Three-storied diversity: Mapping and barcoding crustose lichens and lichenicolous fungi in the Norwegian rainforests (TSD)

Norske regnskoger er kjent for å ha et rikt naturmangfold, av blant annet epifytter.  NTNU Vitenskapsmuseet skal kartlegge og DNA-strekkode lav og lavboende sopper i norske regnskoger, fra Rogaland i sør til Troms i nord. I motsetning til lav, så har lavboende sopper nærmest ikke vært kartlagt i Norge.

Prosjektet vil blant annet registrere relevante miljøvariabler som kan brukes til å klassifisere disse skogstypene i henhold til naturtypeinndelingen "Natur i Norge" og gi ny kunnskap om taksonomi, utbredelse og økologi for denne gruppen av skogepifytter. Midtveis i prosjektet vil det bli arrangert en «LichenBlitz» hvor forskere og amatører i løpet av et døgn skal registrere så mange arter av lav og lavboende sopper som mulig. I samarbeid med Forskerskolen i biosystematikk vil det bli arrangert et feltkurs i artsbestemmelse for studenter og forskere for å sikre videre kompetanseoppbygging på disse økologisk viktige gruppene av epifytter.

In this three-year project we will map, for the first time, the overall species diversity of crustose lichens and lichenicolous fungi in the boreonemoral and boreal rainforests along the Norwegian west coast. We will monitor these forests for new and cryptic species at three different levels of diversity by using established and new sampling methods.

The project’s work schedule includes (1) field inventories in 60 randomly selected stands of boreonemoral and boreal rainforests and 30 adjacent stands of age class IV managed forests along a S/N-transect from Rogaland to southern Troms, (2) DNA barcoding of 300-350 species of lichens and 50-100 species of lichenicolous fungi, and (3) application and evaluation of DNA-metabarcoding for mapping total and cryptic species diversity. We expect to discover at least 30-50 species that are new to science or new to Norway, and considerably improved data on distribution and habitat ecology for 350-450 species. We aim at linking species habitat requirements to NiN, thus connecting epiphytic rainforest communities to the NiN system.

We will disseminate our results via (1) our own projects webpage, (2) established repositories for biodiversity data such as BOLD, GBIF, NLD, and UNITE, (3) social and traditional media, and (4) scientific publications. We will exchange and transfer competence through workshops on lichens and lichenicolous fungi, teaching on student courses, and institutional co-operation. To achieve our goals, nine institutions related to biodiversity research and education from Norway and Sweden will be involved in the project. 

WP1 – Mapping crustose lichens and lichenicolous fungi

We will sample total species richness of crustose lichens and lichenicolous fungi in 60 stands of non-managed rainforests along a south – north transect from Sokndal municipality in Rogaland to Målselv municipality in southern Troms. We will additionally sample 30 randomly selected mature (age class IV) managed forest stands. We will perform random sampling of presence / absence data for epiphytic lichens and lichenicolous fungi from all constitutive tree species. In a second step, we will monitor all stands for additional species not recovered by the random sampling. In this step we will also monitor substrates such as dead wood, rock walls and underhangs, and the forest floor. We will additionally map all red-listed or diagnostic foliose and fruticose lichens. We will further utilize the extensive collections and databases available from botanical institutions in Norway and Sweden (e.g., BG, O, S, TRH, UPS) and our collaborators for gathering additional data on the ecology and distribution of species for field work planning. Vouchers will be collected for species identification and DNA barcoding. Species identification includes standard light microscopy and the identification of secondary lichen compounds with thin layer chromatography (TLC). We will use standard ordination methods for analyzing relationships between species and the recorded NiN variables.

WP2 – Barcoding crustose lichens and lichenicolous fungi

The specimens for DNA barcoding will be collected in WP1 or received from public and private herbaria. We will further barcode foliose and fruticose lichen species that are either red-listed or diagnostic for rainforests when sequence data are missing or insufficient. Additional new or rare species collected during the project period outside the monitored forest stands in WP1 will also be considered. DNA barcoding will follow the NorBOL, and take place within the OLICH project. Direct PCR and metabarcoding techniques will be used for specimens too small for successful DNA extraction. This will mainly apply to the lichenicolous species that often are difficult to separate from their hosts. As for WP1, WP2 will be active/ongoing throughout the whole project period.

WP3 – Metabarcoding for mapping cryptic biodiversity

Sampling will take place in three boreonemoral and three boreal rainforest stands in parallel with the field work for WP1. We will select stands which were found to have high species diversity in WP1. In each forest stand, three randomly selected Populus tremula, Alnus incana, and Sorbus aucuparia trees will be sampled. For each tree, presence/absence data for lichens and lichenicolous fungi will be recorded for four sampling plots 40×20cm in size placed on opposite sides of the tree, two each at tree base and at breast height. All visible lichens and lichenicolous fungi will then be scraped off the plot area without harming the tree and placed in a plastic bag. Samples from each tree will be pooled and subjected to metabarcoding. We will use standardized protocols for wet-lab DNA work and various DNA sequencing techniques: Sanger for the one-by-one sequencing and Illumina for the DNA metabarcoding. Various statistical tools will be used for estimating diversity.