// Twitter Cards // Prexisting Head The Biologist Is In: The Color of Beans 4

Friday, February 10, 2023

The Color of Beans 4

In my last post (https://the-biologist-is-in.blogspot.com/2023/01/the-color-of-beans-3.html), I shared a couple figures illustrating the flavonoid/anthocyanin pigment pathway in plants and in common beans (Phaseolus vulgaris) specifically. A couple days later, I found some additional evidence which led me to feel the need to update my figures somewhat.



Starting section from the top: phenylalanine to cinnaminate to 4-coumerate to p-coumaroyl-CoA (+ 3x malonyl-CoA) to naringen chalcone to naringen. Naringen goes left and right to eriodictyol and pentahydroxy flavone. Eriodictyol goes left to flavan-4-ols and then to phlobaphenes (highlighted in red). Eriodictyol goes right to tricetin. Naringen goes right to apigentin (highlighte light brown). Pentahydroxy falvanone goes right to luteolin (highlighted pale yellow). Naringenin goes down to dihydrokaempferol. Eriodictyol and dihydrokaempferol go left to dihydroquercetin. Pentahydroxy flavanone and dihydrokaempferol go right to dihydromyricetin. Dihydroquercetin goes right to quercetin (highlighted in yellow). Dihydrokaempferol goes right to kaempferol (highlighted in yellow) and then down to astragalin (highlighted in yellow). Kaempferol goes to a series of question marks highlighted in a gradient from white to brown. Dihydromyricetin goes right to myricetin (highlighted light brown). Dihydroquercetin goes down to leuocyanidin. Dihydrokaempferol goes down to leucopelargonidin. Dihydromyricetin goes down to leucodelphinidin. Leucocyanidin goes down to cyanidin and then cyanin (both highlighted red). Leucopelargonidin goes down to pelargonidin and pelargonin (both highlighted orange). Leucodelphinidin goes down to delphinidin and delphinin (both highlighted blue). Leucocyanidin, leucopelargonidin, and leucodelphinidin go left to 2,3-trans-flaven-3-ols (catechin) (highlighted in in a gradient from white to brown). Cyanidin, pelargonidin, and delphinidin go left to 2,3-cis-flaven-3-ols (epecatechin) (highlighted in a gradient from white to brown). Catechin and epicatechin go down to proanthocyanidins (highlighted in a gradient from white to brown). Luteolin, apigenin, and tricetin have a group label 'flavones'. Myrictein, kaempferol, and quercetin have a group label 'flavonols'.  The figure has enzyme labels at most steps.  In the top starting section: PAL, C4H, 4CL, CHS, and CHI. Naringenin to eriodictyol is F3'H. Naringenin to pentahydroxy flavanone is F3'5'H. Eriodictyol, naringenin, and pentahydroxy flavanone to tricetin, apigenin, and luteolin are FNS. Eriodictyol, naringenin, and pentahydroxy flavanone to to dihydroquercetin, dihydrokaempferol, and dihydromyrecetin are F3'H. Eriodictyol to flavan-4-ols is DFR. Dihydrokaempferol to dihydroquercetin is F3'H. Dihydrokaemperol to dihydromyricetin is F3'5'H. Dihydroquercetin, dihydrokaempferol, and dihydromyricetin to quercetin, kaempferol, and myricetin are FLS. Dihydroquercetin, dihydrokaempferol, and dihydromyricetin to leucocyanidin, leucopelargonidin, and leucodelphindin are DFR. Leucocyanidin, leucopelargonidin, and leucodelphindin to cyanidin, pelargonidin, and delphinidin are ANS. Cyanidin, pelargonidin, and delphinidin to cyanin, pelargonin, and delphinin are GT. Leucocyanidin, leucopelargonidin, and leucodelphindin to catechin are LAR. Cyanidin, pelargonidin, and delphinidin to epicatechin are ANR.
Certain combinations of published genes lead to production of a brown pigment when an excess of yellow astragalin would be expected. Here I've made up an enzyme called FNR (FlavoNol Reductase) leading to production of the pigment, modeled after the production of proanthocyanidins by ANR (Anthocyanin Reductase).

The evidence for a brown pigment derived from the yellow pigment pathway comes from the gene B changing a bean color from yellow-brown to mineral/dark-brown when present. B is thought to be a transcription factor that enhances the expression of other pathway genes. Yellow can only arise when red and blue pigment branches are absent, so the brown pigment produced with B can't be the brown proanthocyanidins derived from the red and blue pigment branches. Instead it must be an analogous pigment produced from kaempferol and/or astragalin. I haven't found any research papers discussing this pigment pathway branch, but all the evidence seems to point to it being there anyhow.

This version of the flavonoid pigment pathway is trimmed to be limited to the main pigments (cyanidin, pelargonidin, delphinidin, & astragalin) along with the core of the pathway leading to them and the metabolites derived from them.
The figure trimmed to the metabolites and pigments significant in common beans... now includes pelargonidins. This version of the figure has different thickness arrows in places to illustrate when one branch has a higher priority, when one branch of metabolites is more likely than another.

Dihydrokaempferol is a central metabolite for all the different pigment branches in the pathway. F3'H and F3'5'H leading to red and blue pigments have the strongest branches, followed by FLS leading to yellow pigments, and then last is DFR leading to orange pigments. For the orange pigments to dominate the final color, it looks like the other three branches have to be disabled or significantly reduced.

pale tan beans with orange-toned eyes.
So. Are there orange varieties of common bean? Maybe. Instagram user @g3netic_lottery shared a mixed variety of beans they grew this year and some of them had a distinctly orange-ish color, especially in the hilium ring. Now, I'm not entirely sure that is the color of pelargonidin, but it is the closest to orange I've seen in common beans. It's enough of a suggestion that I decided to rework the pathway figure to include it for this species.

I still think it would be really cool to find (or breed up) a variety that had distinctly orange color all over the seed. These beans at right hint that it is feasible if one has the right sort of luck. @g3netic_lottery on Instagram grows beans and other crops in South Africa. I'll be keeping my eye out for useful seeds more accessible to me that might help me get to a nicely orange common bean.


The other domesticated bean species will have very similar pathways as the common bean, but the specific available mutations are going to be different. Maybe we can find a good pelargonidin orange color in one of them. I spent some time looking around for orange seeded varieties of the other species, with limited luck.

  • Runner beans (P. coccineus): This photo of "Ayacote Mexican" beans includes some very orange looking seeds, but I have no idea how much that reflects reality.
  • Lima beans (P. lunatus): "Pima Orange" has seeds that look yellow-brown or orange depending on the photo. Again, it is hard to say what they really look like.
  • Tepary beans (P. acutifolius) : There are some photos around of very orange tepary beans, but I can't find any varieties available for sale that look anything near orange.
  • Year-long beans (P. dumosus) : This species is interesting, but only grown in a limited area. I found several articles about it, but I have yet to find anyone selling seeds.
Mix of dark purple and black beans.
I ordered the runner and lima bean varieties above, so I can see what their colors actually look like. It is so very easy to intentionally or accidentally tweak the color of a photo, so the color of photos online isn't always something you should trust.

The runner beans turned up first. Unfortunately, there was nothing like orange among them. I did get the lovely dark purple beans at left though, which are perfect for another project I'll describe in another post. After further looking around, I found a different vendor selling ayacote beans that are at least advertised with some nice orange tones. As they too are selling the beans intended for food use, there's no guarantee any yellow seeds will turn up.

Mix of orange colored beans with variable dark markings.
The lima beans arrived a few days later. When I opened the package, I was greeted by seeds with a surprisingly nice orange color! Now, this isn't anything like the orange you get from beta-carotene in some carrots and tomatoes, but this might just be what a pelargonidin orange looks like.

Alternately, this could be a mix of a yellow and a brown pigment. There may be some simple basement-lab tests I can do to help me identify the pigment, but that will take some further research.

In either case, I think it is entirely reasonable to expect a similar orange colored common bean should be possible. It's just a matter of finding the right mutations and crossing them into the same plant.

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