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Paraburkholderia species promote Broccoli growth in a cultivar-dependent manner
Root tip inoculation of the two Broccoli cultivars with strains of three different Paraburkholderia species led to changes in leaf color (deep green leaves), shoot biomass, root biomass and root architecture (Fig. 1a). Percent change in biomass was used as a measure to assess the growth-promoting effects of the Paraburkholderia species in the two Broccoli cultivars. Two-way analysis of variance (ANOVA) was conducted to assess the influence of the two independent variables (strains of Paraburkholderia species and Broccoli cultivars) on both shoot and root biomass. The Paraburkholderia species included three levels (Pbg, Pbh, Pbt) and the Broccoli cultivars consisted of two levels (Coronado, Malibu). For shoots, all interactions, except Pbt-Malibu, resulted in significant increases in biomass relative to the non-treated control plants, while for roots all three Paraburkholderia species significantly increased the biomass in both Broccoli cultivars (Fig. 1b). In general, the relative impact of Paraburkholderia species was up to 3 times higher for root biomass than for shoot biomass (Fig. 1b). Two-way ANOVA showed highly significant interactions between the strains of Paraburkholderia species and Broccoli cultivars regarding the percent changes in shoot and root biomass (Supplementary Table S1). Overall, for cultivar Coronado the percent change in shoot biomass was about 40% compared to the control, and not significantly different between the different strains of Paraburkholderia species, whereas in cultivar Malibu the percent change in shoot biomass was significantly higher for Pbg (~ 70%) and Pbh (~ 90%) as compared to Pbt. Furthermore, inoculation with Pbh led to a significantly higher increase in shoot biomass in cultivar Malibu than in Coronado. Regarding the percent change in root biomass, only inoculation of Pbt showed significant differences between the two Broccoli cultivars. As indicated above, the shoot biomass of cultivar Malibu inoculated with Pbt was not significantly different from the control plants (Fig. 1b). Over a period of 11 days, both Pbg and Pbh-treated Broccoli cultivars showed significantly higher shoot and root biomass from 7 days post inoculation (dpi) onwards, while Pbt-treated plants showed higher shoot biomass in Coronado from 9 dpi onwards (Fig. 1c).
Figure 1

Biomass and phenotypic changes in Broccoli cultivars in response to root tip inoculation with strains of three Paraburkholderia species. (a) Pictures of MS agar plate with two Broccoli cultivars (Coronado and Malibu) at 11 days post inoculation with strains of three Paraburkholderia species (Pbg: Paraburkholderia graminis PHS1, Pbh: P. hospita mHSR1, and Pbt: P. terricola mHS1). (b) Percent changes in shoot and root biomass (mean ± standard error, n = 4 (shoot) and n = 6 (root)) of two Broccoli cultivars inoculated with the strains of the Paraburkholderia species. Treatments sharing the same letters are not significantly different (Two-way ANOVA, Tukey’s HSD post hoc test, P  More

Focal species
The Lesser Prairie-Chicken is a medium-sized grouse endemic to, broadly, the shortgrass prairie ecosystem of the south-central United States, where it is found only in southwestern Colorado, western Kansas, northwestern Oklahoma, the Texas panhandle, and eastern New Mexico. As with almost all open-country grouse of temperate environments, the Lesser Prairie-Chicken forms leks at which a cluster of males (in this species, usually 5–12 individuals) display vigorously and female visit to assess males with an intent to secure sperm to fertilize her eggs, which she will lay and raise without male help. Outside of male lekking (mid-March to mid-May) and female nesting (late April to early July), birds congregate is small flocks to forage, at times on grain remains in farmed fields but typically, as through the rest of the life cycle, restricting themselves to native prairie. Accordingly, this species has three distinct aspects of habitat selection: general occurrence, lek placement, and nest placement.
Data
Lesser Prairie-Chicken were tracked at two study sites, one in Roosevelt County in east-central New Mexico, U.S.A., the other in Beaver, Harper, and Ellis Counties in northwestern Oklahoma, U.S.A. Birds tagged with VHF transmitters were tracked from April 1999–March 2006 in New Mexico and from March 1999–July 2013 in Oklahoma11,12,13,14. Study periods differed chiefly because of funding, which for New Mexico was insufficient after 2006. The study sites differ markedly in land tenure history. Parcel size in New Mexico averages 1300 ha versus 180 ha in Oklahoma11. The difference largely stems from settlement patterns over the past two centuries. New Mexico was part of the Spanish land grant system, which tended to yield huge parcels. In our study area, parcels approach 8 km2 ( > 1900 acres). By contrast, during the “land rush” era of the late 19th Century most of northwestern Oklahoma was parceled into 65-ha (160-acre) plots as part of the United States’ Homestead Act. Smaller parcels translate to a higher density of roads, fences, buildings, and powerlines11.
Radiotracking typically yielded a triplet of coordinate readings, from which we had to triangulate a grouse’s location. We estimated latitude and longitude using a maximum likelihood estimator (MLE), although it some cases the MLE algorithm failed to converge. If it failed, we instead used the Andrew and Huber methods15. R code for the estimation procedure can be found at https://github.com/henry-dang/triangulation/blob/master/lenth_triang.R.
From these data we used kernel density methods (R package ks16) to estimate annual home ranges (235 in New Mexico, 263 in Oklahoma). Tracking data included lek (12 in New Mexico, 23 in Oklahoma) and nest (122 in New Mexico, 128 Oklahoma) locations. For home range centroids, the outer contour of home ranges, leks, and nests, we estimated distance to seven anthropogenic features: roads (highways, primary, and secondary roads only; small farm roads or one-lane gravel roads were excluded), powerlines (overhead only, with buried or trunk lines excluded; https://hifld-geoplatform.opendata.arcgis.com/datasets/electric-power-transmission-lines), oil wells, gas wells (for both types of wells, http://www.occeweb.com and http://www.emnrd.state.nm.us/ocd), outbuildings (barns, grain silos, poultry houses, and similar large structures; chiefly the TIGER database), and fences (Bureau of Land Management) in both states, plus private houses in New Mexico and railroad tracks in Oklahoma. We placed 2000 random points on each study area to estimate distances to each of these same anthropogenic features, which provided an estimate of feature density on the landscape.
Analyses
The initial step was to estimate the probability of a grouse occurring a certain distance from a feature. We treated New Mexico and Oklahoma data separately, giving us a replicate assessment because these populations have been isolated from each other for  > 100 years17 and, as noted above, land tenure history differs strikingly between the states11. We estimated probabilities of grouse occurrence, πi, via a Bayesian model with binomial likelihood and flat prior (i.e., no assumption of the central tendency of occurrence probability at a given distance from a feature):
yi ~ binomial (πi, n) with yi the cumulative count of grouse at distance i (i.e., the data) and n the total number of home ranges, leks, or nests. Distances, i, were binned to the smallest extent possible, from 10 to 100 m, to allow the Markov chain Monte Carlo (MCMC) algorithm to converge in a reasonable number of iterations (e.g.,  More

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