Ancient DNA extraction from mid-century-

old increment cores of Scots pine unlocks

new avenues for research and discovery.

Changes in Scots pine genetic diversity 48 years old

across the past centuries

Yok Wongkaew, F. Gil Muñoz, & M.R. García Gil 216-708 years old

https://commons.wikimedia.org/wiki/File:Tall_trees_at_Rolley_Lake.jpg

This project aims to study the original genetics of the Northern Swedish Scots pine forest prior to
19th-century seed imports. The study will determine the impact on genetic diversity and locally
adapted alleles. The findings will inform the preservation of the forest’s unique genetics.

How can DNA be extracted from forests that no longer exist?

https://pxhere.com/sv/photo/487663?utm_content=shareClip&utm_medium=referral&utm_source=pxhere https://vastmanlandslansmuseum.se/pa-gaende-fot/ https://commons.wikimedia.org/wiki/File:Tall_trees_at_Rolley_Lake.jpg

INTRO
• Forest products have been essential resources in Sweden after the ice glacier retreat
• Scots pine seed imports began in the early 19th century due to a shortage of domestic supplies
• Most imported seeds came from Russia, Finland, Norway, Germany, Poland and France
• Lack of information on original genetics and seed imports in the Swedish forest

METHODS
Scots pine samples:
45 DNA samples from natural forest Jukkasjärvi (1474-1608)
Kalix (1315-1595)
Bygdeå (1318-1494)
Innertavle (1551-1763)
66 samples increment cores Tiondebod (1411-1737) Balfors (1553-1803)
Luleå (1432-1558) Snöan (1608-1807)
18 samples ~ 48 years old Öjebyn (1530-1785)
Åby (1521-1773) Östersund (Natural forest)
48 samples ~ 200-700 years old Bureå (1542-1743) Arjeplog (Natural forest)
Lövånger (1368-1505) Jokkmokk (Natural forest)
https://commons.m.wikimedia.org/wiki/File:Map_of_Sweden_Cities_(polar_stereographic)_Sv.png#

1. Three different DNA extractions methods bp M 1 2 3 4 5 6 7 8 9 10 11 12 PC NC M

• Macherey-Nagel NucleoSpin Plant Kit 5000

• DNeasy Plant Mini Kit (Qiagen, The Netherlands) 1500

followed Akhmetzyanov et al., 2020 modifications

500
•
75
• CTAB DNA extraction
bp M 13 14 15 16 17 18 19 20 21 22 23 24 PC NC M
5000

2. Genotyping through PCR and Sanger sequencing 1500

• Microsatallites (SSR) markers 500

75

3. Data analysis using R programming software A representative image of agarose gel electrophoresis of comparative
amplification of different Scots pine wood core DNA extractions methods.
Lane 1-6: DNeasy Plant Mini Kit (Qiagen, The Netherlands). Lane 7-12:
• SNPs array data from DNA samples of natural forest Macherey-Nagel NucleoSpin Plant Kit method. Lane 13-18: CTAB DNA
extraction method. Lane 19-24: CTAB DNA extraction method on the old
• Microsatellite data from the old increment core samples samples from 216-708 years old. The samples in lanes 1-18 are 48 years
old. PC: positive control. NC: negative control. M: molecular marker.
Created with BioRender.com

ACKNOWLEDGMENT
Grateful appreciation is extended to Maarit Kalela Brundin and Jonas Fridman for their provision
of the old increment core samples of Scots pine.

REFERENCE:
Akhmetzyanov, L., Copini, P., Sass-Klaassen, U., Schroeder, H., Groot, G.A. de, Laros, I. and Daly, A. (2020) DNA of centuries-old timber can reveal its
origin. Sci. Reports 2020 101, 10, 1–10.