We anticipated that endophytes of aquatic plants would represent subsets of otherwise terrestrial taxa which have colonized water independently (prediction six). Lastly, we predicted that aquatic endophytes would be largely distinct from communities of endophytes in proximate terrestrial plants (prediction 7), and that they would be readily recovered from lake sediment and water (prediction 8), congruent with the horizontal transmission that characterizes the vast majority of endophyte associations [7].NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptMaterials and MethodsAquatic macrophytes had been collected from each and every of three microsites along the shore of one natural freshwater lake (Stoneman Lake) and five freshwater reservoirs (Lower Lake Mary, Morton Lake, Mud Lake, Watson Lake, and Willow Creek Reservoir) in northern Arizona, USA, in September-October 2011, and three reservoirs in early June 2012 (Reduce Lake Mary, Watson Lake, Willow Creek Reservoir) (Table 1). Microsites were spaced at ca. 30?40 m intervals along accessible shorelines. Average water depth at every single sampling web site wasMicrob Ecol. Author manuscript; accessible in PMC 2015 Might 01.Sandberg et al.Page33.5 cm in 2011 (variety: four.four?8.four cm) and 29.0 cm in 2012 (variety: 9.4?four.0 cm) (Table 1). Interlake distances ranged from 2 km to 110 km. Water temperatures in Stoneman Lake, Decrease Lake Mary, Morton Lake, and Mud Lake (Small Colorado River Watershed, Walnut Creek and Canyon Diablo complexes) commonly are cooler than these of Watson Lake and Willow Creek Reservoir (Verde River Watershed) [4]. With the 3 reservoirs sampled in both years (Lower Lake Mary, first filled in 1905 [4]; Willow Creek Reservoir, very first filled in 1936 [4]; and Watson Lake, 1st filled in 1915 [4]), Decrease Lake Mary varies most in water volume: it frequently dries absolutely in the course of the summer dry season (Table 1). Each Willow Creek Reservoir and Watson Lake maintain some water volume each and every year, but shorelines fluctuate according to rainfall and snowmelt.4,4′-Diphenyl-2,2′-bipyridine web All are stocked routinely with fish [4].Methyl 5-bromo-3-hydroxypicolinate site Plant collections We focused our sampling on 3 with the most abundant, native species of freshwater plants in the study region (see [27, 60]) (Table 1): Persicaria amphibia (L.PMID:23291014 ) A. Gray (syn. Polygonum amphibium L.; Polygonaceae; emergent), Stuckenia pectinata (L.) B rner (syn. Potamogeton pectinatus L.; Potamogetonaceae; submergent), and Elodea bifoliata H. St. John (Hydrocharitaceae; submergent). We also examined an extra species popular to Arizona [see 27, 60] when it was encountered in our study sites (Myriophyllum sibiricum Komarov; Haloragaceae; submergent). All four species were sampled in 2011 (Table 1). Sampling in 2012 focused only on P. amphibia and S. pectinata, which had been particularly abundant at all sites (Table 1). Submergent plants have been totally uprooted and partitioned into shoots (photosynthetic stems and leaves) and roots, which were placed quickly into sealable bags having a tiny amount of water from the collection web page. Emergent plants were removed in pieces according to tissue type (roots, submerged stems and leaves, and emergent stems and leaves) and were stored with water (submerged tissues) or without the need of water (emergent tissues) in sealable plastic bags. All plant material was transported for the lab inside a cooler for processing within 72 hours of collection. Vouchers of all plants were deposited in the University of Arizona Herbarium (ARIZ: collections DCS001-DCS022, accessions 40.