RESPONDING TO SOIL FUNGAL COMMUNITIES: a look at interactions between arbuscular mycorrhizal fungi and the common yellow monkeyflower Mariah McIntosh

What are AMF? soil-borne fungal symbiont From the vertebrate microbiome to the underground nests of leafcutter ants, symbiosis is a fundamental part of life on Earth. Arbuscular mycorrhizal fungi are a group of highly abundant soil-borne fungal symbionts, which colonize the roots of 80 percent of vascular plant species, providing nutrients, pathogen protection, and more, in exchange for plant carbon. soil-borne fungal symbiont 460 million year old symbiosis associate with ~80% of vascular plants exchange plant C for nutrients (P) provide pathogen protection

MECHANISMS OF INTERACTION AMF Strongly Affect Plant Ecology nutrients pathogens agriculture plant fitness conservation restoration community ecology MECHANISMS OF INTERACTION adaptation specificity dependency soil texture pH soil type plant diversity

MECHANISMS OF INTERACTION AMF Strongly Affect Plant Ecology nutrients pathogens agriculture plant fitness conservation restoration community ecology MECHANISMS OF INTERACTION adaptation specificity dependency soil texture pH soil type plant diversity community composition

MECHANISMS OF INTERACTION AMF Strongly Affect Plant Ecology nutrients pathogens agriculture plant fitness conservation restoration community ecology MECHANISMS OF INTERACTION adaptation specificity dependency soil texture pH soil type plant diversity community composition ecosystem variability

MECHANISMS OF INTERACTION AMF Strongly Affect Plant Ecology nutrients pathogens agriculture plant fitness conservation restoration community ecology MECHANISMS OF INTERACTION adaptation specificity dependency soil texture pH soil type plant diversity community composition ecosystem variability overall productivity **Implications for plant community ecology, agriculture, restoration, and plant response to climate change.

MECHANISMS OF INTERACTION nutrients pathogens agriculture plant fitness conservation restoration community ecology MECHANISMS OF INTERACTION adaptation specificity dependency soil texture pH soil type BIOTIC FACTORS Plant Functional Traits Plant Communities Microbe Functional Traits Microbe Communities ABIOTIC FACTORS Soil Properties Soil Chemistry Climate

Plant Response to AMF is Highly Variable Grassland Species } Benefit The influence of Glomus etunicatum (home) on the growth of 64 home plants (Experiment 2). Bars represent the percentage change in biomass of mycorrhizal plants compared to nonmycorrhizal controls. The mean 95% confidence interval for all the treatments is ±17%. The plant species names are in Table 1 **More about life histroy } Cost Klironomos, 2003

Plant Response to AMF is Highly Variable Grassland Species Crop Species } Benefit Benefit The influence of Glomus etunicatum (home) on the growth of 64 home plants (Experiment 2). Bars represent the percentage change in biomass of mycorrhizal plants compared to nonmycorrhizal controls. The mean 95% confidence interval for all the treatments is ±17%. The plant species names are in Table 1 **More about life histroy } Cost Cost Eo & Eom, 2009 Klironomos, 2003

Plant Functional Traits Predict Dependency on AMF **these are broad trait categories other traits include root traits, specific leaf area Focus on life history Wilson & Harnett, 1998

Plant Functional Traits Predict Dependency on AMF Wilson & Harnett, 1998

Plant Functional Traits Predict Dependency on AMF Wilson & Harnett, 1998 Perennials Live multiple years Slow growing More time to associate Likely to benefit more from AMF

Plant Functional Traits Predict Dependency on AMF **Macroevolutionary scale, highly differentiated taxa Wilson & Harnett, 1998 Annuals Short lifespan Fast growing May be a cost to allocating carbon to AMF Less likely to benefit from AMF Perennials Live multiple years Slow growing More time to associate Likely to benefit more from AMF

Contrasting Mimulus guttatus Populations DUN **To test for differences at a microevolutionary scale, we used to closely related, but contrasting populations of the same species **Color scheme perennial low elevation dunes few large flowers slow to flower thick, extensive roots rhizomatous

Contrasting Mimulus guttatus Populations DUN IM **Reference color scheme **Microevolutionary scale, genetic differences perennial low elevation dunes few large flowers slow to flower thick, extensive roots rhizomatous annual high elevation alpine many large flowers quick to flower shallow, fragile roots reproduce by seed

Both Populations Are Highly Locally Adapted \ IM DUN DUN IM Hall & Willis, 2006

Dependency Do DUN and IM plants differ in how they depend on AMF?

Dependency Do DUN and IM plants differ in how they depend on AMF? Because of its perennial life history, DUN plants will be more dependent on AMF than IM plants, performing better when grown with AMF than when grown without AMF. plant performance DUN IM -AMF +AMF plant type

Local Adaptation Do DUN and IM perform better in their native AMF communities versus exotic AMF communities?

Local Adaptation Do DUN and IM perform better in their native AMF communities versus exotic AMF communities? IM DUN AMF type plant performance Because DUN and IM plants are known to be highly locally adapted, both plant types will be locally adapted to native AMF communities and will perform better with these communities than with other AMF communities.

Common Garden Experiment Full factorial common garden greenhouse experiment

Common Garden Experiment DUN = 12 12 + - x IM + IM GEN NONE 2 perennial annual - - + + + + + + + + x x - - - - - - - -

Common Garden Experiment DUN = 12 12 + - x IM + IM GEN NONE 2 perennial annual - - + + + + + + + + x x - - - - - - - -

Common Garden Experiment DUN = 12 12 + - x IM + IM GEN NONE 2 perennial annual - - + + + + + + + + x x - - - - - - - -

Common Garden Experiment DUN = 12 12 + - x IM + IM GEN NONE 2 perennial annual - - + + + + + + + + x x - - - - - - - -

Common Garden Experiment DUN = 12 12 + - x IM + IM GEN NONE 2 perennial annual - - + + + + + + + + x x - - - - - - - -

Common Garden Experiment DUN = 12 12 + - x IM + IM GEN NONE 2 perennial annual - - + + + + + + + + x x - - - - - - - -

MECHANISMS OF INTERACTION nutrients pathogens agriculture plant fitness conservation restoration community ecology MECHANISMS OF INTERACTION adaptation specificity dependency soil texture pH soil type BIOTIC FACTORS Plant Type Microbe Origin AMF+/- ABIOTIC FACTORS Soil Type AMF greatly influence plant diversity, community composition,ecosystem variability, and overall productivity. Therefore, this important mutualism strongly affects plant ecology, with implications for agriculture, ecological restoration, and plant responses to climate change. Understanding the mechanisms that govern plant-AMF interactions may explain patterns seen at ecological and evolutionary levels. Current research on plant-AMF relationships considers these interactions on a community level; however, these mechanisms remain a mystery at the individual level. **Break down into plants, soil, microbes, interactions and how they affect plant performance

What Biotic & Abiotic Components Affect Plant Growth & Fitness? Growth (aboveground biomass) ✔️ R2= .66 Source Nparm DF Sum of Squares F Ratio Prob > F Soil Type 1 3.4384703 311.0525 <.0001* Plant Type 0.0682288 6.1721 0.0135* Soil Type*Plant Type 0.0003042 0.0275 0.8684 Microbes 8 1.6097469 18.2027 Soil Type*Microbes 0.4752787 5.3744 Plant Type*Microbes 0.1368769 1.5478 0.1403 Soil Type*Plant Type*Microbes 0.1143153 1.2927 0.2468

What Biotic & Abiotic Components Affect Plant Growth & Fitness? Growth (aboveground biomass) ✔️ R2= .66 Source Nparm DF Sum of Squares F Ratio Prob > F Soil Type 1 3.4384703 311.0525 <.0001* Plant Type 0.0682288 6.1721 0.0135* Soil Type*Plant Type 0.0003042 0.0275 0.8684 Microbes 8 1.6097469 18.2027 Soil Type*Microbes 0.4752787 5.3744 Plant Type*Microbes 0.1368769 1.5478 0.1403 Soil Type*Plant Type*Microbes 0.1143153 1.2927 0.2468 **Robust experiment with power Soil type, plant type, microbes, and some interactions between these components accounted for a large percentage of the variance Fitness (potential fruit) ✔️ R2= .56 Source Nparm DF Sum of Squares F Ratio Prob > F Soil Type 1 11245.409 49.8827 <.0001* Plant Type 52112.658 231.1626 Soil Type*Plant Type 2261.687 10.0324 0.0017* Microbes (Origin & AMF+/-) 8 5546.238 3.0753 0.0024* Soil Type*Microbes 2358.622 1.3078 0.2390 Plant Type*Microbes 2185.630 1.2119 0.2915 Soil Type*Plant Type*Microbes 1435.571 0.7960 0.6065

Are DUN and IM dependent on AMF? Growth We did not find evidence that DUN or IM plants are dependent on AMF. P<.0001 P=.0043

Are DUN and IM dependent on AMF? Fitness Growth P<.0001 P=.5691 P=.0043 P=.2339

Are DUN and IM locally adapted to native AMF? Growth We did not find evidence that DUN or IM plants are locally adapted to native AMF. P=.0024 P=.0024

Are DUN and IM locally adapted to native AMF? Fitness Growth P<.0001 P=.0024 P=.0024

Are DUN and IM different in how they interact with AMF? No evidence for dependency on AMF in either plant type

Are DUN and IM different in how they interact with AMF? No evidence for dependency on AMF in either plant type Expected results for IM, but not DUN plants **Some cost to associating with AMF for annuals, like fast growing IM plants

Are DUN and IM different in how they interact with AMF? No evidence for dependency on AMF in either plant type Expected results for IM, but not DUN plants Experiment was robust, and plants manifested their growth forms **We had power to detect differeences, and the plants were allowed to grow long enough to fully mainifest different growth forms It is possible that, had DUN plants been allowed to grow for longer (years), we could have detected differences Other experiements that did detect dependency did not exceed one year

Are DUN and IM different in how they interact with AMF? No evidence for dependency on AMF in either plant type Expected results for IM, but not DUN plants Experiment was robust, and plants manifested their growth forms No evidence for local adaptation to native AMF communities

Are DUN and IM different in how they interact with AMF? No evidence for dependency on AMF in either plant type Expected results for IM, but not DUN plants Experiment was robust, and plants manifested their growth forms No evidence for local adaptation to native AMF communities Many mycorrhizal plant species and few fungal species 300,000 species of plants associate with AMF ~150 microbe species Suggests local adaptation is unlikely

Ongoing Work May Detect Differences in Colonization and Fungal Communities Root Colonization Data

Ongoing Work May Detect Differences in Colonization and Fungal Communities Root Colonization Data Amplicon Sequencing

Davidson Honors College Special thanks to: Lila Fishman Ylva Lekberg Alexii Rummel Findley Finseth Aurora Bayless Hanna McIntosh Patrick Demaree Andrew Demaree Sigma Xi Grant-In-Aid Davidson Honors College I would like to thank Lila Fishman and Ylva Lekberg for being wonderful mentors on this project. It has been challenging in nearly every way possible, but I’ve learned a ton and can’t thank you enough. Also, thank you to Findley Finseth, Aurora Bayless, Hanna McIntosh, and Patfick Demaree for help along the way. Thank you!