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British Columbia, Canada

Hi Matt, I'm just a little curious if you tried germinating a 2328 seed for your greenhouse project?

1/8/2014 11:44:04 PM

NE Kansas

It will be in his diary a month after it happens.

1/9/2014 8:23:40 AM

Massachusetts

lol

1/9/2014 12:51:40 PM

New England

ahahaha

1/9/2014 1:51:21 PM

Torrance, Ca.

It's still not in the ground,,,,, just ask Scott.....

1/9/2014 2:10:49 PM

Connecticut

Quick Summaries:

1.)    Germination of 2328’s does not look good
2.)    I may have found a media that could help increase the germination of older seeds
3.)    Stay tuned for continued diary updates;-)

For those that want the details read further…

I did receive the very rare 2328 seeds and upon inspection I did not give them very good odds of germination as they were mostly empty even though the outer coat looked complete. So, I decided to run multiple trials with different media blends and recipes on other compromised seeds that I had in an effort to determine which one (if any) would work better. After determining the trial media blends, everything had to be prepared. Once the media preparations were completed the test seeds were run through a cleaning procedure before being plated on the different medias.

Germination rate was monitored and documented for each media type. Of all the ones tested there was one that seemed to produce a slightly higher germination rate on compromised seeds. This was exciting as this proved the concept. So, now with this information in hand I selected for just the better producing media to use on the 2328 seeds.

The media creation process had to be recreated, now with just for the chosen recipe. So, this requires gathering the materials, prepare, autoclaving so it is sterile, pour and then let set. After going through the proper treatment procedure the 2328’s were plated and set in the incubator. During the plating process it was clear that there was not much to the inside of the seeds, there appeared to be at best, just the embryo.

1/9/2014 3:09:50 PM

Connecticut

Time elapsed and the ideal germination window of opportunity came and went with no root formation. So, I decided since there was no obvious contamination to take a risk and add in some grow lights to the embryos even though there were no roots. Well, one of the embryos did begin to respond by developing a nice green color. This provided hope as the color only got deeper and what appeared to be seed leaves were expanding. However, over this past week the growth has stopped and upon inspection in the lab (yesterday) the growing tip area is begging to get some discoloration which is not a good sign.

While I did restick the embryo back in the media where the root should have initiated it seems to have done little good. I am still keeping the plant under lights but it looks like if I want to get a plant now I would have to go the route of developing callus tissue and propagating it that way which would put me out of the winter project “season”. So, if it does not develop soon I will have to take a loss despite a long and trying effort.
This is the reason you have not seen any pictures yet in my diary because there are many steps that can get confusing but I assure you there will be pictures in my diary. Hopefully the two seeds I have chosen will provide some good winter reading. Yes, I know my diary is a little behind but my diary is intended to tell a story and be used as a resource so I spend time going through all the pictures I take to provide the best diary I can for everyone. Working through all of the little challenges in the lab is what I enjoy but this is where the “Mad Scientists” idea comes from and it does not make for logical step-wise reading and my goal is to have the growing community understand what I am doing.

1/9/2014 3:10:06 PM

Connecticut

Now, while this may seem like a complete loss, I am seeing a glimmer of hope as all of the different recipes I tried the fact that one could be more effective is very interesting. This discovery may open the opportunity to increase the odds of germination on older seeds as it is often the storage energy that is first to degrade. It may be possible for my media formulation to act as a substitute for this initial energy which could allow for the increased odds of older seeds to germinate. While this may have not been the original intention of this project it is a potential positive outcome from the work that I have done.

I am sure you were just looking for a simple “yes” or “no” and you got a full up to the hour update;-)

1/9/2014 3:10:38 PM

British Columbia, Canada

Alright thanks Matt, keep us posted!

1/9/2014 3:26:33 PM

Whitehall Montana

Matt it is so great of you to undertake all of these challenges and keep us apprised of what you are doing.We really appreciate what you do Thankyou

1/9/2014 3:47:01 PM

Mnt.pleasant ,tennessee

So will we get to here what the special medium is envtually?i got a carrot seed up in 5 days with rabbit manure.sand mix watered with a mix of algi& willow water.algi some great stuff cause of the chloraphyll

1/9/2014 4:40:41 PM

Torrance, Ca.

Very interresting Matt!

Can't wait to hear the . " potential positive outcome " for germinating the harder to germinate or older seeds.

Awesome job Matt!

1/9/2014 5:02:20 PM

Long Island,New York

no agar matt?

1/9/2014 5:47:07 PM

Connecticut

yardman- Your mixture sounds interesting, but for the speed of germination I think the temperature would also play a role. In my case, the materials I am using are measured in milligrams per liter with very low tolerances for error as the materials are in direct contact with the embryo. I am also trying to supplement materials that are found in the endosperm as this component was not present in the compromised seeds I was testing.

LIpumpkin- Agar was a component which was the reason for the need to preform the trials in a sterile environment to reduce the odds of contamination. It is hard to take pictures and maintain a sterile environment but I have one that will show the flow-hood.

1/9/2014 6:48:32 PM

East Jordan, MI

Awesome report Matt, thanks for the information!

1/9/2014 7:01:54 PM

Eastern Pennsylvania

In my line of work, drug development, the study drug may not produce the anticipated results in later phase clinical trials. But what can happen is that unexpected results are observed in enough frequency to re direct the purpose of the drug product. Most notably Viagra, which was origionally supposed to affect blood pressure but the results were minimal at best. But the male subjects sure reported improved performance in other areas! Matt I hope you have stumbled upon something just as great for seed germination. thumbs up lol! Good work- Jim

1/9/2014 7:03:46 PM

Vermont

Very interesting stuff Matt. Sorry to hear the seeds were under developed. Looking forward to catching up at the VGPGA winter meeting!

1/9/2014 9:39:34 PM

Mnt.pleasant ,tennessee

Thanks matt always enjoy your reads.& yes i stayed on top the temperature during germination.

1/10/2014 10:00:47 AM

Springfield, Missouri

Matt,

If you need help with the formula for your growth medium shoot me an email. Try formulas with IAA. NAA and IBA generally didn't work very well.

I have been doing germplasm research with cucurbits for the past few years. The majority of this research has been in the production of callus tissue and the propagation of plantlets.

We noticed that pumpkin callus is harder to differentiate than many other types of plants. (For pumpkins) Once callus has formed the rate of mutagenesis is very high. If the callus is older than 2 weeks at room temp it becomes harder to induce root and shoot formation.

If the growth media you are using currently isn't having the desired effect you can refrigerate callus for several days to keep it "young" while you create new media.

I really really hope you are able to get a plant out of all of this! Best of luck.

1/10/2014 4:00:59 PM

Springfield, Missouri

I have only done embryo rescue 3 times. The embryos I worked with were fresh, and had not been allowed to dry. I used a plain 1/2 MS media mix with sucrose. Two out of the three matured into plants.

I had the embryos under florescent lights the entire time. Shoots grew in the second to third week. Roots took an additional week and a half.

1/10/2014 5:44:03 PM

Connecticut

Thanks to everyone for their replies.

Logan- I appreciate your insight into the methods you have used in the past. I am fortunate that the two plants that I have growing (1734.5 Steil + 335 Scherber) are looking great so the winter project will still go on. While the 2328 was the goal to incorporate into the winter breeding project, I think I have some great genetics to work with in hopes of being able to produce quality seeds by spring time.

1/11/2014 11:09:45 PM

Owensboro, Ky.

Matt, I prbly will sound like a dummy here...but 2328 is a legitimate AG? Serious? Peace, Wayne
PS...I have been away from this sight for a while!!!

1/18/2014 3:56:22 AM