Phenology, the Seasonal Appearance of Species, and Nitrogen-Fixing Microbes at the GLORIA Site in Southwestern Montana, USA

Martha Apple, John Warden, Josh Prince, Sergio Morales, Tyler Pullman, Charlie Apple, and James Gallagher GLORIA (Global Research Initiative in Alpine Environments) www.gloria.ac.at

Questions:

1. Can individual species be detected throughout the growing season?

2. If GLORIA sites are surveyed at different times in the growing season, are different observed?

3. Do lichens at the GLORIA site have nitrogen-fixing Cyanobacteria as their phycobiont, and, do the legumes have nodules with N-fixing bacteria?

4. Which functional groups of microbes are found in cryptogamic crusts near the SW Montana GLORIA site?

SITE: • The Southwestern Montana, USA, GLORIA Site, US-PIO, is east of the North American Continental Divide:

• On Mt. Fleecer, 2873 m (9425 ft.), (45°49’36”N, 112°48’08”W) between the Pintlar and the Pioneer Mountains,

• On Mt. Keokirk, 2987.3 m (9801 ft.), 45°35’37” N, 112°57’03” W, south of Mt. Fleecer in the Pioneer Mountains.

METHODS: We collected a series of photographs at the three Mt. Fleecer sub- summits at the corners of 3m2 grids placed 5m below the high summit points at N, S, E, and W

Between the latest (July 3) and earliest (August 30) snows: • Fleecer Treeline and Lower Alpine - July 9-12, Aug. 3 & 28 • Fleecer Upper Alpine - July 23, Aug. 22

We then compared the photographs for differences in species presence, flowering, fruiting, and seed production, and also noted new species not found during the baseline survey.

We also collected and examined: The legumes, (Lupinus sp.) and Oxytropis campestris, for root nodules with N-fixing bacteria, Lichens and Cryptogamic crust for N-fixing Cyanobacteria

Fleecer Treeline, South 11:July 9, August 3, August 28 2010 FTLS117910 FTLS118310

FTLS1182810 Fleecer Treeline South, July 9, 2010 Fleecer Treeline South, August 3 2010 Fleecer Treeline South 8-28-10 Mt. Fleecer Lower Alpine Fleecer Lower Alpine, East, July 12, 2010 Fleecer Lower Alpine, East, August 3, 2010 Fleecer Lower Alpine, East, August 28 2010

Fleecer Upper Alpine, August 22, 2010 Fleecer Upper Alpine, South, July 23 2010 Fleecer Upper Alpine, South, August 22 2010 New Species Found in 2010 at the Mt. Fleecer Treeline Sub-summit In July: Ranunculus glabberimus, Ranunculaceae Claytonia lanceolata, Portulacaceae Heucheria cylindrica, Saxifragaceae Cryptantha sp., Boraginaceae (only one in bloom).

In August: Woodsia scopulina, Dryopteridaceae, a cliff fern calycosa, Gentiana amarella, Gentianaceae Solidago multiradiata, Asteraceae Agoseris glauca, Asteraceae Campanula rotundifolia, Campanulaceae Boletus sp., Boletaceae

Key Points on Phenology and Species Appearance: • Structural and life history differences among species play an important role in when they are visible.

• All of the new species were found at Treeline

• Greatest and fastest change in species presence at Treeline.

• By late August, flowering still in progress at Treeline but mostly completed at Lower and Upper Alpine.

• Little change in species during the summer at Upper Alpine.

• Grazing in late August at Lower Alpine (mountain goat, elk, and the hay-gathering pika) and probably a bear digging and turning rocks at Treeline.

Some species can be detected throughout the growing season, while others are visible for only part of the season. Some species are more visible at certain times, but can still be detected earlier or later because their new and/or remnant leaves, stalks, or seeds are visible.

Examples: Dodecatheon pulchellum absent by late August, even though it is abundant and showy in early summer bloom and mid-summer fruiting. Phlox sp. visible throughout summer, leaves from previous year(s) remain visible although Phlox seeds are unobvious. Gentiana sp. only apparent in August but then very showy. Grass and Carex sp. leaves and stems visible long before and after flowering. Cassiope mertensiana and other woody shrubs are always visible.

Of the 42 species found at Mt. Fleecer in 2008, all but 4 were found in 2010. Of the remaining 38 (which include 2 trees, 3 shrubs, 1 Lycopod, and 32 herbaceous angiosperms):

5 were only visible on specific dates in 2010: 3 only in early season – Dodecatheon pulchellum, Lewisia pygmaea, Dryas octopetela 1 only in mid-season – Arabis drummondii 1 only in mid-to-late season – Erigeron simplex

Of the 8 new vascular plant species found in 2010, 3 were visible on all dates (Heucheria cylindrica, Woodsia scopulinum, Solidago multiradiata) 3 were only visible early (Claytonia lanceolata, Cryptantha sp., Ranunculus glabberimus, 2 were only visible later (Gentiana calycosa, Gentiana amarella).

Fine-Scale Phenology and Photographs: Are Useful in Comparisons of: Summers Perennials, annuals, biennials, Length of time from: Leaf emergence to flowering, Flowering to seed production Seed production to seed dispersal Appearance of early or late species Availability of flowers to pollinators

In mountain regions with short summers, even a few weeks can result in changes in presence and phenological phases of plant species.

Nitrogen-Fixing Microbes

The Search for Nitrogen-Fixing Microbes: Root nodules with N-fixing bacteria present in Lupinus sp., not yet observed in Oxytropis campestris. The lichens had Chlorophyta, green algae, but not N-fixing Cyanobacteria. The search continues for Cyanobacteria in cryptogamic crusts. Schematic of major nitrogen transformation steps in soils and microbial genes involved in them. Dashed lines=Biological incorporation, Solid lines=transformations. Microbes can be sensitive biomarkers of climate Boxes=specific genes involved in step change in alpine environments - and targeted by our study. We plan to assess temporal and spatial changes in microbial communities by measuring bacterial abundance and key enzymatic gene expression for a suite of nitrogen and carbon cycle related genes in cryptogamic crusts on Mt. Fleecer.