Popular Post Auxin Posted August 7, 2017 Popular Post Share Posted August 7, 2017 Quote Screening of Hallucinogenic Compounds and Genomic Characterisation of 40 Anatolian Salvia Species Seda Damla Hatipoglu,a,b Burhanettin Yalcinkaya,c Muslum Akgoz,c Turan Ozturk,b,d Ahmet C. Gorend* and Gulacti Topcue** ABSTRACT: Introduction – Salvia, an important and widely availablemember of Lamiaceae family. Although comparative analysis on secondary metabolites in several Salvia species from Turkey has been reported, their hallucinogenic chemicals have not been screened thoroughly. Objective – This study provides LC–MS/MS analysis of 40 Salvia species for screening their psychoactive constituents of salvinorin A and salvinorin B. 5S–rRNA gene non-coding region of Salvia plants was sequenced, aligned and compared with that sequence of Salvia divinorum plant. Methodology – Targeted molecules of salvinorin A and salvinorin B were quantified, using LC–MS/MS, from all aerial parts of 40 Salvia species, collected from different parts of Turkey. Regions of 5S–rRNA gene from different species were amplified by polymerase chain reaction and DNA sequences were aligned with Salvia divinorum DNA sequences. Results – Very few of the Salvia species (S. recognita, S. cryptantha and S. glutinosa) contained relatively high levels of salvinorin A (212.86 ± 20.46 μg/g, 51.50 ± 4.95 μg/g and 38.92 ± 3.74 μg/g, respectively). Salvinorin B was also found in Salvia species of S. potentillifolia, S. adenocaulon and S. cryptantha as 2351.99 ± 232.22 μg/g, 768.78 ± 75.90 μg/g and 402.24 ± 39.71 μg/g, respectively. The sequences of 5S–rRNA gene of 40 different Salvia species were presented and it was found that none of the Salvia species in Turkey had similar DNA sequence to Salvia divinorum plant. Conclusion – This is the first report of screening 40 Salvia species in Turkey according to their psychoactive constituents, salvinorin A and salvinorin B and their genomic structures. It is possible that some of these Salvia species may exhibit some psycho activity. Thus, they need to be screened further. DOI 10.1002/pca.2703 15 Quote Link to comment Share on other sites More sharing options...
Strontium Dawg Posted August 8, 2017 Share Posted August 8, 2017 Cryptantha may be promising then. 2 Quote Link to comment Share on other sites More sharing options...
MeanGreen Posted August 18, 2017 Share Posted August 18, 2017 I bought S. recognita seeds after reading this article last week. 5 Quote Link to comment Share on other sites More sharing options...
Dicko Posted August 23, 2017 Share Posted August 23, 2017 Hey all. Could some one please explain the little plus/minus symbol part? '212.86 ± 20.46 μg/g' The way i read it (probably wrong) is 212.86 micrograms give or take 20.46 micrograms per gram, like a margine of error type thing? 1 Quote Link to comment Share on other sites More sharing options...
DualWieldRake Posted August 28, 2017 Share Posted August 28, 2017 (edited) plus minus is the inaccuracy so yea you are right for some more in depth expanation. it's when for example the measuring equipment has a certain precision, like a digital scale. And also human precision with analogue reading, you can at most decide with a strong certainty if it's more or less than halfway between the marks Edited August 28, 2017 by DualWieldRake 1 Quote Link to comment Share on other sites More sharing options...
Dicko Posted August 28, 2017 Share Posted August 28, 2017 Thanks Dual Ok so approx 4.7g of recognita holds 1mg salvinorin?? Ordered seed, hope it gets through. Quote Link to comment Share on other sites More sharing options...
Dicko Posted August 28, 2017 Share Posted August 28, 2017 How do the levels of salvinorin in recognita compare to divinorum? Quote Link to comment Share on other sites More sharing options...
MeanGreen Posted August 29, 2017 Share Posted August 29, 2017 It's quite a bit higher in Salvia D. "The retention time for salvinorin A was 23.09±0.95 min and the measured concentrations ranged between 8.32±0.65 and 56.52±3.77 mg/g dried leaf." https://www.ncbi.nlm.nih.gov/pubmed/25710779 1 Quote Link to comment Share on other sites More sharing options...
Inyan Posted August 30, 2017 Share Posted August 30, 2017 Still, one might consider making hybrids between these species that contain different genetics yet, still contain the active particulars. https://www.ncbi.nlm.nih.gov/pubmed/28722248 Quote Link to comment Share on other sites More sharing options...
Inyan Posted August 30, 2017 Share Posted August 30, 2017 (edited) http://www.hardyplants.com/seeds/SA81-A3.html Salvia recognita seeds are only $2.95 at this place and they might indeed make for some interesting hybrids with Salvia divinorum if one made the cross both ways. An interesting article on hybridization of Salvia. http://journal.ashspublications.org/content/136/1/41.full Edited August 30, 2017 by Inyan 1 Quote Link to comment Share on other sites More sharing options...
Halcyon Daze Posted September 1, 2017 Share Posted September 1, 2017 Nice paper From what I've read there is only 1 other likely candidate for a hybrid, Salvia venulosa. It's a rare species from Columbia that is not readily available for purchase. I think it mainly came down to the number of chromosomes. Quote Link to comment Share on other sites More sharing options...
Dicko Posted September 1, 2017 Share Posted September 1, 2017 Would the recognita not be usable for a norm extraction like one would follow to concentrate salvia d? Instead of trying to just breed a desirable hybrid? Quote Link to comment Share on other sites More sharing options...
Anodyne Posted September 2, 2017 Share Posted September 2, 2017 Dicko - I assume one point of trying to hybridise divinorum would be to get a plant that produced viable seed? Since S.divinorum is sterile and has to be propagated from cuttings. That's just a guess though. Other times hybrids are bred to tolerate different climates or pests better, for example an active species with no frost tolerance can be bred with a hardier (but inactive) species to create an frost-tolerant plant that still has some activity (eg. Psychotria "nexus"). This can be useful when trying to grow plants outside of their natural habitat, but you're right - it isn't necessary. If they grow happily where you are, then you can just follow normal procedures, no fancy hybrids required But another reason to cross plants together is just to see what happens! Quote Link to comment Share on other sites More sharing options...
Dicko Posted September 2, 2017 Share Posted September 2, 2017 Heheh yeah I understand now, many whisky's were had last night Lol. Wasn't looking at the bigger picture. Cheers Anodyne Quote Link to comment Share on other sites More sharing options...
Dicko Posted September 6, 2017 Share Posted September 6, 2017 Sweet. Seeds arrived today :D 1 Quote Link to comment Share on other sites More sharing options...
MeanGreen Posted September 7, 2017 Share Posted September 7, 2017 My first Salvia recognita seedling 5 Quote Link to comment Share on other sites More sharing options...
Auxin Posted September 8, 2017 Author Share Posted September 8, 2017 S. recognita is a turkish species, before trying to cross it with a species from another continent it would be worthwhile to simply breed it for potency through 7 or 10 generations. Quite a few uncrowded plants could probably be grown to first seed set in just 3 square meters of garden space Even if it were eventually used in hybridization schemes, wouldnt it be better if it were brought up to 1 mg/g or higher first? ;) 3 Quote Link to comment Share on other sites More sharing options...
Inyan Posted September 8, 2017 Share Posted September 8, 2017 8 hours ago, Auxin said: Even if it were eventually used in hybridization schemes, wouldnt it be better if it were brought up to 1 mg/g or higher first? ;) Why could you not go along both routes at the same time? To selectively try to breed for a better S. recognita utilizing a large batch of seed which may show some variability could indeed take many generations and while it would be cool to do so with a completely different species on its own, what would it hurt to try to do the same through hybridization efforts at the same time? When S. divinorum has a status in many places that is less than legal getting those genes into another species via hybridization would be a boon for those wishing to work with that species where they can't as they would effectively be working with a hybrid. Introducing S. divinorum genes into S. recognita one could then do your standard F2, f3, f4, f5 sibling crosses to select out the strongest from the batch each generation. Many serious hybridizers not only work these f2, f3, f4, etc. type crosses, but they would also breed back to each parent as well to stabilize traits from either parent. The idea being to make a cross like this... (S. recognita x S. divinorum) x S. divinorum. While also making the cross (S. recognita x S. divinorum) x S. recognita. One could very easily then go on to create 2 more lines with these backcrosses to each parent. Never mind the reverse crosses (S. divinorum x S. recognita) x S. recognita and (Saliva divinorum x S. recognita) x S. divinorum could also be done as mitochondria and chloroplasts are donated from the maternal side I think we would be remiss if we did not also count those reciprocal crosses as well as good potentials. With that being said, we could in theory have 5 good working lines of of hybrids going trying to selectively improve and introduce new traits into the mix while also working with each species separately trying to increase the desirable traits there as well. So, all told we would looking at 7 different lines. Given how hard it is to get some hybrids started... I would think the sooner the better to get started on any of these lines. Then there is the logistics of testing the material from each of these 7 different lines to determine which ones to breed with. If I can grow 10K Brugmansia from seed to maturity in a small space... I can only imagine how many more Salvia could be grown in that same small space. An important question though... do the two species have the same chromosome number or can they be easily made to have the same chromosome number? If not as one member suggested... another species with the appropriate number of chromosomes either the same or double would need to be used to most easily facilitate a hybrid cross outside of protoplast fusion techniques. 1 Quote Link to comment Share on other sites More sharing options...
Inyan Posted September 8, 2017 Share Posted September 8, 2017 On 9/1/2017 at 8:44 AM, Halcyon Daze said: I think it mainly came down to the number of chromosomes. Chromosome numbers can be changed via the application of colchicine to the plant itself or seeds. There are other chemicals available that will also do the same so what one should be looking for in chromosome counts is anything with either the same number of chromosomes or double those chromosomes. With Hemerocallis there have been crosses made with triploids after many many attempts. What one has to understand is that even when the chromosome counts don't match up... there is always the possibility of unreduced pollen and many other variables that can come into play. Things like using a mentor pollen, donor pollen, pollen cocktails, and the like. The bottom line is I'd be excited to see any species mixed in with S. divinorum and given just two seeds of a viable hybrid such as this I could extend those two seeds into many plants and produce a small field of f2 seedling followed by a larger field of f3 seedlings. Breeding cold hardiness into a S. divinorum type hybrid... would that not be nice? 1 Quote Link to comment Share on other sites More sharing options...
MeanGreen Posted September 8, 2017 Share Posted September 8, 2017 (edited) A recognita x divinorum hybrid would definitely be nice! Just doing selective breeding on S. recognita would be great too, but it would require ongoing access to GC/MS since the salvinorin content is so low, we're not going to be able to do selection for potency through bioassay (unless maybe by doing standardized extracts but even then it is quite subjective). But a cold or drought-resistant divinorum hybrid that retains at least a good part of the plant's activity would be amazing, for sure! Edited September 8, 2017 by MeanGreen 1 Quote Link to comment Share on other sites More sharing options...
Auxin Posted September 8, 2017 Author Share Posted September 8, 2017 (edited) In the case of recognita x divinorum, the chromosome counts wouldnt be helped by colchicine as they are 2n = 16 and 22, respectively [1]. You might get a Triangle of U type situation after several thousand pollinations yielding a 2n=38 hybrid but I have yet to see anyone with several thousand divinorum flowers. I'm unclear how much mentor pollen and hormone applications might help cause a triangle of U scenario. Protoplast fusion could be an option if someone were set up for that. Those options could be done by very few people but, realistically, I see no trouble in several people growing several dozen plants each year and making a standardized extract of each for bioassay, and then using the best four to eight plants as the parent for the next generation. Every six years or so they could exchange genetics to diversify their breeding pool. Sure, there would be subjective error and possibly a significant procedural error but on the whole things would be moving in the right direction. Estimating from pictures I've seen, 45 uncrowded plants could be grown to seeding in a 1 meter by 3 meter plot [which by my gardening standards is small but non-trivial] the harder part would be finding gardeners that would want to do 45 acetone extractions every year and bioassay 45+ samples every year. Salvia always slapped me back into the cradle, doing that 45 times each winter would be stressful As a side note, as near as I can see the salvinorin content of Salvia tiliifolia is unknown. S. tiliifolia has the same chromosome count as S. divinorum and is native to a region just like 200 to 400 kilometers north of S. divinorum. Edited September 8, 2017 by Auxin 3 Quote Link to comment Share on other sites More sharing options...
Inyan Posted September 9, 2017 Share Posted September 9, 2017 http://salvia-hybrids.blogspot.com I present the above for those interested in how to hybridize Salvia. Just because it has not been done with Salvia divinorum does not mean it can not be done. The more varieties that are tried the more likelihood that a compatible species may be found. Just because a single pollination attempt does not garner any seeds it does not mean that the 1000th time you will not get a seed. The more people that try the more likelihood that someone will be successful. For those for whom this instruction is too complicated.... simply planting your S. divinorum in a bed of a different type or species of S. divinorum may create a hybrid if the bees decide to do the work for you. Most importantly, you must collect the seed from your S. divinorum and grow them out. Of course, if the cross can only be done with S. divinorum as the pollen donor.... you might want to have a bed of S. divinorum with a single specimen of another species in the middle of your bed of S. divinorum. For the record, I think emasculation is the much better route to go, but I leave no option unchecked in such a goal.... 2 Quote Link to comment Share on other sites More sharing options...
Inyan Posted September 9, 2017 Share Posted September 9, 2017 "In Salvias the pollen must be recognised and accepted before the female makes moisture available for the pollen grain to then imbibe and germinate. This is a very selective process and many interspecific crosses fail at this point." This is where mentor pollen comes into play. Dead S. divinorum pollen is presented to make the pistil receptive. This dead pollen is mixed with the desired species pollen one wishes to use to pollinate. This is of course just one way of doing this or getting around a barrier to pollination. 2 Quote Link to comment Share on other sites More sharing options...
Sallubrious Posted September 9, 2017 Share Posted September 9, 2017 I don't know if bees can pollinate Salvia d. flowers, their anatomy isn't really that bee friendly. Maybe native bees would have a better chance because they are much smaller. Their long narrow flowers seem to more like something that's pollinated by a moth of some sort. Some moths feed on nectar successfully with other elongated flowers when bees can't because they can sometimes have a tongue almost as long as the moth that can reach all they way in.Hand pollination with a brush worked for me before but the seeds never grew. Maybe the application of an irritant like the cement trick could help the flowers become more receptive to pollen from a different species. 1 Quote Link to comment Share on other sites More sharing options...
Inyan Posted September 9, 2017 Share Posted September 9, 2017 While we are talking about tricks to get different species to pollinate... mutilated or cut style approaches also sometimes works. I say leave no trick un-tricked in the quest for a hybrid until of course ones patience or resources run out. 1 Quote Link to comment Share on other sites More sharing options...
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