Seed Mothers vs. Clone Mothers

Blused said:
Hi Chana how are you, are you looking for testing equipment or lab equipment for micropropagation, if micropropagation, a Laminar flow hood would be a great benefit in cutting your explants, a autoclave would be very useful but you can get away with a good pressure cooker, if you so preferred a glove box could be used instead of a laminar flow hood for a small portable sterile area. A assortment of beakers would be nice to mix your hormones and prepare your agar for sterilization and a collection of baby bottles for your explants is required, a handheld microscope with 200 - 400 x will get you to a cellular level, there are a few nice sites for new and refurbished lab equipment with Lab X scientific marketplace being a good one, hope this helps.
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B!!!!!!!!!!!!!!!!!!!

How are you my friend?

You are the man I was looking for... I am not sure you had a chance to read the thread a bit but some of the other cultivators were discussing viruses in Cannabis and if one can isolate a virus and cure a particular genotype by taking new cuttings of an infected plant or taking molecular culture of same and being able to by-pass or skip the virus by utilizing a fresher more vigorous copy.

Can you comment with regard to some of your work you have done on the micropropagation front. Also what viruses if any, you have encountered and if in fact you were able to isolate a plant, make a copy of it and not have that virus carry over to the new copy.

Blu, I was not referring to micro propagation equipment but the gas chromatograph and/or a spectral machine.

I am interested in using the equipment with regard to my herbarium project but I noticed that the equipment would also be able to help with regard to identifying viruses in plants and re-testing with the equipment once a treatment has been tried.

Thanks for stopping in my friend and it was nice to meet you in person the other day.

Take care,

Chana
 
Chana Masala said:
I was just wondering what fellow cultivators prefer when keeping their mom or dad's around.

Do you think it is better to keep a the mother/father from original seed around, or is it better to keep the clone of the mother/father instead.

Chana
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Clones are great IF Im sure I want to do a "repeat" grow--doing the same strain twice. But if I want to skip one or more grows...a supply of seeds is handy. My ultimate fantacy is to create BONSAI Moms of every strain I really love. The small size makes themm easy to keep--they dont take up much room!

The one thing I hate tho...is UNfeminized seeds! OH THE HEARTBREAK of having a bunch of males show up! aaagghh! This past summer I had six WW's on the deck rail about two feet tall, still vegging. The dog jumped against the rail to bark at a squirrel...the plants fell two stories, got broken and mangled,...the two I was able to save, guess what? MALES!

Dixie
 
hi all
hi Dixie

The one thing I hate tho...is UNfeminized seeds! OH THE HEARTBREAK of having a bunch of males show up! aaagghh! This past summer I had six WW's on the deck rail about two feet tall, still vegging. The dog jumped against the rail to bark at a squirrel...the plants fell two stories, got broken and mangled,...the two I was able to save, guess what? MALES!
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Thats why its better to sex the plants as fast as possible..

there are several methods you can do this , one is to start the plants under 12/12 from seed , sex em during their first few weeks , toss males and put the sexed females under veg light regime (ie 18/6). Another method is to start plants normaly with veg but shade/cover one branch of each plant. Soon the branch will show sex.
Another method is to take the top cut of the small/young vegging plants (at ~week 3) and flower the cut to find out the sex of the plant in veg.

If you dont want growing males then you shouldnt keep plants of unknown sex for a long time as you are waisting resources

cheers
l33t
 
L33t said:
hi all
hi Dixie

Thats why its better to sex the plants as fast as possible..

there are several methods you can do this , one is to start the plants under 12/12 from seed , sex em during their first few weeks , toss males and put the sexed females under veg light regime (ie 18/6). Another method is to start plants normaly with veg but shade/cover one branch of each plant. Soon the branch will show sex.
Another method is to take the top cut of the small/young vegging plants (at ~week 3) and flower the cut to find out the sex of the plant in veg.

If you dont want growing males then you shouldnt keep plants of unknown sex for a long time as you are waisting resources

cheers
l33t
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Hi Dixe and L...

D, fems obviously have there place, keeping the mothers alive in cut form is a proven method for preserving your genetics but working and creating the F2 is important as well. Maybe you will consider it given more resources... sorry about the squirrels... I had a battle with black squirrels in my area. Ended up cost me over 5000 in damage when it was all said and done and also her off-spring took a couple of my prized F1 Free Leonard beans along the way...

For me it has become very rewarding working a line with both male/female specimens but I am humble by the process and fortunate to find the resources to continue the journey.
Cheers,

Chana
 
cabron said:
ok i'm pleased to see this thread back on track...
My apologies for taking out the trash...
It was needed...
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wow cabron you are a real piece of work, you make fun of someones physical appearence and not even worry that they might have a medical condition causing a deformity, that doesnt make you look smart, that makes you a low life scum

the trash that needs taken out is you !

P.s. A quick google search as someone mentioned earlier has indeed came up with answers you post almost word for word, only problem is they were originally posted by others
 
brothers and sisters...

This is not TY or ICMAG...

Please, please and please...

Can we cool...

This thread sucks now...

Chana
 
Hey Chana you are a very kind person and it is great to know you man,
truly as you know i been gathering alot of resources and theory on the subject,
as i take it there are micropropagation of leaf or stem, with stem explants being more durable and desired because of sterilzation and ease of propagation, now there is micropropagation on the cellular level also requiring use of electron microscopes and Dna manipulation and even genetic splicing. Alot of this revolves around whether we believe the Dna code for each pheno type within a genotype is there on a cellular level, if it is then why would micropropagation of stem tissue change the desired traits, when we clone traditionaly we are making stem tissue grow roots which is in a medium of choice, when we clone with micropropagation we are making stem tissue grow either shoots or roots in a medium of choice being Agar, most all of the plant industry uses micropropagation very effectively and they also have many traits to perserve with there explants, colour, fragrence, growth structure, vigour and even sometimes taste, so i believe it will preserve the traits that are in the Dna code of the given explant, but with viruses that i cannot say, if the virus is in the cells we will most likely transfer it along, but in he book Plants in test tubes they support research that explants taken from the apical meristem will be of such new growth rendering it a disease free, i will pose the question to someone much more experienced then i in tissue culture, and post back, just to throw it in there i think it is best to replace your mothers every year with a new cut, take your chosen strain to breed with and do open pollination with all the males and females to ensure as much genetic preservation and keep a stem based tissue culture for backup, very soon we shall test out some of these posing questions in the very near future, the dream shall be 100 or so mothers and dozens of male specimens on a 6x8 foot shelf system under simple low level light, hope me taking this thread off track gets it back on track

Love yall
 
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This is great stuff,and cutting edge...just the way I like it...

Breeder Steve got really excited with this tech but never did get a chance to realize it...



Blused said:
Hey Chana you are a very kind person and it is great to know you man,
truly as you know i been gathering alot of resources and theory on the subject,
as i take it there are micropropagation of leaf or stem, with stem explants being more durable and desired because of sterilzation and ease of propagation, now there is micropropagation on the cellular level also requiring use of electron microscopes and Dna manipulation and even genetic splicing. Alot of this revolves around whether we believe the Dna code for each pheno type within a genotype is there on a cellular level, if it is then why would micropropagation of stem tissue change the desired traits, when we clone traditionaly we are making stem tissue grow roots which is in a medium of choice, when we clone with micropropagation we are making stem tissue grow either shoots or roots in a medium of choice being Agar, most all of the plant industry uses micropropagation very effectively and they also have many traits to perserve with there explants, colour, fragrence, growth structure, vigour and even sometimes taste, so i believe it will preserve the traits that are in the Dna code of the given explant, but with viruses that i cannot say, if the virus is in the cells we will most likely transfer it along, but in he book Plants in test tubes they support research that explants taken from the apical meristem will be of such new growth rendering it a disease free, i will pose the question to someone much more experienced then i in tissue culture, and post back, just to throw it in there i think it is best to replace your mothers every year with a new cut, take your chosen strain to breed with and do open pollination with all the males and females to ensure as much genetic preservation and keep a stem based tissue culture for backup, very soon we shall test out some of these posing questions in the very near future, the dream shall be 100 or so mothers and dozens of male specimens on a 6x8 foot shelf system under simple low level light, hope me taking this thread off track gets it back on track

Love yall
Click to expand...
 
hi all

from the papers I 'm reading on plant viruses it seems that not all of the differnt sanitation methods (in vivo and in vitro Heat treatement(thermotherapy) / Meristem tip culture / Somatic embryogenesis / Chemotherapy / Cryotherapy / electrotherapy) are equally effective when it comes to viruses:
http://www.sipav.org/main/jpp/volumes/0306/030608.pdf

These methods are used alone or in combination.

Info from a paper on hops and virus elimination:
"A maximum of 4 sub-clones were generated from meristems for each hop. None of
the 182 plants regenerated from heat-treated meristems tested positive for any of the 6 viruses. A single sub-clone was selected to replace each original virus-infected clone in the NCGR core collection."


from:http://www.ars.usda.gov/SP2UserFiles/person/4630/Contaminants/2.%20Acta%20Hort.%20668%20143-148%20%282005%29%20Humulus%20viruses.pdf

Virus-free hop plants can be produced from material infected with hop mosaic carlavirus (HMV), hop latent carlavirus (HLV) and apple mosaic ilarvirus by meristem culture (Adams, 1975; Kremheller et al., 1989). Thermotherapy is necessary in addition to meristem culture for the elimination of some strains of ApMV. Freedom from ApMV can be achieved by growing the infected plants at 35-38°C for 4-10 days followed by excising shoot tips about 5 mm long. These can be rooted in potting compost without resorting to sterile culture procedures. It may be possible to obtain viroid-free plants from HLVd-infected material by cold treatment followed by meristem culture (Adams et al., 1996).
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From:http://webcache.googleusercontent.c...on&cd=12&hl=el&ct=clnk&gl=gr&client=firefox-a
 
Blused -"Plants in test tubes they support research that explants taken from the apical meristem will be of such new growth rendering it a disease free"

I suspect and hope that the above is fact. All of these guys stating the disease is always there are getting me worried though.

Here is how I THINK I cured my plants from TMV.

I treated my infected plant(s) first with sunlight. A week later they were treated with Harpin proteins. The next day i took cuts from the meristems. They rooted slowly, but most rooted. This was also done outside. The healthiest appearing cuts were transplanted and placed in direct sunlight and treated with Harpin at 3 week intervals. This process was repeated three times from May to September when the sun was intense and growth was rapid. Eventually cuts rooted as fast as I thought they should and moms appeared to have NO viral load.

The most effective way to deal with invaders (pest or viral) is to understand them. Once you understand how they work you can interrupt/kill their defense mechanisms and eliminate(?) them with a little hard work.

Hey Blused do you ever use Harpins or anything that induces SystemicAcquiredResistance (SAR response)? Do you believe these can help battling viruses by stimulating the immune response of plants?

Legally speaking not one of those little test tubes is considered a plant until it has roots. People who live in medical cannabis states with plant limits may be able to use microprop to maintain a wide selection of genetics in their veg rooms without concern of police interference.

Also because of the preperation Blused mentioned. None of the Agar tubes contain plants that have bugs. So if you have been battling the borg tissue culture might also give growers another way to eliminate pests.

Love Learning About New Cultures,

KB 8)_~
 
Kyle Bro you got some very interesting finds with immune response in plants, i have not come across this and i am by far no expert, immune response treatment has been put to good use in human treatment. Leet those are some great studies thank you for that information. I can't say for myself that apical meristem micropropagation will defeat TMV but will say that there is research to show that it results in disease free sterile cuttings that can be placed in a dormant state kept in a growth or divided in to many explants over a short period of time and kept in a low light intensity in very compact storage
area, wins me over, back to my pressure cooker i go, where is that agar pack, anyone got some bap, lol
 
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Peace friends,

L33T, in the grapevine study, to quickly quote "However, regeneration through somatic embryogenesis always gave rise to plants free from RSPaV. Among the tested techniques, somatic embryogenesis seems to
be the most promising procedure for GRSPaV elimination, despite the technical problems related to the use of this technique and the theoretical possibility of somaclonal variation."

The scientific term of the day for Mr. Masala is "Somalonal variation".

Curing virus with stem culture seems to be possible, but it would appear unlike other fruit and floral cultivators, it would be easiest to just order fresh stock of healthy plants.... but for the normal farmer, unless they are able to address the actual issue of cause (environmental, hereditary or just man) and like Kyle mentioned identifying through species screening, will they be able to deal with there crop issues. But the new stock is a quick fix and I am sure we can all agree that a quick fix is never the answer for a long term or serious cultivator/breeder.

We get cuts and seed from around the world but I have never screened my new specimen, the most I have done with the case of a cut is isolate for a few weeks and spray with chemical... seed is worst, I trust in the breeder and never considered virus until it was mentioned in this thread.

Maybe a little to much, but it would be amazing to screen your genetics before dedicating years to developing a particular cultivar.

Kyle, I got a little scared to with the talk of tobacco virus as I smoke cigarettes and will try to avoid smoking in gardens but, I honestly dont believe I have tobacco virus. Knock on wood my cultivars are actually doing well. Maybe a little heat stress from time to time, or root bound, but mostly grower related issues... thank god... I would really be difficult to start from scratch again. But by sharing my cultivars and making f2's of proven stock, if disaster did strike, it would not be that difficult to be back to it in no time. Provided I dont share a virus that can not be treated and cured lol.

Fuck, scary world... computer virus, plant virus, people virus. I know it is a fine balance though and too much or not enough is an underlying theme in life.

Blu, this is home brother dont go to far you have some insight in our world that will help us cross over to academia... back to the word of the day "Somalonal variation" and Exodus Cheese. Please consider doing a thread on how it goes... this would be a great explant study and indeed if the stem cutting will hold the phrenology of the Skunk#1 phenomenon. Tissue plant culture has pros and cons. If a pheno varies from the original donor, then I can see why many cultivators are turned of building their tissue library but on the other side, as Kyle mentioned for us legal growers, having a vault that can quickly be grown out is wonderful in the world of preservation and plant numbers.

I still think seed is the best way to preserve genetics but your work Blu is very interesting.

As to the thought of putting the practice of viral infection, and isolating of an outbreak, I am affraid that this is out of my scope and I do not have the ability to take the samples and later test. But hopefully you can continue on this path and I can look to you for research. Thanks for sharing.

But it would seem that a few of us are going to get there, that is for sure, keep up the great work everyone, I think that time is finally on our side...

Chana
 
Well from looking into things abit and asking some questions. R.F White found that when using meristems of tobacco for explants that the tobacco mosaic virus didn't transfer to new cultures. He actually wanted the virus to transfer because he was studying it. Gee, he wanted contamination and didn't get it. That's plant tissue culture for you. It always has a way of throwing a curve ball at ya. Nice little useful discovery he made. I like using meristem for difficult to disinfect plants growing outside. It is not hard to get "clean" explants from them. now as far as variation in tissue culture it looks as if leaf culture and callus culture is not desireable and may lead to issues, now from a friend "depends on a few things; the type of explant used, the type of plant growth regulars used and the amount of time spent in tissue culture. If shoot tip, apical buds or axillary buds are used then the plant is more likely to keep its traits then using leaves for explants or developing callus for multiplication. When it is important to keep the traits of a plant shoot tip, apical buds or axillary buds are is used. Even with shoot tips, apical buds or axillary buds traits such as variegation may not be passed on. That depends on what layer or layers in the meristem the variegation is present. It also depends on the plant. Some plants are more susceptible to mutations than others. Generally though, if one wants the traits to remain the same they would use shoot tips, apical buds or axillary buds as explants. So from a expert looks like there easily could be some variation in the Exodus cheese explant, i quess we need to blaze the new frontier and see how various genotypes respond to apical meristem tissue culture.
 
Kyle
I hate to piss on your parade but a lil info..
I used Harpin Proteins to stimulate an elevated
immune reaction in my plants as well..

This was a product called messenger and is now defunct..

I still have a nice quantity in my lab fridgerator..

This will not work as the plant is already infected,,and the elevated
immune system while aiding the plant to appear healthy does not remove
the virus,,only masking it.


Same theory behind immunizations ...you cannot expect to get the shot
after being infected and have it work..


The symptoms will occur again after the stimulant has worn off...
Even placing them outdoors reduces stress and sunshine helps to revitalize
the plant and appear disease free..
Try taking a few cuts and bringing them Back indoors and see how they react.



This observation is of course with my experience and the virus I had..
It may not be applicable to your situation solely...
I don't expect to see the virus I encountered by others very often..

This was not your typical Mosaic virus as in the ChemD cut I had,,,,
that was a manageable situation..


Mine wasn't...



best of luck to you...
 
Hey Cabron,

Harpins last for three weeks after application.

My thoughts in regards to immunization was that the uninfected tips (which I was taking my new cuts from) would also be immunized. Wouldn't you protect the cuts?

I too have read about how things are impossible to treat(usually in HighTimes mag), like root aphids were at first. But guess what Cabron, you can drown root aphids with smart pots and water. You don't even need to buy ANY chemicals. Simple, and I figured it out with trial and error, instead of listening to people telling me to get rid o ALL my plants. Horrible advice, one of my plants is 22 years old, get rid of it? I'm glad I did not follow my co-workers advice. He was a know it all grower with three years experience growing and 20+ smoking. Heh.

Most of the people on this site are not into perpetuating ideas we have read. Most of us here share things WE have personally observed, you could probably be more helpful if you did the same thing.

KB 8)_~
 
kylebroflovski said:
Hey Cabron,

Harpins last for three weeks after application.

My thoughts in regards to immunization was that the uninfected tips (which I was taking my new cuts from) would also be immunized. Wouldn't you protect the cuts?

I too have read about how things are impossible to treat(usually in HighTimes mag), like root aphids were at first. But guess what Cabron, you can drown root aphids with smart pots and water. You don't even need to buy ANY chemicals. Simple, and I figured it out with trial and error, instead of listening to people telling me to get rid o ALL my plants. Horrible advice, one of my plants is 22 years old, get rid of it? I'm glad I did not follow my co-workers advice. He was a know it all grower with three years experience growing and 20+ smoking. Heh.

Most of the people on this site are not into perpetuating ideas we have read. Most of us here share things WE have personally observed, you could probably be more helpful if you did the same thing.

KB 8)_~
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i agree 110% , but no 1 ever listens to me , so i just chuckle as they all fuk up
 
Ooops. Maybe I lost my cool but "piss on my parade?" Pretty condescending IMO. Most forum members want to just share VALID info around here and maybe make a couple like minded friends. Yes sometimes we disagree (many times it is just a language issue though) but most of the time we try to contribute our experiences in a non critical way.

I'm not so sure that's why you are posting but I'm willing to treat you with respect, hopefully you can do the same.

More Of A Zookeeper Than Parade Leader,

KB 8)_~~
 
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