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Friday, January 13, 2023

The Color of Pineapple

Clear glas bowl filled with chunks of cut pink pineapple. The pineapple we all grew up with is a bright yellow color. The pineapples of today isn't necessarily the same shade. Del Monte is now selling a variety with a distinctly pink flesh, called PinkGlowTM or Rosé pineapple. This is a bio-engineered variety, first conceptualized way back in 2005. A patent for the variety was issued in 2012 and the US FDA deregulated the variety in 2016, deciding the variety was essentially the same as other varieties with regards to safety and regulatory concerns.
The variety started as an extra sweet variety grown in Hawaii called MD2. This pink version shares the extra sweet and low acid traits of that original variety. I think it is a worthwhile product, even though (in my limited experience) most people's reactions to seeing the pink cut pieces at left was to think they looked like pieces of meat.


Figure depicting the carotenoid biosynthesis pathway in plants. Starting at top: Acetyl-CoA -> Isopentyl pyrophosphate -> Geranyl pyrophosphate -> Farnesyl pyrophosphate -> Geranylgeranyl pyrophosphate -> Phytoene. An arrow also goes from Geranylgeranyl pyrophosphate to Phytol -> Chlorophyll ->->-> Un-colored metabolites. From Phytoene -> Phytofluene -> Ksi-carotene -> Neurosporene -> Prolycopene -> Lycopene -> Delta-carotene -> Alpha-carotene -> Lutein. A second branch from Lycopene -> Gamma-carotene -> Beta-carotene -> Beta-cryptoxanthin -> Zeaxanthin -> Antheraxanthin -> Violaxanthin -> Xanthoxin -> Abscisic Acid aldehyde -> Abscisic acid. A side brance from Gamma-carotene -> Torulene. A side brance from Violaxanthin -> Neoxanthin -> Xanthoxin (already in the pathway described). The patent is a bit of a pain to read, as they generally are. The "DETAILED DESCRIPTION OF THE INVENTION" section is where they describe the details of the alterations they made.

A figure representing part of the carotenoid pathway described in the previous image. A larger arrow goes from Geranylgeranyl pyrophosphate to Phytoene. A large X covers each arrow leading away from Lycopene. At left is a sketch of the carotenoid pathway in pineapples. There is limited published information about the specifics of the pathway in pineapple, so this diagram was constructed from more general research in tomatoes, peppers, and other species. At right is a closeup of the pathway altered to illustrate the changes that were made in the pink pineapple, as described in the patent.

The first modification was to introduce a second copy of the phytoene synthase gene, driving increased amounts of metabolic energy through the carotenoid pathway. This is represented in the figure by a larger arrow at the top. The added gene was combined with a pineapple fruit flesh specific promotor, so the rest of the plant doesn't have its carotenoid pathway messed around with.

The second modification was to shut down two enzymes, lycopene beta-cyclase and lycopene epsilon-cyclase, normally responsible for converting lycopene into the next steps in the two branches of the carotenoid pathway after lycopene. The consequence of this is all the metabolic energy passing through the pathway is stopped at lycopene. Shutting down these genes was performed by RNA interference (RNAi), also driven by a copy of the same fruit flesh specific promotor. Again, this prevents the modification from interfering with the carotenoid pathway elsewhere in the pineapple plant.

The carotenoid pathway is important for a plant's stress response and other systems. It is likely a pineapple plant would survive more dramatic alterations to the carotenoid pathway that impacted the entire plant, but doing so would throw off the existing balance. The efforts they've taken to limit the pathway tweaks to only happen within the fruit flesh were important to ensure the plants generally are as productive and healthy as the pineapple they started with.



A third modification was atempted, but how the patent is written indicates they're not exactly sure the alteration worked. Commercial pineapple production relies on precision planning. To get a pineapple crop to mature at a specific planned time, the plants are treated with a hormone which induces flowering. In pineapple, the hormone that triggers flowering is the simple gas ethylene. Either ethylene or the similar shaped molecule acetylene is used to induce a crop to start blooming at a specific time. The problem is, pineapple plants will initiate blooming all on their own, when the growers may not want the plants to do so. This is called "natural flowering" and interferes with the plans of the growers.

So, to try and reduce the rate of natural flowering, the third modification was to try and supress the ACC synthase gene important for normal ethylene biosynthesis. They again used RNAi for this, targeted to growing meristems where the gene enzyme activity is important for normal flower induction. I suspect the reason the patent expresses uncertainty about this modification working is because at the time of patent filing, they didn't have enough experience with growing the new pineapple in field conditions to be able to see a reduction in the rate of natural flowering. By now they'll know for sure if it worked.

References:
  1. Marketing piece: https://specialtyproduce.com/produce/Pinkglow_Pineapple_17105.php
  2. Patent: https://patents.google.com/patent/USPP25763
  3. FDA statement: https://www.fda.gov/food/cfsan-constituent-updates/fda-concludes-consultation-pink-flesh-pineapple
  4. Carotenoids in tomatoes: https://the-biologist-is-in.blogspot.com/2014/04/the-color-of-tomatoes.html
  5. Carotenoids in peppers: https://the-biologist-is-in.blogspot.com/2015/11/the-color-of-peppers-2.html
  6. Pineapple flower induction: https://www.echocommunity.org/resources/f0e9cfeb-ba1d-435e-a515-7705ca79b409

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