Darklight

Torsten contracted me to undertake micropropagation of this species back last millennia. It was a crapshoot, as the species hadn't been done in tissue culture ( that I could find ) at that stage and seed numbers were limited. I had two clients with seeds, T was the one who sorted the germination riddle, which is why it was his seed which produced the Rifat clone in-vitro. We got it! It's quite the saga. We were going to do a presentation on the process at EGA but decided to do the NMT presentation instead. Had to abandon the species entirely after the TGA scheduling came into effect in 2005. At the time we weren't aware of the intricacies of TGA scheduling and believed they wouldn't be so stupid as to S9 schedule- no known medical use- an entire plant with such medical and clinical potential ( it was the height of the heroin epidemic in Australia ) as a 'potential future drug of abuse'. They were. We learned a lot from that, it was horrible, stupid, ignorant and cruel. Many long stories, much happiness, learning, letdowns and lots of generousity on the part of T. Much snipped from here. One example of one very small part: Torsten and I were discussing retail price for this species once we had an established protocol. As it was highly sought after I suggested a general release, with first adopters and collectors paying a higher price for limited release, and subsequent releases of increasing numbers of plants each year at lower cost. We knew there were a few collectors who would pay good money, and I was impatient to capitalise on our significant investment ( his funds, my time and experience ) I figured the plant would leak out sideways after a few years anyhow, retail price would drop and any delay to the general public would be limited He nixed it straight out. His take was that it is an important potential anti-addiction plant and should be available to try by as many people as possible as soon as possible. This attitude of his is why I adore working with him :D As it was, a bunch of potential clients offering megabucks for clonal material had expressed real interest. And as soon as they realised it was in tissue culture they demanded we supply it to them at $0.30c a plant. Many demanded the micropropagation media alongside it. That price was set in 1976 and is for mass run easily produced plants with no research input. And only a fool gives their IP away so cheaply. When we refused sale at that price and made a counter offer all the clients vanished. So we did a general public release instead. Over time the performance in micropropagation dwindled ( happens in many species ) and the TGA scheduling coincided with the decline. We dropped it. We were too well known for producing the plant and the risk of continuing wasn't worth it. I had a feeling the authorities would have considered us tasty headline clickbait if we had continued and it would have endangered other conservation and medicinal work. We also had to abandon significant plans to undertake more phytochemical research, including food safety and storage It may have been possible to apply for federal compensation for future losses against the value of our investment. It was a fair sized figure by then. T also nixed that on the grounds that supplying information about our investment could leave us open to a requirement to divulge personal details of all purchasers of the clone in Australia. That wasn't acceptable to either of us So we took it on the chin, and we're still recovering. At least it's now available for consumption in many other countries and has a track record as an effective taper/ substitution/ remedy for opiate abuse for many people. That was our initial aim and even though we couldn't get it to happen here there will be data out there at some point which will hopefully one day support a re-scheduling in .au There are many stories left out of this post for brevity. Hope it gives you an idea of it My firstborn. Or one of 'em. So excited I was Full production The decline and fall Really kind of you. I loved working with the species too, I'm glad it's still bringing people joy and comfort

13 Most generous of you- thank you!

Pending. I did mine in-vitro and am having trouble deflasking them

Hahahaha would be hilarious if it were Griffonia suckers after all and it meant that I suck at growing both Griffonia and Ephedra You could be right. I'll take more pics as they grow

Mate, it's a hard task and a big ask. But it will be really worth doing as a single document. It'll prolly also help formalise a lot of the notes into new ideas. And when it's finished it will be gorgeous I write lab notes like this, all good

I have a Griffonia simplicifolia growing in a pot at home, and these popped up last summer. Until last week I thought they were new shoots from the vine and was waiting for them to develop leaves. After 15 months waiting I had a closer look last week and they look like bloody Ephedra! Not sure which one. I recycle my propagation mix and throw a lot of old prop mix into different pots to top them up. They could be *really* old E. gerardiana from 20 years ago, E. major or E. sinica from seed that didn't germinate over 5 years ago. Whoever they are they put a huge smile on my face. Never give up Thanks for keeping us updated, this is great! Sorry I didn't get to mention this thread at T's and my EGA presentation on research. It was in my notes to do but I got nervous and left heaps of major points out :/ Hope I'm not interrupting here, lemme know and I'll move it

l-dopa SAB sells an almost hairless strain of Mucuna which is easier to process. The hairs on the plant are a PITA, if you grow it I'd advise growing this type: http://www.shaman-australis.com.au/shop/mucuna_pruriens_v_utilis_velvet_bean_seed_packet_pr_598.php

Yup- sleep and constipation are huge barriers in PD Melatonin is excellent for sleep for a lot of ppl Melatonin ref from 2011 I know the kefir sounds like too simple a constipation solution to be effective, but please let me know how it goes for you. It's such an easy place to start and if you can't make your own, local Farmers Markets often sell them, or find a good quality local fermented foods person on FB. Is a bit of a thing right now so locating someone with experience could be easy

Now that is a shame Are there any informal dispensaries in Perth you know of? I would def acquire a script for melatonin for your husband, it's becoming increasingly recognised as beneficial for Parkinsons. Unlikely to interact AFAIK but please do your own checking of anything I say Wrt Lions Mane I'd advise caution and small doses. In fact given the Sifrol/ Mucuna etc etc I'd advise against it. I read something about it potentiating psychosis in some late stage Parkinsons patients. I lost those refs a while back and am chasing them up again. There are a bunch of variables around that statement above which do need checking Have you used it earlier in your husband's PD progression? With you in hope

Further, given the gut bacteria changes implicated in Parkinsons progression, have you tried either water kefir or milk kefir? In addition to slowing gut bacterial changes it can improve gut motility and decrease constipation in healthy individuals. There is potential for it's regular consumption in Parkinsons patients but I have yet to see any academic data on it. As gut bacteria studies are a relatively new field the absence of formal data doesn't surprise me too much. Because it is unlikely to interfere with pharmaceutical drug actions I'd be giving it a go for a week or so. This, however is just my untested opinion My relative with longer term Parkinsons will not try it. My relative with the rapid-progression form cannot tolerate water kefir at all- we have yet to try them on milk kefir We make our own water kefir from fresh grains. It all sounded terribly hippy-ish to start with back in the day, but there are a few of the fam who are deriving benefit from it and the taste is pretty good so it's a winner

Hi Paulinepie, so sorry you both have to work with Parkinsons and thank you so much for the info. I find it really valuable to hear about direct experience with treating Parkinsons with natural products May I ask what stage your husband's Parkinsons is at, and how long he has been taking Mucuna ( Velvet bean ) for? Hadn't heard of using Withania ( Aswagandra ) for Parkinsons but I have used it as a general health product previously and found it really well tolerated and effective- thank you again A few of my close relatives have Parkinsons- one with the longer term form ( +20 years since diagnosis, now late stage + dementia ) and two with the faster term form ( one has passed, the other diagnosed recently and progressing rapidly ) My family member with the longer term form is remarkably resistant to trying anything not prescribed by the GP or specialist, which is frustrating but is their choice. When I saw them last they were willing to try medical cannabis ( informally sourced ) high CBD tincture ( unsupported by formal analysis but a widely distributed form ) to combat the frozen state and hopefully reduce tremors. However his tremors and frozen state are quite advanced, and the tincture wasn't suitable to titrate a dose with in my absence- it was too slippery to measure small amounts and would require some dilution. His partner was not at all supportive of the treatment and informed me that they would not continue with it in my absence as his tremors would not allow him to dose alone reliably. We ended up not trialling the product at all. I've heard high CBD products can be effective for Parkinsons tremors, freezing etc and if you are comfortable trialling and able to source some I'd recommend beginning with a low dose and set a comfortable amount as soon as possible after diagnosis. If your clinician is comfortable with CBD co-administration with pharmaceuticals I'd confer with them. My relative is off all anti-Parkinsonians at this late stage but is taking antipsychotics for the delusions- they have been helpful, but potential contraindications with the antipsychotics were another reason for not undertaking the CBD even at small doses and they do not wish to confer on this matter with their clinician. Thank you so much for your valuable insights into potential non-pharma treatments. I wish you both, and your family, all the best on this difficult journey. If you are able to share further experience I'd love to hear of it, either online here or via PM

Bloody excellent to see you around here again. How's tricks?

O M G thank you EGA crew for another excellent weekend. So much fun and so very professional all at once <3 Righto, I didn't see many actual presentations cos I was largely freaking out trying to finesse my own ( shared ) upcoming workshops, but that's OK as a bunch of them are coming up on the EGA YouTube channel Any time I was outside I'd walk ten paces and run into someone I hadn't seen in five years and we'd get talking Which is perfect for me as that's exactly what I wanted to do. Personally EGA is, for me, about networking and catching up and trying to pitch in at that level of excellence that the other EGA crew show Also I got to hang out with Kaz at the EGA info tent which is always a goal for me But there's always something for everyone at an EGA, and everyone's EGA is accordingly different. Special thanks to Obtuse for sharing the cloning workshop facilitation ( halfa beta blocker rocked for me and was the first time I'd ever felt comfortable presenting anything ), mate you rocked and I would love to co-present another if you'd have me. The people who came along were wonderful and involved and we kinda went overtime but thankfully only into lunch break Special thanks to Torsten for saving my stuttery arse at our Experiment Design workshop when I was nervous even tho we had rehearsed it so many times. Same on the co-presentation, next time I'll do it better and actually get all the crucial points across Melbs crew, EGA crew, you rock. I had the best time. Your professionalism, compassion, humour, comprehensible cohesiveness and sense of fun... whoa... What a great weekend. Also: while in transit I musta eaten halfa VIC. The entire place is delicious. Wagon Wheels the size of yer head at Yarck, off-the-track pubs providing exquisite burgers for $$fuckall. Even the round trip is fun So: how was your EGA?

No wuckas, is a pleasure walking with you on the path, your thorough note taking, good questions and sharing results here is an inspiration Sorry i shoulda checked yr earlier responses This is very true. It also has a use by date. I know this because I used some really old stock once past the date and it wasn't effective. Lesson learned This is also true IME It often is, but not always. Depends on circumstance and your experience with the workflow for whichever species it is Ye gods it all sounds arduous, I feel your pain. Scattergun approach is probably best at this point and at least you're taking good notes and making sure you have backup material. It's hard starting cultures for a species new to TC when you have very little parent material, and heartbreaking. Worth it when it works tho No rush, let it grow back and be strong That's completely unscientific of you and is exactly what I do all the time too Nup, not down to me at all. I had a few years of having a good crack at aseptic culture, as did a well known TC local who had similar luck. Another young fella took the population pressure off the phlebs by identifying other easy to cultivate Australian acacias with a similar profile- so there was no longer a need to hunt the phlebs specifically in the wild. Then a third young fella found a rhizobium inoculant which promotes survival past the critical time point where most of the people cultivating it were experiencing progressive dieback which was ultimately fatal. Point being this project is a community effort. Not deliberately, but it didn't happen in isolation. And some outcomes ( like the reduced population pressure from wild harvesting ) were unanticipated and secondary to other research. And a good many people posted and shared results both formally and informally which allowed patterns to emerge and be followed. Lots of things didn't work- until a few did. I'm not 100% confident the species is out of the woods yet ( har har ) but it's looking to be in better shape wrt numbers and cultivation than it was 20 years ago. Keep an eye on it, research where you can, and understand that collective direct community input has the capacity to trump government grant applications for actually making things happen

Just confirming- those aseptic Acacia in tissue culture are Acacia obtusifolia. Dunno about priceless, they're as valuable as you make them Practice your deflasking, run simple tissue culture experiments on 'em, mutate the hell out of em, do what you like

I used sulphur burners on a timer with good efficacy in commercial planthouses. It's not 100% but it did reduce the amount of spraying I needed to do by 80%. Sulphur gas is toxic tho, you need to be sure once your closed room is treated the gas vents safely somewhere no-one is going to breathe it ( ie- innocent bystanders ) , and vented entirely before you enter the room. There may be species sensitive to the topical application of sulphur- I never worked with any. Over time ( a decade ) it may degrade some metals, like plant stands, slightly The presence of residual sulphur on plants may negatively impact some experiments, but so will a lot of other sprays ( residual fungicides and embryo work in some species is anecdotally one of these ). For TC parent explants everything is washed off prior to culture anyhow so that's not an issue here Look, nothing beats good planning, cleanliness and getting onto things quickly before pathogens go exponential ( which happens over about 36 hrs under optimal conditions for the pathogen ) It sounds too simple to be true and is often ignored. I did TAFE chemcert recertifications a number of times and they did emphasise this heavily as a preferred solution to pest management and they were spot on. Once I started Integrated Pest Management strategies my spraying times went to about 20% of previous A crowded plant space with a dirty floor, a clogged drain, poor airflow and overly nutritious propagation mix for your species is a bug buffet, and much of your effort will need to be redirected into sorting out pest problems. Also: Trichoderma. I know I rave about that shit, but the plant immune response is ( empirically ) worth it for me. At least I know which pathogen I'm fighting off at sterilisation time too. Double sided yellow sticky tape over anything which touches the floors or leads onto the benches. Like this: https://bugsforbugs.com.au/product/sticky-traps/ The yellow attracts some bugs, and the sticky prevents them climbing. Easy to manually inspect, if you see a bunch of bugs adhering to the trap, you have an entry point for them. Work out where it is and close the loophole. Inspect the plants for signs of infestation ( if the tape gets overloaded it will lose it's stickiness and become a ladder for more bugs ) Keep your plants from touching the walls or floors when using these, and wrap tape around any power leads which could let bugs walk onto your babies ( easy to miss if you are running heat mats )

That's the spirit! And yeah, that last bit sucks. It's hard to resist that temptation I know, but in the long run it's sooooo much better for both your work and the plant. Taking parent material from weak plants rarely results in a win for either of you Hey I can't remember, but were you initiating new cultures into 1/2 strength media with low sugar? Always a good start, low ( 50% or less ) sugar doesn't feed contaminants anywhere near as much as full strength. Gives your species time to recover. I've always believed explants retain some residual immune system after excision ( this is just a belief- I've never looked deeply into whole-plant immune response ). I've seen a few species carry residual contamination for yeeeeeeears without effect as long as there is PPM in the media- and have that contam re-emerge a decade later in the absence of PPM ( positive controls were in place, so yes, it's a thing ). PPM is a biotstat- not a biocide. I no longer always run batches of media with PPM, it's exxy. And while I was working this out I did lose batches of plants, so I got cautious and decided PPM needed to go into everything. Then I isolated the compulsory PPM requirements to a few species only, and also always keep a few of every species in PPM in case of equipment or operator error.

Experiment abandoned. I can get a kilo of Trich ferts for about $60, and if I aliquot it under sterile conditions that'll last for 2-3 years I don't need the bacto elements in the commercial blends, previous experiences with different commercial fungal/bacto blends have shown some deleterious consequences under some weather conditions with my target species ( long story snipped ). Have worked successfully with Trich nutes in these species previously- a fungal blend is overall better but I never did comparisons at a level to determine which one- I just know that without the Trich added to ferts the results were much poorer and I have the dead plants to prove it. The Trich is the constant Is gunna be quicker to get $60 together than it is to work out whether the home Trich LC mix is set back/ viable due to shock when diltued into non-sterile working strength solution, and whether that solution is producing comparable outcomes wrt plant growth Plus I'd need to buy the Trich nutes anyhow to run as a positive control

Am a huge fan of commercial Trichoderma spp blends as a liquid fertiliser addition to my home garden. The commercial products are usually sold as dry powders- spore mixes treated for storage. I have run right out of these and need some soon. I have Trichoderma spp cultures in my library, and I can easily do liquid cultures based on my standard fertiliser blend. Would the shock of throwing a growing but otherwise sterile culture, even at log phase, into the wild to compete with other organisms potentially any advantage? What sort of optical density of the LC Trichoderma would I need as a fertiliser base, to add as 10x, 100x or 1000x. Can I wing it or do I need to switch the spectrophotometer on? Anyone had any experience trying this?

So glad to hear tickets news, this is gunna be a corker. So excited. Thank you to the whole EGA team for your level of professionalism and commitment to the event. I'm seriously impressed by the amount of work, passion and consideration going into this. If you're sitting on the fence about a ticket like Ronny said I reckon you'd be insane to miss this. Something for everyone, every level of knowledge and interest in the fields. Everyone.

Thanks heaps for the clarification, it def adds weight and meaning to the numbers And thank you again for all your excellent, quality, long-term dedication to the species and issues around them. Brilliant.

Nice- must try that sometime I'm prolly not thinking this through properly, but I figure the commercial products w encapsulation directly penetrate leaf surface- once sprayed on they germinate. I have no idea if this is accurate. Spraying live unprotected log phase colonies onto plants even in low light might not be as effective as the light shock, sudden exposure to air etc. could kill them Might work straight into soil tho. Hmmmm... Duckweed... nice idea, thank you! That way I can avoid all those tedious petri dishes and centrifuging and resuspension and optical density measurements and just go straight to the guts of it- ie does it give visible growth results I don't mind re-application as that way I can be certain that precious plants will be OK, especially if I can make an ongoing supply for practically nothing. So competitor organisms aren't an issue Positive control will be commercial blend treatment. Negative controls will be untreated Duckweed, and the fert blend minus the Trichoderma. Space limitations mean the number of repeats per variable will be limited to 2-3 and I don't want to be running more than 10-15 duckweed containers for more than 2 months. Dammit I'll need to randomise their placement to make sure that light differences aren't fudging results :/ Hmmm, won't have time to do this before EGA. Still open to other improvements or teks while we're thinking about it Thanks heaps Crop!

Trich and other microfungal/ bacterial components have, for me, increased yield and decreased disease in a few species. Also used them for increasing strike rate with unknown success- I don't have a controlled climate seedling house so am at the mercy of the elements which tend to be a bigger factor overall I've used a few commercial Trich blends with great success but it's getting exxy. There have been a range of other components from Azotobacter to fucked-if-I-can-remember-I-lost-the-labels. Trich is by far my favourite ingredient and has been the constant. Bunch of mechanisms, many of which I don't understand. A couple which stand out in my specific experience are: I have a species with a strong tendency to die of fungal disease when stressed. When I foliar fertilise regularly with a Trich blend, the mortality rate for this species drops to almost 0%. Just an hypothesis, but I'm figuring the parts of the plant prone to environmental fungal infection are asymtomatic if infected with Trichoderma. The effect wears off if I fail to fertilise for a month or so- it's not a one-off treatment, there are competitor organisms ( I've got a locally isolated slime mould in-vitro which has the most fascinating battles with Trichoderma- when it's cool the Trich takes over the substrate and the slime mould retreats or is eaten- when it warms up the Trich is eaten and the slime mould grows super rapidly- fascinating to watch, happens over hours ). My tentative conclusion is the Trich in the soil succumbs to the slime mould and similar at low temps, allowing the local bloody pestilence to attack my plants if overwhelming amounts of Trich isn't re-introduced regularly Trich apparently evokes a mild stress response in plants, just enough to make them grow in response. No references, just something I've read Then there's the whole microfungi nutrient breakdown, saving plant roots some work in accessing them I would totally recommend trying this on at least some of your species- just a sample, no more than 30% of your stock on any species so you have some left if they don't like it

Anyone? Orbital shaker is on, I can easily run a scale-up on what I have, maybe make 100ml using malt extract + some sterile liquid fert components so it's not as much of a shock to the culture when I dilute it with non-sterile liquid nutes to spray Or does Trich need to be added as encapsulated spores ( ie commercial preparation ) directly into liquid nutes cos it only works as a foliar spray that way- throwing log phase sterile cultures into a non-sterile mix could potentially shock em to death? I can't even imagine an experiment around this which would be facile and robust

Yeah kill the fucken thing. I like the flowers but if they go into yr gutters the tank water tastes foul Time lapse video it, it'll be like Megashark vs Giant Squid I have to trim the vine here every few weeks, but it's just used a hedge. If I left it the thing will grow up a tree and make a dash for the powerlines in six weeks during the growing season Not sure if it'd do that where you are, depends on how much water it can get