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Gabe15
02-10-2008, 10:31 PM
M. velutina, about 15 days old.
http://www.bananas.org/gallery/watermark.php?file=7940&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=7940&ppuser=5)

M. balbisiana, about the same age, seem to be slower to germinate and grow in the same medium.
http://www.bananas.org/gallery/watermark.php?file=7941&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=7941&ppuser=5)

Ensete glaucum subcultured from a single embryo, 25 explants.
http://www.bananas.org/gallery/watermark.php?file=7942&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=7942&ppuser=5)

microfarmer
02-10-2008, 11:30 PM
Looks good! Can you give us the Cliff's Notes version of the procedure? What tools would I need?

bikoro child
02-11-2008, 12:44 AM
wow!!! that's great !!! they are in agar agar?

Las Palmas Norte
02-11-2008, 01:27 PM
I've never been able to germinate any of the store bought banana seeds (Thompson & Morgan etc). Is there a trick beyond soaking and a warm growing medium? Have these store bought seeds lost their viablity with age? If so, how fresh do the seeds have to be, and where would one source them out?
Sorry for all the questions ... thanks in advance. Cheers, Barrie.

51st state
02-11-2008, 07:03 PM
Wow, Gabe thats some awesome photos, loving the Glaucum.

Can you germinate from seed?

I may have a source for some 'special' plants which it might be fun to propagate.

Let me know and I'll PM you.

Kev

bigdog
02-12-2008, 12:05 AM
That's cool, Gabe. Now do that with some Musa ingens, and you're in business!

Gabe15
02-12-2008, 12:17 AM
I should have written more down, but I was in a bit of a hurry at the time. The pictures I posted above are of embryos germinating on a tissue culture medium. Each banana seed contains an embryo which is the actual plant that grows from the seed, the rest of the seed is there to feed and protect the embryo as it germinates and grows. The technique is called embryo rescue germination, the embryos are removed from the seed and (if all goes well) forced to germinate. This is a common method for germinating seeds from hybrid plants which may not germinate normally (usually due to lack of a fully developed and/or functional seed), it is also good for plants that are normally difficult to germinate as well.

The 3 species I cultured were just for practice, they are not rare or necessarily difficult to germinate normally. The Ensete glaucum was originally a single germinated embryo which I induced shoot proliferation on (made many meristems, instead of just one), and then subcultured (divide and re-culture) once to produce 5 new plants, and each of those was then subcultured later to produce many new plants each, I had so many (about 50) that I had to throw half of them away, I kept 25 of them which are now growing. They are genetically identical and suitable for subculture an additional 3 more times if I wanted to create more. They are very young right now, which is why they just look like a mass of green tissue instead of full plants.

I have also put into culture various other Musa and Ensete seeds, but have had limited success with some of them. I will be trying out new and different culture mediums and methods in my trials to come.

I am lucky to have training in tissue culture and full access to a tissue culture lab and all of the equipment and materials necessary. However, I think it could be done with those kitchen culture kits for home tissue culture, or if you do your research on basic tissue culture you can probably rig something cheap and easy. The materials are not very expensive themselves, but the equipment like an autoclave and hood is where you would need a real labs help, but like I said earlier, with enough experimentation it can probably be done at home.

Gabe15
02-12-2008, 12:20 AM
That's cool, Gabe. Now do that with some Musa ingens, and you're in business!

I actually did! Unfortunately I only had one seed left and it seemed to be contaminated on the inside (it had been in soil at one point), so the culture was over taken by mold within a few days. But if/when I get more I will try it again. The same thing happened with E. perrieri. Embryos of M. beccarii var. hottana, M. ornata, M. laterita, Ensete sp. 'Thai Superbum' and Musella lasiocarpa are all in culture as well but have not shown any signs of life yet, but that could be due to a number of different variables.

inkcube
02-12-2008, 12:42 AM
embryo rescue itself might be difficult in a kitchen lab but with a pressure cooker for an autoclave, fruit jars/babyfood jars, and a fish tank on its side as your hood plus a little ingenuity seed sowing can easily be done at home. i started back in the early '80's with orchid seed in a kitchen lab.

getting chemicals may be tough since they are usually sold in quantities that would be large for hobbyist plus some are quite expensive and others can be difficult. also buying a good scale would be necessary and that might put you on a DEA watch list, good precision scales are considered drug paraphernalia.

mrbungalow
02-12-2008, 04:41 AM
Try this one for chemicals and equipment: Home Page - PhytoTechnology (http://www.phytotechlab.com)

For bananas, go for the "Banana multiplication media". Basically, just add (sterile) water, meristem, and a drop of PPM (wich you buy somewhere else.)

I have not been sucessfull yet, but I also lack a sterilizer. (Microwave or pressure cooker)

The race is on to produce TC-plants!!

BTW, Gabe, could you elaborate on the differences between TC and embryo-rescue?

Cheers
Erlend

Gabe15
02-12-2008, 05:46 AM
Your best bet is to probably use a pressure cooker to sterilize the media containers, with the media inside of it (before it gels), the PPM is supposed to reduce contamination but in my trials with it, it cannot handle the mold alone, it needs extra sterilization.

You can get fancy with the medium, but what I am using is basic MS media, sugar, thiamin HCl and gelrite, with minute amounts of hydrochloric acid or sodium hydroxide to achieve a pH of 5.6-5.8. I am experimenting also with levels of shoot proliferation hormones, this essentially makes the embryo either germinate into one or many plants at a time. If anyone is serious about trying and wants more detailed info, PM or email me.

Tissue culture is just a basic term for the culturing of tissue in a controlled environment, in vitro. Embryo rescue means you use the embryo from the seed as your source of tissue, in "regular" tissue culture, you would likely be using the meristematic tissue taken from a shoot which has not flowered yet, although other parts can be used for different reasons.

I also do normal micropropagation tissue culture (standard, as what Agri-Starts and such do), and will hopefully be able to send some of the interesting banana plants here back to the mainland so people can grow them.

inkcube
02-12-2008, 10:17 AM
BTW, Gabe, could you elaborate on the differences between TC and embryo-rescue?

Cheers
Erlend

if you go to this link:
http://www.bananas.org/f30/germination-2992.html

and down to post #3 you will see my post on the basics of embryo rescue. it was basically developed to TC difficult to germinate hybrids and perfected in the day lily labs.

use a pressure cooker, it works on the same principle as an autoclave. ppm is a waste of money. when working in a home lab the main detail is to pay attention to sterility.

mrbungalow
02-12-2008, 01:21 PM
PPM is almost more expensive than gold by weight! As a novice to TC, I bought some just to make sure. Is it possible to be sterile enough as a home TC-experimenter? Or would you reccomend other antimicrobials?

Also, do you have any tips on using explants etc.?

Erlend

inkcube
02-12-2008, 01:58 PM
a pressure cooker will adequately sterilize, they use the same principles of pressure plus heat as an autoclave - pressure cookers will reach a temperature of 250F/121C, @ 15lbs of pressure you need 20-30 minutes to be effectively sterilized. also longer time do not equal more sterile - 15lbs of steam pressure for 15 minutes will kill all bacteria, viruses, spores, and other living things. the key with a pressure cooker is to monitor its temp by watching the release valve (the weight you set on the nozzle) on top which rocks as the steam is let off. be sure set the weight on 15, the weight is designed so that it will lift and hiss at the desired pressure. you want it to rock slowly, about 10 rock per minute. put about 3cm (~8 oz) of water in the cooker to generate your steam. always let the cooker get cold before opening it if it is hot. the hot steam inside containers condenses to a liquid and will suck in one volume of non-sterile air. if you let the cooker get cold, the dust in the room air collects on the surfaces it strikes and no non-sterile air gets sucked inside the containers. you also do not want to tighten the lids of the containers you sterilize (screw them on loosely) - the pressure change will make it difficult to open.

when i used to do this in the kitchen PPM did not exist and i never had problems with contamination - good technique is always the ket to success with TC.

sandy0225
02-29-2008, 06:05 PM
I'm going to have to try that next winter when I get bored. This time of the year is already starting to get busy. That really looks like fun!

microfarmer
03-02-2008, 12:58 AM
embryo rescue itself might be difficult in a kitchen lab but with a pressure cooker for an autoclave, fruit jars/babyfood jars, and a fish tank on its side as your hood plus a little ingenuity seed sowing can easily be done at home. i started back in the early '80's with orchid seed in a kitchen lab.

getting chemicals may be tough since they are usually sold in quantities that would be large for hobbyist plus some are quite expensive and others can be difficult. also buying a good scale would be necessary and that might put you on a DEA watch list, good precision scales are considered drug paraphernalia.

That's the same setup I used for a mushroom culture experiment. I sterilized the grain, with water, in the jars, in the pressure cooker. Worked like a charm. Very low tech. I might just might dig out the equipment and try it again. Maybe some Chanterelles this time...

mrbungalow
03-04-2008, 05:01 AM
Gabe, How did you sterilize the embryos before putting them in the media?

Gabe15
03-04-2008, 05:09 AM
I sterilized the seeds in bleach for a few minutes, then cut them open on a sterile surface. The seeds are sterile on the inside if unplanted, I have had mixed results with seeds that have been in soil prior to culture, some showed instant contamination and others appeared to be clean (but have yet to germinate).

Alternatively, if you are working with fresh fruit, you can simply sterilize the fruit in bleach and then cut in open on a sterile surface and everything inside will be sterile, I have had good results with this and you also get the freshest embryos.

mrbungalow
03-04-2008, 12:43 PM
So here's the million dollar question: How on earth do you get the embryos out??

Erlend

bigdog
03-04-2008, 03:07 PM
So here's the million dollar question: How on earth do you get the embryos out??

Erlend

I have used a simple vice to crack the seeds. Once you crack it, pry it open with something sharp, being careful not to injure the embryo. Then just pluck the little guy out with some tweezers!

Gabe15
03-04-2008, 03:44 PM
For most Musa seeds, if they are fresh or soaked in water for a few days prior to cutting, the seeds will cut easily with a scalpel. If they are dry, I use tweezers to hold down the seed while I spear the seed next to the micropyle with the tip of the scalpel, once its in deep, I press down on one side to form a crack in the seed and it usually splits nicely and the embryo either falls out or is easily dug out.

For Ensete, Musella, Musa ingens and other thick shelled species, I haven't found a good reliable way yet, but Frank seems to have a good idea with the vice clamp, I bet that would make opening those types really easy.

mrbungalow
03-05-2008, 09:33 AM
Well,

Tried this yesterday, and now some more or less sterile embryos are lying on banana multiplication medium.

In sikkimensis I couldn't even find the embryos, all that came out of the seeds was white powder. So no sikkimensis are tested this way.

A little more easy with ensete ventricosum. There I could find the embryos pretty easy. I guess time will show if it works.

How long should I give them?

Gabe15
03-05-2008, 01:38 PM
You should know whether they are growing or not in 2-4days. They will first begin to swell and after a week or so you may start to see the beginning of a leaves and a root. Can you post some pictures of your cultures?

microfarmer
03-05-2008, 10:36 PM
Do you have these on a heat cycle like normal seed germination, or a steady setting? I'm thinking about 85 degrees?

mrbungalow
03-06-2008, 02:29 AM
I keep them in constant 27 degrees C, as I've heard this is the ideal temp. for cell-activity in most musa species. I wouild think this is the naked embryo and doesn't contain the other seed structures that inhibit germination. I doubt changing temps is neither necessary or ideal for getting cells in an embryo to start dividing. (Gabe might know more)

Gabe, will post pics soon! Holding my breath to see what will happen!

Regards
Erlend

Gabe15
03-06-2008, 03:19 AM
As long as they are not in a cold room, they will be fine, temperature does not matter much, just keep them somewhat warm.

marksbananas
08-31-2008, 01:20 PM
For most Musa seeds, if they are fresh or soaked in water for a few days prior to cutting, the seeds will cut easily with a scalpel. If they are dry, I use tweezers to hold down the seed while I spear the seed next to the micropyle with the tip of the scalpel, once its in deep, I press down on one side to form a crack in the seed and it usually splits nicely and the embryo either falls out or is easily dug out.

For Ensete, Musella, Musa ingens and other thick shelled species, I haven't found a good reliable way yet, but Frank seems to have a good idea with the vice clamp, I bet that would make opening those types really easy.

I ma after some advice what inside the seed is the embryo ?

Chironex
10-13-2008, 11:08 PM
I have used a simple vice to crack the seeds. Once you crack it, pry it open with something sharp, being careful not to injure the embryo. Then just pluck the little guy out with some tweezers!

Frank, how do you keep contamination out of the embryo when using this method? Do you sterilize the vise first? Is it inside of the hood? I have a small wooden vise that is used for seal stamp carving (Chinese seals) if I can find it, I may use that under the hood since it is very small, already has a V groove to hold the seed, and uses a small screw to keep from over-cracking the seed. I would think a small C clamp might also be good since it can be sterilized easily. I might also get some fine tipped needlenose pliers to pull apart the seed hull pieces once they crack. Another idea would be to use a rotary tool (Dremel) with a fine saw blade - this might be where the vise would come in handy.

So far no contamination in the 5 embryo rescues I have going. Looks like the embryos are swelling some, but they have only been in culture since 10/5.

Chironex
10-14-2008, 01:19 AM
For most Musa seeds, if they are fresh or soaked in water for a few days prior to cutting, the seeds will cut easily with a scalpel. If they are dry, I use tweezers to hold down the seed while I spear the seed next to the micropyle with the tip of the scalpel, once its in deep, I press down on one side to form a crack in the seed and it usually splits nicely and the embryo either falls out or is easily dug out.

For Ensete, Musella, Musa ingens and other thick shelled species, I haven't found a good reliable way yet, but Frank seems to have a good idea with the vice clamp, I bet that would make opening those types really easy.

I never thought of using the spear technique, I have been trying to use the scalpel like a steak knife. I will have to try your method, but I also think the seed cracking vise has some merit. Thanks Frank and Gabe for the ideas.

Chironex
10-30-2008, 08:24 PM
Well, the ingens embryo has contamination on the latest one that I attempted to rescue. I threw caution to the wind, opened the jar and sprayed it with Physan 20 to see if I can halt the contamination. We will see if that works.
I am going to wait on any further embryo rescue attempts until I can make some Nitsch & Nitsch medium modified for embryo rescue specifically. I am also going to use sterile coconut water and activated charcoal to see if I can get the endogenous contaminants out of the embryos. Could sterilize with silver nitrate too, might try it.

Chironex
01-21-2009, 02:02 AM
The second trial is underway. I prepared the modified N&N medium with only 2 exceptions, first I added PPM and secondly, I used sucrose in place of maltose.
The biggest challenge I have is getting embryos out intact, those guys are tiny! The seeds I have are quite fresh, too, so it may be that the embryos are not fully mature. I have read something about embryo maturation. From what I recall, this explains why some of the older seeds actually germinate and fresh ones do not. This is contrary to what I understood, but I only scanned the article, so don't take this for gospel.
This medium is quite dark due to the activated charcoal used in preparation. I will post some photos in the gallery.
Keep your fingers crossed!

http://www.bananas.org/gallery/watermark.php?file=15323&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=15323)

http://www.bananas.org/%5Bhttp://www.bananas.org/gallery/showphoto.php?photo=15323%20http://www.bananas.org/gallery/watermark.php?file=15323&size=1&filefix=.jpg%5D%20%20%20%28credit%20%5Bhttp://www.bananas.org/member.php?u=2761%20Chironex%5D%29

Gabe15
01-21-2009, 02:25 AM
The other day I put in embryos of M. sikkimensis, M. nagensium, M. cheesmanii, M. 'Darjeeling Giant', M. 'Helen's Hybrid' and Ensete perrieri. I aimed for 5 of each but some of them were bad, damaged or lost during excision. I am using normal MS for the time being until the N&N ingredients come in. No germination yet, but its only been 3 days. I've had them start to grow after 2 days and take as long as 7. It seems that if they do not begin showing even small signs of growth after about 1 week then they might not be viable, this however is just based my own (few) observations.

Since almost all of these species have very thick and hard seed coats, I used the pliers on a leatherman to crack them open. It worked very except that some are so hard my hand would hurt after a while and I would have to take a break!

With enough practice I find it easy to find the embryos because they are always a different color from the endosperm, usually an off-white to slightly yellowish color and shaped like a mushroom.

Some embryos actually do well with being placed into TC media while immature. In some hybrid experiments (I know in particular with papaya), embryos would not mature properly so the seeds could not be planted. However the immature embryos could be rescued and would grow in vitro.

griphuz
01-21-2009, 04:35 AM
So is the idea that one embryo gives one plant, or do you want to devide the growint cells afterwards before producing plants?

And do you sterilise the embryo's before putting them in the culture?
Kind regards,
Remko.

Chironex
01-21-2009, 04:53 AM
So is the idea that one embryo gives one plant, or do you want to devide the growint cells afterwards before producing plants?

And do you sterilise the embryo's before putting them in the culture?
Kind regards,
Remko.

First we get the embryos growing, then they are divided once they start developing shoots. At that point, they are like normal TC plants.

I sterilize the seed surfaces first, then crack them open. The embryos are supposed to be sterile while inside the seed. I do as much of this under the hood as possible to minimize the chances of contamination. Sometimes you win, sometimes you lose.

I was not pleased with the first jar of embryos, so I did another set of them in another jar. This time I was pleased with the embryos I was able to extract. I do not recommend soaking them as this seems to make it more difficult to dissect the embryos. It seems easier to find them and get them out if the seeds are dry.

Richard
01-21-2009, 11:06 PM
Some embryos actually do well with being placed into TC media while immature. In some hybrid experiments (I know in particular with papaya), embryos would not mature properly so the seeds could not be planted. However the immature embryos could be rescued and would grow in vitro.

Ah! Exactly the information I need about my papaya seeds.

Chironex
01-21-2009, 11:44 PM
Just put the rest of my E. perrieri seed embryos onto the new medium this evening. These are much easier to find than ingens embryos. Big difference when they are dry versus soaked! Once the water hydrates the endosperm, I found it very difficult to locate the embryos, even though I knew where they were supposed to be.
I believe I had 12 E. perrieri embryos that made it to culture. I am sure Gabe has not mentioned how many go flying across the room when you crack them sometimes! (hahaha!) I probably have 4 embryos in carpet medium now! Not holding much hope for their survival, although the carpet does have plenty of dirt, who knows!
I will try to post some pics, but my camera does not have quite the resolution that Gabes has.
Well, back to the TC's, I have 16 plantlets to divide, but first I need to prepare more medium and fill some jars. Guess I will be here for awhile.

griphuz
01-22-2009, 03:17 AM
Nice Scot!

So you get 1 plant out of 1 embryo usually, OR you have to invoke shoot/sucker formation in a special medium or something?
Is that more difficult with Ensetes compared to Musas (because Ensete normally doesn't do so as a plant)?
Kind regards,
Remko.

Gabe15
01-22-2009, 03:34 AM
Nice Scot!

So you get 1 plant out of 1 embryo usually, OR you have to invoke shoot/sucker formation in a special medium or something?
Is that more difficult with Ensetes compared to Musas (because Ensete normally doesn't do so as a plant)?
Kind regards,
Remko.
In my trials, depending on the species and how much hormone is in the medium, sometimes they will germinate normally or sometimes they will form many shoots right away. Either way, depending on the level of hormones you put into the medium you can control how much they sucker. I have germinated embryos that could be transferred directly to soil but I prefer to subculture them and create more plants before planting them in soil in case anything happens to them. It's always good to have backups.

I haven't had much experience tissue culturing Ensete (just do to availability), however when I got an E. glaucum embryo to germinate in vitro, it grew very strangely and I could never regenerate any plants from it. I grew some E. ventricosum from meristem tip culture for a short period of time and they seemed to grow normally but I would have to try more to really compare them. The tissue culture process can be made very difficult from the differing habits of different varieties in culture. Some banana varieties are very easy and "behave" well while other varieties are very slow, grow out of control, or grow in strange and hard to deal with patterns.

Chironex
01-22-2009, 01:33 PM
In my trials, depending on the species and how much hormone is in the medium, sometimes they will germinate normally or sometimes they will form many shoots right away. Either way, depending on the level of hormones you put into the medium you can control how much they sucker. I have germinated embryos that could be transferred directly to soil but I prefer to subculture them and create more plants before planting them in soil in case anything happens to them. It's always good to have backups.

I haven't had much experience tissue culturing Ensete (just do to availability), however when I got an E. glaucum embryo to germinate in vitro, it grew very strangely and I could never regenerate any plants from it. I grew some E. ventricosum from meristem tip culture for a short period of time and they seemed to grow normally but I would have to try more to really compare them. The tissue culture process can be made very difficult from the differing habits of different varieties in culture. Some banana varieties are very easy and "behave" well while other varieties are very slow, grow out of control, or grow in strange and hard to deal with patterns.

Gabe, I have some protocols for TC'ing Ensete that I came across in research articles. Let me know if you want them.

Chironex
01-29-2009, 06:56 PM
The embryo rescues continue with Ensete and Musa. I am experiencing a lot of what seems to be endogenous contamination. Next I will try sterilization using 1% silver nitrate instead of bleach solution. Dr Frederic Bakry has sent a research article about his procedure and reports little to no contamination using this technique. Another Musa researcher has also corroborated this finding.
Some are still in good shape and a few appear to be doing something, but it is still a bit soon to expect significant changes in the embryos.

griphuz
01-30-2009, 04:05 AM
If you have any info on that Scot, atricles of some kind. On the Silvernitrate sterilisation,...I would be very interested. Neder heared of people using that for sterilisation.
Kind regards,
Remko.

Gabe15
01-30-2009, 04:16 AM
I have one E. perrieri beginning to grow on 1/2 strength MS with 1µl/L BA. It's too early to tell if its going to germinate normally or grow callus, I have had both happen depending on the species and medium. If they germinate fairly normally, then they are easy to deal with but if they grow callus then it can be hard to regenerate plants.

Chironex
01-30-2009, 12:39 PM
If you have any info on that Scot, atricles of some kind. On the Silvernitrate sterilisation,...I would be very interested. Neder heared of people using that for sterilisation.
Kind regards,
Remko.

Remko, send a PM to me with your email and I will attache the research article in my reply. Being a heavy metal salt, special disposal laws may apply, so be cautious before you use it.

I have one E. perrieri beginning to grow on 1/2 strength MS with 1µl/L BA. It's too early to tell if its going to germinate normally or grow callus, I have had both happen depending on the species and medium. If they germinate fairly normally, then they are easy to deal with but if they grow callus then it can be hard to regenerate plants.

Gabe, that's great news. Was that 1 microliter/liter or 1 micromole/liter. Just being sure. I think a few of the surviving, uncontaminated E. perrieri are showing signs of growth, but not much!

I may take some photos of contaminated cultures to post so everyone can see what it looks like. Some of them are unique patterns.

Gabe15
01-30-2009, 01:07 PM
Gabe, that's great news. Was that 1 microliter/liter or 1 micromole/liter. Just being sure. I think a few of the surviving, uncontaminated E. perrieri are showing signs of growth, but not much!


1 microliter per liter.

In our lab, we use 6 different mediums. Depending on the variety, they will get either 1/2 strength MS (which is standard) or full strength MS (if they are too slow in the 1/2).Within those, we use either 5µl/L of BA for shoot multiplication, 1 µl/L for shoot elongation, and 0µl/L for rooting.

For embryo rescue, I haven't done enough experiments but have done enough to see that each species responds differently in different media. Sometimes the 5µl/L was too much and they multiplied like crazy with no way of regenerating plants easily, and sometimes the 0µl/L did nothing. So I chose the 1µl/L because it is not extreme like the 5µl/L but still has a little bit in there to help induce some growth hopefully.

Gabe15
02-02-2009, 12:22 AM
I have one E. perrieri beginning to grow on 1/2 strength MS with 1µl/L BA. It's too early to tell if its going to germinate normally or grow callus, I have had both happen depending on the species and medium. If they germinate fairly normally, then they are easy to deal with but if they grow callus then it can be hard to regenerate plants.

It looks like its germinating normally right now. I will transfer it to a medium with no BA in it tomorrow, I don't want to mess with it anymore hormonally while its germinating. After it gains some size in the vessel, I will subculture it to produce backups before regenerating any to plant out.
http://www.bananas.org/gallery/watermark.php?file=15550&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=15550&ppuser=5)

Chironex
02-03-2009, 08:15 PM
Still nothing to report on the kinetin/ethrel formula. The seeds are still sitting there but not germinating as yet.
The embryo rescue cultures continue, but progress is slow. Frustrating, but I press onwards.

Gabe15
02-23-2009, 01:47 AM
updated photo of the E. perrieri, its a little slow but appears to be growing fine.

http://www.bananas.org/gallery/watermark.php?file=15827&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=15827&ppuser=5)

Chironex
02-23-2009, 01:48 AM
I am green with envy! Nice job Gabe, it looks great! Wish I could get mine to do something.

Gabe15
02-23-2009, 02:20 AM
I didn't do anything special, and for every successful embryo rescue I've had there have been about 15 more that didn't work! So just keep trying, eventually you will be rewarded.