Tutorial

#TissueCulture #BananaPropagation #InvitroBanana
In this tutorial I'm going to show you how I perform tissue culture in banana. Im using a protocol by Dr. Frederic Bakry with slightly modifications.
Enjoy!
Tissue cultures were initiated from shoot tips taken from banana suckers. For sterilization, banana suckers were reduced to explants of ˜10 cm in length and 2-4 cm in diameter; they were then immersed in bleach 1% and then 95% (v/v) ethanol, followed by flaming. Flaming
was repeated two times.
Burned tissues are removed and then discarded, in sterile conditions,
up to innermost intact tissues. Shoot apices are slice in two and place in a solid proliferating medium containing:
MS macro- and microelements, vitamins from Morel, 88 mM sucrose, 22.2 µM 6-benzylaminopurine (BAP), 651.5 µM adenine sulphate
and 7 g·L-1 agar.
The pH of the medium is adjusted to 5.8 before autoclaving. Cultures
are kept at 25 - 27 °C in darkness for 45 days.
Active multiplication occurred following
subdivision of bud clusters and subculture
of individual pieces on proliferating medium or rooting medium

[@smex1942]

Admired by this video.... good job...wishes from Tamil Nadu, India....

[@mashikc3no]

Tons of thanks for sharing your knowledge and expertise! 🙏🙏🙏🙏🙏🙏🙏

[@ferchuckygarcia]

Thank you so much 🥰

[@plantasaquaticas3483]

Qual hormonio pra divisao Celular usou. E em qual proporcao desse hormonio?

[@starGirl-dl1rx]

How fascinating

[@ferchuckygarcia]

thanks a lot

[@figman4890]

Hi Fernando sorry but I don’t see the link to Dr Bakrys protocol. Can u please post it. Many thanks from New Zealand

[@figman4890]

Also how long u u keep the explants in the dark for? Thanks

[@JazzMaquilan]

so we can put in the put the source of the corm? or wen can bag it? not necessary of these explant to be planted on field?

[@JesusBarrios-w4k]

Que contiene el envase donde colocas el meristemo y si está totalmente cellado

[@Redbull357]

Great video! Please make a video explaining and showing how to make the media. Thank you!

[@ferchuckygarcia]

Good idea i will do that.

[@benjaminnyirenda7336]

​@@ferchuckygarciadid you do it?

[@martinachvalova7691]

Hi, i buy a litle Banana in vitro. It is 3cm in a test tube. What Is The next setup pleas? Should i plant it in a Jiffy And cover? Or find jim on the tube. Thank you ❤

[@FloraWithAkash]

Thank you sir 🙏 , l like your all videos for helping in my practical 😊

[@ferchuckygarcia]

All the best

[@slstats7948]

Thank you❤️😇 From Sri Lanka🙏❤️

[@ferchuckygarcia]

my pleasure greetings from a Colombian researcher in the Netherlands

[@khinmarcho7225]

Thanks so much for sharing this informative video and explanation.

[@ferchuckygarcia]

You are so welcome!

[@dlxofficialentertainment]

I liked the video

[@figman4890]

Hi, really appreciate ur videos. I have tried this but got necrosis of the explants. Do u use citric acid or potassium citrate to reduce the possibility of necrosis?

[@kingsleynangantani1831]

We use citric acid

[@physics3632]

Gracias por el vídeo, es muy informativo. Por cierto, sabes si se podría usar un método parecido para intentar crear unas plantas de semillas que tendrían una probabilidad baja de germinar solas ?

[@ferchuckygarcia]

Claro que si, se llama embryo rescue (rescate de embriones) this has the potential to regenerate multiple plants from one single seed.

[@czarnyheniek]

Great material 👍

[@ferchuckygarcia]

thank you very much

[@valencia81750]

Thanks for the great video. You did not explain how to divide the explant after it grew in 45 days. Where do you cut it? Has anyone ever tested the final product banana to see if it contains traces of bleach?

[@ferchuckygarcia]

So nice of you. No traces are found after, no worries. Plants are divides using again a scalpel. And transferred directly to soil or new media if you want to keep multiplying them.

[@kathirineviolet8773]

Very informative.. 👍

[@ferchuckygarcia]

my pleasure, thanks a lot

[@sandipsanas8100]

It is helpful viode for us thank you so much😊

[@ferchuckygarcia]

thank yo uso much

[@Toskyval]

In the very last part you say "many plants are coming from this half meristem.... and from here you continue propagating as many plants as you want", which mean you still "duplicate" them by using the 'plant' hosted inside the little container or you must chop an adult banana plant again and again?

[@ferchuckygarcia]

Yes exactly, from the half you can continue propagating plants for many cycles. You don't need a new adult plant.

[@christophermasterton9346]

Thank you ever so much for sharing. However, I like to to know what substance is in the jar before placing the banana tissue inside.

[@ferchuckygarcia]

MS macro- and microelements, vitamins from Morel, 88 mM sucrose, 22.2 µM 6-benzylaminopurine (BAP), 651.5 µM adenine sulphate and 7 g·L-1 agar. The pH of the medium is adjusted to 5.8 before autoclaving. Cultures are kept at 25 - 27 °C in darkness for 45 days.

[@خيراتالارض-ص6د]

Goed bezig, bedankt Heb jij ervaring in het telen van dadels?🌴

[@PatilDipakD]

Very nice video. How plants come out from one tissue and how to separate those plants?

[@ferchuckygarcia]

Is easy using the scalpel. It has to be done under sterile conditions.

[@PatilDipakD]

I mean to say, how many plants come out of one tissue? Is it possible to setup a lab at home? Where can I get all these equipment and specially that gelly in which we plant tissue?

[@ferchuckygarcia]

from one plant you can get in an average of 1 to 10 little plants in the first round and then is exponential. It is possible but it requires a little investment. I'm planning to make a tutorial to make this at home for bananas and other plants

[@PatilDipakD]

Wonderful, I will wait for your video. Thank a lot 😀

[@r.guerreiro140]

@@ferchuckygarcia I'll be waiting too :)

[@zahedhossain6736]

We saw in the video that you collected all the tissue from a small mother plant. How can we make sure the mother plant is free from disease and pathogen?

[@ferchuckygarcia]

By looking at possible external symptoms of the disease. But in general, most diseases can be eliminated by tissue culture techniques. No viruses, although there is evidence that suggests that viruses can be gradually removed from tissue culture plats.

[@Tomy2705]

Good

[@user-iw8kd8vy2h]

Super thanks for vidioes 👌👌👌👌👌👌👌👌👌👌👌👌👌

[@ferchuckygarcia]

your are welcome

[@jaylar1485]

what is the name of jelly look like chemical did you put the last process?

[@ferchuckygarcia]

All the information and the recipe is in the information box

[@dr.iftikharahmad8274]

Breliant thank u sir

[@nancyma1332]

can this be done with other parts of the banana plant?

[@ferchuckygarcia]

Unfortunately, not. Only corms

[@askeruchu1495]

@@ferchuckygarcia ĺ

[@joy-dc]

It's a monocotyledon. Check on Wikipedia.

[@Duke_Scanlan]

Extremely interesting.

[@ferchuckygarcia]

Thank you very much.

[@nataliaarbelaez9857]

Thanks Dr. big banana!!!

[@myhanslombard]

Thank you :) Why do you put in the dark for the first 45 days?

[@ferchuckygarcia]

It is something i learned at Cirad in France but i later figured out that is not really necessary for all the accessions. so i put them directly to light now. it works well

[@AppEditorTutorial]

Nice

[@ferchuckygarcia]

Thanks

[@bobjob3632]

It looks cool!!

[@ferchuckygarcia]

Thanks a lot my friend

[@vineleak7676]

Great can you make a video on how to do tissue culture starting with fruit cells?

[@cexxy]

You cant do tissue culture from fruit cells

[@agvocateqc379]

What model of forceps to you use? Thank you!

[@ferchuckygarcia]

I dont know i will check and i will let you know

[@edwins2595]

The very informative discussion was ruined by the blasting music.

[@ferchuckygarcia]

sorry about it

[@widinugraheni4209]

thanks fercho..it helps me a lot for teaching my students 👍🏻😍

[@alianatasha8577]

Thank you for your sharing! ✨

[@ferchuckygarcia]

You are so welcome!

[@anelekhethwa637]

Thank you so much for sharing with us, Sir.

[@ferchuckygarcia]

my pleasure!

[@HikaruKn01]

Hello! I would like to know what measurement μM means and what it would be equivalent to in milligrams? please Thank you very much for your video!😊

[@miguelangelsimonfernandez5498]

I think he means molar weight. uM is micro-mol. one millionth of a mol. mM is mili-mol, one thousandth of a mol.

[@mohirjonturakulov1968]

Здравствуйте, рецепт среда можете сказат пожалюста.

[@maracachucho8701]

Hold on, I missed the part where you made so many shoots. Did they all come from the two last little pieces or did you use everything you had cut up?

[@ferchuckygarcia]

Depends on the initial material, but most of the time you get multiple shoots from one piece indeed.

[@pablocingciang3310]

Cant we grow callus from leaves?

[@ferchuckygarcia]

as far as i know. it is not possible. we have tried different parts of the plant but so far only the meristem work well (either form corm or flower)

[@GusTakdir]

Botles from plastic or glass?

[@ferchuckygarcia]

Does not matter as long as is sterile material

[@rubiohoyosysugrupoideal]

Que más ferchito te habla Hardy ome q gustazo saudarte

[@ninaflora5484]

How long the explant immersed in the ethanol?

[@ferchuckygarcia]

not more than 1 - 2 mins

[@silverianjannvs5315]

What are the results? GMO banana?

[@ferchuckygarcia]

Not at all. Just clones

[@KHH595]

What will happen if the environment is not sterile? Will it just not work?

[@ferchuckygarcia]

it won't. Because the media gets contaminated and everything (microorganisms) can grow in that media and make the plant rot.

[@alfredoanitjr.6480]

Im your new subscriber, thank you for sharing Sir, i have a plan to have own plant nursery in the Philippines if i can go home, i want to do tissue culture and have LAB

[@ferchuckygarcia]

Thanks and welcome

[@GauravGupta-tu9pk]

What had you used as proliferation media

[@ferchuckygarcia]

componets are described in the info box :D

[@imeldaontolan1010]

Want to know more about the solution

[@ferchuckygarcia]

Hello Imelda you can check the information box all the components

[@marvinmarba6133]

How to make media?

[@ferchuckygarcia]

Hello Marvin, all the components are in the information box of this video, if you want a tutorial for media preparation I can include it for a future video.

[@evillajosie]

What solution used?

[@ferchuckygarcia]

you can find all the information in the info box of this video :D

[@jonathanelcamel9125]

What are those chemical you use ser?

[@ferchuckygarcia]

Check description

[@tranqi]

but you use the dirty knife after flaming? shouldnt you clean the knife in between?

[@ferchuckygarcia]

The cleaner the better 👍

[@ธนศักดิ์มีเพียร-ฏ2ฑ]

What bl ??

[@whatthefunction9140]

Did you just destroy one plant to grow one plant...?

[@ferchuckygarcia]

Technically yes, but: 1st. Banana plants die after producing bunch. 2. with micropropagation, you obtain unlimited number of plants from a single one (cloning)

[@apoloniobacasmas4667]

Hello sir may I ask the ingredients of tissue culture please

[@ferchuckygarcia]

Check description

[@autisticguitar666]

I really wouldn't recommend using plastic gloves, Ethanol and high heat in the same place. Use your bare hands, wash them well before you start and desinfect them. That's enough. Having plastic fused to your skin is no fun

[@erdbeerzwiebel3404]

People who work with agar do it all the time ;)

[@Wavy_Gravy]

You use 70% iso, so it retards the evaporation rate (to aide in the iso's ability to destroy or permeate, the cell walls of microbes) . The vapors ignite all the time. The water, aka 30%, coats your hand if you ignite vapors. If you light the lighter with iso wet gloves, you're fine. Iso's danger is in the vapor increasing fluid pressure in its vessel, thus causing a physics flamethrower. We did jacka** stunts as kids with the rationalization that it required science to not hurt ourselves.

[@Wavy_Gravy]

Also gloves retard the evaporation of iso's water content. If you used iso on bare hands, it won't clean them. Also, yeast live under skin cells that haven't flaked off. You'd get yogurt culture every time.

[@Wavy_Gravy]

So everyone is im agreement. The use of retarded and retard are in the sense of timing. No hate, now or ever.

[@autisticguitar666]

@@Wavy_Gravy We work without gloves and it works very well. You just need to know how to handle your material properely

[@ahamedwazeer4511]

👍

[@jamesbarton3165]

Piff method is quicker

[@kurreembukusnathalie3561]

Why bleach

[@ferchuckygarcia]

Works better

[@miguelangelsimonfernandez5498]

it is cheap, easy to find, well tried and effective

[@aaminahwaseem9728]

the sound is not good I am quite disappointed

[@rulisapocilps3317]

Hawa

[@dustyflair]

what if you dont have a lab?

[@ferchuckygarcia]

Then you can use the macro propagation method. Check my other tutorial :)

[@MickesVideo]

You sell me thoose plants becouse it was like a scamvideo😂

[@mkanwar3208]

The music volumne is annoying😏

[@Abdanthedestroyer]

Dooe