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Restoration Notes Restoration Notes have been a distinguishing feature of Ecological Restoration for more than 25 years. This section is geared toward introducing innovative research, tools, technologies, programs, and ideas, as well as providing short-term research results and updates on ongoing efforts. Please direct submissions and inquiries to the editorial staff (ERjournal@ aesop.rutgers.edu). Riparian Soil Seed Banks and the Researchers have tested an array of methods to determine their effectiveness on controlling giant reed with varying Potential for Passive Restoration of Giant success. Current leading methods for control include fire, Reed Infested Areas in Webb County, herbicide treatments (Spencer et. al 2008), mechanical Texas removal, and utilizing biological control agents from giant Amede Rubio (corresponding author: Department of Biology reed’s native habitat which ranges from the Mediterranean and Chemistry, Texas A&M International University 5201 to Southern Asia (Goolsby et al. 2009). University Blvd., Laredo, TX, [email protected]), There is little information on the potential for passive Alexis E. Racelis (Department of Biology, The University of restoration in riparian areas where giant reed has been Texas-Pan American, Edinburg, TX), Thomas C. Vaughan removed or controlled, based on persistent soil seed bank. (Department of Biology and Chemistry, Texas A&M Inter- In this study, we conducted a seed bank study following national University 5201 University Blvd., Laredo, TX) and the selective removal of above ground biomass of giant reed John A. Goolsby (United States Department of Agriculture- along the Rio Grande in Webb County, TX (Racelis et. al 2012). The aim of this was to evaluate questions about Agricultural Research Service, 2413 E. Hwy. 83, Weslaco, TX). the composition and viability of seeds at various depths ative landscapes like riparian areas continually and to utilize different methods to identify collected seeds. Nundergo changes through land management prac- We tested two hypotheses: 1) the greatest number of seeds tices, exotic species invasions, and/or natural processes and highest viability is in the leaf litter-0 cm layer; and (Poff et al. 2011, Racelis et al. 2012). Overtime, heavy 2) seed count and viability decreases significantly at 5 cm disturbances and/or exotic species invasions could deplete and 10 cm. soil seed banks (Reid et al. 2009). As a consequence, soil Prior to seed bank collection, a survey of the vegetation seed banks become an increasingly important ecosystem surrounding the sampling plots was conducted, and when component that can prevent extirpation of native plant available, seeds were collected from each plant. The seeds species (Moody-Weis and Alexander 2007) and facilitate collected consisted mostly from tall grasses, cool season survival of rare species for future generations (Coteff and forbs and late succession trees and shrubs characteristic of Van Auken 2006). Tamaulipan scrubland and riparian types. The information Currently, remote sensing studies by Yang et al. (2011) gathered was used for the identification of seeds collected have shown that Giant Reed (Arundo donax), an exotic in the soil seed bank study. invasive grass, has invaded the banks of a major portion Soil seed bank sampling occurred in May 2010, after the of the Rio Grande River in Texas and especially in Webb spring growing season and following the completion of a 27 County, where the study took place. Giant reed, is receiv- month giant reed removal study that began in December ing attention in the United States and Mexico due to the 2007 and was completed in January 2010 (Racelis et al. deleterious effects it has on native riparian plant com- 2012). We sampled a total of eleven 25 m2 plots within munities (McGaugh et. al. 2006, Goolsby et al. 2009). In four sites previously used in the giant reed removal study. optimal conditions the above ground shoots of giant reed Latitude/longitude coordinates for the four sites are as can grow up 10 cm per day (Iverson 1994, Bell 1997) and follows: A) 27°36.200 N 099°34.874 W; B) 27°36.681 N form a dense underground network of woody rhizomes. 099°32.969 W; C) 27°34.744 N 099°31.658 W; and Once established, giant reed has the ability to competitively D) 27°29.195 N 099°28.758 W. Samples within these exclude native vegetation for water, space, and nutrients, plots were extracted using a soil corer with dimensions as well as rapidly diminish the presence of a persistent seed of 13.5 cm (diameter) and 15 cm (chamber length). Five bank (Herrera and Dudley 2003, Watts and Moore 2011). sampling locations were chosen randomly in each plot; soil was collected within each plot at depths of 0 cm-leaf Ecological Restoration Vol. 32, No. 4, 2014 litter, 5 cm, and 10 cm, and placed in brown paper bags. ISSN 1522-4740 E-ISSN 1543-4079 Samples were cold stratified at 4°C for approximately ©2014 by the Board of Regents of the University of Wisconsin System. 60 days (Coteff and Van Auken 2006). Following cold December 2014 ECOLOGICAL RESTORATION 32:4 • 347 color (Perez et al. 2009). We incubated seeds in the dark at 23°C with 1 ml TTC solution for 24–48 hours, in a small plastic container with a removable lid. Some hard coated seeds were sanded or nicked with a scalpel, in order for the assay to contact the respiring tissue. The remaining soil from the sieving process of each sample was placed on 50.8 cm × 25.4 cm × 5.08 cm growing flats with the same potting mix. Initially, pots and flats were on a three day a week watering schedule. Upon germination watering was scaled back to two days a week or as needed. The seed bank analysis yielded a combined site total of 5,798 seeds, with 65 different morphotypes. Data show that 0 cm-leaf litter yielded the highest seed count with 3,508, 5 cm 1,617, and 10 cm 673 (Figure 1). Total viable seed count for the TTC assay and germination were 1,242, Figure 1. Comparison of total seed count and total seed viability at three soil sampling depths in four sites which was approximately 21% of the total seed count (eleven 25 m2 plots were sampled). A G-test for inde- (Figure 1). We used a G-test for independence to test the pendence was performed and a significant difference in differences in viability different depths. The model was viability was encountered at different depths (p < 0.001). significant in that there was a decrease viability at lower soil depth (G test for independence; G = 31.12, df = 2, p stratification all soil samples were processed with a tower < 0.001). Eight seeds germinated in the shade house, but sieve (Hubbard Scientific® Chippewa Falls, WI) with mesh only two grew enough to be positively identified. Twelve sizes 4 mm, 2 mm, 0.5 mm, 0.25 mm, and 0.063 mm. All seed morphotypes were positively identified to species visible seeds were collected and separated by morphotype. and one to genus, through germination and comparison Seed identification followed two paths; 50% shade house methods (Table 1). Seeds of the native trees mesquite (Pro- germination and voucher seed comparison. In the voucher sopis glandulosa), palo blanco (Celtis laevigata), and retama seed comparison phase we identified seeds based on samples (Parkisonia aculeata) were found in the study and all are gathered in the preliminary plant survey (voucher speci- considered to be keystone species of the riparian corridor mens kept at Texas A&M International University her- in South Texas (Ewing and Best 2004). Furthermore, native barium). In the shade house germination phase we selected woody species can potentially compete with aggressive and sowed 5–10 seeds of each unidentified morphotype exotic invaders like giant reed, which is important for in pots with Earth Gro® potting mix and set the rest aside long term restoration efforts (Palenscar and Holt 2010). for viability testing. The remaining seeds representing the Light trampling throughout the 27 month removal study unidentified morphotypes (not used in the germination could have affected seed count and viability in conjunc- phase), and all known species were tested for viability tion to other factors like granivory, seasonal flooding, and using Difco™ 1% Triphenyltetrazolium Chloride solution desiccation of surface seeds (Yenish et. al 1992). These (TTC), which stains the metabolically active tissue a red preliminary results re-emphasize the importance of seed Table 1. General information of thirteen species identified in the seed bank study that was initiated in May 2010 at restored sites along the Rio Grande in Webb County, TX. Fifty-two morphotypes were not identified. Method of Scientific Name Common Name Type Identification Life History Exotic or Native Cenchrus ciliaris buffel grass Grass Comparison Perennial Exotic Helianthus annus common sunflower Herbaceous Comparison Perennial Native Melia azedarach China berry Woody Tree Comparison Perennial Exotic Vicia ludoviciana deer pea vetch Herbaceous Germination Annual Native Celtis pallida granjeno Woody Shrub Comparison Perennial Native Datura stramonium jimsonweed Herbaceous Comparison Annual Exotic Prosopis glandulosa mesquite Woody Tree Comparison Perennial Native Sphaeralcea angustifolia narrow leaf globemallow Herbaceous Comparison Perennial Native Celtis laevigata palo blanco Woody Tree Comparison Perennial Native Descurainia pinnata pinnate tansy mustard Herbaceous Comparison Annual Native Parkinsonia aculeata retama Woody Tree Comparison Perennial Native Panicum spp. panicum Grass Comparison — — Setaria leucopila plains bristle brass Grass Comparison Perennial Native 348 • December 2014 ECOLOGICAL RESTORATION 32:4 rain from established native vegetation to replenish the Moody–Weis, J. and H.M. Alexander. 2007. The mechanisms soil seed bank. and consequences of seed bank formation in wild sunflowers Habitat restoration projects can use seed bank infor- (Helianthus annuus). Journal of Ecology 95:851–864.