# sowing multies



## polyantha (Sep 21, 2011)

Hi everyone

I know how to germinate orchids, but I never tried sowing seed of multifloral paphs because I never had something to sow . Perhaps someone could tell me if they (species) are difficult or not.
If someone has a suggestion what media I should use, please tell me...

thx polyantha


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## Pete (Sep 27, 2011)

meh. pretty much the same as anything else when your sowing the seeds. just shake/sprinkle them around in a thin layer.
i like the phytotech orchid maintenance media P668 and i dont go quite full strength, add 10g banana per liter.. pH to around 5.2 i think. i gotta check my notes.


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## polyantha (Sep 29, 2011)

Thx for your answer. I have a lot of p668 media, but is it relly a good idea to add banana?
What is better: p668 or western medium?


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## likespaphs (Sep 29, 2011)

how do you prepare the banana? 
how ripe should it be?
just the fruit or the skin too?


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## Roth (Oct 1, 2011)

Actually multis germinate much better in liquid media, but it requires skills ( stupid Vacin and Went with 1g/L peptone and 15g/L sucrose, pH 5.7, this does not work if you add agar, believe me...).

For solid media, I would go for a MS base at 1/4 strenght, 10g/L sucrose ( not more), 1g/L peptone, pH around 5.7-5.9. High pH kills many protocorms, the seeds will expand, but the embryo will die, believe me... For the banana at 10g/L I used to use 30g/L for the germination of ripe banana, but it is a bit tricky, as you must replate as soon as the seeds germinate, or for some crosses the protocorms will never give a leaf.

Multis are easy, 

the easiest to begin are Maudiae pot plant types, and the brachypetalum species, even leucochilum etc.... They can give many thousands seedlings for one seed capsule. 

Parvis (except vietnamense and delenatii), and many things like mastersianum, volonteanum, etc... are much more temperamental and require specific care...


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## polyantha (Oct 1, 2011)

Interesting that you mentioned vacin and went, roth. I also noticed that it does not work, even if you add Na-Fe-EDTA and/ or tryptone. What do you use as replating media?


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## Roth (Oct 1, 2011)

polyantha said:


> Interesting that you mentioned vacin and went, roth. I also noticed that it does not work, even if you add Na-Fe-EDTA and/ or tryptone. What do you use as replating media?



For the Vacin and Went with 1g/L peptone and 15g/L sucrose, pH 5.7, it works perfectly well, as a liquid media ( no agar). You have to change the liquid twice month about, or whenever it turns an amber color. With agar, it does not work at all. Do not use tryptone, I got problems with it several times depending on the supplier... Peptone is much safer overall.

Donald Wimber used actually a liquid Tsuchiya, which is a modification of VW in fact, for all the paph seed germination, with excellent success.

I suspect first because the seeds get wet in the liquid media, second, it dissolves any metabolism byproducts, etc... from the germinating seeds, in agar the metabolism products would be close to the seeds and kill them. Some media formulations makes for heavy waste products production ( said to be phenolics, but they are absolutely and definitely not).

Last replate, I use a proprietary stuff I developed myself ( same for seed sowing) to get very fast results, however, I used to use half strenght MS with 5mg thiamine 1mg pyrixdoxine 1mg nicotinic acid, 18g/L sucrose, 120g green banana, gelrite 2.5g, and 200mg peptone, pH 5.7.

One thing you must remember, and that's why most labs cannot be trusted too. The seeds must be germinated, then replated anyway when they are white protocorm, then when a leaf tip is coming, then the last replate when there is a leaf and a root. If you delay the plants slow down dramatically. That's in fact the biggest secret in the lab, very frequent replates and timely replates... Multiflorals from seed to 8-10 cm seedlings takes about 9-10 months if properly replated on time. Most commercial labs will wait until a batch from various customers is ready to replate, by that time some plant will have slowed down their growth speed and that's a mess then...


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## Brian Monk (Oct 2, 2011)

You use sucrose @ 1%. I was under the ompression that Paphs did not use sucrose well, and used Fructose much better. Any thoughts? Asn what about D-inositol? Do you have any thoughts on BAP?


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## Roth (Oct 3, 2011)

Brian Monk said:


> You use sucrose @ 1%. I was under the ompression that Paphs did not use sucrose well, and used Fructose much better. Any thoughts? Asn what about D-inositol? Do you have any thoughts on BAP?



Yes, but a fair part of the sucrose is dissociated during autoclaving according to some chemists... I tried actually sucrose vs fructose/glucose, and sucrose made a better germination. Plus, autoclaving pure fructose is quite risky depending on what is inside the media, its reaction products can be phytotoxic...

See this:

http://www.jstor.org/pss/2482086

BAP I never use it for anything at all, there are always better, safer ways. BAP produced plants have a very high rate of mutation, and furthermore the BAP takes ages to be evacuated from the tissues once it enters. Most high standard labs avoid the use of BAP nowadays... Myo Inositol, I used to use it, but I did not find any real difference. 

One more very important thing, I always make the media from scratch myself, I never buy salts preparations etc... 

The MS etc... sold as a powder are not the same as the formulation, or the result of making the MS as an example by dissolving the salts 1 by 1.

As a stupid example, mix calcium nitrate and FeEDTA as powders, let the powders stand for an hour, then dilute it. After a day, you get a precipitate. If you dissolve the Calcium nitrate on one side, the FeEDTA on the other side ( never forget to boil it to dissolve it and bind it...), and add them together, it's stable. There were a few studies showing that media preparated from concentrates had a subtantial amount of precipitated things.

The second reason, the manufacturers make huge batches, and split them. As they are salts of various size, densities, etc... even if you homogeneize it for hours, it is very difficult to guarantee that a small part of the bulk will have everything in the same proportions as the bulk. That's why the suppliers provide analysis for this batch, or general formulations, but that's quite clear that, let's say sodium molybdate will not be in every 'ready to make 1L' jar sold, because the quantity for 1L will be one crystal or two... You get the same with the fertilizers too, 1g of fertilizers very, very rarely contains the proportions indicated on the bag. 1kg, most likely, 10kg, pretty sure, 25kg, definitely you should be close to the formulation.

Anyway, for the media, there are a lot of variations, how long is it sterilized, what is its composition to start with, how it is prepared, what kind of agar or gelrite, quantity per flask... All of this makes variable media ( hence I use only a few types of flasks, about 5 different ones, same for the plugs/caps, because indeed different flasks give different results and different media composition once they are properly autoclaved).

I expect too that the microwave sterilization of the media, which becomes affordable and popular, will make even more problems, because our formulations were made before with an autoclaving of 10-25min and 115-132 degrees celsius, which definitely made the final product different from the prepared media. With those new technologies like microwave or microfiltration, the final results are way different...


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## TyroneGenade (Oct 3, 2011)

Hi Roth, 

Thanks for the useful info. I guess I have to get moving and replace my thaianum protocorms!


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## NYEric (Oct 3, 2011)

Very fascinating reads on things I will most likely never try.


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