Composition and decomposition of rhizoma peanut (Arachis glabrata Benth.) belowground biomass
Experimental siteAll procedures for the experiment involving animals were carried out in accordance with relevant guidelines and regulations and they were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Florida (protocol #201509019). The experiment was conducted at the University of Florida North Florida Research and Education Center (NFREC) located in Marianna, FL (30° 52ʹ N, 85° 11ʹ W, 35 m asl) during 2018 and 2019.The study site was an existing mixed RP-bahiagrass grazing study where ‘Ecoturf’ RP was strip-planted into ‘Argentine’ bahiagrass on 12 June 2014. Rhizoma peanut strips were approximately 2-m wide, making it possible to harvest RP forage, roots, and rhizomes free of bahiagrass contamination3,4. The RP was collected from a nursery at the University of Florida—NFREC, whereas the bahiagrass seeds were bought from a seed company. All plants were collected, purchased, managed, and the research was conducted in compliance with relevant institutional, the corresponding national, and international guidelines and legislation.Soils at the experimental site were classified as Orangeburg loamy sand (fine-loamy, kaolinitic, thermic Typic Kandiudults24. At the beginning of the study, soil pH was 5.7 and soil OM was 15.4 g kg−1. Additionally, Mehlich-I extractable soil P, K, Mg, and Ca concentrations at the beginning of the experiment were 26, 99, 43, and 224 mg kg−1, respectively. Total annual rainfall and average annual temperature at the experimental site were 1889 and 602 mm, and 19 and 21 °C, for 2018 and 2019, respectively, and their monthly averages are shown in Fig. 5.Figure 5Monthly weather conditions at North Florida Research and Education Center (NFREC) Marianna, FL, during the experimental years.Full size imageTreatments and experimental designTreatments were two defoliation regimes applied to RP, continuously stocking and 56-days interval between clipping harvests. At the continuous stocking, stocking rates were variable to maintain similar herbage allowance among pastures, which was assessed every 14 days as described by Sollenberger et al.25. Two tester Angus crossbred steers (Bos spp.) remained on each pasture throughout the experimental period. Put-and-take cattle were allocated as needed to maintain a target herbage allowance of 1.5 kg DM kg−1 bodyweight3. Treatments were situated adjacent to each other (i.e., paired sites) in monoculture strips of RP within each of three 0.85-ha pastures. Each pasture was considered a block, thus the experiment consisted of three replicates of each treatment in a randomized complete block design. Within each replicate, treatments had three repetitions (pseudo replicates). To prohibit animal access to the non-grazed treatment, three 2 × 2-m exclusion cages were placed on RP strips in each pasture. Rhizoma peanut herbage mass was determined at both the grazed and non-grazed sites three times each year, at days 56, 112, and 168 of the experimental period by using a 0.25-m−2 quadrat. Two quadrats were collected in each repetition by clipping all the biomass within each quadrat at 2-cm stubble height. After each herbage mass sampling, the non-grazed residual dry matter inside the cages was clipped to a 2-cm stubble height using a weed eater and the herbage removed by raking. On average, across sampling dates and years, herbage mass at the grazed and non-grazed sites was 1050 and 1810 kg of organic matter (OM) ha−1, respectively.Long-term and short-term decomposition studiesThere were two types of root-rhizome decomposition trials. The first is referred to as the long-term decomposition study, and the second is the short-term decomposition study. The long-term study had an incubation period of 168 days, with a single in-situ incubation per year starting in May. The short-term study had in-situ incubation periods of 56 days and there were three incubations per year, occurring in May, June, and August. In all cases, only roots and rhizomes attached to the plant were used in both trials.Long-term studyOn 26 Apr. 2018 and on 23 Apr. 2019, right after RP emergence after breaking dormancy, RP roots and rhizomes were collected from an existing mixed RP-bahiagrass grazing study where RP had been planted in strips into bahiagrass (Paspalum notatum Flüggé) in 2014. Rhizoma peanut strips were approximately 2.75-m wide, alternating with similar wide bahiagrass strips. A pure stand of RP had been maintained in the strips during previous years using herbicides3, making it possible to harvest RP roots and rhizomes free of bahiagrass contamination. Roots and rhizomes were collected at 24 different points in each of three blocks of the original experiment. Roots and rhizomes were collected at 20-cm depth using shovels. As defoliation treatments had not being applied at this time of the year, the same material was used to perform the incubation inside and outside the exclusion cages. After harvesting, excess soil was removed by shaking from the root-rhizome mat using a 1.4-cm diameter sieve. Thereafter, the existing aboveground material was clipped, and the roots and rhizomes were then washed over the same sieve to remove the remaining soil. After washing, roots and rhizomes were dried to constant weight in a forced-air drying oven at 55 °C.To perform the decomposition study, approximately 12 g of dry roots and rhizomes were placed in Ankom bags (10 by 20 cm, 50 µm porosity; ANKOM Technology) and sealed17. Roots and rhizomes were aimed to be placed intact into Ankom bags, nonetheless, when they could not fit inside the bags, they were cut in the middle before being placed. On 2 May 2018 and 1 May 2019, the incubation period began. For each treatment, bags were incubated in situ in the field at 10-cm depth in the same blocks from which they were collected. Bags were removed from the field after 0, 3, 7, 14, 28, 56, 112, and 168 days. For each treatment within each block, three bags were incubated for each incubation time. Additionally, empty bags (one bag per treatment per time per block) were placed in the field. After removal of the in-situ bags from the field, samples and empty bags were dried at 55 °C for 72 h, cleaned with a brush, and weighed. Thereafter, samples were ground to pass a 2-mm screen using a Wiley Mill (Model 4, Thomas-Wiley Laboratory Mill, Thomas Scientific) and analyzed for DM and OM. Subsamples of the 2-mm ground samples were ball milled in a Mixer Mill (MM 400, Retsch) at 25 Hz for 9 min. Ball-milled samples were analyzed for C and N by dry combustion using an elemental analyzer (Vario Micro cube, Elementar). Additionally, samples ground at 2-mm were used to determine ADF in aboveground samples26. The N concentration in the ADF was determined using the above protocol to obtain the ADIN.Short-term studiesThe short-term studies were performed following the same procedures as the long-term study, except that the incubation period was only 56 days, and these studies were repeated three times each year. Roots and rhizomes were incubated in situ on 2 May, 27 June, and 23 Aug. 2018 and on 1 May, 26 June, and 21 Aug. 2019, following the same protocol as described above, except that bags were removed from the field after 0, 3, 7, 14, 28, and 56 days of incubation. The incubations occurring in May, June, and August will be referred as early, middle, and late season, respectively.The early-season incubation period uses the data from the first 56 days of the long-term study described above. For the middle- and late-season incubations each year, roots and rhizomes were harvested approximately 7 days days prior to incubation. Approximately six points in each repetition were collected at 20-cm depth using shovels. For the grazed treatment, roots and rhizomes were collected in the grazed area nearby the exclusion cages, whereas for the non-grazed treatment, the material was collected inside the exclusion cages. After removal of the bags from the field, they were processed and analyzed for DM, OM, C, and N following the protocol described above.Statistical analysesLong-term studyRemaining biomass, remaining N, C:N ratio, ADF, and ADIN were analyzed using the PROC GLIMMIX from SAS27, with treatment and days of incubation as fixed effects, and years and blocks as random effects. Days of incubation were considered repeated measures. Means were compared using the PDIFF procedure at the 5% significance level. When treatment or the interaction of treatment × day of incubation were statistically significant in the ANOVA, nonlinear models were tested to fit the data for each variable and treatment. Nonlinear models were selected for a given response based on data distribution and type of response. If only days of incubation was significant, the same model was applied for all treatments.Remaining biomass (OM basis), remaining N, and C:N ratio were explained by the single exponential decay model14,17,28. The equation describing this process is:$$X=B0, {exp}^{-kt},$$
(1)
where X is the remaining biomass, remaining N, or C:N ratio at day t, B0 is the disappearance coefficient, and k is the relative decay rate (g g−1 day−1). The model used to describe ADF and ADIN was the two-stage model “linear plateau”15,29. The equation describing this process is:$$begin{gathered} Xt = A + b1 times t, {text{if t }} le {text{ T}}, hfill \ {text{and}},{ } Xt = A + b1 times T, {text{if t }} > {text{ T}}, hfill \ end{gathered}$$
(2)
where X is the concentration of ADIN, t is the day of incubation, A is the initial concentration, b1 is the rate of increase in concentration from the beginning of incubation until plateau is reached; and T is the day in which concentration reaches the plateau.Short-term studiesThe single exponential model was applied in the remaining OM and remaining N, for each experimental unit, to obtain individual values for B0 and k. The data for initial N concentration, initial C:N ratio, and B0 and k for remaining OM and remaining N were analyzed using the PROC GLIMMIX from SAS27, with treatment and period as fixed effects, and years and blocks as random effects. Means were compared using the PDIFF procedure at the 5% significance level.Arrive guidelinesThis is study is reported in accordance to ARRIVE guidelines. More
