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Tissue Culturing & Other Propagation Techniques of Banana Plants This forum is for discussing propagation techniques of banana plants. Tissue culturing is the popular process of creating clones from a source plant. There are other techniques to propagate banana plants however, such as nicking corms or dividing corms. Learn more inside.

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Old 05-02-2011, 10:13 AM   #1 (permalink)
 
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Smile First time hardening TC rooted plantlets...

Hello!

On Friday,i received an order of TC rooted plantlets which arrived individually packed in sealed cultu bags. First thing i did was mix a 60:40 mix of commercial peat based soil and perlite. in sufficient quantity and pour boiling water on it. After that,i put a lid on the container with the soil to retain heat and moisture and left it cool. The reason of treating with boiling water was to lower the microbial content of the soil and mainly of the perlite as i had it exposed to the elements before.

After the soil cooled,the great transplant began! After cleaning and preparing a table in one of the cleaner(more plant pathogen free) rooms i had available for the transplanting process and a thorough hand wash,i began carefully taking the rooted plantlets out of the cultu bags and the culture medium and after removing it as best as i could from the roots and washing the plantlet under the tap,i was setting it in a clean soak bowl. Then,i was getting the pot ready,filling it with soil up to where the lowest roots would be,sprinkling mycorrhizae and then planting the rooted plantlet on top. After that, i watered and settled the soil well,leave it to drain till i plant the next plantlet and then cover the now potted plant with a freezer bag for lack of a proper box. I treated and planted each rooted plantlet individually to minimize contamination and yesterday that i checked them twice in the day,they grew some within 6h after placing most of them in a PP box to allow for better visibility and airflow than with the individual bags

Here are some photos of them in the PP box! Please excuse the awful quality of the photos,they are taken with my phone,under fluorescent lighting which my phone's camera doesnt seem to like,and the box was full of condensation due to the high humidity. I didnt want to disturb them again.









Some of the plantlets,upon arrival,they had yellowing and blackening leafs. To avoid contamination if this was due to a pathogen,i chose to leave those with individual freezer bags. Here they are!




A question i have about those is why the leafs of certain plantlets were yellowing and blackening while still in the culture medium and with only a day in transit. Was this due to contamination by a pathogen(bacteria?)or something else? Those yellowing leafs are now blackened and dead till the petiole while the leaf base and petiole seem to remain healthy. The newest leaf that wasnt yellowing is nice and green. Whatever and if it was,even these plantlets are showing active growth with only 1 or 2 functional leafs. They seem to be regaining vigor already!

That was how i did it and i really hope things go well and they all survive! I was in a rush to read the most i could before planting them so i am sure there may be better protocols to stick to when transplanting rooted plantlets from the culture medium to soil. What i was trying to achieve is provide a low contaminant growing medium and area for the plantlets to adjust from the sterility of their culture medium to the outdoor environment slowly. I added mycorrhizae as well which will help their root systems adjust faster and better. I plant to keep them enclosed in the box for 1-2weeks and then creating a little gap between the lid and the container and slowly increasing it till humidity drops to the levels of the room i grow them in(60-80% depending on how often the door opens,which i try keeping to a minimum) within a period of another 4 weeks,depending on how their leafs respond to the change.

I checked them today again and they certainly are growing nicely and fast! I am amazed with how well they are growing already,just a couple of days after the transplant! I really look forward to see them grow big!!!!!!

If you see anything wrong in what i did,please let me know as well as what and how i can treat them better from now on till they are fully hardened. I would really appreciate your help in helping these plantlets succeed in life!


Thank you very much in advance!

I hope you liked it!
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Old 05-11-2011, 04:43 AM   #2 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Just a quick update on the above mentioned plantlets,they are all doing very well and many are on their second new leaf already, after soil planting!
They have doubled in height and seem to be hardening successfully! I increased the ventilation of their almost closed growing environment a few days ago and they havent been negatively affected by it so far. The ones in the bags have had their humidity fall enough for condensation to barely form on the clear bags and leafs while the ones in the plastic container had much less of a humidity drop and i will be farther increasing their ventilation soon.
All of them continue their fast growth and have had no older leaf affected from the humidity drop.

Looking forward to their full hardening!!!!
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Old 05-18-2011, 10:09 AM   #3 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Yesterday, i moved the plantlets that were covered individually with freezer bags,in the PP box along with the rest of the acclimating plantlets as they grew too tall and had problems unfolding their leafs in the bags from pushing up the top of the bags. They were now totally healthy looking and robust so i felt safe to keep them all together.

Today i made the first significant change in ventilation of the PP box as i raised the cover half a cm above the box and it now has about 1-1,5cm gap all around,with 3-4cm gap at part of the small sides. I had gradually been raising the cover more and more so this was the immediate next increment but made the gap several times bigger. The plantlets have been guttating a lot in there so i am confident this wont affect them negatively. They have to hurry up and acclimate fast as they wont fit in the box comfortably for much longer,many are just a few cm from touching the cover!!!

Looking forward to their full acclimation!!!
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Old 05-18-2011, 10:17 AM   #4 (permalink)
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Default Re: First time hardening TC rooted plantlets...

Given your climate and the time of year, I would put them outdoors on a table underneath a canopy, umbrella, or 50% shade cloth for about a week. If they are drying out (if they need water) every day, then transplant them into 1 or 2 liter pots.
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Old 05-18-2011, 10:39 AM   #5 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Thank you very much for your reply Richard!

At this stage,i dont feel comfortable getting them outside unprotected as there are lots of things that can happen to them like snail nibbling,breaking by the wind,animal damage or their leafs wilting from not being able to cope with the dry noon and afternoon conditions we have on hot(yet normal to below normal temperature) days we get from time to time. Also,due to this being a really cool and much below average,spring,i am not getting very fast growth from my bananas yet which would prolong their acclimation and growing up time. So i thought i would save me time and worry and acclimate in my warm basement first,under controlled,safe conditions till they are big enough to be able to survive the cruelty of the outside world Then i plan to do as you describe to acclimate to more sun and real life conditions The weather should improve by then and they will continue to be in active growth mode for more months!
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Old 05-18-2011, 10:49 AM   #6 (permalink)
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Default Re: First time hardening TC rooted plantlets...

At this stage, providing them with indirect natural light and air circulation is key. If you can accomplish that in your basement, then hooray!
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Old 05-18-2011, 11:35 AM   #7 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

I can provide them with air circulation(and i am increasing it as they are producing less humidity needy leafs) but certainly not natural light unfortunately! They are under about 50W(i think) fluorescent lighting(the pictures are dark due to my phone camera's dislike of fluorescent lighting,the conditions are quite bright!). I plan to first grow them to about 30cm tall and then start adjusting to natural light. The bigger ones are currently about 15cm tall with only 2-3 new,hardy leafs and a tiny pseudostem diameter. Its also quite convenient not having to water them for the time being
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Old 05-18-2011, 12:29 PM   #8 (permalink)
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Default Re: First time hardening TC rooted plantlets...

Fluorescent light is a good choice -- if they are fluorescent plant bulbs (tubes), even better. At the present size, you'll want about 3.7 Watts per plantlet. Consider giving them a tiny amount of an inexpensive iron supplement.

In my experience, 10cm would be the maximum height before introduction to natural light. The natural light is going to cause the plants to grow faster, and a larger plant will be under stress to develop a root system to accommodate it.
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Old 05-18-2011, 04:20 PM   #9 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Thank you very much for your reply and suggestions Richard!

It seems am a little below in wattage from the recommended but at least the bulbs are of the right spectrum,the tube is for aquarium use and favors plant growth while the compact fluorescent is ''natural daylight''. I plan to fertilize with fish fertilizer once i get them outside along with a little slow release for fruit bearing plants(i remember its the recommended proportions for bananas but dont remember the exact nutrient value). At the present,the soil is saturated and i cant use any liquid till enough evapotranspirates. I may add a little of the slow release though. Its organic and 3month release formula so it should be safe but i dont know if its actually needed at the time being?

I understand your reasoning and that would indeed be better in a more controlled environment than what i can offer them outiside. I fear for the unpredicted(snails,cats,dogs,pill bugs,birds,high winds,heavy rain,etc) that has many times ruined some fine young plants of mine in the past,many times to the point to death. So i am overly cautious with those little plantlets....
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Old 05-20-2011, 04:07 PM   #10 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

I recently planted four bare root tc plants i ordered on line. Two of the plants looked similiar in size two yours, while the other two were taller and skinnier.

All four went from bare root to the ground. The first few days after planting we had high temps (95f). And the next few we had wind in the 30mph range along with a few inches of rain.

I had two small plastic square tables that i placed over the plants. I would rotate the tables every day so each plant got one full day of sun and one off.

All four plants have pushed out one new leaf with the two smaller ones working on their second.

Here is some pics of them from 1 week ago (this is the start of their fourth week in the ground)

Photo Album - Imgur


Quote:
Originally Posted by Kostas View Post
Thank you very much for your reply and suggestions Richard!

It seems am a little below in wattage from the recommended but at least the bulbs are of the right spectrum,the tube is for aquarium use and favors plant growth while the compact fluorescent is ''natural daylight''. I plan to fertilize with fish fertilizer once i get them outside along with a little slow release for fruit bearing plants(i remember its the recommended proportions for bananas but dont remember the exact nutrient value). At the present,the soil is saturated and i cant use any liquid till enough evapotranspirates. I may add a little of the slow release though. Its organic and 3month release formula so it should be safe but i dont know if its actually needed at the time being?

I understand your reasoning and that would indeed be better in a more controlled environment than what i can offer them outiside. I fear for the unpredicted(snails,cats,dogs,pill bugs,birds,high winds,heavy rain,etc) that has many times ruined some fine young plants of mine in the past,many times to the point to death. So i am overly cautious with those little plantlets....
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Old 05-20-2011, 05:02 PM   #11 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Looking great Kat!!!

Thanks for sharing your experience here! Even my bigger ones are some leafs away from the size your smaller ones were when you got them. I grew mine straight from in vitro plantlets while what yours were already hardened TC plantlets. I am currently hardening mine to in vivo conditions and look forward to their full hardening!


My plantlets are showing some slight damage on their older leafs from the humidity drop but the newer leafs are unaffected. The condensation has now cleared completely from the box sides so it's time to get some photos for you!
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Old 06-11-2011, 07:47 AM   #12 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

To update on my plantlets,soon after my last update,i removed the cover completely as the new leafs were pushing on it and humidity was already close to what was outside the box. I left them grow some more in my basement,lowering the humidity some more in my basement but not too much as its my propagation area and needs to be humid. Then finally,2 days ago,i moved them to my balcony in a spot receiving morning sun for about 1-2hours.

Here they are today! The white stuff you see on the leafs is dried up chitosan with insect pathogenic nematodes,from last night's application.


They seem to be taking the transition quite well and none has burned so far. The oldest leafs have some yellowing where they get full sun but they aren't affected too much so far!

Looking forward to new,sun adjusted leafs!!!
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Old 07-24-2011, 07:45 AM   #13 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Just an update on my TC bananas,they adjusted fine to morning till noon sun at my balcony after acclimating them by slowly moving them closer and closer to the edge of it. Then,just 2 days ago,i repotted them to bigger pots and set them outside to enjoy more light and soil. I unfortunately lost 1 to rot(it rotted just above the growing point as well as part of the corm and half of the root system,it may or may not start growing again or throw a pup) from the pot sitting in water for too long or from overwatering,and almost lost another one as well to the same(the leafs started dying back but i kept it drier and it seems to be growing). A third one was affected some but not much. All the others are fine. I will try keeping them a little drier now.

Here are the plantlets after repotting!




Looking forward to see them big!!!!
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Old 07-24-2011, 10:13 PM   #14 (permalink)
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Default Re: First time hardening TC rooted plantlets...

Just a few tips:
1. The soil height in the second pot from the left is correct. In the other pots in the "front" row, the soil height is too close to the top.

2. When I receive banana TC's, I plant them in 2 gallon pots almost immediately then put them outside under shade cloth. Two weeks later they start getting full sun.
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Old 07-25-2011, 02:27 AM   #15 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Thank you very much for your reply and advise Richard!

The soil level is going to lower to appropriate after a few waterings. When i fill to the correct level,it always ends up getting too low.

I received my plantlets growing in sterile,nutrient medium directly from the tissue culture so i thought it was a little different from getting tc plantlets already rooted in plugs and thought it was better to root in small pots first in humid conditions and then move on with acclimation
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Old 07-25-2011, 03:47 AM   #16 (permalink)
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Default Re: First time hardening TC rooted plantlets...

Do U know what they are?
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Old 07-25-2011, 05:04 AM   #17 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Of course!
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Old 07-25-2011, 01:53 PM   #18 (permalink)
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Default Re: First time hardening TC rooted plantlets...

Quote:
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The soil level is going to lower to appropriate after a few waterings. When i fill to the correct level,it always ends up getting too low.
When transplanting TCs:
  1. put the soil in the pot, soak it, and let it drain.
  2. press the soil down very moderately.
  3. if it is to low, add more soil and try again. If it is too high, remove a bit.
  4. get a rod of some sort that is the diameter of the plug or rooting.
  5. press the rod down deep enough to accommodate the plant.
  6. put the plant in and lightly press the soil so the hole closes around the plant.

This technique also works in assembly line fashion. So for example, I'll do steps 1-3 on 288 pots. Then steps 4-5 on all of them. Finally, come along with 4 trays of TC's and do step 6.
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Old 08-08-2011, 12:13 PM   #19 (permalink)
 
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Default Re: First time hardening TC rooted plantlets...

Thank you very much for your reply and advise Richard! I incorporated some of your methods in the second batch of plantlets I potted and did it faster this time!


Just a quick update to let you know the bananas I repotted 2 weeks ago are doing great and getting significantly bigger and more robust! Looking forward to see them grow big!!!
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