Who is our resident Tissue culture master?

[englishrick]

No,,,,im attempting to argue that we dont really need deep tc tech as home growers,,I think generally , home growers can rely on initiation and micro cloning for removal of all nasties provided initiation specifically is performed properly!!,,emphasis on the initiation process

micro cloning is literally just surface cleaning clones,,,initiation is the process of witch you introduce genetics into tissue culture and attempt to remove nasties before you go full apical meristem, initiation includes surface cleaning but i felt the need to state the difference between the 2,,,initiation happens first inside the grow room before you go micro


Pm is not systemic,,,it just puts its roots into cells

The best way to remove pm is to Bath clones in h2o2 before you root them,,,when you cut from a plant go directly into a 2ml per l ho2o2 bucket and leaves them spinning for an hour,,,in tc we use a 10 sec iso wash but i really like the h2o2 bucket for non tc,,

If you can use uvc too on cuts, ,like the uvc light in my tc bag

Dunk clones once a week in Bath of 1ml h2o2,,,shake well

 

[Mudballs2.0]

The best way to remove pm is to Bath clones in h2o2 before you root them,,,when you cut from a plant go directly into a 2ml per l ho2o2 bucket and leaves them spinning for an hour,,,in tc we use a 10 sec iso wash but i really like the h2o2 bucket for non tc,,

so a little machine you can clamp the cutting in and then lower it into a solution bath and it starts spinning at a low rpm?
Cant we just leave them submerged? Like a redneck boy just jam a handfull in the bucket im carrying with me in field?

 

[englishrick]

The proper tc kit is a magnetic hotplate ,,it spins the liquid while you bath the explants,, in tc we use isopropyl for 10 secs,,,depending on the protocol for initiation you can add all sorts of measures and steps,,, some people go from iso to water and then to h202,,all depends on what you want to implement

Outside of tc in a semi mass production environments ,I like to use a h2o2 bucket and let them sit,,I dunk rooted clones in a slightly weaker bath when they come out the props too,,,don't be afraid to fully dunk and shake

If you want to get rid of pm and thats the only nastie you need to scrub,,just take clones and dunk them in h2o2,,dump the mother's,,spray the whole room down with h2o2,,try to ozone it, try to uvc it,,,keep dunking clones and then reintroduce at high temps dry humidity,, make sure you run the room empty and turn the heat up to mad levels,, literally burn the room and dry it out, do this repeatedly and you won't have an issue,,don't be afraid to flip mother's in weeks,,there is an advantage to taking clones from clones quickly,,,get rid of old plants ASAP and dunk all new ones

 

[Mudballs2.0]

I read somewhere the exposure to H2O2 efficiency is determined by content percent...like 15mins minimum for the H2O2 to burst the cell walls of all the nasties...but i forget the mix strengths, happy you gave one.
I heard 70% iso is better than 90% iso at ipm cuz the added water in the 70% facilitates reaction more than the 90%.
Idk where to go with this now...dudes crazy smart, pick his brain while he's here

 

[englishrick]

I like h2o2,,,I use it all the time on cuts,,,it makes them root quicker,,

 

[englishrick]

I read somewhere the exposure to H2O2 efficiency is determined by content percent...like 15mins minimum for the H2O2 to burst the cell walls of all the nasties...but i forget the mix strengths, happy you gave one.
I heard 70% iso is better than 90% iso at ipm cuz the added water in the 70% facilitates reaction more than the 90%.
Idk where to go with this now...dudes crazy smart, pick his brain while he's here

I was talking about groth technologies h2o2,,,il need to check the 10%,,,they changed the strength a while back

 

[Mudballs2.0]

Mattbho et.al.
What is the function of haustorium?

The haustorium penetrates the tissues of a host and absorbs nutrients and water.

 

[midwestkid]

Quietly listening from the back

 

[Dequilo]

Quietly listening from the back

and taking notes

 

[Mudballs2.0]

No,,,,im attempting to argue that we dont really need deep tc tech as home growers,,I think generally , home growers can rely on initiation and micro cloning for removal of all nasties provided initiation specifically is performed properly!!,,emphasis on the initiation process

micro cloning is literally just surface cleaning clones,,,initiation is the process of witch you introduce genetics into tissue culture and attempt to remove nasties before you go full apical meristem, initiation includes surface cleaning but i felt the need to state the difference between the 2,,,initiation happens first inside the grow room before you go micro


Pm is not systemic,,,it just puts its roots into cells

The best way to remove pm is to Bath clones in h2o2 before you root them,,,when you cut from a plant go directly into a 2ml per l ho2o2 bucket and leaves them spinning for an hour,,,in tc we use a 10 sec iso wash but i really like the h2o2 bucket for non tc,,

If you can use uvc too on cuts, ,like the uvc light in my tc bag

Dunk clones once a week in Bath of 1ml h2o2,,,shake well

I cant stop myself from asking for clarification on your hypothesis and the procedure of initiation .is it based on probabilities, protocols, and numbers?
The idea being if i microclone enough from a mother, i will get a clean specimen just by process of elimination and playing against the infestation severity in vitro?
Layman, there isn't a viroid every square centimeter, every cell, in the plant, you can out pace it.
I cant resolve how we get the clean vegging tiny plant infection free from a plant to this

 

[englishrick]

Initiation is stage 1 of tissue culture,,, if you read any real books you will see this word being used,,INITIATION

this is where you get your genetics clean,,,micro cloning is part of this process,,,the protocol for micro cloning does a surface sterilisation ,,, after the micro cloning you know the explants are clean of pm and free from viroid spreading bugs,,,this would be the first thing to do,,,

Next, you would grow the micro clones and subject them to an environment where viroids find it difficult to multiply,,ie super hot etc,,this is some personal IP again so won't go into too much detail,,but anyway,,,this environment stops the viroid moving and fresh groth remains free,,once you clone from the tops on this run you should have totally clean plant material,,,

This is an attempt to avoid the full apical meristem work for the home grower,,,im curious to know if plants still test positive after this work

 

[Mudballs2.0]

im not a botanist, im just a pothead who paid attention in highschool more than others. From what i've found you could be right, he has a sound hypothesis that passes initial scrutiny (see what i did there lol). I know heat can do it, but i don't feel comfortable going further into something he considers IP (intellectual property). suffice it to say i think it 'might' work and if properly documented and repeatable may be an incredibly valuable addition to cannabis home growers.
i would have to study how HLVD replicates and all sorts of other ridiculous not fun stuff to play along....i think im gonna defer to the pro's and hope you get that to us at your convenience. i see a 'white paper' in your future

 

[englishrick]

I don't have a problem sharing my personal ip with personal home growers,,when im ready to drop the kit, then list everything,,till then im just not happy with some company's soaking up my unique ideas and pumping them out as there own,,so I'm keeping some ideas to myself, for now,,,,some of the magic ingredients ive found are just too special , the mix ratios are unique, the protocol is novel

a handful of tricks and styles make me unique ,,, I suggest people read the books and take the journey ,, get some unique tricks yourself ,,when I'm ready il drop the kit and explain everything,,,till then I suggest getting some books and take the journey yourself like I did

 

[Mattbho]

Thanks guys!!! Awesome discussion

 

[Mudballs2.0]

Thanks guys!!! Awesome discussion

Hearing one of the first tissue culture pros tell us we probably dont need tc has some wow factor huh lol.

 

[englishrick]

Some of the techniques are priceless,,,tc style surface cleaning is the most effective way of removing bugs and bacteria, pm etc,,But you don't need to go invitro (into glass containers) or even micro to get the desired effect,,,you can basically just do really small cuts and do a tc style surface sterilisation,,

 

[englishrick]

The best way to remove pm is to Bath clones in h2o2 before you root them,,,when you cut from a plant go directly into a 2ml per l ho2o2 bucket and leaves them spinning for an hour,,,in tc we use a 10 sec iso wash but i really like the h2o2 bucket for non tc,,

so a little machine you can clamp the cutting in and then lower it into a solution bath and it starts spinning at a low rpm?
Cant we just leave them submerged? Like a redneck boy just jam a handfull in the bucket im carrying with me in field?

If I was trying to get clones out of a field I might well just walk around with a jam jar of h2o2 mix and take small explants from the tippity tip tops,,,this is probably the best start to initiation in a field of weed plants,,,I'd lable jars do leaf punches, take meristem,,,I'd test it and culture it from the individual jars

 

[GMT]

Hey Rick,
T.C. is fantastic for large scale storage of huge numbers of genomes in relatively small spaces.
But it requires economies of scale to be workable. I agree that this makes it unsuitable for the individual grower.
It seems to be the misjudged belief that the process can mend a broken plant though, as if it's a medical treatment, that makes it popular. Popular, not common.
The confusion between the cleaning process and the storage and reanimation is clear. But even this cleaning process is not a magic cure all. While being able to remove surface molds, pests and surface bacteria, it will not remove a virus from the plant tissue, and the theory of out running a virus is flawed.
I'm glad you're putting T.C. into it's correct perspective, but I think some more of the mysticism still needs to be removed from this particular tool set.

 

[Mudballs2.0]

The fermentation trials showed that HLVd was significantly degraded after 30 days at mesophilic or after 5 days at thermophilic conditions, respectively. However, sequencing revealed that HLVd was not fully degraded even after 90 days. The incubation of hop harvest residues at different temperatures between 20 and 70 °C showed that 70 °C led to a significant HLVd degradation after 1 day. In conclusion, we suggest combining 70 °C pretreatment and thermophilic fermentation for efficient viroid decontamination.

...you can outpace it. Ive done more research and theyve already done heat cleaning hlvd clones.
Remem, you cant delete once you post here.
Btw, mesophilic means normal temps and environ for an organism...if you didnt know...

 

[NEED 4 SEED]

The fermentation trials showed that HLVd was significantly degraded after 30 days at mesophilic or after 5 days at thermophilic conditions, respectively. However, sequencing revealed that HLVd was not fully degraded even after 90 days. The incubation of hop harvest residues at different temperatures between 20 and 70 °C showed that 70 °C led to a significant HLVd degradation after 1 day. In conclusion, we suggest combining 70 °C pretreatment and thermophilic fermentation for efficient viroid decontamination.

...you can outpace it. Ive done more research and theyve already done heat cleaning hlvd clones.
Remem, you cant delete once you post here.
Btw, mesophilic means normal temps and environ for an organism...if you didnt know...

Wouldn't a 70°C treatment for one day destroy the living plant sample?