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Quest for most potent cactus - from seeds with help of pereskiopsis grafting Options
 
pete666
#101 Posted : 12/11/2018 9:51:19 PM

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My grafting success rate is quite high, I would say something around 90%, but the problem is that about 15-20% takes very long to start growing. And because I am paying the electricity bill, I am not willing to wait weeks for seedlings to take off. So how to increase the success rate? If there are enought seedlings, then more of them can be grafted together and the best growing one is allowed to grow, others are discarded. If there are not many seedlings available, the seedling can be split into two parts(horizontally) and they can be grafted together onto one stock.
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Acceptance of the fact that our reality is not real doesn't in fact mean it is not real. It just leads to better understanding what real means.
 

STS is a community for people interested in growing, preserving and researching botanical species, particularly those with remarkable therapeutic and/or psychoactive properties.
 
pete666
#102 Posted : 12/13/2018 10:50:29 AM

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An1cca wrote:
pete666 wrote:

I am targetting just mescaline, so I believe the taste should guide me.


That might be true for Trichocerei, at least that's what we all hope. Although when I tested the full-alkaloid extractions of T. pachanoi, peruvianus and bridgesii, the mescaline concentration of dry alcoholic extract was somewhere between 65 and 85%. Stupidly enough, I never actually tasted the leftover goo after recrystallization to determine whether or not it contained any bitter substances as well. Something on my list...

For Lophophora, I think the screening method based on bitterness will not be valid. It seems many more alkaloids are present that might have a bitter taste as well. That's why I'll be forced to do biopsies of my small collection (65 individuals, 10+ years of age) of LW and do a DCMQ (about half an hour of work for every specimen) to establish the hall of fame Laughing .


pete666 wrote:
If you are interested, you are more than welcome to share your endeavours in this thread. I am aware you were inactive for long time, but I know you from your posts. And I guess I owe you for the nitrazine usage, am I right?


You're welcome, I'm glad it helps. And yes, the title of that thread sounds like our common goal Thumbs up


I am replying here, as original thread is rather about grafting and I don't want to flood it with irrelevant discussion

Yes, the taste method is just a theory. I am convinced it works, especially for peruvianus and likely for pachanoi too. Bridgesii is questionable and lw is out of question. But I don't see why anyone would grow lw just for pure mescaline, leaving other chemicals behind.

I currently have about 400 specimens (all trichos) grafted and growing and next 80 is on the way. The plan is to "analyze" all of them in April-June. Expected time required for analysis of one specimen is about 15 minutes (just the work), so I can hardly do all of them. I will have to do some preselection to reduce the total count and using the taste is the main option. Any information whether it really works would be valuable.

I have some remainders after ipa/tone/mek washes of HCl extract. This was not re-x'ed or anything so it definitely contains some mescaline. I might try purifying it and checking the taste. Have you an idea how to remove as much mescaline from that as possible?
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An1cca
#103 Posted : 12/13/2018 11:11:55 AM

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Probably the most complete removal would be by spotting on a TLC-plate and after developing, scratch away the M-zone, then scratch off the rest, swirl in minimally acidified water (e.g. sparkling water) to reconvert to water-soluble salts, filter, evaporate the filtrate to dryness and taste.

Far easier is to do some repeated recrystallizations. If the remainder only tastes 'slightly' bitter, then it won't influence the overall technique much...
 
pete666
#104 Posted : 12/13/2018 12:02:34 PM

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TLC is not in my toolbox :/

What would you use for the re-x? For sulphate it would be water, but for HCl? Mek, tone, ipa? Mix of all?
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An1cca
#105 Posted : 12/13/2018 12:34:00 PM

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Conversely, HCl is not in mine Very happy ...

I'm a sulphate man and do my re-x by dual-solvent acetone-water. After 2 or 3 times, it's >99%.

TLC really isn't that hard to do. It has many applications in our field of research. A very useful one is to determine the M-content of a crude alcoholic extract, resin or tea. That way, you don't have to further purify to know what's coming your way (Razz or Shocked ). It saves time&effort and creates the possibility of enjoying the 'individuality' of a certain clone. With peyote-extracts, I assume this is of even greater importance. I started with this kit. Works like a charm in combination with ninhydrin.
 
pete666
#106 Posted : 12/14/2018 8:50:49 PM

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One of the scions was growing much less than his brothers and sisters so I checked it and saw some infection near the join. So I cut it and about 1/4 of the cacti was infected, so I decided to taste as usual before discarding and there definitely was bitter taste. Not very strong, but clearly identifiable. Bridgesii - 74 days old
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An1cca
#107 Posted : 12/14/2018 9:09:31 PM

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Nice! Does this hybrid have a name?
 
pete666
#108 Posted : 12/15/2018 6:48:53 AM

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No, it was bridgesii from Koehres.

I have tried to re-x (just once) the peruvianus extraction alkaloid remainder, in hot mek/ipa/tone mix. This remainder is about 5-7% of mescaline HCl extract obtained by multiple washings of the extract with cold/dry mek/ipa/tone in few recent extractions.
About one third of it precipitated, it was not crystalline though. I evaporated the solvents and tasted the remainder. It was very bitter.

I know there still may be some mescaline, but I think the other components are bitter too. The bitterness didn't seem to decrease by re-x.

But if it is just 4% of extracted alkaloids and if it was just as bitter as mescaline (assumption), I don't think it would make the cactus taste noticeably bitter in that concentration. And even if it could influence the taste method, it would have to be in high concentrations before the mescaline, which is not very likely too.

Acceptance of the fact that our reality is not real doesn't in fact mean it is not real. It just leads to better understanding what real means.
 
pete666
#109 Posted : 12/22/2018 8:54:29 PM

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So my next observations from the battlefield...maybe it has been stated before, but regarding my watering scheme...
I am bottom watering every 3 days. The tray is flooded with water(with nutrients) for about 8 hours (overnight) and then the excessive water is removed. I have tried leaving the water there for longer - this led to yellowing leaves (random, both old and new were affected). I have tried to prolong the periods and again, yellowing leaves (again random, both old and new were affected). So it seems both overwatering and underwatering leads to the same result - yellowing leaves.

The best is to have the soil moist, but not overwatered. The soil should never dry off totally, but most of the water (80%?) should be away before next watering. Weight of the tray can give very good picture of current state
Acceptance of the fact that our reality is not real doesn't in fact mean it is not real. It just leads to better understanding what real means.
 
pete666
#110 Posted : 12/22/2018 8:58:15 PM

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Again, picture of current state of section with the oldest grafts...
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An1cca
#111 Posted : 12/23/2018 8:11:30 AM

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Looking good, Pete666!
I have some root-stocks that even lost all of their leaves (drying out of substrate Embarrased ). If the scion is already big enough, it will make up for the lost photosynthetic area and still maintain a decent rate of growth. Looking at your setup, I assume a time will come when the scions will overshadow many of the leaves. Besides, can you raise the ceiling of your growchamber or do you plan on degrafting them when +/- 20cm tall?
 
pete666
#112 Posted : 12/23/2018 8:37:31 AM

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Thanks An1cca
Yes, this is what I think as well. But doing my best to keep the leaves attached as long as possible.

No, the ceiling is fixed. But I have an adjustable double bottom of every section of the growbox, so I can move it about 20cm. Maximum space available for scions is 25cm, then I have to degraft. I hope the scions will have about 100g at that size, which is amount I need for my semi-analysis.
Acceptance of the fact that our reality is not real doesn't in fact mean it is not real. It just leads to better understanding what real means.
 
pete666
#113 Posted : 12/28/2018 5:40:07 PM

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Bottom watering is very convenient, in fact I can't imagine watering so many plants the ordinary way. But it has one very important drawback. This watering method is not washing away the salt and mineral deposits. Their levels have to be kept under control, otherwise the plants may (and likely will) cease growing and even die.
What to do against it? The soil can be flushed with plain water so it will drain out the bottom once a month. Enough to remove most of the deposits, but not too much to remove all the goods from the soil.

It is very convenient to be able to move whole tray with bunch of growing containers somewhere where watering can could be used. But this works only to some size of scions. My section with the oldest scions undergone this procedure today and I think that was for the last time, they are simply too big to be moved. They will have to stay on in the growing box from now and I will have to find some another way how to flush them in the future.
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An1cca
#114 Posted : 12/28/2018 6:27:39 PM

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I use an automated flood&drain setup that pumps the nutrient solution 5 times in the tray and lets it stay there each time for half an hour. This gives the substrate enough opportunity to come into equivalence with the solution. In this hydroponic system I use expanded clay (Seramis) and aim for an EC of around 600 at a pH between 5,5 and 6. I also add 40ml of 12% H2O2 for every 50 L of nutrient solution. I water&feed about once every 5-7 days. Temperature in my growroom is kept at around 26°C just by the heat of 1 Migro and a thermostatically controlled 60W halogen.
 
pete666
#115 Posted : 12/28/2018 7:08:17 PM

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Nice An1cca.
I was considering that as well, as I have quite a lot of experience with hydroponic and aeroponic grow setup used for mj. Unfortunately I am limited by the size of my growbox and I've chosen to use the space for cactuses rather than for the reservoir and tubes.
Acceptance of the fact that our reality is not real doesn't in fact mean it is not real. It just leads to better understanding what real means.
 
pete666
#116 Posted : 12/29/2018 10:26:58 AM

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One of the grafts got an infection, so was cut - Scopulicola x Zelly12, age from grafting 78 days, weight of whole scion 41g. There were signs of bitter taste, though very weak.
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pete666
#117 Posted : 12/31/2018 8:33:53 PM

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An1cca wrote:
I use an automated flood&drain setup that pumps the nutrient solution 5 times in the tray and lets it stay there each time for half an hour. This gives the substrate enough opportunity to come into equivalence with the solution. In this hydroponic system I use expanded clay (Seramis) and aim for an EC of around 600 at a pH between 5,5 and 6. I also add 40ml of 12% H2O2 for every 50 L of nutrient solution. I water&feed about once every 5-7 days. Temperature in my growroom is kept at around 26°C just by the heat of 1 Migro and a thermostatically controlled 60W halogen.


By the way, An1cca, any chance uploading picture of your setup/cacti?
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An1cca
#118 Posted : 1/1/2019 3:29:39 PM

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Of course Pete666, here's the setup I use. As already mentioned, it uses a Migro 100w that is most of the time able to keep the 80x80cm closet at a temperature of around 25-26°. I used Orca-foil to line the closet. 2 small 16w fans keep the air going and the heat well distributed. One of them is aimed at the top of the doors, so that some (hot and humid) air is constantly refreshed through the small crack between closet and door. Humidity stays between 50-65% this way, depending upon temperature and watering schedule. Day-night cycle is 18/6. In the front at the right, you can see the overflow-tube of the flood&drain system. This setup is running about three months now and doing fine. I have active and healthy growth on peres, grafted peres as well as trichs on their own roots. Some of the scions are over 20cm and I'm planning on degrafting them soon (letting a part of it attached to the peres of course).

And while I'm here, there are 2 things that might be of interest concerning the topic of this thread:

1) Wakinian's parafilm-technique works like a charm. Even the grafts I spoke of earlier that weren't well put off-center have started growing again. The technique is very forgiving, probably because of the pressure and humid macroclimate that is maintained through the parafilm. What I would like to add however, is that it helps a lot if you PRE-STRETCH THE PARAFILM. Not only the part that goes over the scion, but the whole length of that piece. This helps in getting the elasticity and stickiness right that you'll need for the moment of truth when you try to pin down that scion.

2) Concerning the bitterness-test: I cut some small bits of the different trichs I'm growing and was glad to have an observation confirmed that I already discovered when doing earlier research, namely that there's an ALKALOID-GRADIENT IN AN INDIVIDUAL TRICH THAT RUNS FROM HIGH-IN-THE-TOP TO LOW-AT-THE-BOTTOM. The same scion is practically tasteless at the bottom while it's definitely bitter near the top. So when sampling, I would advise to always compare between samples taken from the same vertical fraction of each cactus. In my earlier research for example, I found that the DCMQ-technique seemed to predict final extracted M-yield best when the punch-biopt was taken at around 1/5 of the stem length above ground level. Extraction was done here with cores included. I hope to further confirm these findings in my genetically better batch of trichs once I get my lab-stuff back.
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pete666
#119 Posted : 1/1/2019 7:43:07 PM

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Lovely. The setup is good, especially the flood&drain system. I envy that Smile
Yes, the parafilm is working very well. Wakinyan provided very good directions, including what you are writing - stretching is crucial. It is very good prevention against mashing the scion.
Btw I haven't seen Wakinyan for quite some time. I hope he is ok.

Your findings regarding the bitterness are very valuable. I was planning to cut the scions once about 100g of weight and cutting a piece at the bottom, which would be used for tasting. This might fail, if your findings are valid. I will rather use some part closer to the top.

And I've got some positive news - I had to degraft two small grafts of the same hybrid, they were not more than one month old, quite small yet. And believe or not, they vere seriously bitter! Not just a little, but rather like mature specimens I have! I was really surprised, I was expecting no bitterness at all at that size and this had shocked me!
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An1cca
#120 Posted : 1/1/2019 8:15:06 PM

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You mean the Scop x Zelly12? Interesting, these should produce some nice flowers as well according to the breeder. In my little garden I only have Scop x Huar and that's quite a bitter hybrid as well. The future looks bright (with rainbow tracersCool )...
 
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