Acacia phlebophylla TC genetic contamination

[Darklight]

This is just a quick note- I'll expand on it when the main festivities are over.

The Acacia phlebophylla TC project is once again a disaster.

After eight years ( not unusual for TC projects- the Wollemi pine TC project took seven and they have more toys and staff than us) we believed we had a successful replication and rooting protocol and released a batch of TC plantlets to SAB to deflask as a trial.

None of these plantlets had developed adult leaf morphology while in TC- but that's not unusual with plants generally nor specifically with acacia, as during TC the ideal is to keep plants as morphologically young as possible so that tissue can spend energy on replicating rather than development and differentiation. In short, the plants looked great and were consistent in form throughout the years.

SAB nursery staff quickly noticed that morphologically adult development did not evince- in other words, the typical rounder shape of the adult leaves did not develop; they stayed thin and sickle shaped. There was no reason for this to happen, as no hormones had been used some of which might persist and affect morphological development for some time. We waited- and nothing happened. Adult leaf formation occurs pretty rapidly in phlebs.

Torsten and I went back through the seed bank and found that I had inadvertantly included a pack of maidenii seed in with the phleb batches- the label had rubbed off. Must have been happening for about 12 months since I did my last seed bank re-org through here. Boo hiss, but no problem overall as the number of seeds germinated and plantlets kept from each batch exceeded the number of maidenii contaminated genetic stock I used each time ( I germinated 20 each from 5 separate bags of seed and kept 30 of the best for parent stock ). The problem was that the rest of the parent stock- at least ten per batch- showed identical morphological traits and none of those were consistent with phlebophylla.

After germination tests from both mine and Torstens seed stock we concluded that most or all of the seeds we were given were NOT phlebophylla, despite being labelled as such. I've been working from this stock ( received from a single 'reliable' individual ) since 2000/2001. And the implications for the work we've done are appalling. It means that the protocol we have spent so long to develop has to be retried on new and confirmed phlebophylla seed, and may not be successful so we could be in for years more fiddling and hoping.

Protocols for species- especially those as diverse as acacia- and in some case for individuals within a species- can very huuugely. From experience and having worked on 8 acacia species in the past you read the protocol for a related species and then fuck with it for a year or two if it so much as looks like a goer. Or you give up.

We're not giving up. We've put too much work in it. I have no idea as to what we'll do next, because I'm back to the stage of mistrusting any seed source we get as being the correct species ( a common problem in our field as I'm sure many of you are aware ). And I'll be afraid for the fate of any seed we do get in case we have to risk running a wide range of media should the formula we have developed fail. But at the moment I'm still devastated, and I'll want to sulk for a bit until I regroup for another round

Thanks to all who supported and followed this project over the years- I'm hoping 2006 will bring better fortune. I'll fill you in on progress when we have some. But really, I could weep ... https://www.shaman-australis.com/forum/applications/core/interface/imageproxy/imageproxy.php?img=&key=ed93ee4b8a158835e0af19ead9c794b11af03de360911f7858b9da338588c45d

https://www.shaman-australis.com/forum/applications/core/interface/imageproxy/imageproxy.php?img=&key=ed93ee4b8a158835e0af19ead9c794b11af03de360911f7858b9da338588c45d

[Stonehenge]

Don't worry about it, DL, stuff happens. I would not say it was a total loss al all. The experience you gained doing that will stand you in good stead for years to come. Plus, you now have your procedures down pat for maidenii. Your work with a. phleb will go much faster now that you have worked out so many of the intermediate steps. Good luck and keep your chin up.

[Rev]

sorry to hear it DL

might i suggets a winter ski holiday/ summer cool mountain holiday on mt buffalo to recuperate

itd give you a chance to see and feel the environment up close and tune in

get some new ideas over good food with some company

you can also collect soil samples, leaf samples and maybe your own verified seeds if youre lucky

it may be a good idea to take the soil samples from the root zone, cut with moist peatmoss and use that to inoculate both any wholeplant phlebs and also some maidenii as bait plants.

in the soil will be the correct rhizobia as well as VAM fungi spores (hyphal fragments remain viable inoculum for up to 2 weeks)

if these are placed in contact with acacia phleb or related species in pots of a sandy media they will act as hosts

they use this method to capture and breed up VAM spores using clover (T resupinatum) as a bait plant. Clover too may be suitable as most VAM are broadly adaptable

[AndyAmine.]

Sorry to hear about the mix up DL, must be very disenchanting.

What goes around comes around mate, perhaps you have recieved this bad luck because something really good is around the corner?

[planthelper]

of the ashes of disaster, growing roses of success.

sounds to me just like one of those things which do happen, and at hindside will just add hallmark to a legendary, earthshattering story of how dl (or "love" as she preferes to be called by the author

<___base_url___>/uploads/emoticons/default_rolleyes.gif ) gave life to the rarest accacia on our planet.

tragedy+time=comedy!!

[Torsten]

...or "love" as she preferes to be called by the author...........

LOL, I hope you have your asbestos undies on!!

tragedy+time=comedy!!

This is a lot easier when it is your own fuck up, but not as easy when it wasn't your own fault.

[-bijanto-]

'Accept the challenges so that you may feel the exhilaration of victory'

- General George S. Patton -

[Flip]

Dl

You have my most sincere understanding and support

Please keep up the battle!

Don't give up!!!

[Benzito]

How crushing!

You guys think about what you we're doing in 2000, and where you are now, in relation to that.

Now imagine that you had to undo it all and go back to where you were before.

I would still be working in a pet shop, rather than my current job, earning a lot less, and I would still be a very immature young man fresh out of high school, without the loving partner I now have.

This may be an odd comparison, but basically that is what has happened for DL. Everything she thought she knew may now be totally wrong.

Best of luck Darklight, I really hope you get your phleb protocol up and running, and let's hope that whatever seed you were working with, is similar to the phleb's. Maybe you won't have to change anything.

I've got my fingers, and other bits, crossed for you.

[mescalito]

DANG!

..and I get upset when the cat uproots my only Loph <___base_url___>/uploads/emoticons/default_blink.png

Maidenii's not too bad a trade off though.I'm sure I'm not the only one that would love more maidenii's around their gardens!!

I'm with Benz though I just spent 5 hard years working a protocol that should have worked...

but she was mis-labelled too...I reckon my protocol's spot on now so I too am awaiting the real deal <___base_url___>/uploads/emoticons/default_newimprovedwinkonclear.gif

Take it easy mate

[Darklight]

Thanks for the kind words everyone https://www.shaman-australis.com/forum/applications/core/interface/imageproxy/imageproxy.php?img=&key=ed93ee4b8a158835e0af19ead9c794b11af03de360911f7858b9da338588c45d

Maidenii's not too bad a trade off though.I'm sure I'm not the only one that would love more maidenii's around their gardens!!

Back when we started in 1998, we developed a protocol for maidenii. Lots of seed suddenly came on the market so TC replication wasn't economically viable and really, seed grown plants keep the gene pool diverse. So we didn't keep it going. The potential is there to replicate the high yielding maidenii, I'm not sure overall what the percentage increase is ( and whether it is merely related to climate or sampling time ) so maybe it would be relevant for plantation work. But maidenii isn't endangered or rare ( except in Vic ) and endangered and rare is what we do most and best

Because, as I've said previously, TC protocols can vary just as much for individuals within a species as they can between species, and because the number of accepted parent plants was higher than the number of seed grown from each batch, who knows what the fuck did end up growing well. With 25 seed per batch over 4 bags ( total 100 seed ), and 30 of the best selected overall as parent plants you can bet at least 2 species were mixed up in it.

The only way to sort that out was via expensive DNA testing, and really, it wasn't worth it. It's quicker to ditch the lot and start again with confirmed seed than it is to try to identify which of many potential species the labelled phleb seed that went into TC is and hope its useful

I'm with Benz though I just spent 5 hard years working a protocol that should have worked...

The protocol worked, the error was unfortunately human. But if there's anything you need mesc just let me know

Take it easy mate

Thanx, am trying https://www.shaman-australis.com/forum/applications/core/interface/imageproxy/imageproxy.php?img=&key=ed93ee4b8a158835e0af19ead9c794b11af03de360911f7858b9da338588c45d

[RonnySimulacrum]

DL your hart is the right place, you are truly dedicated, thing will work out for you and the plants

Hug

[poo]

Wow that's such a terribly frustrating story... really feel for those involved! So much time...

Really is great to hear that you've made the positive decision to keep going and try again - obviously we all appreciate it!