Joebass said:Interesting treating buds. I have never heard of that before. So are you putting polyploid pollen on these treated buds? Please explain this further.
The science behind what I am trying to do is not new. There have been several research reports published related to trying to produce unreduced gametes in Lilium by treating the buds at an early stage with different methods (colchicine injection, heat treatment, nitrous oxide pressured gas, caffeine solution injection). The idea is to disrupt the normal meiosis process going on inside the bud such that some of the eggs & pollen formed will have 2x the base number of chromosomes. The research is typically done using cultivars that are considered sterile and often the result of a wide cross - like Black Beauty. The reason for this is that if you get any fertility it is very likely that it resulted from unreduced gametes so you know you have been successful.
I started playing around with this a few years ago and it was sort of an off-shoot of something else I was tinkering with. My original project was to mix hormones in with lanolin to treat buds on wide crosses to see if it helped achieve a viable cross - there was a research article written back in the 50's on this by Emsweller. So ended up mixing up a batch of this lanolin paste with some Surflan (Oryzalin) and treated a few different buds of lilies just to see what it would do. I was initially just trying to see if it would kill the buds or have any affect. What I found was that for most treated buds they continued to grow and develop and open but there was a noticeable difference in the petal texture and flower form - that was two years ago. Last year I did a larger experiment and put it on lots more buds and intentionally made crosses with the buds to see what would happen. Based on the results I do believe I am getting some unreduced gametes. The kinds of crosses I made last summer were across the board including:
2N treated x 2N treated (Orientals, Aurelians, & OT)
2N treated x 4N (Oriental x OT)
4N x 2N treated (OT x Oriental)
3N treated x 4N (OT x OT)
4N x 3N treated (OT x OT)
On many of these crosses I did Embryo Rescue as they were wide crosses low probability crosses. For the 2N crosses intersectional crosses I just harvest seed and am growing them normally - the L. henryi var. citrinum treated x L. henryi Ault treated was this sort of cross.
There are a few embryo rescue explants I am excited about that came from this. The first was Casa Blanca treated bud x Mr. Job (Griesbach 4N fertile OT). It is possible this seedling will just be a triploid - either way could be an interesting lily. The other cross is from Scarlet Delight (4N OT) x mixed oriental pollen from treated buds. I had quite a few embryos from this cross but only one grew. Again it is possible that this was formed from diploid pollen rather than tetra pollen but I won't know until it grows out.
In all this, the thing that was very encouraging to me was that I treated many buds on my diploid Black Beauty and it only set pods on the treated buds and only with pollen from treated buds of diploid Orientals - all the other pods didn't even form. I didn't end up with any seed from these crosses but the fact that it tried to set some pods was encouraging. The other thing I noticed is that the pollen on treated buds looked different. I treated a wide variety of OT's that would be considered to have mostly sterile pollen and there was a definite difference in color & texture of the pollen from treated buds. I need to figure out how to look at pollen under my microscope to get some real data. I have been coached how to do this but haven't been able to make it work yet - I need to invest some time in this as it will help validate if this is working.
For the crosses I did between two treated diploids, there is likely a larger portion that would just be normal diploid. I would hope for some triploids and even a few tetraploids from these crosses.
I am excited about the prospects of these experiments. Being able to produce tetraploid seedlings directly from crossing diploid cultivars would help to increase the diversity in the tetraploid breeding lines. The alternative has been doing polyploid conversions on cultivars which is a slow and low probability process and then the converted cultivars tend to be short lived based on feedback from others who have been down that path.
I attached a picture of the pod from Ypsilanti from this cross. I haven't seen a pod this big on any of my henryi crosses before. Pod size is just one indication and not a smoking gun.