Source: CORNELL UNIVERSITY submitted to NRP
GLOBAL CHANGE AND URBAN TREES: PROMOTING TREE GROWTH THROUGH A BETTER UNDERSTANDING OF THE RHIZOSPHERE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1017091
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2018
Project End Date
Sep 30, 2022
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
CORNELL UNIVERSITY
(N/A)
ITHACA,NY 14853
Performing Department
Horticulture
Non Technical Summary
The urban forest will face a fundamental challenge in a changing climate. Stakeholders, including the green industry, land owners and urban foresters have identified significant barriers to tree growth and survival that require advancements in the area of tree response to predicted frequency in drought. Urban trees must survive in unique growing conditions including limited rooting volumes and poor soils that exacerbate abiotic stresses such as water limitation. This project contributes to long-term research on understanding tree processes below-ground and leverages recent work on furthering the green industry tree species palette and tree species root exudate diversity. Characterizing the soil-root interface (rhizosphere) of urban trees that represent different water use strategies allows for a better understanding of water limitation to tree productivity and potential strategies for mitigation.
Animal Health Component
20%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
10206991070100%
Goals / Objectives
Urban environments tend to be inhospitable to tree growth. Poor soils with low water holding capacity and low soil organic matter can alter the growth and physiology of tree root systems. The aim of this research is promote tree productivity by analyzing the effect of quantitative and qualitative root exudate characteristics on the rhizosphere across tree species that represent diverse water use strategies. This research will lead to an understanding plant-rhizosphere interactions by 1) characterizing how root exudates of urban tree species representing diverse water use strategies influence root and rhizosphere hydrology, and by 2) evaluating how the ability of a root system to alter root exudation may function as a mechanism to manipulate the rhizosphere microbiome during dry-wet cycles. Long-term aims are to increase urban tree health and productivity through potential management of the rhizosphere in the face of predicted increases in soil moisture limitation due to climate change.
Project Methods
A Long-term field experiment that mimics reduced rooting volumes via 48 in-ground, hydraulically isolated, tree boxes (1.83m x 1.83m x 1.5m, w x h x d) were recently established at the Cornell Orchards. Boxes were lined and sealed with 40 mil root barrier to prevent water from getting into the sampling area of the root box. Two sides of each planting box were constructed of 2.3 mm thick, 1.2m x 1.2 m clear polycarbonate sheeting to allow for easy viewing of the tree root system and ease of cutting of the material for root access. Subsurface drainage was established during tree box instillation and soil from the site was amended with compost and back filled to attempt to meet city planting recommendations. Researcher/public access boxes with installed ladders in-between each tree allow individuals to view the trees belowground. When the access areas are not in use they are covered with an insulated lid to prevent large temperature fluctuations.We have selected seven tree species based on their water use strategies, i.e. insensitive to water stress, anisohydric and sensitive to water stress isohydric. We have already evaluated several tree species water use strategies in the lab including Acer rubrum, Quercus bicolor, and Tilia cordata. We plan to additionally screen Magnolia, Koelreuteria paniculata, Ostrya virginiana, and Cercidephyllum japonicum. After tree establishment, drought stress will be imposed via precipitation exclusion shelters. These consist of sub-canopy halved pvc troughs covering 50% of the soil surface area that divert precipitation from reaching the soil surface. These methods align with current precipitation reduction experiments at Hubbard Brook (Asbjornsen personal comm.) and are also in line with current precipitation reduction experiments (Asbjornsen et al., in press, Drought- Net, NSF program). These protocol for inducing drought follow recommendations from the NSF funded Drought-NET IDE program which allows for study systems to be compared on a world-wide basis.Minirhizotron tubes will be installed with each tree to allow for root observations during tree establishment and to quantify root life span and root interactions in situ, tubes will be installed at a 30° angle to a depth of 1 m, 50 cm from the base of the tree. Root development will be monitored by measurements taken every other week by a specialized laparoscopic camera (Bartz, Inc, Santa Barbara, CA, USA) during the growing season and once a month during plant dormancy.These sampling times follow the common protocol for minirhizotron imaging (Bauerle et al. 2008). If needed root imaging can be increased to a higher frequency in order to capture particular growth or death events of interest. The acquired pictures will be analyzed using specialized root analysis software (WinRhizo Tron MF, Regents Inc. Quebec, Canada).Time domain reflectometry (TDR) probes will be established through the mylar sheets at three depths monitor soil moisture and temperature. Water retention and conductance of soil samples collected from the rhizosphere will be measured and compared with those of the associated bulk soil. The multi-probe Wind evaporative method will be used in a custom soil column for small soil volumes (~5 ml) with a pair of microtensiometers placed at 1.5 and 3 cm from the exposed surface (Tamari 1993); following Šim?nek et al., water retention and conductivity will be extracted by non-linear parameter optimization coupled with numerical simulations of Richards equation (Šim?nek 1998).After tree establishment tree branches and roots will be sampled for hydraulic conductivity at declining water potentials to construct vulnerability curves. These curves estimate the "vulnerability" of the tree organs or in other words when the organ is likely to form embolisms or air bubble formation in its xylem that potentially reduces the flow of water within the tree. Tree height and shoot length as a measure of plant growth will be collected monthly.In collaboration with the Kessler lab at Cornell, we have established a method for phenolic root exudate analysis. Preliminary results confirm that different tree species show great variation in the quantity and quality of phenolic exudates. Fine roots will be accessed through the polycarbonate windows, cleaned and put in a syringe containing glass beads and a carbon-free nutrient solution. Exudates are collected from the flushed root solution following Phillips 2008. In order to confirm that root exudate quality directly selects for microbial community composition and activity, we will expose native microbial communities to exudates collected from the tree species that were grown under either ideal conditions or water stress in the experiment. To simulate a rhizosphere environment, plastic cylinders (50 mm x 20 mm) with stainless steel mesh bottoms will be filled with 10 g of field soil and covered with a microfiber-glass filter. The membrane filter will allow for even application of exudates to the soil (Falchini et al. 2003).Additional roots will be collected for rhizosphere microbial analysis. For all of the experiments, the rhizosphere will be defined as the soil adhering to the root after gently shaking off bulk soil (Yergeau et al. 2015). Total DNA will be extracted using the NucleoSpin 96 Soil kit (Macherey-Nagel, Bethlehem, PA). We will amplify the 16S rRNA gene region to examine bacterial composition, as well as the fungal internal transcribed spacer (ITS) region, using universal primers recommended by the Joint Genome Institute (Daum 2016). Sequences will be processed following the Brazilian Microbiome Project (http://www.brmicrobiome.org/). Droughted microbiome communities will be compared to community composition in the control treatments (e.g. wet-adapted microbes only) to determine whether current soil conditions, or relative microbiome proportions, are more important for determining tree root microbiome recruitment. We will look at this by comparing both global community distance (e.g. Bray-Curtis distance) and by looking at the relative abundance of individual OTUs, to determine whether specific taxa are disproportionately impacted, regardless of their initial abundance.

Progress 10/01/20 to 09/30/21

Outputs
Target Audience: Nothing Reported Changes/Problems:We were unable to hire staff or students to analyze these images for root dynamics data. Hence a backlog of one-year of data has occurred. We have recommenced the analysis but have not been able to catch-up. Additional analysis time is needed to complete the data set. A backlog of sample processing has occurred that resulted in severely delayed processing of root exudate samples. Additional time is need to take a final sample and analyze the data. What opportunities for training and professional development has the project provided?A postdoctoral researcher and undergraduate student were active on this project until complications from COVID regulations required us to halt our studies. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?We plan to start measurements back up so that the project may be completed.

Impacts
What was accomplished under these goals? I have established a large replicated instillation of 48, 1.8w X 1.5l x1.5d meter in-ground root boxes at the Cornell Orchards. This long-term research site investment allows for the continual viewing, monitoring, and sampling of trees both above- and below-ground. The site serves for both research and educational purposes that simultaneously aim to increase knowledge on the link between tree water use and below-ground processes. Minirhizotrons were established that allow for the tracking of root dynamics and biweekly to monthly imaging has occurred.

Publications


    Progress 10/01/19 to 09/30/20

    Outputs
    Target Audience:Target audiences for this reporting period include undergraduate students and interns through the use of the site as a teaching laboratory. Ten students participated in a formal class, Arboriculture PLHRT 3050, for an experiential lab based on tree and soil management. One undergraduate researcher participated in an internship program on root and soil processes through the Bauerle Lab. Changes/Problems:Due to COVID-19 an the inability to visit campus we were unable to meet some of our sampling goals. However, we are now able to visit the site again and continue our data collection. While there is little we can do for the loss of data within the spring and summer of 2020 we hope that our current dataset will be adequate to meet our goals. What opportunities for training and professional development has the project provided?This project has provided training for undergraduate students in a formal class setting. Educational materials and soil plant interactions were developed as part of an arboriculture course module. Students were able to utilize the unique experimental set-up for course demonstrations and experiments. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Continued measurements of root dynamics including root lifespan and growth in relation to soil parameters will be recorded. Root exudates will be collected in the spring of 2021 for comparison on tree species target exudate profiles. Total organic carbon (TOC) content will be determined with a TOC analyzer. The metabolome of the captured exudates will be analyzed for both primary and secondary compounds using Liquid Chromatography Mass Spectrometry (LC-MS). The data will be correlated with species- and site-specific characteristics to deduce a carbon storage capacity profile for the studied species considering soil type and especially plant available soil water in the respective depth layers. Based on these profiles, practical guidelines will be developed.

    Impacts
    What was accomplished under these goals? We installed 60 cm-long transparent minirhizotron (MR) tubes in the soil allowing individual fine roots to be tracked from "birth" to "death" using specialized digital imaging. Each of the tree plots consists of one MR tubes (1 per species x 8 species x 6reps) adding up to a total of 48 tubes. Each tube provides 44 pictures per imaging session. Using WhinRHIZO TRON imaging analysis software, root life span, rooting depth, root production, root morality, total standing crop and root morphological characteristics will be estimated. HOBO soil moisture and temperature gauges were installed in a subset of boxes to monitor seasonal discrepancies in soil moisture and temperature as a result of the hydraulically isolated planting boxes. Results show that additional insullation on the sampling access portals allows the tree planting boxes to maintain the same temperature as "field" soil.

    Publications

    • Type: Conference Papers and Presentations Status: Other Year Published: 2019 Citation: Zwetsloot M Mu�oz Ucros J, Kessler A, Wickings W, Sparks J, Wilhelm R, Buckley D, and Bauerle TL. 2019. A slow path towards measuring root exudate effects on rhizosphere functioning and soil biogeochemical cycles Wageningen Soil Conference, August 27-30, Wageningen, Netherlands.
    • Type: Conference Papers and Presentations Status: Other Year Published: 2019 Citation: Zwetsloot M, Mu�oz Ucros J, Kessler A, Wickings W, Sparks J, Wilhelm R, Buckley D, Bauerle T. 2019. The unexplored role of secondary metabolites in root priming: phenolic root exudates cause striking changes in soil organic matter decomposition and microbial community structure. 7th International Symposium on Soil Organic Matter, October 8-11, Adelaide, Australia.


    Progress 10/01/18 to 09/30/19

    Outputs
    Target Audience: Nothing Reported Changes/Problems:The trees required a longer period of establishment and so some soil measurements were delayed by one season. However, this delay should not impact are ability to complete the project on time. What opportunities for training and professional development has the project provided?The site is serving as a training location for an Arboriculture course at Cornell. It is also providing a site for sample collection for graduate students. How have the results been disseminated to communities of interest?The project is ongoing but has been included in talks to both academics and growers. What do you plan to do during the next reporting period to accomplish the goals?Soil and tree instrumentation will be established to allow for continual monitoring of the site. Root and soil microbial samples will be taken in the spring to initiate a seasonal look at root- soil interactions.

    Impacts
    What was accomplished under these goals? The trees will require an additional year of establishment, yet, we were still able to garner some preliminary data that will help us to address our initial questions on root and rhizophere interactions. For the following experiments we utilized Salix sp. as an initial test species due to its fast growing time and availability. We investigated the role of soil microbes on the drought legacy effect in a two-phase experiment. First, we tested whether plants could select for a different microbial community in their rhizosphere while undergoing drought stress. Then, we used those assembled communities to evaluate whether they had an impact on the performance of a new plant under similar drought stress. Using clonal willow material selected for consistent performance allowed us to more cleanly evaluate the changing microbial community as a mechanism of phenotypic plasticity and adaptation for plants under water stress.

    Publications