With the lima beans, I acquired this pink/magenta variety (from https://trueloveseeds.com/products/ceceilia-davis-family-lima-bean) and then produced this unmarked yellow variety (after starting from https://www.nativeseeds.org/products/pl011) to cross w/ hopes of getting me closer to the goal.
So far, I have failed to cross the two.
That I was only able to recover the combinations of the red and blue genes seen in my starting varieties tells me that the genes for the two important enzymes (F3'H & F3'5'H) are tightly linked.
The "V" genes (F3'5'H) is on chromosome Pv06 of the common bean genome.
F3'H, responsible for producing red anthocyanins, may or may not have a specific gene label associated with it.
When doing real research, you very quickly find unknowns, and you have to forge your way forward anyhow.
It turns out that F3'H is important for how plants respond to UV stress. (https://www.mdpi.com/2223-7747/10/1/118)
Thus, essentially, every plant species will maintain an active copy of this gene.
Beans with this gene inactive in the seed coat have to be doing so via a different mechanism than breaking the gene.
Since I've already found that "R" is tightly linked to "V", maybe there is a transcription factor responsible for activating F3'H in the seed coat that just happens to be linked to the V/F3'5'H gene.
As I puzzled over how to identify such a transcription factor gene, I found myself rereading the 2022 paper that identified the location of the V/F3'5'H gene. (https://pubmed.ncbi.nlm.nih.gov/35432409/)
In that paper, as an aside, they mention finding a 2nd copy of F3'H adjacent to the F3'5'H gene, with both resting within a heterochromatic region around the centromere. Such regions show dramatically reduced crossovers, thus recombination.
I may still slowly move towards getting these final steps completed, but already, I have a pretty good answer to the question that started this all.
I know that many of you have been following this story over the last years. You don't have to worry, I will continue posting bean content.
I'm just feeling quite pleased with coming to a solid biological answer for this, admittedly quixotic, quest of mine.
With the runner bean project, I may take a different direction to get there.
The two species are very closely related. Their genomes have largely the same order of genes.
I already made one cross between my blue common beans and a purple runner bean with black speckles.
The result is that F1 hybrids are easily made by transferring runner bean pollen to the common bean flower, but not the other direction.
Thus, I can use such hybrids to introgress the blue seed trait into runner beans!
The ideal cross would be between my blue bush beans and the "white" seeded runner beans I found in the F2 population I described above.
I use quotation marks because the seeds aren't actually white.
Instead, they're a pale brown color caused by the oxidation of colorless flavonoids into brown tannins.
The result of this cross will produce blue seeds, and some of them will look almost 100% like runner beans.
@thebiologistisn a blue butternut squash would bring interesting contrast to the winter holidays meals 🤩