Distinguished and Nurtured Kind

@CannaRed - I'm really not sure why I've started doing that. I read it some where as the SOP for cloning recently so I made a change. I thought it was in a study I had read here, but that one didn't address lighting variations and used a 18 hour cycle, so I had to remove the earlier edit.

I will say I notice this. I can take a tray of cuts. Dark cycle. Couple days of light - if there is a weak link in the tray, it shows it self. I've had two cuts of the go completely hollow and limp in the tray. The other 4 cuts of Sis/PK are fine. The other 6 cuts of Cobalt are fine and the cuttings of the male are fine.

So, I lifted the dome and removed those two weaklings from the tray. I'd not be surprised if everything left roots.

What I'm making is certainly a false correlation that can't be attributed to a single factor - ie - a brief dark period after cutting, however, I am going to be keeping notice of it now to see if I do in fact see such a pattern developing.

------------

I had to clean up the canopy again this evening. I removed branches from 42 different internodes. 15 of those, were "main" branches longer than 18" but had simply been overgrown by more dominant branches, whose secondary growth above the screen was outpacing this lower growth and filling in the canopy directly above it anyway.

This happened last grow and I just assumed things would catch up as they do under a 1k, but instead, the growth in the prime zone just dominated. I'm working much more diligently to keep any useless growth out of the canopy this round. If it was thinner stemmed and a bit wispy it got cut out.

The canopy looks a lot cleaner now. I can see more airflow moving through out and more light reaching the lowers on the larger colas that are shaping up. I think from this point forward I'll just let things fill in and grow upward only removing the first internodes on longer secondary branches, to ensure there is enough air flow within the lower portion of the canopy and main stems.

It's shaping up nicely. The room is starting to define itself and the colas are really starting to take shape as individual towers rather than a mass of green growth. Cobalt #2 is starting to produce the faintest hint of resin on her leaves already. I'm honestly excited to see what she is capable of in the resin department with the better air flow and proper temps this round.



dank.Frank

Also...this: Tissue Culture. It's from 2016, but I'm still trying to catch up.

https://www.sciencedirect.com/science/article/pii/S221478611530019X?via%3Dihub

Nodal segments containing axillary buds from selected mother plants were used as an explant for initiation of shoot cultures. Explants were surface-disinfected using 0.5% NaOCl (15% v/v bleach) and 0.1% Tween 20 for 20 min. and washed in sterile distilled water three times for 5 min each, prior to inoculation on the culture medium.

Disinfected explants were inoculated on Murashige and Skoog's medium (Murashige and Skoog, 1962) containing 3% (w/v) sucrose, 0.8% (w/v) type B agar (Sigma Chemical Co., St. Louis, MO) and 500 mg L−1 activated charcoal supplemented with various concentrations of ... MS + mT ranging from 0.05 to 5.0 μM adjusted to pH 5.7. Sterile medium was dispensed (25 ml) in glass culture vessels (4-cm diameter × 9.5-cm height) with magenta B caps.

All cultures were incubated at 25 ± 2 °C with 16-h photoperiod under fluorescent light with a photon flux of ∼52 μmol m−2 s−1.

Well-rooted plants were carefully taken out of the medium, washed thoroughly in running tap water to remove all traces of medium attached to the roots. These plants were pre-incubated in coco natural growth medium (Canna Continental, Los Angeles, CA) in thermocol cups for 10 days before transferring in sterile potting mix-fertilome (Canna Continental) in large pots. All these plantlets were kept under controlled environmental conditions (light, ∼700 μmmol m−2 s−1 with 16-h photoperiod, temperature 25–30 °C and relative humidity ∼60%) in an indoor cultivation facility. Mother plants and well acclimatized tissue culture raised plants were transferred ex vitro for further cultivation.

Eight ISSR primers were screened with the DNA of mother plant and nine randomly selected tissue culture raised plants. Each tested primer produced clear and scorable amplification products ranging in size from about 155 bp with UBC 845 to 2000 bp with UBC826 with an average of 6.6 bands per primer generating monomorphic patterns across all 10 plants analyzed (Table 2). No ISSR polymorphism was detected showing genetic integrity of micropropagated plants.

4. Conclusion

In summary, the use of mT (2.0 μM) provides a one-step protocol effective in promoting adventitious shoot formation and root induction in the same medium. The maximized regeneration protocol is simple and cost effective considering the high shoot proliferation rate, the ease of rooting, and the 100% survival frequency of acclimatized plants. To the best of our knowledge, this is the first report on the successful application of mT on in vitro propagation of C. sativa . Our results confirm the clonal fidelity of micropropagated plants and reveal that current protocol using mT is safe for large-scale production of true-to-type C. sativa plants.

Click to expand...

dank.Frank

tissue culture is so neat. to have a library of clean vigorous cuts ready to go would be a game changer for the modern farmer.


after reading a few of the TC articles on the sterilizing the plant material prior, i have personally found using a similar 10% bleach an distilled water to rinse an soak the cuts one time prior to cloning have increased my success rates. i do not rinse the solution, they go right into the rooting hormone an pucks/rockwool wet. Once they root the clones seem abit more vigorous right off the bat.... but thats me.

i seem to get less overall problems with random fungal or mold growths inside the humidity dome. i also add 15mill to a gal of m-pede insecticidal soap to that bleach solution in hopes of stopping any hitchhicker bugs, if any. that will also suffocate any spores. doesnt hurt to be preventive, that is certain. not that trying to turn this into a cloning hijack. but it works well.



you planning at trying your hand at some TC there DF?

I've been looking at TC now for a couple of years. Not so much for cuts but rather to start seed embryos. This particular format is a MUCH simpler approach. I have old seed that I know the embryos are viable in, but they don't have the energy left to kick shells and elongate cellular growth. For me, I see tissue culture as a way of extracting embryos and placing them in sterilized containment until they are able to germinate.

The other interesting thing about this particular tech outlined in the article: full acclimation back to the greenhouse and the tech produces zero somaclonal variants, which means they are true to the parentage. (which is a problem in the past with other techs)

I was talking with DLB the other day in chat and he linked me to a microbial product he was using. While browsing their other options, I came across this VERY interesting combo of PGRs: Agra-Rouse

Cytokinin, as Kinetin .............. 0.05%
Gibberellic Acid (GA3) ............ 0.05%
Indole-3-Butyric Acid (IBA) ..... 0.09%
"Inert Ingredients"" ................ 99.81%

This product, would actually be quite useful in TC applications.



dank.Frank

Thats interesting about the dark cycle once you take cuts. I'm going to give that a try.. So basically clone then leave them in the dark 3 days then lights back to 24/7?

I wish I had this thread back when I tried SCROG. I thought it was just get the screen filled and your golden for that many colas. Boy is it a lot more work then that.. Just cleaning up the under screen is a job in itself along with clearning out anything not getting sufficient light...

That screen looks great tho.. I can't wait to see a pic in 30 days once they are blown up..

Im over here rooting for that Sis x PK to show something special..

No. Not 3 days. 24-36 hours. Day...day and a half at most. Again, not sure where I picked this up from, but when I figure it out, I'll post it. I'm still not sure there is any benefit to it either so...grain of salt for sure. It just happens to be what I'm doing at the moment.

You and me both on the Sis/PK. She's looking good. Still only throwing 5 blades per fan leaf. Center blade a bit larger than the other, but not nearly as bulbous or exaggerated as you'd expect to see on a heavy Chem leaning plant. I really think she's pretty much a 50/50 type expression as she continues to mature.



dank.Frank

Only problem I've had with cloning of this pheno run is they have all rooted. Which means instead of 80 I now have 160..ugh these males gotta go.

My cloning is high tech,
Cut clone
45 angle
light scrap
dip in powder hormone
into homemade areocloner with just clonex liquid added

cloner
PH - never checked
Temp - never checked

Like I said High tech lol

Americangrower said:

Only problem I've had with cloning of this pheno run is they have all rooted. Which means instead of 80 I now have 160..ugh these males gotta go.

My cloning is high tech,
Cut clone
45 angle
light scrap
dip in powder hormone
into homemade areocloner with just clonex liquid added

cloner
PH - never checked
Temp - never checked

Like I said High tech lol

Click to expand...

That's pretty hi tech compared to mine.

Cut clone
45 angle
light scrap
dip in powder hormone
into solo shot glasses with perlite for medium.
Those are set into a rockwool plug tray where they sit in a 1020 tray full of water with liquid kelp and 1 ml of superthrive on my seedling mat set for 81* F.

If the cuttings remain turgid for 48 hours, they'll root. Matter of fact this last time around I took 18 cuttings and now have 18 clones. First time I've never lost one or two.

trichrider said:

but blame it on the camera. lol

Click to expand...

dank.frank said:

But MOSTLY - it is this stupid 3.5mp camera. I'll have to borrow a camera and prove my point. LOL

Click to expand...

ABF Seeds Cobalt Haze @81 days






This is what I've been smoking on. I told you it was the camera!!!



dank.Frank

lookin' good

:jawdrop:WOW!

How does she smoke?

Looking mighty proper broski much respect. I know it feels good to be blowing on some of your own head stash again.

Thanks for the kind words!

@bigtacofarmer - she smokes. You can feel her climb a bit more with each puff...she's very expando in the lungs. The first toke is like some just punched you in the sternum. People who aren't heavy smokers just take a puff or two and they walk away eyes winced, fist balled, held up to the mouth while cough from the hips and still trying to walk straight. Second puff you feel it crawling from your shoulders and into your neck. By the third hit, you're already high, but the third hit will get you stoned. She doesn't have any mental laziness to her, but it does smoke like a sativa leaning HYBRID, so it's not that crystal clear soaring high either. There a good bit of body to it, it's just not couch lock, in how it translates. It is a strong plant though.

I'm not sure why she hits so hard. The 9wk sibling was very smooth and like sucking air. But the 12wk'er - she has a lot more resin and a completely different smell. I honestly wouldn't be surprised if she goes 13 or even 14wks when grown properly. The trichs on her were surprisingly just getting cloudy in most places when I chopped her. There isn't an amber any where to be found and it's still probably at 80% cloudy after cure.

It's probably more honest to say, I don't know how she REALLY smokes at this point. I'd not be surprised to test her and find out she is in the 27-30% THC range, which honestly, I think is part of the reason she rips your lungs out and laughs at you like a lil bitch...

@Prodigygrower - Thanks! Good to see you around again. Indeed, it is. Nothing better than your own. We all know that.



dank.Frank

That new camera really brings out the beautiful colors of the bud.. Really beautiful bugs DF..

Now show us some pics of that screen all filled lol

Beautiful garden, my friend, I love your large soil beds! I seriously thought about going that way for my flower room but I stuck with the 20 gallon fabric pots, mostly since I already had them. We're running the same cmh lights, though. Great minds think alike, right?

Keep up the great work and spreading the good word!

Here's a peek into my room--hope you don't mind! Stank Bros Booberry Kush is in the center alongside my purple haze bagseed keeper.

dank.frank said:

View Image





ABF Seeds Cobalt Haze @81 days

View Image

View Image


This is what I've been smoking on. I told you it was the camera!!!



dank.Frank

Click to expand...

Looks really delicious!

Current cycle @ 20 days. The tall lanky beast in the front is the Chem Sis x PK. That whole side has stretched like mad. The 9wk Cobalt is every bit as big as it was last time. It's stretched about 32 inches above the screen. The green stakes sit right at 6' from ground height, so that gives you an idea of how big they have managed to get still.

These girls are going to STACK HARD.

Unfortunately, the borrowed camera had to be returned after a couple days use. It did give me an idea of what to look for though before I decide to make that purchase. I know I HAVE to do so for the Karma Genetics testers that will be coming soon.





dank.Frank

The camera does a nice job but it is your green thumb that impresses. Those plants are a picture of health just beautiful! Love the structure. Nice job!

Nice smoke report I felt high after reading it!

I wasn’t expecting them to be that huge!! that soil bed is killing it. They look super happy.
This is going to be nice to see in 30 days from now