Peas in the lab – results

Principal coordinate analysis of old Danish peas
(Click on picture to see more clearly)

Finally I finished writing a small report (in danish) about the old peas in the laboratory.
The report is located here: 2010 Resultat af ærteforsøg (DNA)
I had google translate this version, with a bit of my help: 2010 Pea results

When reading the results, so it is vital to understand that this investigation can determine whether some of the peas are genetically different, but can not determine whether they are genetically identical! We have investigated only some small bits of the total DNA.
The peas that look similar, are either identical or different, we do not know!

At the top is shown the principal coordinate analysis. It gives a good visual picture of the distance between our varieties. Some old Swedish varieties was blended in, as they have been studied previously. It gives some reference points to past studies.

The principal coordinate analysis is based of course, on some very concrete data, namely the length of the micro-satellites we tested.

Table. Displays length of each micro-satellite.

Micro-satellites we have measured have letter names. A9 – D21 – AC58 – C20 – AA5 – AC75.
We investigated 6 different, but one would not work, the second was identical in all varieties, and therefore uninteresting. The last four showed differences. Each micro-satellite was measured twice.

I end up concluding:

So there are varieties that we now know is different from all others in this sample.
There are also groups of varieties we should check for phenotype differences. It is a great advantage that it is now clearly delineated the varieties to be compared by cultivation experiments. There is probably also varieties that are identical with each other. It is not something that can be resolved by this trial, which is only suitable to detect differences in the 4 areas (micro-satellites), we got useful results from. Whether they are identical in many other micro-satellites, we can not know before we make a new attempt!

Read in the google translated report: 2010 Pea results

If you can take a bit more, this is the excel file we were sent after the course: Diversity analysis Pisum

 

Heirloom peas in the laboratory

Pee shoots of heirloom varieties in the laboratory at Department of Agriculture and Ecology, Copenhagen University (Copyright Ingrid Nolde)

Last weekend I joined the Danish seed savers at the laboratory at the Department of Agriculture and Ecology (Copenhagen University). Gunter Backes and Jihad Orabi led us through a study of our heirloom pea collection. It was a fantastic and educational weekend!

In preparation, we had grown peas for 3-4 weeks, so they were ready for analysis. We had to use leaves that had not been given too much light. Chlorophyll can make it difficult to extract DNA material from the rest of the leaf tissue. For safety’s sake, we were two cultivating the same peas. It went well and we only used one set.

Test tubes on ice (Copyright Ingrid Nolde)

We were divided into four groups. That way we could manage to work with all the varieties we had brought. After a brief introduction to laboratory work, to use gloves and gowns – take them off when you leave the lab, we started slicing the pea leafes.
Once cut, the natural enzymes in the leaves activates. To inhibit them, we set all tubes in a tray of ice. Millimetre small pieces were placed in the tubes, a solution of caustic soda were added and the tubes were boiled for 1 minute to denature the proteins. Then we blended the boiled leaves with the tip of a plastic stick and added a resolution that brought the pH down to a desired level. To separate DNA from the rest of the leaf broth, test tubes were centrifuged.

Pea leaf broth bottom, DNA located at the top in the clear part of the liquid (Copyright Ingrid Nolde)

Now we could move the purified DNA into new tubes, still set in ice.
Then it was time to mix liquids with primer for the DNA fragments we searched for. We looked at 6 different micro satellites. Micro satellites are good to tell the difference between varieties of a species. A primer is the DNA sequence immediately before the micro satellite, leading the DNA strand to be cut at the desired location. We used a primer for each end of the micro satellite, because it must be “cut” at both ends. To the liquid is also added Taq DNA polymerase, an enzyme that replicates DNA at specific temperatures.

We used pipettes for several hours. Even with multi pipettes there was plenty to do (Copyright Ingrid Nolde)

Next the test tubes are undergoing repeated temperature changes between
94 °C, where DNA strand separates
64-55 °C, where the primer binds to the DNA strand, and
72 °C, where Taq DNA polymerase copies the DNA strand from the primer and forward

By the repeated changes in temperature, we multiplied the selected micro satellite exponentially. The first cycle gave very little DNA material, but for each cycle, there was more copies. Just like reproduction of yeast in a bread dough.

Finally, we let an electric field pull micro satellites through a viscous liquid in capillary thin tubes. A dye was coupled to micro satellites, for a photocell at the end of capillaries to detect how quickly they passed by. Small micro satellites migrate faster through capillary tubes than larger.

First presentation results

The photo shows the horizontal bands, each representing a pea variety. The small vertical bands are micro satellites. The red vertical band marks a scale, so we know how long the micro satellite strands are.
Jihad and Gunter did the analysis work Sunday morning, before we all met up – thank you!

It was best to group the peas in 7 groups

The results will soon arrive in my Inbox. I look forward to delve deeper into the results. I might have a more to write about then.

Big thanks to both Gunter and Jihad for a learning and rewarding weekend 🙂

PS: I’m just a happy amateur. Feel free to comment if I’m wrong in some of the techicalities 🙂

 

Pea and Barley Field

Naked barley ‘Hora’ and graypea “Errindlev ært”

As I some years ago started growing graypeas (oldfashioned fieldpeas) I soon started to wish growing them together with barley or oat. I remember this kind of fields from the homesteads around where I grew up, back when others enjoyed the sixties in other ways 😉

Asking around, I was kindly informed, that this co-growing was harvested green for fodder, not for human consumption. But I still cling to the dream of this co-growing.

I want the barley to be naked, so I can cook and eat it straight forward. Ordinary barley is for malting and brewing, unless you have special equipment.

The varieties of graypea and naked barley must support each other in one way or the other, to compensate for the competition. Naked barley supports the peas by their straight straw, graypea fix nitrogen and share it with the barley. Last year I mixed graypea Lollandske Rosiner and naked barley Hora. The high number of tendrils in Lollandske Rosiner was almost strangulating the barley, being too short for a fair competition. It was OK, but with room for improvement.

This year I have mixed graypea Errindlev Ært with naked barley Hora, and it seems to be a perfect match. Spikes of barley raise well above the graypeas. The gentle tendrils doesn’t strangle the barley, and the leaves suppress weeds in the bottom.

For graypea Lollandske Rosiner I try to find another naked barley variety. This year I’ll try Gängel, a swedish variety, said to grow tall.

Naked barley is one of the good flavours that for some generations has been neglected. It’s a pity, as naked barley boiled and added a lump of butter has a heavenly taste, similar to sweet corn (much more demanding to grow in cool Scandinavia)

Tendrils of graypea Lollandske Rosiner

Ps. My “field” is just 1 m2

 

Grey Peas

Grey Pea ‘Lollandske rosiner’

At last I had a chance to boil the grey peas. Grey peas was stable food in northern europe before the potato made its way into the kitchens. Since then the use of grey peas have been forgotten. To learn how to cook them, I looked them up in a danish cookbook from 1847 written by Madam Mangor. You might be able to translate it with BabelFish or Google.
But an anarchist live somewhere inside of me, always making me do things a bit different to the recipe.

Grey peas in their own gravy

I soaked the grey peas overnight well covered in tapwater. Changed the water, brougth to boil and added a pinch of baking soda, then gently boiled for two hours. Now the grey peas was tender and had developed their own gravy. We dined on some of it. Next day the gravy had turned in to jelly, but melted quickly on boiling. They tasted even better the second day. Taste was superior to the yellow peas, and to my surprise with no gas problems.
I hope grey peas will one day be awailable in the ordinary supermarkets – ithey are delicious.

PS. Thanks to Poul, who saved ‘Lollandske rosiner’ from extinction, and to Kirsten, who grew and presented this batch of grey peas to me.

 

Update – Color variation in Golden Sweet snowpea

Dried pea seeds of Golden Sweet

After five purple pea seeds found in a single pod last year, I only had a single pod full of purple seeds. That pod was attacked by pea moth, I could only rescue a single seed. Plants were very true to type.

Preliminary conclusion is that the purple trait in this color variation in Golden Sweet probably depends on environmental factors.

I probably can’t resist sowing the last purple seeded Golden Sweet next spring, just to see what happens. I don’t expect to find any purple seeds in that future batch, but who can tell without trying it out themselves?

 

New seeds from the seed meeting

Fieldpea ‘Biskopens gråært’

At the seed meeting there was a major seed exchange on the first evening (and during breaks the following days) I would like to introduce some of my new friends.

‘Biskopens gråært’ is one of the rare solid purple seeded pea varieties. It is a greypea, the oldfashioned fieldpea being a stable food in northern Europe in very old times. Now with famine well at distance the deserve a revival. I hope a famous chef will take courage to dicover it and serve it in modern ways. We have saved a treasure of these old peas.
This particular greypea is a swedish heirloom, passed on by SESAM, the swedish seed savers.

Russian heirloom ‘Goroh’

The pea ‘Goroh’ is an old russian heirloom from Kalmutskaya region in Russia. It has white flowers and rather small round green seeds, drying yellow. Originally from Dr. Tatiana Veronina (Moscow), via Seed Savers Exchange and a norwegian seed saver to Denmark. It can be eaten both as a snowpea and as a delicious and quickly boiling soup pea.

Wax pole bean ‘Gold of Bacau’

This wax pole bean has long broad yellow pods, and is an early romano-type. It’s an heirloom from the Bacau in northern Romania, passing Seed Savers Exchange on its route to Denmark. It’s said to have a gorgous taste, I look forward to the season.

Russian bush drybean ‘Bean 04-2006’

UPDATE february 2011 – I had a look in the original seedsamples, and noticed, that this and the following photo had been misplaced, now they show the correct cultivar.

Russian bush drybean ‘Bean 05-2006’

These two russian heirlooms Frøsamlerne has got from Lothar Juffa in East Friesland, Germany. They originates from volga-german families, settled near Omsk in Sibiria. Zarina Katharina the Great called in a lot of german peasants to settle on land she gave them on the Volga river. Later Staling executed a lot of them and resettled the rest in Sibiria, where they have lived since.
Both are early and prolific drybeans, the latter a bit earlier and slightly taller. I’m about to develop a weakness for these Volga-german varieties, as I have only good experiences with them – they have stand the test of time and hardship.

 

Shelling the first dry peas

Today I’ve been shelling the first drying peas.
It’s an old variety for heavy clay soils, Lollandske Rosiner. Hundred years ago it was a
staple food for the working class. It is a field pea, to be sown in very early spring.
It is boiled in soups, where it after two hours of boiling look like a raisin, and very nutricious.

I grow it, because it must survive to the day a multimedia gourmet taste it and make it famous around the world. Since it is very early, it’s also nice to eat fresh.
I have a dream of growing it intermingled with oath, as I remember this kind of fields from my childhood.
Because Lollandske Rosiner is so early, it didn’t get affected by the larvae of pea moth.