View Full Version : step by step Tissue Culture Pineapple
novisyatria
01-24-2010, 08:05 AM
I did special Tc today for you org. step bus tep tissue culture pineapple. hope can help you interested with tc pineapple
step 1. prepare the crown of pineapple. sometime crown has few baby.
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2736step1.jpg
step 2. throw out the leaf until the last/youngest leaf
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2738step2.jpg
step 3. soak in bleach 10 % for 10 minute
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2740step3.jpg
step 4. cut vertically become 4 if the stem big enough and become 2 if little or do not cut if too little.
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2739step4.jpg
step 5. put in the jars with media N5B2 (MS with 5 mg NAA and 2 mg BAP) to get alot shoot. but if want they growth (elongation) moved in media MS with 2 mg BAP and 1 mg IBA
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2742step5.jpg
step 6. after 6-8 weeks explant will growth become alot shoot. you can do subculture with same media N5B2
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2743step6.jpg
step 7. in 2 next month shoot will growth. very beautiful. isnt it?
http://i909.photobucket.com/albums/ac298/novisyatria/IMGP2746step7.jpg
regards
xavierdlc61887
01-24-2010, 08:07 AM
wooooooo very interesting novi!!!! i will have to do this some time!
Very cool! Thanks for posting this. I may have to try this now someday.
Caloosamusa
01-24-2010, 09:03 AM
Thank you for posting this Novisyatria!! That is good information. :2239:
CValentine
01-24-2010, 02:59 PM
WONDERFUL NOVI!!!
Thank you!!! :D ~Cheryl
zoloff
01-24-2010, 06:29 PM
Yes very cool, I would love to try it Thanks Novi
Blake09
01-24-2010, 07:07 PM
awsome, thanks for sharing
What do you do after step 7... Do you cut the growth off at its base and put it in rooting hormone, then soil, Or do you cut off the growth with a little bit of the base left and put it in rooting hormone...
Thanks for helping me :D
novisyatria
01-25-2010, 07:24 AM
blake, i can do both. move the shoots in to rooting media and than in soil. and also can cut the shoot and put in same media for shoot multification. so i can still have shoots stock.
kamayoi
01-30-2010, 10:55 PM
Great!!
Thanx 4 sharing pak novi :)
knowledge is power!!
Frankco
02-07-2010, 09:52 AM
Thanks Novi! This is great. I'm going to try this today!
NANAMAN
02-07-2010, 04:05 PM
Thank You Novi! I have several varieties that I will be mass propagating very soon. Your pictures have helped me understand the process much better than I did after reading about it. I will also start my tc's using the same media formulations that you prescribe here. Thanks again!
Frankco
02-07-2010, 04:59 PM
Cool!!! It's just like TC'ing Bananas!
I'm working on my pineapple right now. I'll post some picts in a few minutes.
Thanks again Novi!!! This is awesome!
novisyatria
02-07-2010, 05:40 PM
Thank You Novi! I have several varieties that I will be mass propagating very soon. Your pictures have helped me understand the process much better than I did after reading about it. I will also start my tc's using the same media formulations that you prescribe here. Thanks again!
Cool!!! It's just like TC'ing Bananas!
I'm working on my pineapple right now. I'll post some picts in a few minutes.
Thanks again Novi!!! This is awesome!
your welcome and post your progrees soon ok
Frankco
02-07-2010, 06:08 PM
Okay, I'm done cutting the pineapple up. And to think there is a ham in the freezer :nanadrink:
I had this growing in a pot outside. I cut the top off of a pineapple and stuck it ontop of of some dirt in the pot. The frost got to it. I thought it was a goner. The outer leaves died but the inner leave survived. Here is the sad looking pineapple. At least it gave me 10 explants! :woohoonaner:
Sad Pineapple
http://www.root66nursery.com/pineapple/pineapple_in_pot_2.jpg
Cleaning of the dirt
http://www.root66nursery.com/pineapple/pineapple_clean-roots.jpg
There's something alive under all of that:goteam:
http://www.root66nursery.com/pineapple/pineapple-cutting_0.jpg
Hacking away at it.
http://www.root66nursery.com/pineapple/pineapple-beginning-to-trimming_0.jpg
It tastes like chicken:ha:
http://www.root66nursery.com/pineapple/pineapple-trimming.jpg
Into the disinfecting solution
http://www.root66nursery.com/pineapple/pineapple_in_paa.jpg
10 Explants:woohoonaner:
http://www.root66nursery.com/pineapple/pineapple_explants.jpg
:drum:
Thanks Novi!!!!!
novisyatria
02-07-2010, 07:56 PM
cool pic Frank. good luck. and post after the explantgrowth ok
Frankco
02-07-2010, 08:07 PM
Hi Novi,
Will do. That was fun! Now to see if they grow :drum:
Like you said, it is a thing of beauty to see plants grow from little pieces.
Now I'm going try caladiums. Your pictures of them look fantastic. :goteam:
Greenie
02-07-2010, 11:01 PM
This is a great thread!.....keep up the good work guys:bananas_b
Sorry everybody. I´m still out at the countryside: The Great Savannah!
I´ll post some pics next week. Great job Novi! Now I´m in a rush to get home ( that´s where I have my products ) Thanks a lot.
pineapplefarmerwannabe
07-27-2010, 01:27 PM
I wanna do tissue culture of pineapples and was wondering were to buy the little jars and from what I could find the media that novi refers to is different kind of hormones.
Please help me in successful tissue culture
thanks Brian
Frankco
07-27-2010, 03:19 PM
Hi Brian,
Canning jars work well.
As for the media and plant growth regulators, you can get them from Carol at Home Tissue Culture* Group (http://www.hometissueculture.org)
I think you will find her shipping charges much lower than the TC supply houses. You will find her email address on the site if you have any questions.
Frank
pineapplefarmerwannabe
07-27-2010, 08:15 PM
Hi Brian,
Canning jars work well.
As for the media and plant growth regulators, you can get them from Carol at Home Tissue Culture Group (http://www.hometissueculture.org)
I think you will find her shipping charges much lower than the TC supply houses. You will find her email address on the site if you have any questions.
Frank
Have you successfully used any of these products for pineapple tissue culture?If not which would you or anyone that has used any of there products
suggest?Other than the hormones would you or anyone recommend anything else?With the jars what's the best method of keeping everything sterile from start to finish?
Thanks for EVERYONE'S help to a newbie,and yes I've done a little research first.
jmoore
07-28-2010, 02:21 PM
You can buy a lot of the growth regulators from this guy Benzylaminopurine (http://www.super-grow.biz/BenzylAminopurine.jsp) they come as powders so you need a bit of know how as far as chemistry goes and getting them to dissolve is a bit of a pain in the backside, but it's by far the cheapest way of doing things.
The agar is a vegetarian alternative to gelatine and can be purchased from health food shops or on line. As for the containers, yes canning jars or Kilner jars are a good thing.
john_ny
07-28-2010, 02:34 PM
I have used baby food jars.
pineapplefarmerwannabe
07-28-2010, 08:18 PM
do i need anything other than the hormones,I dont understand agar either?
Frankco
07-28-2010, 08:42 PM
Hi Pineapplefarmer,
I'm following Novi's protocol using MS, Murashige & Skoog Basal Medium, that's the nutrients, along with 5 mg/L NAA and 2 mg/L BAP.
NAA is 1-Naphthaleneacetic acid. BAP is 6-Benzylaminopurine. These are the two plant growth regulators you need to add to the media. You can get this already in a solution so adding them to the media isn't very difficult.
Agar, like jmoor said, can be found in health food stores. I purchased 56 grams from Amazon for less than $5. You only need to add 7.5 grams of powered agar per liter of media so it will make a fair amount. It's pretty difficult to go through all of the steps to make the media in a post. These are getting old but I did put some videos on YouTube about my experience getting started with tissue culture. Maybe they would help.
YouTube - fbt2007's Channel (http://www.youtube.com/user/fbt2007)
I use canning jars for bananas and other large plants. They work nice because of the extra headroom. If you use them, drill or puch a small 1/8th inch hole in the lid and cover it with a round band-aid. This allows for air exchange. Cover the lid with a wrap of aluminum foil to help keep contamination from settling on the band-aid and getting in the jar.
http://www.root66nursery.com/bananas/Bananas-01_s.jpg
http://www.root66nursery.com/bananas/Bananas_03_s.jpg
http://www.root66nursery.com/bananas/band-aids 2_s.jpg
http://www.root66nursery.com/bananas/band-aids_s.jpg
pineapplefarmerwannabe
07-28-2010, 09:43 PM
Thanks so much
Hi Pineapplefarmer,
I'm following Novi's protocol using MS, Murashige & Skoog Basal Medium, that's the nutrients, along with 5 mg/L NAA and 2 mg/L BAP.
NAA is 1-Naphthaleneacetic acid. BAP is 6-Benzylaminopurine. These are the two plant growth regulators you need to add to the media. You can get this already in a solution so adding them to the media isn't very difficult.
Agar, like jmoor said, can be found in health food stores. I purchased 56 grams from Amazon for less than $5. You only need to add 7.5 grams of powered agar per liter of media so it will make a fair amount. It's pretty difficult to go through all of the steps to make the media in a post. These are getting old but I did put some videos on YouTube about my experience getting started with tissue culture. Maybe they would help.
YouTube - fbt2007's Channel (http://www.youtube.com/user/fbt2007)
I use canning jars for bananas and other large plants. They work nice because of the extra headroom. If you use them, drill or puch a small 1/8th inch hole in the lid and cover it with a round band-aid. This allows for air exchange. Cover the lid with a wrap of aluminum foil to help keep contamination from settling on the band-aid and getting in the jar.
http://www.root66nursery.com/bananas/Bananas-01_s.jpg
http://www.root66nursery.com/bananas/Bananas_03_s.jpg
http://www.root66nursery.com/bananas/band-aids 2_s.jpg
http://www.root66nursery.com/bananas/band-aids_s.jpg
novisyatria
07-29-2010, 10:19 PM
Frank
can you upload new pic of your pineapples Tc? btw i have 8 type now in jars
pineapplefarmerwannabe
07-29-2010, 10:36 PM
Frank
can you upload new pic of your pineapples Tc? btw i have 8 type now in jars
you have 8 different types of pineapples?
I want one of each type
pineapplefarmerwannabe
07-31-2010, 08:34 PM
Does it matter if you the concentrated media or one with or without nutrients?
theres like a whole page of them...
link
MS-Based Salts & Media | PhytoTechnology Laboratories (http://www.phytotechlab.com/searchresult.aspx?categoryid=52)
Frankco
09-11-2010, 09:06 AM
Hi Novi!!!
I had a hard time finding a nice "green" pineapple. I bough a couple that were too ripe and didn't realize it until I tried to trim them.
I really want to say THANK YOU!!! :bananas_b for your help with this. My pineapple explant has started to put on new leaves. I probably should have sliced it in half or quartered it like you did. Next time. For now I'll just enjoy watching this one develop.
http://www.root66nursery.com/pineappleTC/pineapple.jpg
novisyatria
03-14-2011, 10:49 PM
Franck: congrats. please upload new pic
pineapplefarmerwannabe: i want sell to you a pineapple plantlet but hard to send it to overseas
An experienced pineapple grower that lives nearby told me that rather than planting the 'crown' that you cut off the pineapple, get one of the plantlets that develop between the upper leaves just below the fruit. Once those are planted, they develop into a mature plant much more quickly than those grown from the crown.
Frankco
03-15-2011, 07:53 AM
Hi Novi,
Here's some picts of my baby pineapples. When I divided the original one up I put a couple of young leaves in jars. Those started to develop callus and multiply too. I read it was done before but was happy to see it.
These are still in the multiplying stage. After I get enough I'll put them in the shoot enlongation stage.
http://www.root66nursery.com/pineappleTC/pineapple_08.jpg
http://www.root66nursery.com/pineappleTC/pineapple_05.jpg
http://www.root66nursery.com/pineappleTC/pineapple_06.jpg
http://www.root66nursery.com/pineappleTC/pineapple_07.jpg
http://www.root66nursery.com/pineappleTC/pineapple_04.jpg
Thanks Novi!!!
novisyatria
05-25-2011, 12:10 AM
you did it Frank. congrats.
Frankco
05-25-2011, 08:28 AM
I would have never figured out how much NAA to use without your help.
Thanks so much for your help. :nanadrink:
Andrico20
01-05-2012, 04:48 PM
Bravooo.. and thanks for your pics... i’ll try to TC pineapple first:0517:
tmc2009
01-17-2012, 10:42 PM
I watched a youtube video of a pineapple grown from seeds. What type of pineapple do you get from a seed. Is it the same as the mother plant?
novisyatria
01-17-2012, 10:57 PM
if from seeds could be different from its mother plant
Werner Stiegler
08-01-2012, 08:46 PM
Trying pineapple tissue culture. The bottom of the explant turns brown, and top green is this normal? The samples are only 2 weeks old.
<img src= "http://img827.imageshack.us/img827/3341/20120801132206.jpg">
miss adillina
03-01-2013, 02:44 AM
which one is it is the meristem. im doing tissue culture right now and im confuse which part of the pineapple to be cultured in the media.
alahamm
03-31-2016, 11:01 AM
LOOKS easy, HOPE so! THANKS:
Juicy Bananas
03-31-2016, 04:44 PM
Aloha, this was a great read. Starting with something simple like pineapple is a great way to learn and encourage others to start their own TC practice.
My wife has experience in Mycology, but we have never had the chance to try other flora. I would like to set up a station but we are not yet at that point.
Seeds. We bought a white pineapple at our market. One segment contained several seeds. These next few days I was going to make time to plant them. Curious question. I understand it may take 5 to 8 years to fruit. Also, assuming the other around it are seedless... Could they be from stress? Or maybe an old variety? Or just a case of the plant trying to live on?
Anyway... everyone should TC. It would make the world a better place. My goal is to some day TC my Fe'i... so I can see those little plants grow.
Mahalo :nanadrink:
fortuemuf
04-10-2017, 07:13 AM
Hey Novy
Do you use any preservative in your media. l am having bacteria which l cant get rid off each time l initiate. Have you ever used PPM or give me a tip of how to get rid of bacteria
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