) Pour the liquid medium into glass bottles to about 2 cm in height.
5) Steam the bottles in a big container for about 2 hours to kill bacretial, place the cap on top of the bottle, leaving a small
opening. Do not close the bottle. (Pressure cooker is used in commercial lab).
6) After 2 hours, turn off the fire and quickly close the bottles. Do not over tighten the bolltels as they may not be opened once
cooled down.
7) Take a transparent plastic bag and spray alcohol in it, place a bottle in it and tie it with a string. So, the bottle is kept in
a clean environment.
(Alcohol can be purchased from any chemical lab, it costs about $20 per 5 litre. It is too expensive to buy it from local pharmacy,
as it costs about $3 for 200ml).
8) Leave the bottles for about 1 week, discard those that have bacterial growth in it, use only the good one for seed sowing.

Please share your suggestion to improve the produres.

I wii continue the Seed sowing procedures in my next post.
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#22 User is offline   sthh 

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Posted 07 January 2016 - 12:58 PM

Very interesting. Look forward to more pictures of your home flasking.
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#23 User is offline   AhSeng 

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Posted 07 January 2016 - 11:04 PM

My flasks sometimes fail. More fail than success. But it is quite fun actually. How is the results of your flasking experiments so far ?

Did you use pH paper to measure the pH of your medium ? I have used banana pulp too and realised that it can lower the pH of the medium. Previously, I use citric acid to lower pH, but now I exclude citirc acid. yes 60g banana sounds about right.

I'm using 8g agar for 1 liter of water. I've a small electronic weighting device to weigh the ingredients, such as sugar, banana etc..

Some of my protocorms in flask will turn white or turn brown in flask. Maybe due to something call hydricity.It usually happens at about 2 months from flasking, so that's the main problem I face right now. but some flasks actually made it. The most success is with Cymbidium finlaysonianum. now growing Tolumnias which seed pods only take 2 months to mature, but it is more sensitive than cymbidium.

What is the light system you are using ?

The more difficult technique is cloning orchid shoots. I read that the growers clone the phals using the root meristem. I'm thinking of trying this since the phal roots grow quite easily. However, it is easy to get contamination in flasks, and so it is more difficult than sowing seeds.

This post has been edited by AhSeng: 07 January 2016 - 11:17 PM

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#24 User is offline   Yeong 

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Posted 10 January 2016 - 03:17 PM

Hi Ah Seng, Thanks for your information.
Yes, the pH of the medium should be around 5.5 to 6.0 as banana pulp provides an acidic condition to the gel.

In commercial lab, 0.25g/litre of potassium dihydrogen phosphate (KH2PO4) is added to bring down the pH to 5.3 as the salt gives
a good buffer effect. Also other inorganic salts like calcium phosphate, potassium nitrate, magnesium sulfate, managanese sulfate,
EDTA etc.. are added to provide trace elements.

Since it is not practical to buy all these chemicals and they are only used in small amount, we can use fertilizer as a substitute.

In fact, we can also add about 0.25g (1/4 teaspoon) of a 15:15:15 fertilizer to the 1000ml gel system I mentioned earlier.
This will give a pH of about 5.5.

Sunlight: I place my bottles in the balcony, at about 2 feets away from the window where sunlight reaches.
Never place them under sunlight.

Cloning of Orchid:
Meristem Tissue culture is one of the methods in propagating orchid. It is commonly done in commercial lab, especially for
dendrobium, where a young stem (about 2-3 inches in height) is cut and young shoots/bud is taken from internal part of the stem
and placed in liquid medium (usually Vacin & Went with some modification is uesed) with continuously agitation to allow protocorms to develop.
For phalaenopsis, the shoot is taken from the node of a flower stem.
Carrying out Meristem tissue culture needs a very clean and sterile environment, so a laminar-flow fume hood is requried. I don't recommend
this be done at home.

There are 6 ways to propagate orchids, ie Seed Growing, Meristem Tissue Culture, Division, Back Bulbs, Keiki and Aerial Cuttings.
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#25 User is offline   Yeong 

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Posted 10 February 2016 - 02:58 PM

Sowing seeds from green Pod :
1) Items required:
A metal plate, a piece of forceps, a cutting blade, a spray bottle of alcohol and
a 2-ft fish tank.
https://c2.staticfli...3437a1e22_c.jpg
https://c2.staticfli...8ac83a027_c.jpg

2) Procedures:
a) Harvest the seed pod when it is about 80% ripened (about 3 months after
pollination)
B)/>/> Place the metal plate, forceps, cutting plate and bottles of gel medium
prepared earlier inside the fish tank. Spray the tank thoroughly with alcohol,
make sure that the entire tank is filled with alcohol vapour tp provide a sterile
environment. Seal off the tank by covering it with a piece of plastic sheet and
leave it for at least 3 hours. (The sterile tank is used to replace Laminar-flow
fume hood)
c) Soak the seed pod in a solution containing 9 parts water + 1 part sodium
hypochlorite (Clorex) for 30 minutes, remove it and wash it with water, then
soak it in alcohol for 30 seconds, remove it with a forceps and burn
off the excess alcohol by passing it through a flame (be careful not to burn the
pod)
d) Remove the plastic sheet from the sterile tank, place the seed pod on the
metal plate. Wear a pair of glove and spray alcohol on the hands. Cut open
the seed pod and transfer the seeds into the medium bottles. Seal off the
bottles and place them back to plastic bags filled with alcohol vapour. (Try to
complete this step as fast as you can to prevent contamination).
e) Place the bottles about 2 to 3 feets away from sunlight. Wait for germination to
occur, it may take from a few weeks to a few months.

3) Successful Rate:
For gel preparation: It is quite successful, about 70 to 80%, ie no fungus grow
after leaving the medium for more than a week.

After sowing the seeds : I don’t have much information on as I
have only done 2 seed pods, the first one was done on a mature seed pod and
was unsuccessful. The second one was done on a green pod and was good.
. Protocorms developed.
.
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#26 User is offline   adrianlau 

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Posted 04 January 2017 - 03:08 PM

Thank you Yeong for sharing the agar recipe. Do you do tissue culture of variegated banana plant?
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#27 User is offline   Yeong 

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Posted 05 January 2017 - 10:48 AM

Hi Adrianlau,
I don't do tissue culture of variegated banana. The method is quite similar to Orchid Meristem Culture.
Suckers from mother plant are usually grown in Murashige & Skoog-based media with hormones (Cytokinin and Auxin)added.
Please share with us if you have results from banana tissue culture.
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