CONSERVATION AND RESTORATION
HORTSCIENCE 54(3):445–451. 2019. https://doi.org/10.21273/HORTSCI13600-18 Seedbed preparation is one of the most important actions affecting PR establishment success because of its long-term impacts on Pollinator Refuge Establishment and weed management (Aldrich, 2002; Martin, 1986; Wilson, 1992). Previous studies indi- Conservation Value: Impacts of cate that weed competition can be a critical factor limiting PR success (Aldrich, 2002; Seedbed Preparations, Seed Mixtures, Howell and Kline, 1992; Norcini and Aldrich, 2004; Perry, 2005). Although seedbed prep- aration recommendations for PR installa- and Herbicides tions vary (NRCS, 2007, 2012; Vaughan Gina M. Angelella, Laura Stange, Holly L. Scoggins, et al., 2013), a review of the literature sug- and Megan E. O’Rourke1 gested repeated tilling, herbicide applications, or both are better methods for perennial weed School of Plant and Environmental Sciences, Virginia Tech, 185 Ag-Quad management than other strategies such as graz- Ln, Blacksburg, VA 24061 ing, solarization, or burning (Aldrich, 2002). Studies comparing the results of a no-till/ Additional index words. wildflower, no till, imazapic, graminicide, sethoxydim, perennial, preemergent herbicide method with repeated pollinator tillage have yielded conflicting results (Ahern Abstract. On-farm pollinator refuge habitats can supplement floral and nesting resources et al., 1992; Corley, 1991; Corley et al., 1993; to support wild and managed pollinator communities. Although the popularity of Skousen and Venable, 2008), but a recent installing these habitats has grown, and federal programs provide technical and financial study showed that in the first establishment advice to participating landowners, recommendations regarding habitat establishment year, a mixture of annual, biennial, and peren- and species composition vary. We examined the effects of seedbed preparation, seed mix nial wildflowers planted on cropland is more composition, and herbicide applications on pollinator refuge establishment and pollina- successful in a no-till than a tilled system tor visitation in a controlled experiment across 2 years. Seedbeds were prepared either by (Angelella and O’Rourke, 2017). a no-till method with glyphosate herbicide or by repeated conventional inversion tillage. After preparing a PR seedbed, a critical Seed mixes contained either nine annual, biennial, and perennial forbs (mix AP); seven question is which species of wildflowers to annual, biennial, and perennial forbs that are tolerant to imazapic herbicide (mix IT); or plant. Pollinator habitat mix composition nine perennial forbs (mix P). Mixes AP and P were grown with and without application of may affect both wildflower and weed estab- the graminicide herbicide sethoxydim and mix IT was grown with application of the lishment as well as pollinator conservation. herbicide imazapic. Seedbed preparation methodology had a strong impact on pollinator Annual wildflowers can reduce weed com- refuge establishment. A no-till approach generated greater wildflower and lower weed petition during the critical initial phases of cover relative to tillage, leading to a greater number of blooms. In particular, there were establishment (Aldrich, 2002) and can pro- more Indian blanket, purple coneflower, slender mountain mint, and wild bergamot vide floral resources in the first year, but blooms following a no-till seedbed preparation, indicating that certain species are more species should be selected so as to minimize vulnerable to the effects of tillage than others. The AP and IT treatments displayed more the risk of becoming weeds themselves if wildflower and less weed percent cover than the P treatments during the first year, but in seeds get dispersed outside PRs (Menz et al., the second year wildflower and weed cover were similar across all mixes grown with and 2011). The benefits of perennial wildflowers without herbicide. Overall pollinator abundance, which was dominated by native bees, include that they compete well with an- correlated positively with wildflower bloom counts, suggesting that habitat establishment nual weed species once they are established methods that increase wildflower blooms can positively affect the pollinator conservation (Lulow, 2006) and that they do not re- value of the habitats. This research indicates that establishing on-farm wildflower quire frequent overseeding (Aldrich, 2002). habitats can be most successful with no-till seedbed preparation, a mixture of annual, Moreover, some research suggests bumble- biennial, and perennial forb species, and that herbicides applied after planting bees and butterflies prefer perennials over wildflowers may not be worth the costs of application. annuals (Feber and Smith, 1995; Fussell and Corbet, 1992), although other studies report a lack of preference (Carreck et al., 1999), or Habitat installations rich in floral pollen, participating farmers by, respectively, the U.S. that a combination of annuals and peren- nectar, and nesting resources can support local Department of Agriculture’s Natural Resources nials can work complementarily to attract pollinator populations on farms, thus provid- Conservation Service (USDA NRCS) and the more pollinators (Williams et al., 2015). ing a farmscaping tool to alleviate native and Farm Service Agency (Vaughan and Skinner, Although PRs will certainly be influenced managed pollinator declines and the accom- 2008, 2015). Pollinator refuge habitats (PRs) in by the species-specific preferences of growers, panying declines in pollination ecosystem these programs are designed to provide blooms more can be done to develop general recom- services (Vaughan and Skinner, 2008, 2015). throughout the entire growing season and may mendations about the benefits and trade-offs Pollinator habitat installation is supported by also contain grasses depending on regional of species mixes that consist of all peren- federal programs such as the Environmental recommendations (FSA, 2013). As interest in nials or a combination of short- and long- lived species. Quality Incentives Program and the Conser- these programs develops, there have been calls Managing weeds after planting wild- vation Reserve Program, through which tech- for research into the factors affecting PR nical and financial assistance is provided to flowers is another major challenge that establishment and effectiveness. Specifically, growers and advisors must address during how can wildflower establishment success be the PR establishment process. Some studies maximized to obtain the full diversity of the indicate that herbicides improve PR establish- Received for publication 24 Sept. 2018. Accepted mix planted (Williams et al., 2015)? Further- ment, whereas others indicate that it is an for publication 18 Dec. 2018. more, how does PR establishment success and unnecessary expense. The selective herbicide We thank Velva Groover, Chris McCullough, and species composition affect pollinator commu- imazapic, for example, can enhance the estab- Mike Graham for data collection contributions. nities (Venturini et al., 2017)? We conducted a lishment of tolerant wildflower species (Beran This research was supported by USDA Agroeco- study to address these questions, investigating system Management Grant 11664133 and start-up et al., 1999), but effects can vary by site and funds provided to M.E. O’Rourke by the College of PR establishment and pollinator conservation species (Frances, 2008) as well as by weather Agriculture and Life Sciences at Virginia Tech. value relative to seedbed preparation, seed mix conditions, soil quality, and seed source 1Corresponding author. E-mail: [email protected]. composition, and herbicide treatments. (Norcini and Aldrich, 2004). Similarly, the
HORTSCIENCE VOL. 54(3) MARCH 2019 445 graminicide sethoxydim enhanced wildflower plots on 13 Apr. 2016, making two passes with and three times in 2017 (6 June, 25 July, and establishment and richness in two studies the tiller down the plots. All plots were packed 11 Sept.); total grass percent cover was (Hitchmough et al., 2008; Trowbridge et al., with a homemade roller (132-cm-long barrel quantified similarly only on 8 Sept. 2016 2017), but was found to be ineffective in holding 378.5 L water) pulled by a tractor and on 11 Sept. 2017. We also counted total another (Frances, 2008). Further investigation immediately pre- and postplanting to ensure blooming plants per wildflower species at into the consistency of herbicide effectiveness good seed to soil contact. Seed mixes were three time points in the first year (13 June, 20 applied during the early stages of PR growth developed with USDA NRCS technical advice July, and 8 Sept. 2016) and total number of would enhance our ability to make recom- (B. Glennon, personal communication) for blooms at eight time points in the second year mendations regarding weed management native or naturalized species appropriate for (24 May, 13 June, 28 June, 18 July, 31 July, practices for successful PR establishment. Virginia that would provide bloom coverage 14 Aug., 28 Aug., and 27 Sept. 2017) within To address questions of how to best from late spring through late summer. One mix two randomly dropped square meter quad- establish PRs on cropland and the subsequent (mix AP) contained annual [partridge pea– rants per plot. To quantify the number of impacts on pollinators, we examined the Chamaecrista fasciculata (Michx.) Greene, blooms in the second year, the number of effects of seedbed preparation, seed mix plains coreopsis–Coreopsis tinctoria Nutt., individual stems with open inflorescences on composition, and herbicide use on PR estab- and indian blanket–Gaillardia pulchella them was tallied. Composites and umbels lishment in a controlled field experiment over Foug.], annual or biennial (blackeyed Susan– were considered one flower; e.g., lanceleaf two years. We hypothesized that 1) wild- Rudbeckia hirta L.), and perennial forbs coreopsis with multiple blooming branches flower establishment would be higher and [lanceleaf coreopsis–Coreopsis lanceolata L., off of a stem were individually counted, weed establishment would be lower follow- purple coneflower–Echinacea purpurea (L.) whereas multiple blooms on an unbranched ing a no-till than a tillage seedbed prepara- Moench, maximilian sunflower–Helianthus stem such as partridge pea or maximilian tion; 2) herbicide applications following PR maximiliani Schrad., wild bergamot–Monarda sunflower were counted as one. planting would suppress weeds and enhance fistulosa L., and narrowleaf mountain mint– Pollinators. We recorded pollinator abun- wildflower growth; 3) mixes containing both Pycnanthemum tenuifolium Schrad.]; a second dance and floral visitations through visual annuals and perennials would have more mix(mixIP)consistedofanimazapic-tolerant observations during 2017. We divided polli- wildflower growth, more bloom coverage, subset of mix AP (C. fasciculata, C. tinctoria, nators into the following categories: honey and less weed growth than a mix containing G. pulchella, R. hirta, C. lanceolata, E. pur- bees (Apis mellifera L.), bumblebees (Bombus only perennials; and 4) pollinator abundance purea,andH. maximiliani); and a third mix spp.), other bee species (Apiaceae, excluding would correlate positively to bloom density. (mix P) contained only perennial forbs, in- Bombus spp. and A. mellifera), syrphid flies cludingalltheperennialsinmixAPandfour (Syrphidae), wasps (insects within the sub- Materials and Methods additional perennial species [pale purple order Apocrita), and butterflies and moths coneflower–Echinacea pallida (Nutt.) Nutt., (insects within the order Lepidoptera). Polli- Experimental design. We established ex- spotted bee balm–Monarda punctata L., showy nator observations were made in the mornings perimental sites at the Virginia Tech Kent- evening primrose–Oenothera speciosa Nutt., after dew was dry on seven dates throughout land Agricultural Research Farm, in and tall white beardtongue–Penstemon digitalis the second year roughly coinciding with Montgomery Co., VA (lat. 37 11#59118$N, Nutt. ex Sims] (Supplemental Table 1). For bloom counts (9 June, 22 June, 6 July, 18 long. 80 33#51.8688$W). The farm is in the bulking purposes, we mixed seeds in a 20:1 July, 2 Aug., 16 Aug., and 30 Aug. 2017). USDA Plant Hardiness Zone 6b, and the pelletized lime (Soil Doctor, Haines City, FL) Floral visitations—that is, the wildflower spe- experimental site was on fine-loamy, mixed, by seed weight ratio, at a rate of 30–140 seeds/ cies on which each pollinator landed—were active Ultic Hapludalf soil. The fall before m2 and hand-broadcast them in treatment plots additionally recorded on the three observation our study, the experimental site had been on 26 Apr. 2016. Treatments AP-h and P-h dates in Aug. 2017. Pollinator observations planted with annual rye (Lolium multiflorum were sprayed with the grass-selective, broad- were conducted by dividing each plot in half Lam.) and hairy vetch (Vicia villosa Roth) spectrum, postemergent herbicide sethoxydim and observing all pollinators interacting with cover. The experiment was designed as a at 1.17 L·ha–1 on 18 July 2016. Mix IT, flowers for three minutes in each half of the randomized complete block experiment with containing imazapic-tolerant species only, plot. Care was taken to not count the same four replicate blocks and 10 experimental was sprayed with a postemergent treatment of individual pollinator twice within a particular treatment plots per block. Each plot mea- imazapic at 0.29 L·ha–1 during the first year of plot-half. Plots were observed in random order sured 6.10 · 2.74 m. The plots consisted of a establishment on 18 July 2016; imazapic was on each sampling date. combination of seedbed preparation, seed also applied as a pre-emergent herbicide during Data analysis. We examined the effects of mixes, and herbicide treatments in an un- the second year of the study on 8 May 2017. seedbed preparation and mix-herbicide treat- balanced factorial design. Specifically, we Sethoxydim and imazapic treatments were ments on total wildflower cover, total weed tested the effects of seedbed preparation (till applied with a backpack sprayer (Solo, New- cover, grass weed cover, total wildflower and no till) fully crossed with three wild- port News, VA). Vegetation was mowed in an blooms, and numbers of individual species’ flower species mixes: an annual and peren- effort to control weeds in the P and P-h plots on blooms. Each of these factors was analyzed nial mix (mix AP), an imazapic-tolerant mix 12 July and 8 Aug. 2016 to a height of 10 cm by fitting fully factorial generalized linear (mix IT), and a perennial-only mix (mix P). with a rotary mower (Woods 121; Oregon, IL). mixed models to the data, with seedbed We also examined the effects of herbicide Vegetation within all plots was mown to 30.5 preparation as a fixed effect, the five mix- treatments, but they were not fully crossed cm with the Woods rotary mower after wild- herbicide treatments as a second fixed effect with the other treatment factors because flower senescence on 16 Sept. 2016 and again (i.e., treatments AP, AP-h, IT, P, and P-h), an herbicide management was specific to the on 31 Oct. 2016. All plots were mown only interaction between seedbed preparation and wildflower mix. Mix AP was tested with one time at the end of the growing season in mix-herbicide treatments, and block as a (treatment AP-h) and without graminicide 2017 on 4 Oct. to a height of 30.5 cm with the random effect. Analyses were conducted for (treatment AP), mix IT was tested just with Woods rotary mower. each year of study. Data were averaged over imazapic (treatment IT), and mix P was Wildflowers and weeds. Within the PR plot subsamples and sample dates within tested with (treatment P-h) and without gra- treatments, we quantified wildflower and years before analyses. Total wildflower cover minicide (treatment P). weed biomass and percent cover, and num- was cube-root transformed for both years of To prepare seedbeds before planting, we bers of wildflower blooms during the first data, total weed cover was arcsine square- applied glyphosate to no-till plots at 4.68 L·ha–1 (2016) and second (2017) years of establish- root transformed for 2016 data, percent grass using a tractor-mounted 6.1-m boom sprayer ment. We recorded total wildflower and total cover was log+1 transformed for 2017 data, (Reddick, Williamston, NC) on 13 Apr. and 26 weed percent cover per square meter by and total wildflower blooms were log trans- Apr. 2016. Concordantly, we rotary power- taking two random subsamples per plot three formed for both 2016 and 2017 data be- tilled (Kuhn EL 43-190; Broadhead, WI) tillage times in 2016 (13 June, 20 July, and 8 Sept.) fore analysis to meet model assumptions of
446 HORTSCIENCE VOL. 54(3) MARCH 2019 normality and homogeneity of variance. Be- (AP, AP-h, and IT) than in treatments con- cause of scarcity, slender mountain mint and taining perennials only (P and P-h) (Fig. 1). indian blanket bloom counts from the second This changed in the second year in that, year were converted to presence/absence data although the mix-herbicide treatment effect and analyzed with binomial regression and a was significant overall, differences in wild- logit link. Bloom count data for perennial flower cover between particular mixes were species were not analyzed in the first year not statistically different by Tukey’s HSD post because they were not yet blooming. We hoc analysis (Fig. 1). Wildflower cover was performed Tukey’s honestly significant dif- consistently greater following the no-till ference (HSD) tests of sample means when method than the tillage method in both years model factors were significant at the 0.05 (Fig. 2). level, and there were more than two levels for Only the plots with annual species the factor. bloomed during the first year, whereas all Fig. 1. Proportions of wildflower and total weed Pollinator data were analyzed to examine wildflower plots bloomed during the second cover by mix in 2016 and 2017. Annual and relationships between total blooms and total year (Table 1). Black-eyed Susan provided perennial mix (mix AP-h) includes nine species pollinator abundance, treatments and total the greatest bloom counts overall, followed of annual and perennial wildflowers and was treated with a sethoxydim herbicide, mix AP is pollinator abundance, and specific species’ by indian blanket, partridge pea, and plains the same wildflower mix without herbicide, blooms and specific pollinator groups. To coreopsis. Two species in the perennial-only imazapic-tolerant mix (mix IT) contains a sub- examine the relationships between total wild- mix, pale purple coneflower and tall white set of seven AP wildflower species and was flower abundance and total pollinator abun- beardtongue, never bloomed during the 2 treated with imazapic herbicide, perennial-only dance, we averaged wildflower bloom and years of study (Table 1). Three of the four mix (mix P-h) contains nine perennial wild- pollinator abundance data across all sample annuals and biennials increased in blooms in flowers and was treated with sethoxydim her- dates in 2017 and conducted a linear regres- the second year relative to the first: black- bicide, and mix P is the same wildflower mix 2 without herbicide. Tukey’s honestly significant sion with mean total pollinator observations eyed Susan [c (1,48) = 31.50, P < 0.0001], c2 difference tests were performed to evaluate per square meter as the response variable and plains coreopsis [ (1,48) = 5.34, P = 0.021], # 2 significant differences at the P 0.05 level. total wildflower blooms per square meter as and partridge pea [c (1,48) = 35.31, P < the predictor variable, with herbicide-mix as a 0.0001] (Table 1). Bloom counts peaked at blocking factor. To evaluate the effects of the the mid-July sample dates in both years for experimental treatments on pollinators, total treatments AP, AP-h, and IT. The perennial- pollinator abundance was averaged over sam- only treatments (P and P-h) peaked in Sep- pling dates, log transformed, and analyzed tember in the second year driven by spotted with generalized linear mixed models, in bee balm blossoms. which herbicide-mix and seedbed preparation Although there were no treatment effects and their interaction were fixed effects and on total wildflower blooms during the first block was a random factor. To identify vari- year, in the second year total wildflower ations in floral visitation across pollinator blooms were affected by mix [F(4,27) = groups, we first divided the total pollinator 21.55, P < 0.0001] and seedbed preparation data into five categories: honey bees (A. [F(1,27) = 4.93, P = 0.035]. In 2017, mixes mellifera), bumblebees (Bombus spp.), moths AP-h and AP had the largest, mix IT was and butterflies (lepidopteran spp.), wasps intermediate, and mixes P-h and P had the (narrow-waisted Apocrita spp.), ‘‘other bees’’ smallest bloom counts. Bloom counts were (all other hymenopteran spp.), and syrphid larger overall following a no-till preparation Fig. 2. Proportions of wildflower and weed cover by no-till and till seedbed preparations in 2016 flies (Syrphidae spp.). Only wildflower spe- than a tillage preparation (Supplemental and 2017. Tukey’s honestly significant differ- cies visited by at least one pollinator per block Fig. 1). Individual wildflower species were ence tests were performed to evaluate signifi- were analyzed. Pollinator visitations were also affected by seedbed preparation with cant differences at the P # 0.05 level. 2 averaged over the 3 Aug. 2017 sample dates, more indian blanket in the first [c (1,24) = 2 and simple analyses of variance with pollina- 9.22, P = 0.0024] and second [c (1,24) = 4.37, tor group as a fixed effect and block as a P = 0.037] years, and more purple coneflower chickweed (Cerastium sp.). Total weed cover 2 random factor were used to test the differences [c (1,24) = 19.26, P < 0.0001], slender moun- was significantly affected by mix-herbicide 2 in pollinator visitations among pollinator tain mint [c (1,24) = 11.60, P = 0.0007], and treatment in 2016 [F(4,27) =12.26,P < 0.0001] 2 groups for each flower species. Tukey’s HSD wild bergamot [c (1,24) = 4.78, P = 0.029] in and was greater in the first year in the tests were used in pollinator analyses to assess the second year in plots with no-till than perennial (P and P-h) treatments than treat- significant differences among treatment tillage seedbed preparation (Supplemental ments with annuals, biennials, and perennials groups. Table 2). (AP, AP-h, and IT). However, there were Weeds. Total weed cover decreased over no differences in weed cover among mix- Results the 2 years of study, with large concomitant herbicide treatments in the second year (Fig. 1). shifts in the weed communities. In the first The effects of seedbed preparation on weed Wildflowers. Total wildflower percent year, grasses dominated the weed community cover were similar in both years, with the total cover increased over the 2 years of study, and averaged 89% cover by September weed cover being lower following a no-till with an average across all treatments and across all the treatments. However, by the preparation than a tillage preparation across sampling dates of 9% in 2016 and 49% in end of the second year, the weed community all mix-herbicide treatments [2016: F(1,27) = 2017. Total wildflower cover was signifi- had shifted toward more broadleaf species 100.23, P < 0.0001; 2017: F(1,27) =9.58,P = cantly affected by the mix-herbicide treat- such that broadleaf weeds made up 55% and 0.0045] (Fig. 2). ment and seedbed preparation in both the first grasses made up 15% of the cover, on average Pollinators. Pollinators were more abun- year [mix: F(4,27) = 33.70, P < 0.0001; (Supplemental Fig. 2). The primary weed in dant in treatment plots with more blooms seedbed preparation: F(1,27) = 62.00, P < the first year was large crabgrass [Digitaria (Fig. 3). Their abundances also varied by 0.0001] and the second year [mix: F(4,27) = sanguinalis (L.) Scop.] with additional mix-herbicide treatment (but not seedbed 2.81, P = 0.045; seedbed preparation: F(1,27) = patchy outbreaks of hairy galinsoga (Galin- preparation) with significantly more pollina- 12.68, P < 0.0014]. In the first year, wild- soga quadriradiata Cav.). In the second year, tors sampled in the AP-h and AP treatments flower cover was greater in treatments con- the major weeds were smartweed (Polygonum than the P treatment [F(4,30) = 5.62, P = taining annuals, biennials, and perennials pensylvanicum L.), clover (Trifolium sp.), and 0.0017; Fig. 4]. Honey bees were observed
HORTSCIENCE VOL. 54(3) MARCH 2019 447 Table 1. Mean overall bloom counts/m2, bloom period, and peak bloom time for each wildflower species.
Mean blooms/m2 (±SE)x Species Common name Mixz Life cycley 2016 2017 Bloom period Peak bloom Chamaecrista fasciculata Partridge pea AP A 0.99 (±0.21) ab 30.24 (±7.43) a Late June–Sept. Mid-July IT Coreopsis lanceolata Lanceleaf coreopsis AP P 0.00 (±0.00) c 0.13 (±0.032) gh Late May–mid-June Mid-June IT P Coreopsis tinctoria Plains coreopsis AP A 0.74 (±0.13) ab 2.52 (±0.93) bcdef Late June–Sept. Late June/July IT Echinacea pallida Pale purple coneflower P P 0.00 (±0.00) c 0.00 (±0.00) h NA NA Echinacea purpurea Purple coneflower AP P 0.00 (±0.00) c 0.25 (±0.049) fg Late June–Sept. Mid-Aug. IT P Gaillardia pulchella Indian blanket AP A 0.41 (±0.082) b 0.53 (±0.22) defg Mid-June–Sept. Sept. IT Helianthus maximiliani Maximilian sunflower AP P 0.0084 (±0.026) c 2.64 (±0.48) bc Mid-June–late Aug. Late-July IT P Monarda fistulosa Wild bergamot AP P 0.00 (±0.00) 2.02 (±0.35) c Late June–mid-July Mid-July P Monarda punctata Spotted bee balm P P 0.00 (±0.00) 7.15 (±1.24) ab Mid-Aug.–Sept. Sept. Oenothera speciosa Showy evening primrose P P 0.00 (±0.00) 1.00 (±0.13) bcde Late May–late Aug. Mid-June Penstemon digitalis Tall white beardtongue P P 0.00 (±0.00) 0.00 (±0.00) h NA NA Pycnanthemum tenuifolium Narrowleaf mountain mint AP P 0.00 (±0.00) 0.39 (±0.12) efgh Late June–Sept. Late-July P Rudbeckia hirta Black-eyed Susan AP B 2.66 (±0.56) a 17.37 (±2.99) a Mid-June–Sept. Mid-July IT zAP = annual perennial mix; IT = imazapic-tolerant mix; P = perennial-only mix. yP = perennial; A = annual; B = biennial. xValues within the same column by year are not significantly different when marked by the same letter at the P # 0.05 level; values are analyzed by the Steel– Dwass nonparametric test for multiple comparisons. Data are the number of blooming plants per species in 2016, and the number of blooming stems per species in 2017 in which the number of blooming branches off of the main stem was counted individually, but umbels and composites were considered a single flower. Bloom data are averaged over all plots per block and over three dates in 2016 (13 June, 20 July, and 8 Sept.) and eight dates in 2017 (24 May, 13 June, 28 June,18 July, 31 July, 14 Aug., 28 Aug., and 27 Sept.).
initially high weed pressure relative to wild- 2001). Seeds prone to wind distribution, such flower growth yielded to relative parity be- as indian blanket and purple coneflower, tween weed and wildflower cover in year 2 might also benefit from shelter by residue (Figs. 1 and 2). This dynamic of high weed on the surface of no-till fields. In fact, wind- cover in the first year followed by increases in distributed weeds tend to be more prevalent wildflower establishment during the second in no-till than in tilled systems (Derksen year is not uncommon, and concurs with et al., 1993; Froud-Williams et al., 1981; studies suggesting that high weed pressure Tuesca et al., 2001; Zanin et al., 1997). during the first year does not necessarily However, lanceleaf coreopsis seeds also have affect wildflower establishment as long as flattened extensions that may help catch the perennials are able to survive (Love et al., wind, and we did not observe a seedbed 2016; Skousen and Venable, 2008). preparation effect on lanceleaf coreopsis Seedbed preparation proved to be a sig- bloom counts. Overall, although the effec- nificant factor affecting weeds and wild- tiveness of no-till preparation relative to flowers, and the effect was still apparent tillage contrasts with some previous research Fig. 3. Linear regression of total pollinators by total 2 after two growing seasons. The percent cover (Ahern et al., 1992; Corley, 1991; Corley wildflower blooms/m in 2017. Total pollina- of wildflowers was higher and that of weeds et al., 1993), it supports the results of a tors include the sum of syrphids, wasps, lepi- dopterans, honey bees, bumblebees, and ‘‘other was lower following a no-till compared with previous study conducted on the same re- bee’’ categories of species. a tilled seedbed preparation in both years of search farm that showed an overall benefit of study (Fig. 2). Enhanced establishment cul- no-till soil preparation for PR establishment at very low numbers within all pollinator refuge minated in more blooms in the second year in (Angelella and O’Rourke, 2017). plantings, with a much higher number of bum- plots with a no-till preparation compared Pollinator refuge establishment was un- blebee and ‘‘other bee’’ species observed rela- with a tillage preparation. Seedbed prepara- affected by herbicides applied after planting. tive to all other pollinator groups. The various tion differentially affected different species We did not see a difference between pollinator groups showed markedly different of wildflowers. Indian blanket, purple cone- herbicide-treated mixes (AP-h, IT, and P-h) affinities for different flower species. Black- flower, slender mountain mint, and wild and nontreated mixes (AP and P) in wild- eyed Susan, maximilian sunflower, and purple bergamot had more blooms in the no-till than flower cover, total weed cover, or grass coneflower blooms were visited mostly by bees the till plots. No species displayed more cover. Although postemergent sethoxydim in the ‘‘other category’’; partridge pea was visited blooms following tillage than following no- and imazapic have enhanced wildflower es- mostly by bumblebees and spotted bee balm was tillage. Purple coneflower and slender moun- tablishment in some studies (Beran et al., most frequently visited by wasps (Fig. 5). tain mint seeds are lightweight with a small 1999; Hitchmough et al., 2008; Trowbridge diameter, whereas wild bergamot and indian et al., 2017), our work is not the first to report Discussion blanket are larger, heavier, and chaffier seeds. the absence of an effect (Frances, 2008; Love Smaller seeds may be vulnerable to falling et al., 2016; Washburn and Barnes, 2000). The overall composition of the PR habi- through cracks in the soil to depths at which Imazapic phytotoxicity and effectiveness are tats changed dramatically between years. An germination will not occur (Benvenuti et al., affected by site-specific factors such as soil
448 HORTSCIENCE VOL. 54(3) MARCH 2019 quality and weather in addition to wildflower by postemergent imazapic applications in annuals and biennials. The slow establish- species (Bahm and Barnes, 2008; Frances, cases of high but not low weed pressure ment of a perennial-only mix could also be a 2008; Norcini and Aldrich, 2004). In fact, (Beran et al., 1999). Sethoxydim also has a concern if participating landowners want to wildflower species reported as tolerant to variable record of effectiveness, but in cases see blooms and as few weeds as possible in imazapic, including indian blanket and pur- where it enhanced establishment, it has been the first year (Junge et al., 2009). However, in ple coneflower, have been shown to actually because of alleviating competition with pe- future years, we might expect perennials to respond negatively to the herbicide with rennial grasses (Hitchmough et al., 2008; eventually outcompete with and displace decreased establishment in some cases Trowbridge et al., 2017). None of the primary annuals, which could result in floral species (Norcini et al., 2003; Washburn and Barnes, weeds in our study were perennial grasses. richness and evenness declining in the plots 2000), and there have been variations in Moreover, decreasing grass cover with her- containing annuals and biennials (Steffan- species tolerances noted among seeds from bicides such as sethoxydim can have detri- Dewenter and Tscharntke, 2001; Tramer, varied sources (Norcini and Aldrich, 2004). mental effects on lepidopteran pollinator 1975). The degree of weed pressure may also predict diversity and abundance (Feber et al., 1996; Although all mixes bloomed from late the effectiveness of herbicide treatments, as Russell and Schultz, 2010) because many May through September, the different spe- wildflower establishment may be increased species of butterflies rely on grasses as larval cies included in the mixes resulted in varied host plants (Haaland et al., 2011). Consider- peak bloom times. In particular, the AP ing the possible negative impacts of the and IT mixes containing annuals/biennials/ herbicides tested and the general progression perennials peaked in mid-July, whereas the of all the plots to have less weed coverage in P mix peaked in late August/September. These the second year than in the first year of peaks were largely driven in mixes AP and establishment, we do not recommend using IT by black-eyed Susan and partridge pea, herbicides to manage PR habitats after plant- and by perennial spotted bee balm in mix ing wildflower seeds. P. Peak bloom times are important because Mix composition also proved to be an growers should select mixes that will flower at important factor in wildflower establishment. key times to complement gaps in floral re- During the first year of establishment, mixes sources provisioned by the surrounding land- containing annuals, biennials, and perennials scape (Carreck et al., 1999; Venturini et al., outperformed those containing perennials 2017). The bloom phenology expressed by only: they generated blooms, had higher our annual/biennial/perennial mixes may be wildflower percent cover, and had lower more suitable than the perennial-only mix for percent total weed cover than the perennial- addressing and mitigating the gap in floral only plots. These results support other rec- resources in temperate rain-fed agricultural Fig. 4. Total numbers of pollinators in each mix- ommendations that annual wildflowers can systems. herbicide treatment plot. Data are averaged serve as a cover crop to reduce weed estab- Pollinator abundance correlated posi- over seven sampling dates from June through lishment (Aldrich, 2002; Elmhirst and Cain, tively with total blooms, evidencing the Aug. 2017 and represent pollinators observed during 6-min periods. Means are not signifi- 1990). However, the differences in weed premise of pollinator refuges (Fig. 3). We cantly different when marked by the same cover among the different flower mixes also found evidence that floral preferences letter, as analyzed by Tukey’s honestly signif- disappeared in the second year, suggesting vary starkly among pollinator groups and that icant difference post hoc tests at the P # 0.05 long-term establishment might not have been although bumblebees and other native bee level. enhanced by weed-suppressive effects from species were frequent visitors to the wild- flowers we planted, honey bees were very infrequent visitors. Although our observa- tions provide a small snapshot of the entire season, they strongly suggest that wild bees are using PRs and that we can craft refuge bloom phenology and wildflower composi- tion to enhance conservation effectiveness. For example, a recent study in North Dakota found only 13% of the honey bee floral visitations were to native plant species, whereas 77% of native bee visitations were to native species (Otto et al., 2017). This demonstrates that incorporating native wild- flower species can be important for native pollinator conservation efforts. However, studies conducted in other regions of the United States have reported native wild- flower foraging by honey bees compara- ble with that of wild bees (Morandin and Kremen, 2013; Tuell et al., 2008). Other studies have also noted greater foraging by wild bees relative to honey bees on wildflower strips and seminatural habitats (Balzan et al., 2014; Rollin et al., 2013), whereas honey bees preferred foraging within mass-flowering crops (Rollin et al., 2013). Thus, the scale Fig. 5. Proportions of floral visitations by different pollinator groups to individual wildflower species. Data and spatial configuration of our wildflower are averaged over three sampling dates in Aug. 2017. BS = black-eyed Susan; MS = maximilian plantings may have been more attractive to sunflower; PC = plains coreopsis; PCN = purple coneflower; PP = partridge pea; SB = spotted bee wild bees than honey bees as well. Moreover, balm. Tukey’s honestly significant difference tests were performed to evaluate significant differences pollinators have also been shown to exhibit among pollinator groups per wildflower species at the P # 0.05 level. differences in general preference between
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HORTSCIENCE VOL. 54(3) MARCH 2019 451 Supplemental Fig. 1. Mean total wildflower blooms in 2016 and 2017 by (A) mix and (B) seedbed preparation. Annual and perennial mix (mix AP-h) includes nine species of annual and perennial wildflowers and was treated with a sethoxydim herbicide, mix AP is the same wildflower mix without herbicide, an imazapic-tolerant mix contains a subset of seven AP wildflower species and was treated with imazapic herbicide, perennial-only mix (mix P-h) contains nine perennial wildflowers and was treated with sethoxydim herbicide, and mix P is the same wildflower mix without herbicide. Pairwise means comparisons were tested across mixes and within each year by Tukey’s honestly significant difference post hoc analysis, and between no-till and till methods within each year with Student’s t tests. Means within the same year are not significantly different when marked by the same letter at the P # 0.05 level.
Supplemental Fig. 2. Proportions of broadleaf weed, grass weed, and wildflower cover in different mix-herbicide treatments. Vegetative cover sampled on 8 Sept. 2016 and 11 Sept. 2017. Pairwise means comparisons tested across mixes and within year by the Steel– Dwass nonparametric post hoc analysis in 2016 and by Tukey’s honestly significant differ- ence analysis in 2017. Grass cover in 2017 was log + 1 transformed before analysis. Means within the same year are not significantly dif- ferent when marked by the same letter at the P # 0.05 level.
HORTSCIENCE VOL. 54(3) MARCH 2019 1 Supplemental Table 1. Species and seeding rates of pollinator refuge plantings. Mix Species Common name Family Pollinator valuez Life cycley Seeds/gz PLSx/m2 AP, IT Chamaecrista fasciculata (Michx.) Greenew Partridge pea Fabaceae High A 143 32 AP, IT Coreopsis tinctoria Nutt.w Plains coreopsis Asteraceae Low to moderate A 7,103 75 AP, IT Gaillardia pulchella Foug.w Indian blanket Asteraceae High A 525 48 AP, IT Rudbeckia hirta L.w Black-eyed Susan Asteraceae Low B 3,473 75 AP, IT, P Coreopsis lanceolata L.w Lanceleaf coreopsis Asteraceae Moderate to high P 487 48 AP, IT, P Echinacea purpurea (L.) Moenchw Purple coneflower Asteraceae High P 234 48, 95v AP, IT, P Helianthus maximiliani Schrad.w Maximilian sunflower Asteraceae High P 433 48, 95v AP, P Monarda fistulosa L.w Wild bergamot Lamiaceae High P 2,805 30 AP, P Pycnanthemum tenuifolium Schrad.w Narrowleaf mountain mint Lamiaceae High P 13,334 140 P Echinacea pallida (Nutt.) Nutt.w Pale purple coneflower Asteraceae High P 234 48 P Monarda punctata L.w Spotted bee balm Lamiaceae High P 3,245 75 P Oenothera speciosa Nutt.w Showy evening primrose Onagraceae Moderate P 6,700 75 P Penstemon digitalis Nutt. ex Simsu Tall white beardtongue Plantaginaceae High P 882 48 zB. Glennon, personal communication, U.S. Department of Agriculture’s Natural Resources Conservation Service, Wildlife Seed Mix Calculator V3.2_May2016. xlsx. yP = perennial; A = annual; B = biennial. xPure live seed. wPurchased from Ernst Conservation Seeds (Meadowville, PA). vThe IT mix only was planted at higher density. uPurchased from Roundstone Native Seed Company (Upton, KY). AP = annual perennial mix; IT = imazapic-tolerant mix; P = perennial-only mix.
2 HORTSCIENCE VOL. 54(3) MARCH 2019 H
ORT Supplemental Table 2. Wildflower bloom counts by mix, seedbed preparation, and a mix-by-seedbed preparation interaction effect, for 2016 and 2017. Counts are the number of blooming plants per species in 2016, and the number of blooming stems per species in 2017, in which the number of blooming branches off of the main stem was counted individually and umbels and composites were considered a single flower. S z CIENCE Mean (±SE) Seedbed prep Mix Fixed effectsy Species Common name Yr No-till Till AP-h AP IT P-h P Seedbed prep Mix Mix* seedbed prep
V Chamaecrista Partridge pea 2016 1.18 (±0.31) a 0.80 (±0.24) a 0.89 (±0.35) A 0.89 (±0.28) A 1.18 (±0.41) A F = 0.70, P = 0.4, df = 1,18 F = 0.13, P = 0.9, df = 2,18 F = 1.81, P = 0.2, df = 2,18 OL fasciculata 2017 33.15 (±8.34) a 27.32 (±8.48) a 23.89 (±7.81) A 45.59 (±14.09) A 21.23 (±5.17) A F = 0.067, P = 0.8, df = 1,18 F = 1.17, P = 0.3, df = 2,18 F = 0.33, P = 0.7, df = 2,18
43 M 54(3) . Coreopsis Lanceleaf coreopsis 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — lanceolata 2017 0.18 (±0.059) a 0.084 (±0.033) a 0.094 (±0.061) A 0.070 (±0.042) A 0.16 (±0.11) A 0.18 (±0.084) A 0.14 (±0.082) A F = 0.94, P = 0.3, df = 1,30 F = 0.25, P = 0.9, df = 4,30 F = 0.75, P = 0.6, df = 4,30 Coreopsis tinctoria Plains coreopsis 2016 0.60 (±0.14) a 0.87 (±0.21) a 0.69 (±0.21) A 0.90 (±0.29) A 0.62 (±0.16) A F = 1.40, P = 0.3, df = 1,18 F = 0.18, P = 0.8, df = 2,18 F = 0.77, P = 0.5, df = 2,18 2017 2.27 (±1.09) a 2.77 (±1.14) a 2.03 (±1.32) A 1.38 (±0.75) A 4.14 (±1.73) A F = 0.14, P = 0.7, df = 1,18 F = 1.15, P = 0.3, df = 2,18 F = 0.67, P = 0.5, df = 2,18 Echinacea pallida Pale purple 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — —
ARCH coneflower 2017 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — Echinacea Purple coneflower 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — purpurea 2017 0.48 (±0.10) a 0.022 (±0.022) b 0.38 (±0.20) A 0.13 (±0.066) A 0.18 (±0.10) A 0.35 (±0.20) A 0.23 (±0.090) A F = 47.99, P < 0.0001, df = 1,30 F = 0.79, P = 0.5, df = 4,30 F = 2.46, P = 0.07, df = 4,30
2019 Helianthus Maximilian 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — maximiliani sunflower 2017 2.47 (±0.48) a 2.82 (±0.89) a 2.16 (±0.75) A 2.77 (±0.91) A 3.12 (±1.08) A 1.80 (±0.51) A 3.35 (±1.98) A F = 0.061, P = 0.8, df = 1,30 F = 0.14, P = 1.0, df = 4,30 F = 0.051, P = 1.0, df = 4,30 Gaillardia Indian blanket 2016 0.65 (±0.47) a 0.16 (±0.19) b 0.50 (±0.54) A 0.35 (±0.40) A 0.37 (±0.38) A F = 9.42, P = 0.007, df = 1,18 F = 0.21, P = 0.8, df = 2,18 F = 1.36, P = 0.3, df = 2,18 pulchella 2017 0.94 (±0.43) a 0.11 (±0.042) b 1.06 (±0.60)x A 0.34 (±0.31)x A 0.18 (±0.053)x A F = 7.86, P = 0.01, df = 1,18 F = 4.02, P = 0.04, df = 2,18 F = 0.13, P = 0.9, df = 2,18 Monarda fistulosa Wild bergamot 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — 2017 2.80 (±0.71) a 1.26 (±1.01) b 1.49 (±0.52) A 1.53 (±0.48) A 3.39 (±1.31) A 1.70 (±0.47) A F = 4.44, P = 0.05, df = 1,24 F = 1.47, P = 0.3, df = 3,24 F = 0.98, P = 0.4, df = 3,24 Monarda punctata Spotted bee balm 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — 2017 7.43 (±1.32) a 6.88 (±1.90) a 8.27 (±1.62) A 6.03 (±1.54) A F = 0.054, P = 0.8, df = 1,12 F = 0.88, P = 0.4, df = 1,12 F = 0.21, P = 0.7, df = 1,12 Oenothera speciosa Showy evening 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — primrose 2017 1.19 (±0.17) a 0.82 (±0.25) a 1.11 (±0.23) A 0.90 (±0.21) A F = 1.37, P = 0.3, df = 1,12 F = 0.45, P = 0.5, df = 1,12 F = 0.68, P = 0.4, df = 1,12 Penstemon digitalis Tall white 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — beardtongue 2017 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — Pycnanthemum Slender mountain 2016 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) 0.00 (±0.00) — — — tenuifolium mint 2017 0.76 (±0.23) a 0.012 (±0.0085) b 0.063 (±0.063) A 0.11 (±0.086) A 0.046 (±0.29) A 0.91 (±0.39) A 0.91 (±0.39) A X 2 = 12.87, P = 0.0003, df = 1,24 X 2 = 6.92, P = 0.07, df = 3,24 X 2 = 4.91, P = 0.2, df = 3,24 Rudbeckia hirta Black-eyed Susan 2016 2.80 (±0.67) a 2.05 (±0.52) a 2.81 (±0.81) A 2.50 (±0.89) A 1.96 (±0.51) A F = 0.82, P = 0.4, df = 1,18 F = 0.20, P = 0.8, df = 2,18 F = 0.24, P = 0.8, df = 2,18 2017 20.78 (±5.17) a 13.96 (±2.45) a 24.46 (±5.89) A 13.84 (±5.38) A 13.80 (±2.77) A F = 0.56, P = 0.5, df = 1,18 F = 1.36, P = 0.3, df = 2,18 F = 0.69, P = 0.5, df = 2,18 zMean values are calculated over three sampling dates in 2016 (mid-June, late-July, and early September) and nine sampling dates in 2017 (about every 2 weeks, 24 May–27 Sept.). Significant differences are highlighted in gray. Values within the same row and year are not significantly different when marked by the same letter, analyzed by Tukey’s honestly significant difference test. Lower case letters reflect means variation between no-till and till factors, and upper case letters reflect means variation among mix-herbicide factors. yAll species’ bloom count data were square-root transformed before analysis, except for the following: spotted bee balm was normally distributed and not transformed; indian blanket and slender mountain mint were transformed to presence/absence values. Bloom counts were analyzed in models fitted with fully factorial mix and seedbed preparation fixed effects. Indian blanket and slender mountain mint blooms were analyzed with a binomial regression and logit link, and the remaining species’ blooms were analyzed with a least squares analysis of variance. xMeans compared with the Steel–Dwass nonparametric analysis. AP = annual perennial mix; IT = imazapic-tolerant mix; P = perennial-only mix. 3