Friday, September 12, 2014

Journal September 12, 2014 - Using an Ultrasonic Cleaner On Explants

My post of August 7, 2014 concerned an attempt to micro-propagate single node cuttings from a Chemlali olive tree.  It was my intention to use protocols with slightly different components and compare the results.  

The Chemlali was part of an order of four varieties that I had recently received from a grower in Florida.   All of the trees were covered in a film of debris that looked like it may have been fine dust that had gotten wet and dried on the plants.  That said, I cleaned the trees as best I could before I took the cuttings, sterilized and placed them in vitro.

To my dismay, all of the explants developed contamination within a few days.  Several attempts to rescue the cuttings failed, except for one.

Another trial was started today with a different variety, Empeltre, that came from the same grower  in the same shipment.  However,  I am taking a different approach in processing the cuttings prior to placing them in vitro.

 As you can see by the photo of the cutting above, this plant also has a film on the leaves that has resisted cleaning.

The leaves were  first wiped using a Q-tip with water and a mild detergent, then processed in an ultrasonic cleaner.   Using 500 ml of water, 25 ml of H2O2 and 2 drops of Dawn detergent to act as a surfactant, I processed the cutting for eight minutes at the unit's highest setting.

The liquid and cutting were then transferred to a Mason jar with a screen top and processed normally for one hour.  

After rinsing in sterile water the cutting was cut into single node sections, which were then placed in vitro.  

It should be interesting to see if the ultrasonic unit makes any difference in the outcome.
Last, but not least, the Bougainvillea seed shown in the time lapse video is making progress.  Although the upper portion of the plantlet is growing very slowly,  the tap root is an altogether different matter.  It is the most unusual looking root system I have seen to date.  The two dark circles resemble eyes, giving the root a most sinister appearance.


3 comments:

Corey W said...

Do you have any updates on how adding the ultrasonic cleaning may have helped? I'm looking into this for some of my own home TC, and I'm trying to figure out what ultrasonic cleaner to give a go.

Jack said...

Cory,
The cleaner did not help with these particular explants as they were in pretty dismal condition to begin with. I am still using it on seeds though, it has become part of my regular routine just to be safe.

Nona said...

I found your blog through C. Stiff. Thank you so much for sharing your procedures and adventures. I will be using this method for seeds, and am so very thankful for your site!!