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  #1  
Old 10-17-2007, 11:36 AM
korxi korxi is offline
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Default Why doesn't it work for me? :(

Hi, I've been giving propagation a try lately with seed pods from three phals I pollinated last year.

I did a whole lot of reading up on medium preparation, sterilization etc. and found a solution that seemed to work for me sterilization and sowing wise.

The media I tried were: One based on potato and honey, and one based on regular sugar and orchid fertilizer. I followed the recipes completely for both of them and had perfectly sterile jars after two weeks of waiting. I sowed - still no infections.

However I hardly got any germination! From the first seed pod that matured I only got one protocorm, from the second seed pod I only got three protocorm and from the third I haven't gotten any protocorms at all! They were sown in June, July and august respectively.
Now the few protocorm I did get have browned up and died. They didn't even really develop to be larger that 0,5-1 mm

I had the jars on my windowsill where they have had some direct sun, at least in June and July. During the last month or so the temps in the day time have been around 20-21 degrees celcius during the day and 18-19 during the night.

I really want to get this right and I'm not stopping before I do! So what should I do??

Here's some things I think could be the reasons:

Perhaps the pH levels werent right - I'm using some pH sticks, but they are kinda hard to read of I think.. (it should be pH 5-5,5 right?)

The temps should perhaps be higher, like 25 degress celcius, and be constant throughtout the day?

Perhaps they got to much light?

There's a chance that the seeds just weren't viable, but since it's from three different pods I'm thinking that it is highly improbable.. They were all three noid hybrids though, I don't know whether or not complex hybrids are more likely to produce infertile seeds?

...

I have actually build a box now with light fixtures in it where the temp will be aprox. 23-25 degress celcius when the light is on. Now I'll just have to get some "cool white" lighting since I've read that ít's the blue light of around 450 nm that starts of growth.. How does that sound?? Is it ok that the temp drops to 18-19 degress celcius at night when the light is off?

I hope some of you can help! I see so many of you having jars filled with protocorms and seedlings - and I REALLY want that to!

Christian

Oh btw. I did ventilate the jars after "protocorm" development. However I didn't replate since they were so small..

Last edited by korxi; 10-17-2007 at 11:39 AM..
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  #2  
Old 10-17-2007, 02:12 PM
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Tindomul Tindomul is offline
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Direct sun on glass jars/containers spell disaster. Can you say runaway greenhouse effect. I am willing to bet the temps inside the jars were very very high.
I also thought you should check pH.
Now tell me what you did to get sterile media? The last and first time I tried I got fungal media
Well good luck next time. Did you save any seeds? Usually you do this incase something goes wrong.
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We must not buy their fruits:
Who knows upon what soil they fed
Their hungry thirsty roots?"

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  #3  
Old 10-17-2007, 04:07 PM
newflasker newflasker is offline
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Why doesn't it work for me? :(
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Some comments:
Light: direct sun light could kill your seeds/protocorms. Germination doesn't not required much light.

pH: 5 - 5.5 is ok. I use pH paper before without any problem.

Temperature: 20 - 25 C degree is ok. 18-19 C at night is a little bit low but still ok (Seedlings may grow slow).

Sterilization: Rise seeds with distiled & sterilized water after sterile with hypo-chloride or peroxide hydrogen.

Medium: what kind and how much fertilizer do you use? mine is 1.5 g/L fertilizer 20-10-20 (with minor elements such as: Cu, Mo, Fe..., No urea).
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  #4  
Old 10-17-2007, 05:10 PM
korxi korxi is offline
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Thank you guys for the replies!

Quote:
Originally Posted by Tindomul1of9 View Post
Direct sun on glass jars/containers spell disaster. Can you say runaway greenhouse effect. I am willing to bet the temps inside the jars were very very high.
I also thought you should check pH.
Now tell me what you did to get sterile media? The last and first time I tried I got fungal media
Well good luck next time. Did you save any seeds? Usually you do this incase something goes wrong.
I pressure cook to sterilize, and I have only had two or three contaminated jars, so the problem does not come from that. To sow I use the syringe mnethod with peroxide. In this way the seeds are actually sown in peroxide that turns in to water. Now I think of it I was actually wondering whether it would be a problem if the seeds were lying in water or if that will work just as fine?? I have read about liquid and semi liquid media, so my guess would be that it's ok.. Furthermore this technique is well known and described all over the web, which must mean that other people have had success with it!

Oh and I didn't save any seeds from the first two pods, but I did form the third. However I'm pretty sure those seeds are infertile. Well I couldn't see any embryos through a microscope - but it is a really cheap toy kinda microscope.. And another thing was the fact that the seed pod only contained very few seeds compared tot he other two phal pods.. In any case from now on I'm waiting till the seeds are dry and I'll save some in the fridge.. Lesson learned

Quote:
Originally Posted by newflasker View Post
Some comments:
Light: direct sun light could kill your seeds/protocorms. Germination doesn't not required much light.

pH: 5 - 5.5 is ok. I use pH paper before without any problem.

Temperature: 20 - 25 C degree is ok. 18-19 C at night is a little bit low but still ok (Seedlings may grow slow).

Sterilization: Rise seeds with distiled & sterilized water after sterile with hypo-chloride or peroxide hydrogen.

Medium: what kind and how much fertilizer do you use? mine is 1.5 g/L fertilizer 20-10-20 (with minor elements such as: Cu, Mo, Fe..., No urea).
I'll be using my "germination box" from now on with the cool white fluorescent bulb then, and make sure that the temps don't drop below 20 degrees!

In the media I made I used the amount of fertilizer that is recommended for watering the orchids.. I don't know if that was a good move cause I kinda made that media up my self (though heavily inspired from posts and articles of the web).. It was:
1 l of rain water (I used rain water since I had it around.. Could this be a reason for the lack of gerimination?? My thought was that if the orchids like it the seeds probably will to - and that's what they get in nature!)
25 grams of sugar
1/5 tsp of fertilizer (this is in powder form, the guide says to use one tsp. for 10 l of water.. This fert has micro nutrients included)
½ multi vitamin pill (it actually had ginseng - could that do any harm??)
8 grams of agar

I adjusted pH with citric acid, but as I said I don't know if I got it right.. I think those red colors look very much alike on that scale
I'm considering buying one of those cheap digital meters - found one called "milwaukke instruments PH600" for something like 35-40$


Christian

Last edited by korxi; 10-17-2007 at 05:15 PM..
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  #5  
Old 10-18-2007, 12:12 PM
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Tindomul Tindomul is offline
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8 grams of agar was enough? Usually it takes 15 grams per liter to make anything solid.
I also used the syringe method, but in the end what did me in was cheap flasks that were not air tight.
__________________
"We must not look at goblin men,
We must not buy their fruits:
Who knows upon what soil they fed
Their hungry thirsty roots?"

Goblin Market
by Christina Georgina Rossetti
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  #6  
Old 10-18-2007, 01:47 PM
newflasker newflasker is offline
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Why doesn't it work for me? :(
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Quote:
Originally Posted by Tindomul1of9 View Post
8 grams of agar was enough? Usually it takes 15 grams per liter to make anything solid.
Tim,
I use only 5grams/liter agar, enough to make it solid and soft enough so root can get inside of medium.

Christian,
You may use inositol (100mg/l, at health store) and fresh young coconut water (50 ml/l) to enhance germination.
Nguyen
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