Micropropagation of wild-type Vaccinium Vitis-idaea

A few years ago my friend introduced me to a tasty and edible berry while we were on a hike up mount Washington. I had no idea what it was, and much later I learned that they were lingonberries, which are related to a lot of other shrubby berry plants like blueberries.

Recently I learned that plants are very sneaky and do not always come from seeds. In fact, most plant cells seem to be totipotent (capable of creating any other kind of plant cell) and capable of creating undifferentiated cell mass (callus culture) if you feed them the right stuff. I had previously considered a plant (particularly woody plants like trees) to be single monolithic organisms with parts that fall off that are dead/not useful. It seems like it is more the case that any part of a plant can turn into another copy of the plant, under the right conditions. Think about that next time you are looking at cabbages in the supermarket.

It turns out that most ornamental plants, and many commercial plants are grown not from seed, but basically from cuttings that are carefully grown in lab conditions to maximize growth and multiplication of the plants. This is called micropropogation, and its useful for getting (usually) genetically identical copies of plants. This is helpful for preserving desirable mutations.

You can probably see where this is going: tasty plant + new technique = a small project that will likely take many years to complete. I want to grow our local variety of lingonberries!

Media Prep:

these blueberries were growing right on top of the lingonberries!

After weighing my options on media I decided to go with a media that was actually recommended for blueberry bush micropropagation from “Plants From Test Tubes: An Introduction to Micropropagation”. While there was more specific media recommended in some papers on propagation, it was not clear to me (as a neophyte) that “modified ms media” was actually a specific modification of MS media, not MS+2ip. It was possible to have ordered that media, but a lot of items were backordered for months, so I ended up with regular MS media, and 2ip. At this point it was too late to get the specific modified MS, and blueberries and lingonberry plants grow right next to each other in the wild, so it seemed like it might work.

The media I made was not exactly per the book either, since I think my MS mix already had inositol. The final mix for initiation media was:

  • 1L distilled water
  • 2.3 g MS basal media
  • 5ml 2ip @ 1mg/L (roughly .25 umolar)
  • 20 g sucrose (refined white sugar)
  • 6 g agar

The 2ip concentration was in line with what I had read was ideal for initiation from Jakkola et al. 2001, which was encouraging. The media was not checked for pH since I didnt have a meter or strips, but I would have liked to adjust it down to 4.8, which is a reasonable pH for a lingonberry plant.

One nice thing about this media is that it does not seem particularly rich (surprising given the sugars). I intentionally exposed a plate to contaminate it and it took weeks for some mold to show up. With LB, I would have expected it to be disgusting almost immediately.

Explant Collection:

Explants were collected from a population near the alpine garden trail on mount Washington. only a few grams of plants material was collected over about a hundred yards of trail, in order to minimize impact on the environment. Explants were stored in sterilized falcon tubes for transportation.

Explant Preparation and Plating:

Explants were placed under running water in a strainer with a bowl beneath it in order to create an agitated water bath. The plant tissue was washed for 10 minutes under running water to remove dirt. explant material was then sterilized in 70% IPA for 30s, and then washed in 1:10 bleach solution for 25 minutes. a drop of dawn soap was added to the bleach as a surfactant. After sterilization, the plants were washed three times in sterilized water.

Plants were handled inside of a plastic bin that was sprayed regularly with bleach solution. Tools were sterilized/stored in bleach as well in between uses. I would recommend a clear bin for better lighting. It was challenging to cut the plants in a dark box, but after roughly three weeks, there was only one obviously contaminated sample.


I’d like to call this section something more optimistic, like “success”, but I never really got anything to root. However, there was evidence that the media is sufficient to support the plants. In the photos above, the left photo (with new leaves) was grown from the small browned sprig on in the right photo. The right photo was taken on October 5, and the photo on the left was taken on the 19th, two weeks later. New leaves are clearly visible, and it seems like the plant is producing chlorophyll.

I’m not sure if this is strictly useful, but the lack of contamination and the plant being alive is a good first step.

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