Some of you might remember back in April that I had read about microwaving pollen to kill it and putting it on the stigma of a related plant, to break down the resistance of the stigma to pollen that it would normally reject? And in my mad enthusiasm I had put microwaved pollen from Aech. flavorosea onto a dark banded Aech. chantinii, with the somewhat confused notion that this might help it to accept pollen from Bill. Domingos Martins? I've included the link below if you want full details of the original lunacy.

Well, there have been some results, but, er .... what are they???

The pic below shows the chantinii flower spike in late September, approx. 5 months after I did the pollinations.

I had chopped it off the plant because the wind had broken the stem and although it was heavy with a nice fat berry from every flower, not a single one had shown any sign of changing colour to indicate that it was fertile. I was just about to throw it in the bin when I thought I'd have a look inside a couple of the berries - not because I expected to find anything, but more out of a vague interest to see what the inside of a berry was like. Anyway, I pulled off one of the berries that I had pollinated with only microwaved flavorosea pollen ( I had done a few of these as a control, to see if this had any effect by itself) and dissected the berry apart carefully so I could get a clear picture of what it was like inside. And, tucked away in one compartment of the ovary was .... a single seed. I thought "you beauty, a seed!". Then I thought " *&%^@, it was from a flower pollinated only with the nuked flavorosea pollen! %#^$, *&%#@, $#%^! Ah well, I might as well check them all out." So, I dissected every single berry apart carefully to find out what was going on.

Back in April I had pollinated some flowers with nuked flavorosea pollen only (as a control as noted above), some with nuked flavorosea pollen plus live Bill. DM pollen, and some with live Bill. DM pollen only. Taking all of these together, I had ended up pollinating 13 flowers in total out of the 61 on the flower spike. So, five months later there were 13 berries from flowers that I had pollinated, and 48 berries from flowers that I had not touched. Of the 13 berries that I had pollinated, five contained at least one seed, while of the 48 berries that I had not pollinated, not a single one contained any seeds at all.

You could argue that my pollination attempts showed this level of success purely by chance, but to have success with 5 out of 13 flowers when none of the other 48 produced results? Getting that sort of difference purely by chance is pretty unlikely - a bit less than one chance in ten thousand using some standard calculations. So, it is probably a pretty safe bet that the presence of seeds really was a result of me doing something to the flower, but what? I did a few different things, but what was it that I did that produced the results?

Looking just at the berries from flowers that I had pollinated, 4 had been pollinated with nuked flavorosea pollen only and 2 of these had seeds, while 9 had been pollinated with live DM pollen and 3 of these had seeds. So, 2 out of 4 compared to 3 out of 9? Any difference between those results could easily be due to chance. So, just based on whether or not the berries contained seeds, there really is no good reason to suspect that the DM pollen was any more effective than the nuked flavorosea pollen.

Bummer! Maybe a few of the nuked flavorosea pollen grains were still live, and they produced the seeds! Or, maybe my interfering with the flowers was enough to allow a few pollen grains from the chantinii to succesfully self-pollinate! (I was very careful to avoid selfing the chantinii and there were never any visible signs of any pollination activity apart from mine in the fully enclosed shadehouse, but ....). Or, maybe maybe maybe, a few of the DM pollen grains were actually successful as well? Or, maybe all three!

Hmmmm. Looking then at the numbers of seeds, the two "fertile" berries from nuked flavorosea pollen only had 1 and 2 seeds respectively, while the three "fertile" berries from the DM pollen had 4, 3 and 3 seeds respectively. So, 1 & 2 for the nuked flavorosea pollen compared to 4, 3 & 3 for the DM pollen. You could argue that getting that much difference could well happen just by chance, but it is starting to get a bit unlikely - a bit less than one chance in twenty using some standard calculations. So, although the numbers are small, there is some reason for suspecting that, maybe maybe maybe, the DM pollen resulted in more seeds being produced! And, maybe maybe maybe, some of them are actually chantinii x DM!

So, I planted the seeds in late September and here they are approx. 6 weeks later.

From the grand total of 13 seeds, there are five vigorous little seedlings on the way plus another two slow-pokes. I just wish I knew which berries they were from! When I found seeds in the nuked flavorosea-pollen berries I was so disappointed that I didn't keep the seeds from the different berries separate. Still a silly boy! Ah well, I guess I'll just have to wait until they grow through anyway before I get some clear idea of what they really are. I'll let you know in a couple of years.

Cheers, Paul

You're juggling three different species here, Paul, and that makes for a lot of variables. To do a more conclusive experiment, you'd need to have more controls: 1) left alone, 2) emasculated but otherwise untampered with, 3) intentionally self-pollinated minus any outside pollen, 4) nuked flavorosea pollen plus self pollen, 5) nuked flavorosea pollen only, 6) nuked flavorosea pollen plus fresh DM pollen, 7) fresh DM pollen only.

I think you're right to assume that it's probably not mere coincidence that only the tinkered-with flowers set any seed at all, but 1 or 2 seeds vs. 3 or 4 is really not statistically significant. If it was 1 or 2 vs. 20 or 30, that would be more conclusive. Since the purpose of the nuked pollen is supposed to be to trick a self-incompatible plant into accepting its own pollen, the DM element is kind of a red herring anyway. If you had a control with just the Bill pollen vs. one with both Bill and nuked pollen, and saw significantly more seed set with the addition of the nuked pollen, then I'd say you were onto something. Right now you don't really know if the simple act of emasculating the flower might have been enough to put just a few grains of self pollen in the right place at the right time either. I often see that with plants I have used as pollen parents. Sometimes they're fertile, sometimes not.

And yes, you silly boy, after going to all of that trouble it might have helped to mark which seeds came from which berries! Oh well, you can always try again next year. In the meantime we'll look forward to a possible xBillmea domingotinii!

I would have thought that you may have done better than 13 seeds, it will be interesting to see what they grow out like.

Cheers,

Rick

It's a very interesting post and the only way you're gonna know for sure is to grow them out and see. Meantime, enjoy the anticipation during the waiting period.

Long live the hybridizers!

All the best, Nev.

Interesting experimental work for sure, and well-recorded - although too complex for my non-mathematical brain! It seems to me that you HAVE written it all down. Pity about not knowing which of the solitary seeds came from which recorded experiment - but time will prove your suspicions.
Good luck with the results down the track. I hope you get a potent xBillmea 'Martinii' - cheers!

K :)

Lisa, you are absolutely spot-on as usual :))). As a scientific experiment it is a pretty good example of the confusion you get when you don't plan it out properly before you start. The whole thing has been a bit of a giggle, even wheeling out some old statistical analysis tools that I haven't used for the last thirty years or so. I never thought that I'd find a recreational use for those, let alone have some fun doing it!

But, fun or not, I must admit that I was really annoyed with myself within seconds of mixing those seeds up. What a nong! Jack, I am normally an absolute stickler for keeping track of things like that as you suggest and keep telling myself over and over. What a case of brain-fade!

And Rick, I agree, the very small number of seeds certainly does have me wondering - along a few different lines as Lisa indicated. Nick, it is interesting that you've found that to happen as well, and Nev, you're right, the fun continues with the anticipation. Nothing like having a few un-answered questions to make life interesting!

Thanks for your comments, and Lisa, I just love the name. I think you have just claimed naming rights if anything worthwhile eventuates. I'll let you know!

Cheers, Paul

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