Fertility

Continuing on this discussion from here:

http://davesgarden.com/community/forums/t/920110/

Betty, figure we could start on this thread, so that would leave Eric's thread open for his questions and hopefully he and anybody else who wants can join in on this one.

Ok, went hunting back. The one day when when I was out tryign to hybridize and mind you my main work is with Daylilies, I asked asked on my other stomping grounds about a pistil that was spilt into several sections and was wondering if it could be pollinated and if it would take.

This what one of our mentors Admmad from Canada wrote back to me.

"In the normal pistil the three sections are fused. This is important as the pollen tube does not grow through any tissue. When the three 'styles' fuse during development a special channel is left open in the middle and it is through this channel that the pollen tubes grow.
Unless something quite different happens when the styles remain separate it is unlikely that the pollen will be able to reach the ovary and fertilize the ovules. That is, to do so there would have to be an open channel within each of the three stylar sections.

I think you were correct about the extra styles. They are probably part of a developmental problem or an environmental effect, etc. Or perhaps they are related to polytepals genetically.
In any case, just as there are three lobes at the top of the style which apparently lead to the channel, one lobe further down the style that also leads to the channel would work. The qualifier is, does that extra stigma lead to the channel and does pollinating it result in seeds in all four/five, etc chambers and does not pollinating it result in one chamber without seeds (or does pollinating the three stigma lobes at the top cause all the chambers to be pollinated in the ovary because the channel leads to them all.
I know I have pollinated the extra stigmas and found pods with extra chambers and seeds - but in the years before I noticed the stigmas lower on the style I also sometimes found extra chambers with seeds and in those years I would not have pollinated the extra stigma. "

Then I asked, "That is, to do so there would have to be an open channel within each of the three stylar sections.
Would it be common to find an open channel in each one? For future reference.... If I cut open one of the extra styles would I be able to see the chamber or not and would you want to make a horizontal or a vertical cut? What are the chances of having more blooms on the same plant producing the split styles or is it just a once in a while odd thing. I wanted to get the cultivar because of its stules to experiment and play around with, but if it is not gonan continue to make the split styles then it reallly won't do me any good, unless you think a hidden gene for it may pop up somewhere down the road. "

The reply to me was again from Addmad was, "Would it be common to find an open channel in each one
I don't know. Unfortunately there only seems to be one research group active in the field of style/pistil development and they work on Catharanthus (periwinkle). Apparently there are two basic groups of style development in plant species. One is described as 'phylogenetic' and in those species one style grows from the ovary in a compound state. And the other is described as 'ontogenetic' in which multiple styles grow and then fuse. But the process apparently can be different in each species. In Catharanthus the styles fuse after growth and the researchers are interested on what causes the epidermis to fuse and to dedifferentiate. There is not much information otherwise about the anatomy. There are also two different types of styles in plants - more or less solid versus a central channel.
I would expect that Hemerocallis is like Catharanthus and grows separate styles than then fuse. Continuing that line of speculation I would assume that when a flower has separate styles that each style does indeed have its own central channel.
If I cut open one of the extra styles would I be able to see the chamber or not and would you want to make a horizontal or a vertical cut?
You should be able to see the channel but you may need to use a vital stain to see the living cells versus any fluid that may be in the channel. You would need a sharp razor or perhaps a microtome to make slices and perhaps trying in both directions might be worthwhile.
What are the chances of having more blooms on the same plant producing the split styles or is it just a once in a while odd thing.
For some cultivars it may just be a 'random' environmental developmental error. But other cultivars may have a prediliction for reacting to the environment with unfused styles. You would need information about the behaviour of the cultivar over several seasons and many flowers to know which is correct for any specific cultivar. "

So now my question for thr brugs is, what kind of formation is the pistil. Are the brugs phylogentic or ontogentic? I wonder if there is some online source that maybe somebody has done some disecting and documentation of the brug pistil?

Now sometimes it possible and happens for me a lot with seedling and not knowing for sure if they a dip or a tet, that I get the wrong type pollen the first time around. So then I wanted to know about redabbing and again this is what Admmad replied with,

"I would *think* that the initial pollen would have started growing tubes and would block subsequent pollinations.
With a poor pod setter we don't know the reason, so it could be that the stigma is not receptive at the usual time. Pollinating the day before, the day, and the day after the flower opens covers all the possibilities.
When researchers make crosses between species the crosses often fail. But apparently, at least for some of those types of crosses the success rate is higher if the flower is pollinated before it opens or much later in its life rather than at its peak of receptivity.
[Speculation] When the pistil is past its peak receptivity and breaking down there may still be enough moisture and other necessary components that pollen can germinate and pollen tubes can grow. It would then be a race between pollen tube growth and pistil decomposition.
The growth of pollen tubes does not physically block subsequent pollinations. In other words if I were to place five pollen grains on the stigma in the first pollination and then later place another five pollen grains on the stigma all ten could fertilize ovules. I could do this until the number of pollen grains was the same as the number of ovules (which I counted as between 30 and 36 last year). This assumes that all pollen grains are viable, germinate and fertilize ovules. That is what may occur naturally with pollinations. The first pollinator accidentally drops some pollen on the stigma, and later pollinators may also drop more pollen on the stigma, and so on.
However, if the first pollination is successful and an overabundance of pollen is placed on the stigma then it is possible that all the ovules are fertilized by that pollen leaving none for later pollinations. "


Now this might help in the pollination of some of those brugs that folks say are difficult or not producing pods and Betty, might also answer your question.

Also, not sure if yoru able to acess this article or not..

A. G. Stephenson, Annual Review of Ecology and Systematics 12:253-279 1981

Here is an interetsting part of it that relates to seed reduction and pod abortion. The word pod has been injected next to the word fruit.

"In some species, fruits [pods] from self-pollinated flowers tend to have fewer seeds and are more likely to abort than fruits [pods] from cross-pollinated flowers"
"When few seeds are present in a fruit [pod], hormonal activity is reduced and with it the ability of the seeds to attract resources. Consequently, more seeds abort due to limited resources, and a positive feedback system is established which culminates in fruit [pod] abscission."
"Thus the evidence that flower and fruit [pod] abortion are a response to limited resources is prodigious."
"The evidence suggests that fruit [pod] maturation is selective. Depending upon the species, pollinated flowers and juvenile fruits [pods] may selectively mature on the basis of the order of pollination, the number of developing seeds, pollen source, or some combination of these."
"With few exceptions, however, these studies indicate that fruits [pods] abort prior to the midpoint in the period of maturation. In most species, over 90% of the abortions occur during the first third of maturation"
"Fruits [pods] that abort after the midpoint in the period of maturation are usually severely damaged"
"When a juvenile fruit [pod] is going to abscise, the production of growth hormones by the seeds diminishes and the amount of growth inhibitors, abscisic acid and ethylene, increases in the fruits [pods]"
Leaves donate carbohydrates and nutrients to fruits [pods]. Although fruits have the potential for attracting resources from leaves that are a meter or more away, there is a strong tendency for resources to flow into fruits [pods] from the nearest leaves
many plants (a) selectively abscise damaged fruits [pods] and (b) initiate more fruits [pods] than can be developed to maturity with the available resources. The “surplus” fruits [pods] abort.
Unusually high temperatures also promote fruit [pod] abscission, although it is not known whether heat directly or indirectly kills the seeds. Very young fruits [pods] are generally more susceptible to damage than older fruits [pods].
Lastly, Janzen and Chamov have reasoned that if offspring differ in some quality related to fitness (e.g., paternal parentage), and if plants can selectively mature “high quality” fruits [pods], then the production of surplus flowers is advantageous because it provides the maternal parent with a “choice” of offspring to mature. Of course, the increase in fitness due to selective abortion of “low quality” fruits [pods] must exceed the decrease in fitness associated with the costs of producing surplus ovaries and fruits [pods]. This hypothesis is in many ways analogous to mate selection in animals except that it occurs after the male gametes (gametophytes) have been transferred to the female rather than before. Except for studies that show that -- some species selectively mature fruits [pods] based on pollen source or seed number, this hypothesis has not been investigated.

Now in Daylilies you can have a nice firm pod that has progressed normally along, but when you open the pod up, you will find , that usually, not allthe ovules have matured, so some other possible underlying factor has to be at work.

Here is some more from that article,
"When a juvenile fruit [pod] is going to abscise, the production of growth hormones by the seeds diminishes and the amount of growth inhibitors, abscisic acid and ethylene, increases in the fruits [pods]"
Leaves donate carbohydrates and nutrients to fruits [pods]. Although fruits have the potential for attracting resources from leaves that are a meter or more away, there is a strong tendency for resources to flow into fruits [pods] from the nearest leaves
many plants (a) selectively abscise damaged fruits [pods] and (b) initiate more fruits [pods] than can be developed to maturity with the available resources. The “surplus” fruits [pods] abort.
Unusually high temperatures also promote fruit [pod] abscission, although it is not known whether heat directly or indirectly kills the seeds. Very young fruits [pods] are generally more susceptible to damage than older fruits [pods]."

Ok, have I got you totally confused yet? LOL. I get myself all confused and twisted all the time as new thoughts constantly pop up.

I have found the pictures of the pollen tubes being developed and the crossection of the chambers and am going to email the photographer for permission. Sheesh had to go back three years to find em. That ok, it good to bring things back into review. I sometimes think that we learn a procedure, it works and so we don't change it and with things changign so much in the environment and new discoveries being made in hybridization techniques that it good to learn alot of different opinions and ways for havign success with our plants.

Don't know how long it wil take fo rme to get an email back from her, but hopefully it won't be too long.

Well, I think I done hogged enough space up for the moment. hehehehehe

No way hogged anything ...that is really interesting and I understand quite a bit of it ...what an interesting subject. May add that one cross pollination was no good with the blooms exposed to the hot sunny side of the tree and the pods done the same way with the same cross at the same time, but in the shaded side of the tree were very successful. It was done very early in the morning so it was cool when I did the cross.I think high heat does damage the devoloping seeds.
I do recall reading a discussion about different pollen grains not fitting the tube shapes and sizes ...I can't remember where that was sorry, thankyou for starting this thread ...great work!

Starlight, thank you for posting portions of that article. Many of the scientific articles are available for purchase only. I understood it. Just had to rethink some terms and the way they were used

As I understand this portion, it is very similar to a self thinning lemon tree. Retaining only what the plant can support. Eric mentioned that many seedpods containing seeds pollinated by incompatible pollen aborted. The process mention above couldn't be responsible for responsible for the aborted seedpods Eric is seeing. As I read on after this quote, it said those pods aborted after the "midpoint" were due to damage. I wondered if perhaps these "foreign" seeds were somehow unable to produce growth hormones thus being responsible for the abortion. Maybe considered damaged. Then I realized I didn't know if two months was before or after the midpoint for the pod.

Chrissy... Do you happen to recall at about what the temperature was when you made those crosses . I trying to put in my little notebook all kinds of little things that might help me. I'll put somethign heat pod problem, shad emade pod and then if you happen to have temp can make note next to it.

The discussion we were having about pollen grain sizes and which is better more or less pod and the number of seeds is here on Eric's thread.
http://davesgarden.com/community/forums/t/920110/

I have a tendency if you haven't noticed to be chatty at times and get off track, and figured I better move on over to a new thread. : )

bettydee...I get the same inpression you do like about the lemon.

You wrote:

Eric mentioned that many seedpods containing seeds pollinated by incompatible pollen aborted. The process mention above couldn't be responsible for responsible for the aborted seedpods Eric is seeing
As I read on after this quote, it said those pods aborted after the "midpoint" were due to damage. I wondered if perhaps these "foreign" seeds were somehow unable to produce growth hormones thus being responsible for the abortion. Maybe considered damaged. Then I realized I didn't know if two months was before or after the midpoint for the pod.

No, it may not be what Eric is seeing. It could be that maybe what he is seeing is that the genes are such that maybe they most of the pollen is sterile from lack of a chromsome or two or something making it sterile. Have you pelled a ripe pod opened and looked at the order of the seeds?

I wonder where in the pod the seeds are the plumpest and the fullest. I don't have a ripe pod, had to cut the pods all off to save the cuttings from frost. There is alot of seeds in that one pod. In the Daylilies usually the seeds at the bottom of the pod are the fullest and the biggest with the smaller being at the top, all though you cna open a pod up that ha slike maybe 14 seeds in it that look viable and you will 8 or 9 tiny black spots where the seeds started laying down their coat and then aborted.

I gettign from Eric, where we need to get those first few seeds to be compatable pollen that would attract the hormones to the pod and then get a few of incompatable then a few good pollen and kind of hope the weak pollen would be able to kinda suck up some of the hormones and nutrients needed to fully develop which seems to be a very hard and tricky process. Maybe that is why you pollinate with yoru mix, and then come back further up and do it again. Still don't understand that concept.

I have no idea when midpoint is for the pods. Maybe Eric or somebody else has records of timing and development of pods they can contribute.

Think I gonan head and lay down. Midnight is not a good time eveidently to be eating sausage and egg biscuits with chololate milk. LOL Se e ya in a morning.

Ok re the temperature ...it was aprox 85F/90'sF when I first did my crosses. After harvesting the first pods, the ones sitting in a fair bit of sunshine gave me pods which looked great but had many empty corks ...however the ones done on the same tree in the shade a much cooler area ...I don't know exactly how much cooler, were full of big plump happy easily germinated seeds.I found the crosses made on many different trees certainly all took very well in the mid Autumn (in other words cooler temps in general) these pods went through the winter outside ok, even though we had frosts because the pods themselves were not frosted.I would not cross seeds again in warm temps.I think they like moderate conditions for optimal fertility (I speak only from my own personal experience and zone conditions).
I hope this may help you and anyone watching ...my suggestion for full sun plants would be to only pollinate the lower sheltered blooms as the upper ones may not be able to tolerate hot sun and set the seeds.

Brugmansia species/natural hybrids are divided into two groups: The B. aurea Group (aurea, insignis, suaveolens, versicolor, and x candida (aurea x versicolor) and the B. arborea Group ( arborea, sanguinea, vulcanicola, and x flava - (arborea x sanguinea). The species within each group are compatible and readily hybridize with each other. Normally, members of the aurea Group are not compatible with members of the arborea Group. Apparently, with a lot of manipulation and chemical intervention, it is possible to get a few crosses that grow. Now this is where I got in trouble with Eric, by saying that the crosses were sterile. He says they are not very fertile. To make them fertile takes a lot of intervention. As far as most hybridizers and growers go, if the seeds were to grow, they would be sterile because we wouldn't be using the chemicals it takes to make them fertile. In the animal kingdom, if you cross a horse and a donkey, you get a sterile male mule. In fact with most animal interspecies mating, you end up with sterile offspring because of the differences in chromosome numbers. I know, I'm rambling, but I need to relate to what I'm most familiar. What Eric is suggesting/doing is to crossbreed a member from one group with a member of the other. You may be right in blaming the difference in chromosomal numbers as the reason for sterility, or near so. I don't know enough about plant genetics to hazzard an explanation. I could guess, but that's all.

This my only photo of some seedpods (Audrey Hepburn). These were just a few of a whole slew of pods she produced one winter. She must have had almost 20 pods. Notice that the pod at the top is not full. Two of the pods that ripened later had all the corky capsules, but most of them were empty. I didn't pollinate any of those pods. I had bees and a sphinx moth or two in the greenhouse that winter. I"ve never peeled an entire seedpod in order of pod placement to see where the plumpest seeds are. I recall reading an old post in which someone said said size was not an indicator of plant size, health or beauty in bloom.

I read somewhere that it takes three swipes with the brush to ensure pollination.The same article stated that the bees contribute just enough DNA to trigger a response ...I think this is why there is more success with "unopened" blooms ...due to beating the insect triggers.I am only guessing. Perhaps a large insect will carry enough pollen on it to do the job, here in my garden I never saw a pod until I used the artist brush ...success! except for the ones in the very hot area.It is possible the bees got there before me but I don't think so because another time I was too late and there were bees all over the flush of blooms ...they all dropped the ovary though in a few days ...while the ones in the hot spot (that I had pollinated) went on to form perfect pods with perfect corks only to be empty.Since learning from that mistake I have no longer experienced empty pods ...lots of happy seeds instead.
About mixing the genes ...I am not clever enough to understand it all yet but I am learning.

Peeled and perfect

I tried to get back on Dg this mornign and it just wasn't to be either my puter or Dg wouldeat my posts when I tried to send. Cyber space finally must have got full sense I seem to working now.

Crissy. Thanks. That is good info and sure to help us in these warm climates. Could be a factor why some Folks in the South have problems.

When you said the cooler pods had more seeds. I wonder with the pods being inside a shealth, hoep that the right word, if not plese correct me, with the high temps and the heat build up maybe the respiration is high and less seeds woudl mean that the seeds may be starting to lay down the coat , need to find which chemical that is, and then are not able to finish it and so there are the aborted seeds in the pod.

Betty... It a real pain sometimes when you want to try and learn about something and all the articles are in online paid subscriptions. Some I can get too and some I can't. Those then I try and find some friends who might be have subscriptions and access them for me. Nothign more frustrating than readign an abstract that seems promising, paying 35 bucks for it only to find out there nothing you can use in it.

I don't have but a tiny bit of gentics too. It a very complex subject and I constantly having to run to friends with a gentics background to get something explained to me or broken down to a level I can understand.
Guessing is what the world is about. We don't know, we question, we guess and we hope that others may have asked the questions already and will share. With questions and guesses comes discovery. If the question wouldn't have been asked about the white and funny colored seeds alot of would never have known about the chemical that lays down the coat even before the seed processign begins.

Which I wonder if the brug seeds do that. Since they have a hard coat. Does the hard coat have to start developing before the actual seed. The the question would be what factors would interupt this coating and cause the seed to abort.

I know that with some plants folks have a tendency when they are blooming to give them extra nutrients. Some times those extra nutrients are detrimental to seed production.


Betty you wrote. " As far as most hybridizers and growers go, if the seeds were to grow, they would be sterile because we wouldn't be using the chemicals it takes to make them fertile. "

That may or may not be true. I don't know for sure. Somethign a few years back that ws learned back with Daylilies when I was first starting to get albino seedlings and white seeds is that the seedling will of course die without intervention, but that seeds that were white, light brown and even tan in color where the black coating had not covered them germinated and actually when the first leaves emerged they were greener in color than the normal seeds. Til this time folks were generally throwing those seeds out. Now those seeds did grow up and a few years later they did bloom and made pods and seed. Some were fertile and some weren't.

Do brug people generally disgard white or immature seeds? Does anybody go ahead and grow them out? Now there have been cases where folks thought the plants were sterile, but jump starting the first pod with super pollen go tthe rest of the pods going.

If there is problem with that cross, maybe jump starting it with the first compatible pollen would kick the genes in , or then it use Eric's 50/50 mix.

Usually when I have a pod abort it with in the first week, sometimes I have had air pods. The pods will perfectly normal and develop in size but when you open them up there is nothing. You can see the teeny tiny black dots where the black coating was first going to be laid down and then aborted. I stil have not really found an answer as to why the pod didn;t shrivel and fall off like it normally should. The pods were totally healthy and firm in every aspect except no seeds or if you were lucky maybe one or two tiny ones, which you hoped would germinate.

Yep,remember that about a horse and donkey together being sterile. There are thousands upon thousands of plants that you cross this one and that and you cna get them to make seed and they are sterile, which is why they become a PPAf, (propagation prohibited) and are only propagated vegetatively.

But, and this something I had to ask some field experts about. I forget what I was working on at the time. I think I was tryign to take a type of cucumber from South Africa and try and sweeten it up by wanting to cross it with a sweet watermelon. I was told that the probability of doing it would be nearly impossible as the two were too far removed down the genetic line. They also told me though that with cousins and distant cousins there is the chance of getting them to cross. It not easy , take the right combination and you usally have to do some embryo rescue.

In nature in the wild, you see say two different but similar species, here and there. Now things happen in nature, say a fire or some other disaster befalls species one which was very fertile. Species two is left and not very fertile. Species numebr two has two choices. it cna either go into extinction or it can adapt. Plants and animals have the unique ability I belive over time to genetically change and adapt. I would guess that over time even your sterile animal would/or could become fertile.

Bear with me, cause I can ramble and jump around too. This person also told me that while some pollen may be seem incompatiable, not all of it may be because of recessive genes. That to keep trying and keep trying. That at some point you could get lucky and make that seemingly incompatible cross. I know persistance works because of the hybrids they have created from crosses that shoudl have not worked or taken. Which brigns us back to why Eric was sayign to use the 50/50 mix and pollinate alot of blooms and why it important to use and keep the species of plant lines going too. Where two specie s may not seem to work, there could always be that species out there of the incompatible one that has a bit of a different gentic structure and woudlpolinate real easilier than its brother or sister. Some of them seeds from a 50.50 mix will abort in the pod. some may form and never germinate, but a select few may germinate and grow and turn out to be somethign spectacular, a new hyrbid worth introduction.

Crissy. Lots of happy seeds there. Daylily seeds are called black pearls, do brug seeds have a special name? : )

Is there a close up pic of the brug pistil anywheres? If you heard they need three swipes maybe they have three chambers. I do know that the insects that pollinate can't pollinate alot of plants because like ya say they too small and those bee pods show it, because they may get pollen on part of the pistil that would provide some fertility, but not enough and so you have smaller amoutn of seeds and also may not even have that type of flower pollen on their bodies if they been out field hunting. I think like nature sway of policing.

Ahhhh Crissy, you driving me crazy with the pics of all them seeds. LOL I am a seed aholic and now you got me where I gotta go plant something. : ) great job of hybridizing. : )

I had to laugh at the comment. All mules are male.

I had trouble getting on to DG also. Maybe traffic was heavy trying to get on.


The article you quoted said very young seeds were vulnerable to to heat damage. Also developing seeds produce growth hormones which do two things: improve recources procurement and inhibit the production of chemicals that cause the pod to abort. Will you explain what you mean by "lay down the coat". Are "aborted seeds" seeds that didn't form? If so, seeds don't develop because the ovule was not fertilized.


http://en.wikipedia.org/wiki/Seed
I think you are referring to the corky material covering the actual seed rather than the actual seedcoat. Look at the link above. Cork covers the seed so think of the seed as having an extra outer layer. I've cut into some young pods I have had to remove. It looks like both the corky covering and the seed develop at the same time, that is, after the ovule is fertilized. Seeds need to reach a certain level of maturity or the seed is not viable. It's like expecting a 6 month human embryo to survive outside the womb before it is ready. As long as the seed is mature, whether the seedpod is ready to be picked, the seeds should germinate. I don't think I've ever seen a daylily seed, but mention differing seed coat colors. I peel all the corky covering off my Brug seed. The seed coats all appear to be the same beige color.

About seed fertility with regard to crosses made between the two Brug groups. Using ordinary, commonly used seed propagating techniques, the cross between members of the two group doesn't work. I don't know if an embryo fails to form or whether the seed dies young. I think this is why the seedpods are aborting. Go the the first thread in the Brugmansia Forum, the one written by Dave regarding those two groups we are not supposed to mention in our posts. Go to the website of the group that begins with the first letter of the alphabet (The contortions we have to go through.) In the External links box, go to the public information site, then click on Site Map, then scroll down to About Brugmansia and click on it. This is an abreviated explanation about Brugs. There you will find reference to the seven Brug species and two natural crosses. "There are 7 species of Brugmansia which are divided into two groups. These two groups normally do not cross with each other"
While you are in that site, take a look at Brugmansia Anatomy where you can see a photo of a pistil.

These two groups are not compatible like tomatoes and tomatillos are not compatible, but use enough genetic manipulation techniques, such as chemicals or irradiation, and you just might get a viable seed. The question would still remain. Will the offspring be fertile or sterile? This is what I meant by my comment about the importance of such attempts to hybridizers who don't use extraordinary means.

Moths are the pollinators of choice for Brugs, that is why they have their best fragrance at night. You should see the size of those sphinx moths. They are about the size of hummingbirds and sound like them, too.

[quote]In nature in the wild, you see say two different but similar species, here and there. Now things happen in nature, say a fire or some other disaster befalls species one which was very fertile. Species two is left and not very fertile. Species numebr two has two choices. it cna either go into extinction or it can adapt. Plants and animals have the unique ability I belive over time to genetically change and adapt. I would guess that over time even your sterile animal would/or could become fertile.[/quote/
Now we get to a subject with which I am more familiar. Here genetic diversity has to be present if a species is to have a chance at surviving a catastrophe. Also, there has to be enough time available for the selection process to occur. Both have to be present or the species will go extinct as happens so often. A good example: The Chiquita type banana. For centuries, these bananas have been propagated vegetatively because they don't produce viable seed. So they are all genetically identical. If a disease kills one it will kill all.
http://www.plantmanagementnetwork.org/pub/php/management/bananapanama/
Now the Cavendish type banana is also in trouble and it looks like it is running out of time.

Bettydee, surprise, here's something for you about mules from Wikipedia.

"In its common modern meaning, a mule is the offspring of a male donkey and a female horse, which is classified as a kind of F1 hybrid. The much rarer offspring of a male horse and a female donkey is called a hinny. The term "mule" (Latin mulus) was formerly applied to the infertile offspring of any two creatures of different species. The mule, easier to breed and usually larger in size than a hinny, has monopolized the attention of breeders. The chromosome match-up more often occurs when a donkey is the sire and the horse is the dam. Sometimes people let a stallion (male horse) run with a jenny (female donkey) for as long as six years before she becomes pregnant. Mules and hinnies are almost always sterile (see fertile mules below for rare cases). The sterility is attributed to the differing number of chromosomes of the two species: donkeys have 62 chromosomes, whereas horses have 64.

A female mule that has estrus cycles and can carry a fetus is called a "molly" and can occasionally occur naturally as well as through embryo transfer."

There's then a list of some remarkable and "unnatural" breeding results involving different combinations of donkeys, horses, and fertile male and female mules. When it comes to reproduction and the insistence of life, the impossible is sometimes only improbable.

Shushinggrasses, thanks for reminding me about the hinny. I knew, in the back of my mind that there have been some female crosses, but since I was too tired to do some more research before including the info, I left it out.

Just thinking about such a small animal as a female donkey having to haul around such a large embryo, birthing problems must be horrendous. We just had to have two large calves pulled by the vet. Without such intervention we would have post both sets of mother and calf. And this was the genetic crossing of the same species, but of the same breed. The trauma they all went through especially the first calf, who apparently had been under birthing stress for an unknown number of hours before I located the mother. He had to be ratcheted out of the mother. His eyes were rolled back in two different direction. His tongue was so swollen, it was sticking out of this mouth. It took the vet some time to get him to breath and get the mucus cleared out because the tongue was blocking the back of the calf's mouth. It took us a few days to get him to suckle independently. We decided to go with Angus cattle because of the small birth weight characteristics and there hadn't been a problem for over 7 years, but it happens. To think that people would deliberately breed different animal species just to see if it can be done is animal cruelty. To me it's the ultimate in selfishness.

I still here, but losing my posts. I gonna try for the third time. Am tied up tonday and tuesday, but either tuesday night or wed wil post the pics and you may see some intersting things. I finally got emails for permission to us ethe photos from everybody. : )

LOL.. Can ya tell I don't know about animals. When i was a younger went to a county fair and this city girl fell for a bunch of cowboys when they asked us to help milk the cows only they gave us bulls instead. Now I count before i touch. Turnign pink just thinkign abotu that tiem again. LOL

Since puter acting up will post about above posts when I get the pics transfered f rom them to me to upload here. : )

I will look up some more research for you guys, but for now I will just give it to you straight. Crosses between different plant species which vary in chromosomes by much more than just 2 chromosomes difference have been done and proven to be fertile. Some of these crosses will take all of the genetic material from both parents and then suddenly dump most of the extra chromosomes, but still retain 2 or more chromosomes and the result is a fertile plant that differs in chromosome count from both parents. I like to think of things as being improbable, but not impossible. Just as some animal crosses result in mixed levels of fertility, so do many plant crosses. Arborea x Aurea group does produce an embryo. I have many theories on how to get these pods or a few seeds within such pod to develop by allowing the compatible pollen to help regulate. Think of it this way, a compatible seed triggers the right chemicals at all stages of pod growth to ensure that that the pod develops. Incompatible pollen may trigger a right response, but then later development fails to ensue as the chemicals become more complex and the likelihood that a correct or close enough match to fool the pod into continuing to develop is not seen. Brugmansia pods will also mature faster and turn prematurely ripe when the light is turned off. Just a side note there. White seeds are sometimes viable, but they dry out very fast and thus are harder to keep going. Most seeds that are white, but quickly darken in color will have a chance if the pod is not one that was allowed to artificially ripen via the no light technique. I honestly don't know if heroic measures such as colchicine will be needed to produce a highly fertile hybrid, but I do know that the more crosses we have the more chances we will have that one will be at least semi-fertile on its own and without our further intervention. What we need is to create that first bridge plant. Now, if someone creates a purple red aurea x flava... I imagine that someone would naturally get a hold of it and double it just for the heck of it even if it was not done at first as the plant became vegetatively propagated throughout the community. Now, we may find simple tricks repeated give us another cross towards both sides of the fence and thus we have two different lines going in the tetraploid and diploid range with varying levels of genetic infusion. Yes, I think it is worthwhile and what have we lost... nothing but a little bit of pollen as we are simply adding the incompatible pollen to our compatible pollen. As long as we actually want both types of crosses we win as we will have seeds regardless. Imagine the pleasant surprise when one of those peculiar phenotypes shows up that is not present in any group as we have a brand new assortment of genes.

Eric, please clarify some things I may not be understanding correctly. When you say

Are these plants taking ALL of the genetic material instead of the usual 1/2? Have Brugs done this? Naturally or by chemical intervention? Now if these crosses have a different number of chromosomes than the parents, are they a new strain of hybrids? Are dipoids and triploids of the same species capable of cross breeding? With haploids?

Are these growth hormones species specific or do they all start with the same structure then diverge over time according to species? Without this hormone, the pod aborts. Am I correct in assuming that the embryo can be forced to mature by cutting out the light, but the support systems aren't advanced enough at that point to keep the embryo alive long enough for it to handle life on its own? If not, where did I go off?

http://www.jstor.org/pss/2441344
triploid answer above
Cytological Study of a Triploid x Diploid Cross of Sorghum Vulgare Pers1
http://agron.scijournals.org/cgi/content/abstract/49/5/237
more of your answer

As this relates to Brugmansia....
Allaquippa = Snowbank (tetraploid pod parent) x Golden Lady (diploid pollen parent)

Haploids are a special group and will take a bit more time. For what its worth, you have genetic mutations in pollen grains just as you do in individual plant cells or plants in general. These mutations are often what we seek to isolate or recover as they offer a new source of phenotypic expression. This is another reason for allowing all types of pollen grains from a single compatible cross to make it to the ovule which can only be done by minimally pollinating each flower and making several of the same type crosses. Think of it this way: If I have 10,000 pollen grains each of which travels or grows at a set speed which differs slightly or drastically from the others then simply placing 10,000 pollen grains on one pistil will not give me 10,000 different genetic variants as only the fastest 100-200 pollen grains will make it to produce a viable seed. If however, I make enough crosses all with the same 10,000 pollen grains to ensure that each pollen grain will not have a chance to pollinate an ovule as not all ovules are accounted for (not enough pollen to pollenate them all) then each genetic variant of pollen and a few mutations will be assured of making it to an ovule. Whether those ovules abort or not is another question. We are interested in those that survive and we understand that some mutations and recessive genes obtained from such a cross may or may not be readily apparent with the first cross which is why sibling and back crosses are desirable utilizing the same methods. More on this later.

Off topic.... but relevant as grafting does influence the the ability of a cross to take as well as change the phenotypic expression of genes as well... yes... I realize I have not answered all of your questions yet, but please bear with me as I give you a bit more food for thought and continue to search for some of your other answers.


marcotrigiano and Gouin (66) obtained no chimeral shoots of 871 shoots regenerated from chimeral callus, but recovered 3 interspecific mericlinal chimeras out of 209 adventitious shoots produced at the graft union of grafted plants. They stated that this "absence of chime- ras from tissue culture suggests that shoot organization in vitro may proceed in a different manner than that occurring in vivo". It may be that graft union shoots are more likely to arise from a multicellular origin, but the in vitro environment may allow such rapid cell division rates that rapid formation of homogeneous clusters of cells pre- cludes formation of chimeral meristems.

At least one shoot nearly stable in chromosome content and green subline could be obtained possessing only 6 chromosomes of Atropa belladonna and the original chromosome number of Datura innoxia.
http://www.springerlink.com/content/km53567003q80n37/

Glad to see you back Eric.
Finally have a little bit to try and do some posting.

here is some Daylily pollen grain making germinating tubes on an onion skin. Pic is from my friend Sue on the AHS board.

This may or may not help in the pollination of brugs or not, but thought it might be helpful in some cases, or somethign to look for and think about.

In Daylilies, again, cuz that mainly where I hybridize most, here is some of my conversation and the pictures all belong to Mary Ann Pruden of LilyHouse Praire who has given permission for them to be used and i have copied her discriptions of the pictures to me from 3 years ago form a discussion we had.

It sems to me brugs have alot of the same problems, especially with long pisitils that brugs have.

The conversation started out because, I went to dab and found a pistil that was spilt and had missing anthers. I had also wanted to know if these split pistils made extra pod chambers and seed.

" Ella, Sometimes the pistil splits--its normally made up of three sections but usually they are kind of fused together. Each pistil corresponds to a chamber in the pod. Sometimes there is a little stigma a couple inches down on the pistil as well. I always pollinate them too and I sometimes end of with four and five chambered pods. I havent kept any records of the fertility of split pistils vs. the unsplit ones. I'm always finding something new to observe as well. This year I have seen many pistils that were not really split but more like twisted together. I dont think the fertility was very high on these at all--but I didnt keep records.

Here's the best pic of that I could find--its on one of my seedlings (kind of hard to see): "

If you look closely in the pic, you cna see the twisted pistil. Til she pointed this out, alot of us never knew such a thing existed and ws giving us pollinating problems.

This first picture is of a twisted pistil. Has anybody looked closely at the brug pistils? On the hard to pollinate ones, is there anything different about them than on the ones who accept pollen easily?

Another friend Asmmad, reported that,

" just as there are three lobes at the top of the style which apparently lead to the channel, one lobe further down the style that also leads to the channel would work. The qualifier is, does that extra stigma lead to the channel and does pollinating it result in seeds in all four/five, etc chambers and does not pollinating it result in one chamber without seeds (or does pollinating the three stigma lobes at the top cause all the chambers to be pollinated in the ovary because the channel leads to them all."

Then I had these following questions.

Would it be common to find an open channel in each one? For future reference.... If I cut open one of the extra styles would I be able to see the chamber or not and would you want to make a horizontal or a vertical cut?

Or perhaps they are related to polytepals genetically.

If they are perchance related to the polytepals, any idea what genetically might develop from crossing a spit style with a well known polytepal?

What are the chances of having more blooms on the same plant producing the split styles or is it just a once in a while odd thing. I wanted to get the cultivar because of its stules to experiment and play around with, but if it is not gonan continue to make the split styles then it reallly won't do me any good, unless you think a hidden gene for it may pop up somewhere down the road.

Admmad's answer back to me.

Would it be common to find an open channel in each one

I don't know. Unfortunately there only seems to be one research group active in the field of style/pistil development and they work on Catharanthus (periwinkle). Apparently there are two basic groups of style development in plant species. One is described as 'phylogenetic' and in those species one style grows from the ovary in a compound state. And the other is described as 'ontogenetic' in which multiple styles grow and then fuse. But the process apparently can be different in each species. In Catharanthus the styles fuse after growth and the researchers are interested on what causes the epidermis to fuse and to dedifferentiate. There is not much information otherwise about the anatomy. There are also two different types of styles in plants - more or less solid versus a central channel.

I would expect that Hemerocallis is like Catharanthus and grows separate styles than then fuse. Continuing that line of speculation I would assume that when a flower has separate styles that each style does indeed have its own central channel.

If I cut open one of the extra styles would I be able to see the chamber or not and would you want to make a horizontal or a vertical cut?

You should be able to see the channel but you may need to use a vital stain to see the living cells versus any fluid that may be in the channel. You would need a sharp razor or perhaps a microtome to make slices and perhaps trying in both directions might be worthwhile.

If they are perchance related to the polytepals, any idea what genetically might develop from crossing a spit stlye with a well known polytepal?

I don't think split styles are related to polytepals but extra stigmas and extra styles (above three normal) might be related to polytepals. And if you crossed a plant that showed a consistent extra style/stigma with a polytepal I would expect the offspring would have a higher percentage of polytepals then if you crossed the same polytepal cultivar to a normal three sttigma lobe/fused style cultivar.

What are the chances of having more blooms on the same plant producing the split styles or is it just a once in a while odd thing.

For some cultivars it may just be a 'random' environmental developmental error. But other cultivars may have a prediliction for reacting to the environment with unfused styles. You would need information about the behaviour of the cultivar over several seasons and many flowers to know which is correct for any specific cultivar. Hemerocallis 'Purple Storm' seems to develop completely unfused/split styles sometimes and often seems to have a short unfused section at the tip of the style.

Here will start a series of the disected pistil from Mary Ann.

"I found stigma with a 4th stigma section today and disected it to see what could be found. It had a 4 chamber pod already formed:"

P2

P3 "A disection of the style indeed shows a second channel--much smaller than the main channel. This is the top near the stigma."

P4 "And the bottom--near the ovary."

"The channel appears to narrow significantly towards the bottom but not sure if its filled with fluid or just narrows. By this time I had sufficiently mutilated the style and could not determine whethere the channel continued all the way to the overy or not. Considering that I do find seeds in the 4th and 5th chambers of pods--I would assume that the channel does continue on down.

What is interesting is the channel from the 4th style appears to have formed by the fusion to the outside of the main style. But also looking further--it appears that each section of the style wall fuses with the sides of the other sections. If the sections did not come together--perhaps the two sides would join on their own leaving a gap in the center (notice the lighter colored areas as well).

Hope to find a split pistil soon to see if this holds up."

"Here is a cross section of the top of the pod after the style was snapped off. I think I flipped it upside down as you could see the openings better. dont know if its the very precise top or a hair down. This shows two distinct openings. On a normal 3 chamber pod there only appears to be one opening at the top which then opens to three channels that direct the tubes to the ovules of each chamber further down"

Here's another example of the twisted pistil.

A partially split pistil.

"Next is the top of the style after cutting off all the stigmas. There are several openings resulting from the way the style sections fused together--also making a wide pistil."

"Below is a style cross-section midway down. NOtice three distinct openings. The center one is larger but the other two are open (looks clogged because of the angle of the channels)."

"And finally the very top of the pod--with three distinct openings (although the center one has very little tissue in a long area--could pollen tubes also enter there??)"

"Here's a pic of some pollen tubes that I grew last year (these were taken with a Digital Blue computer microscope and they dont get the detail that you can see with a real one"

"Actually I guess I did something like that last year--just found the picture:

Here are two pollen tubes that germinated on a stigma. They are growing off the stigma because the whole thing is in a mix of stigmatic/style fluid."

"Here's another pic of a pollen tube"

That all on th epics for now folks. : )

So, from looking at the sizes and shapes of pollen grains could it not be possible that the pollen grains of the brugs like in Daylilies and other flowers just can't get into the different chambers when they are all those different sizes ?

Would it not be a good idea, maybe to make up Eric's 50/50 mix and see what size pollen grains are making pollen tubes.

I ask what do the pistils all look like between the hard to cross brugs. What size pollen to they have. If cuts were made in brugs and observed like in the Daylilies, what would we find? It may be that if the chamber opening are too small, even with fertile pollen, they are just not going to fit, unless you have a way of getting the pollen inside through a by-pass way , maybe with injections. Don't know how far down a prson can really cut and fetilize and stil have it take with causing damage.

Starlight,
You have loads of questions and many thoughts going on here at once. While I admit not having the time to adequately research and answer your questions in full or even in part for some of them... this might help.

I have cut the style at many different lengths with horizontal and vertical type cuts and gotten good results i.e. seed pods between compatible and incompatible crosses. All incompatible crosses have aborted so far unless a compatible pollen was mixed. Again, minimal pollination is key as to ensure some of your less or incompatible pollen has a chance to engage the ovules without competition. I have done some research on this and pollen grains may be slowed down or aborted due to chemicals in the style/stigma or the absence of chemicals as well. Given that pollen grains do differ in size in a single batch and size may be another factor in slow or fast growing, age of pollen grains as each matures at a different rate as pollen grains are constantly being produced... similar to how our body is constantly producing red blood cells each of which lives for 120 days plus or minus, .... in short.... there are many factors to consider and mutations within a single grain of pollen certainly can't be overlooked as well. In the end, I have devised and continue to devise methods for pollinating that are a complex amalgamation of the many different theories and knowledge with my biggest hope resting on the fact that a bit of incompatible pollen can't hurt, but most important to me is that I utilize or try to utilize fewer pollen grains per pollination and simply make more of each cross as this will give you statistically more variation from the pollen donor side than you could ever hope to get with a maximally pollinated stigma. If a trait is carried in the pollen as well as the ovule it will never pair up in quantities great enough to notice or at all if minimal pollination techniques are not utilized if that trait is also carried on a slower growing pollen grain type. There has been research done on the speed of growth of different pollen grains between compatible crosses and incompatible crosses and if memory serves me there is something like 15% untapped genetic potential that is lost due to the nature of how most people pollinate. 30-35 seeds seems to be a good number to shoot for each pod when it comes to many cultivars of Brugmansia to ensure you are getting that extra 15%. Still, we can't be so naive as to think that the 15% we have gained is going to be noticed in the F1 cross as it may simply remain hidden in the genotype and a further cross is needed to both parents utilizing the same method and sibling crosses as well. This is why a good test cross is nice between them all as it may not be readily apparent in the phenotype of where these recessives or quantitative traits are hiding. With Daylilies, I would expect the number you should be shooting for would be more along the lines of 3-4 seeds per pod which of course would mean a tremendous amount of crosses for something stupendous at first. Once that trait or phenotype has been isolated in a large enough or homozygous enough manner then you could in theory resort to crossing similar phenotypes to each other without fear of losing that new 15% genetic diversity you have worked for. You can also go back and heavily pollinate with Brugmansia to ensure a higher seed count, but harder time in finding the 15% hidden potential by cutting the style back again a few hours to a day later. With your Daylilies you would have to pollinate a day early and cover the flower to help prevent the style from drying out too much. Then go back and pollinate by cutting the style in half. I imagine 7-12 hours later would be fine for your work, but you may need to experiment a bit as I am doing to figure out the best time to pollinate for the second go round. There has also been some research with second pollination attempts and in many plants the ovules become more receptive after the first attempt at pollination is done. Whether this is because the pollen grains have bored out those chambers a bit more or not, I do not remember... but it would make sense the pollen may stimulate chemicals or the physical boring out via the growth of the pollen grains themselves may both play a part in that increased receptivity of the ovules and thus a larger seed count than you could get with a single heavy daubing of pollen. Thanks for the pictures by the way!

Geitmann, A., J. Hudak, F. Venningerholz andB. Walles. 1995.
Immunogold localization ofpectin and callose pollen grains and
Pollentubes of Brugmansia suaveolens: implications for the self
incompatibility reaction. J. Plant Physiol. 147:225–235.

http://www.jstor.org/pss/985361
Albert Blakeslee – this guy has done some of the most important research I can find on Brugmansia and Datura crosses.


http://jhered.oxfordjournals.org/cgi/content/abstract/88/6/545
Flowers of each species were first pollinated with heterospecific pollen. After various time intervals, conspecific pollen was added. Analyses of the patterns of resulting progeny were used to infer whether relative pollen-tube growth rates act as a prefertilization isolating mechanism. In I. fulva the frequency of hybrid seeds increased with increasing pollination interval, suggesting that hybridization is limited by pollen-tube growth rates. Likewise, in I. brevicaulis hybrid seed production increased, but it was high regardless of the pollination interval. Thus it appears that relative pollen-tube growth rates limit interspecific reproduction in both species, but barriers are weaker in I. brevicaulis

http://cat.inist.fr/?aModele=afficheN&cpsidt=1909580

The aim of this study was to investigate whether genetically different pollen donors (Betula pendula clones) differed in pollen-tube growth rate across 11 maternal plants and in vitro, and whether the differences between the donors were consistent across the recipients. To compare the seed-siring success of competing pollen donors, a two-donor hand-pollination experiment with six donors and six recipients was conducted. The experiments were performed at a plastic-house seed orchard. The donors showed significant variation in pollen-tube growth rate on all the 11 recipients. The rankings of the pollen donors were statistically consistent across different maternal plants. A significant positive correlation between pollen tube growth in vivo and in vitro was found. The seed-siring success of two competing pollen donors was unequal in 20 of 29 cases and there was a significant positive correlation between seed-siring success and pollen-tube growth rate in vivo and in vitro. The results show that fertilizations are not random and pollen competition operates in a B. pendula seed orchard population.

I am still looking for some of the other information I have found concerning Brugmansia pollen tube growth, incompatibility issues, speed of growth of various species, etc.

Morning Eric!

Yep, I am a little chatter box and always asking questions and do find myself all over the place, because I find while I may be working on something it will trigger thoughts and ideas for other areas. I have found, at least for myself the more I experiment and work and when you really start trying to get into the meat and potatoes of some issues the questions really start to fly. Sometimes it is gettign the answers to those questions that can help you decide to take a step forward or go back a step.

I know you are really busy and appreciate any time you volunteer, sometimes experiments your working on just have to come first as they are usually a time critical matter and very sensitive .

Lots of food for thought in yoru posts and lots of links to study and pick through. Thank you for finding them for us.


Betty..... Don't know if this will help you or not information stored in my notes.

There are some cultivars that wil set pods whether they be a dip or a tet.

Dip x tet or visa versa nearly always wil set a pod but after time will abort it.

if the pod matures usually no seeds are inside or you may have aveyr very small ones.

Of the remaining seeds, mostthat are not plump or of good formation will die.

Most of the good plimp ones don't germinate.

In rare cases for example 1/1000 crosses you might find yoruself with a viable seed and produce a seedling. But from what I have been told the only way to find actually determine its polidy is to have the pollen measured or a chromosome test done.

Now, in the rare case that you do get rare seedling mentioned up above it could be a triploid or a tetrapold because of what is known as unreduced gametes.

Dip = 22 chromosomes
triploid = 33 chromosomes
tetraploid = 44 chromosomes.

Now this.. "In rare cases for example 1/1000 crosses you might find yoruself with a viable seed and produce a seedling." is what we want to find. This is a goal to strive for.

Need Eric to expnd on haploids. I get myself lost alot of times when getting into alot of the genetic sides.

Took me forever to understand some of the genetic side that causes varigations albino seedlings to be produced.

Tumoral tissue has been found in many cases of embryo abortion in incompatible Datura crosses.
http://www.jstor.org/pss/2437868
What causes this tumoral tissue to develop and arrest the development of the embryo? My guess is that the tissue is seen as foreign similar to how RH negative moms see RH positive babies as foreign. The first baby may or may not make it without a booster of rhogam and subsequent reactions get worse. In a similar manner, perhaps a single seed or a few incompatible seeds may make it relatively undetected when they are mixed in with compatible pollen and the compatible pollen numbers are just sufficient to trigger a pod to respond by growing. Too many incompatible pollen grains however and the whole pod aborts or the seedlings themselves abort.

Style length has also been a limiting factor, but more so from pollen that comes from a shorter pistil type plant when placed on a longer pistil plant type. Reducing the distance either by simply shortening the style of of the longer pistil type plant and or grafting the stigma and shortening it via the use of glass capillary tubes to hold the graft secure. This has even allowed unbalanced chromosome combinations to breed as the gametophytic selection against such did not occur after removing a portion of the stigma and style and pollinating lower down and closer to the ovary. Other methods of course include simply injecting the pollen directly into the ovary which works with some and not all plants. The above is a basic summary of the article linked below in relation to Datura.
http://www.jstor.org/pss/2437905

acetocarmine stains of smeared plant chromosomes, a technique that became universally adopted as an enormous time-saver and also one productive of better microscopic differentiation of the chromosomes in the set.
http://www.amphilsoc.org/library/guides/glass/blakeslee.htm

Other collaborations, going back many years, were with E.W. Sinnott on quantitative inheritance, with I.T. Buchholz on pollen tube growth, with C.S. Gager on the use of radium to produce mutations. By means of exposures to radium or X-rays, 541 different gene loci were identified by mutation, 81 of which were mapped to a specific chromosome. It was also found that there was an increase of mutations during the storage of seeds. With I. van Overbeek, Blakeslee applied the techniques of tissue culture to the study of Datura genetic types.

Datura, Brugmansia, Iochroma = Haploid 12, Diploid 24, Triploid 36, Tetraploid 48
Also of note, 2n+1 which occurs with every single gene pair on rare occasion.
http://www.questia.com/library/book/the-comparative-anatomy-of-extra-chromosomal-types-in-datura-stramonium-by-albert-f-blakeslee-helen-houghtaling-edmund-w-sinnott.jsp

Blakeslee examines the chromosome count of 84 pollen grains of a triploid Datura. As you can see, the variance is staggering. Some nice bits on genetics can be found in here as well concerning some odd combinations...
http://www.esp.org/books/morgan/theory/facsimile/contents/morgan-theory-ch09.pdf

Male bees, wasps, ants, etc. are haploid or haplont.
http://www.esp.org/books/morgan/theory/facsimile/contents/morgan-theory-ch10.pdf

I've done lots of research into this area and others and have looked for it again and again for others. What is important is that we realize that sheer numbers, bypassing physical barriers, genetic diversity within a single batch of compatible or semi-compatible pollen is a valuable resource we are missing if we do not at the least try. Mixing a smidgen of incompatible or semi-compatible pollen can not hurt nor can growing out larger numbers of pods with fewer seeds per pod than we are accustomed to getting. Pollinating a Brugmansia or any plant is an art which involves a lot of luck and numbers. Attention to details in the cotyledons, seedlings, mature plants,... attention at every stage must be paid. Removing a portion of the style allows for more genetic diversity to pass to the ovule even with compatible pollens, but this is more likely to be the case when minimal pollination techniques are used. Things like 2n+1 or other combinations that would not normally be possible to pass on to your offspring are not generally possible without cutting the style and applying minimal amounts of pollen over a maximum load of pods. The trick is to tease out all the genetic diversity that exists in your pollen and not just your ovules and most importantly to allow those genes that have been teased out to quantify enough to express themselves in a discernible manner. Sure, we can wait and hope to get lucky by ordinary means and this will occasionally happen as genes mutate or are brought together by back breeding or crossing siblings, but there is a lot we may be losing along the way or at the very least postponing until we have reached the end to where we are no longer satisfied with our results and we take the extra time and effort to apply to a single cross rather than a great many crosses. It is hard to pick and choose when we still have so many beautiful and worth while crosses showing such potential... I just want to tease out more than what we have and move forward in larger leaps.

A man after my own heart. There is nothign more satisifying than seeing newly formed seedlings with some touch no matter how big or small of a genetic change.

Still workign on the links above, when I get through them all, wil have some questions and maybe a few ideas that may have been done or for consideration.

Thanks Eric! : )