Source: OKLAHOMA STATE UNIVERSITY submitted to NRP
EFFECT OF MANAGEMENT PRACTICES ON SOIL MICROBIAL COMMUNITY AND ENZYME ACTIVITY IN RELATION TO ECOSYSTEM HEALTH AND FUNCTION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1004134
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2014
Project End Date
Sep 30, 2019
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
OKLAHOMA STATE UNIVERSITY
(N/A)
STILLWATER,OK 74078
Performing Department
Plant & Soil Sciences
Non Technical Summary
Soil microflora and enzyme activities play crucial roles in nutrient cycling, modifying ordestroying environmental pollutants, and ecosystem health and function as a whole. In thepast several decades, great progress has been made in agriculture for large-scale utilization oflegume inoculation, and understanding the influence of microflora on the degradation andpersistence of chemicals used in agricultural production. More recently, understanding of theecology, function, and biochemistry of soil microflora has advanced tremendously because ofdevelopment of molecular biology techniques. However, soil is heterogeneous and soilmicrobes are abundant and diverse. Research progress on the subject is hindered by limitedunderstanding of the complex soil systems and the vastly unknown microbes that reside inthe system. As a soil scientist and biologist, my long-term goal continues to be to betterunderstand the underlying basis of biological and microbial activities in soils in relation tosustaining agricultural production, and maintaining and improving soil and environmentalquality. I continue my effort building a program that links basic research to application. Inthe next five years, my research will focus on understanding soil microbes, enzyme activities,and their relationships in relation to soil health and ecosystem functions. Special attentionwill be given to revealing microbial communities involved in fixation of atmosphericdinitrogen (N2) and in soils under various long-term management practices and agriculturalproduction. These include animal waste management and soils from rangeland and cropland.The proposed research is divided into two main subtopics, including (1) impact ofmanagement practices on soil biology, and ecosystem health and function, and (2) diversityand community structure of diazotrophic microorganisms in agroecosystems. The proposedobjectives support the department and DASNR strategic plan by providing information toeffectively utilize our natural resources and sustain ecosystem health while ensuringprofitability of plant-production systems. The academic outreach in this project will alsoensure our continued national and international recognition in research and education toadvance plant and soil sciences.
Animal Health Component
25%
Research Effort Categories
Basic
75%
Applied
25%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
10201101060100%
Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships;

Subject Of Investigation
0110 - Soil;

Field Of Science
1060 - Biology (whole systems);
Goals / Objectives
(1) To evaluate the impact of management practices such as addition of animal waste onsoil biological properties and health indicators, including enzyme activities, andmicrobial abundance, diversity, and biomass. Specific goals are:• To develop standardized methods for soil enzyme assays; and• To determine basic soil physical, chemical, biochemical, and microbiologicalproperties in soils under long-term management practices.(2) To reveal bacterial and diazotrophic community in soils following long-term wheatcultivation under various management practices. Specific goals are:• To determine diversity, richness, and uniqueness of bacterial and diazptrophiccommunity in soils under different long-term management practices,including those under manure and fertilizer treatments;• To evaluate abundance, diversity, and community structure of free-livingdiazotrophic community in enrichment cultures developed using N-free mediaand in soils under different management practices; and• To isolate, identify, and characterize free-living diazotrophs and othermicrobes for potential application to enhance agricultural production, and/orpromote soil health and productivity.(3) To use the acquired data and statistical analysis to reveal drivers and interrelationshipsbetween the measured soil variables to provide insight into the importance of soil parameters in ecosystem health and function; and to identify or develop strategies for enhanced soil health and productivity.
Project Methods
(1) To reach objective one, studies will be conducted to determine soil microbial activity, community structure and diversity,as well as activities of enzymes involved in nutrient cycling in soils under long-term management practicesa. To develop standardized methods for soil enzyme assaysb. To determine basic soil physical, chemical, biochemical, and microbiological propertiesin soils under long-term management practicesSoil samples will be taken from long-term management practices, in particular,those under wheat cultivation and various manure or chemical fertilizer treatments. Anexcellent experiment site is the Magruder Plots, where a century of continuous wheatresearch has been conducted and the soils are treated with animal manure and/or differentfertilizers.Microbial biological properties in soils will be determined using a combination ofculture-based and modern molecular technologies to reveal key microorganismsinhabiting the soil environment.(2) To reveal bacterial and diazotrophic community in soils following long-term wheatcultivation under various management practicesMolecular and DNA analyses will be employed. Soils obtained in objective one will be used.(3) To use the acquired data and statistical analysis to reveal drivers and interrelationshipsbetween the measured soil variables to provide insight into the importance of soil parametersin ecosystem health and function; and to identify or develop strategies for enhanced soilhealth and productivityThis part of the project focuses on data analysis for meaningful interpretations.Comparisons will be made using the least significant difference test (LSD).

Progress 10/01/14 to 09/30/19

Outputs
Target Audience:Scientific communities, agricultural producer, stakeholders, and state/federal government agencies Changes/Problems:The long-term goal is to better understand the underlying basis of the importance and significance of biological and microbial activities in soils in relation to sustaining agricultural production and maintaining and improving soil and environmental quality. Understanding the complex microbial community in the soil environment has proven to be a challenging task because of the vast diversity and enormously large population inhabiting the environment. The recent development of culture-independent molecular techniques has brought us hope for in-depth understanding of the soil black box. Unfortunately, changes in microbial community structure and diversity due to seasonal and temporal variations in nutrient or physical conditions are slow and gradual, making it difficult to interpret the data and obtain conclusive results. Therefore, in addition, to continue our effort in developing and perfecting enzyme assay protocols and methods for evaluating microbial diversity and activities, we also started to isolate microbes that could potentially strengthen our effort in promoting agricultural production and maintaining soil and environmental quality. In particular, we are interested in free-living nitrogen fixers (bacteria that have the ability to obtained atmospheric nitrogen without partnering with other organisms). However, growth conditions are important influencing their nitrogen fixation traits. Thus, the characterization of the microbes is the initial step of this research effort. A new research project was initiated in 2018 to strengthen the understanding of fate and stability of an antibiotic, ciprofloxacin, in soil and sludge. A preliminary study of this project was funded by the Vice President's Office at OSU. A research team has been established with the goal of seeking funding from the Department of Defense to strengthen biosecurity against bioterrorism. There is a global phosphorus (P) crisis. The threat is two-fold, with P being deficient in nearly 67% of crop production land, while P pollution is generating widespread environmental concerns, especially related to water quality. Unfortunately, the global P supply is diminishing over time, and P reserves are finite, non-renewable, and non-substitutable. The only viable solution to the crisis is to improve P use efficiency through effective management practices. P use efficiency, crop deficiency, and pollution are interrelated and directly linked to mobility of P in the terrestrial environment. Decades of research demonstrated that inorganic P has limited mobility, but organophosphates (organo-P) are relatively mobile in the environment. Further, the mobility of organo-P may be governed by microbial activities. If time and resource permit, an effort will be made to reveal predominant microbial members involved in producing mobile organo-P molecules. What opportunities for training and professional development has the project provided? Training and supervising graduate students as a major advisor (4). Training and advising graduate students as a member of graduate committees (14). Training and advising undergraduate students conducting research projects (3) Teaching and sharing knowledge with graduate/undergraduate students in courses taught or in the laboratory (93 students in SOIL4483, 35 students in SOIL5383, 37 graduate students in the lab, 26 students in SOIL5483, and 12 freshmen visited the lab during orientation) Training and supervising visiting scientists (7 visiting scholars) How have the results been disseminated to communities of interest? Attending and presenting at national and international conferences Publishing in peer-reviewed scientific journals Classroom teaching and discussions As an invited speaker to present at higher education institutions As an invited speaker to present at an international conference in the United Kingdom As an invited speaker to present at national/international conferences, i.e. the Tri-society meeting in Tampa, FL. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Impact of management practices on soil biology. Evaluated microbial communities in different ecosystems, such as Native rangeland, land under Conservation Reserve Programs (CRP, formerly abandoned cropland), and Wheat as well as wheat fields following various length of cultivation period. Highlights of key findings: Bacterial community was more diverse in soils under continuous wheat cultivation than Native or CRP. If cultivation leads to soil degradation, bacterial diversity may not be a sensitive indicator of soil degradation. CRP led to the predominance of Actinobacteria, while continuous wheat cultivation promoted Firmicutes and Chloroflex. Cyanobacteria (many with the ability to fix atmospheric dinitrogen) was the highest in Native, followed by Wheat, and the least in CRP, suggesting the nutritional resilience of the natural ecosystem in nitrogen acquisition. Bacterial composition was more similar in soils under 14-46 years of wheat cultivation than that under >100 years of cultivation. Impact of management practices on the diversity and community structure of diazotrophic microorganisms. Based on the evaluation of 60,797 16S rRNA and 42,931 nifH gene sequences, richness and diversity of microbial communities was enhanced by Manure but reduced by NPK, with the impact more prominent at the genus and species than phylum to family levels. Highlights of key findings: The community abundance was predominantly influenced by carbon availability while the richness and diversity of a community were governed by pH. Following century-long cultivation of winter wheat, Cyanobacteria comprised 2.6% of the microbial community in Check (without fertilization), whereas they were 0.19% and 0.05% in Manure and NPK, respectively. The observations were further supported by data obtained at location 2, where the abundance of Cyanobacteria in Native was 3.75%, in CRP was 0.01%, and in Wheat was 1.95%. The detected diazotrophic communities were constituted by a distinct proportion of nifH clusters I and III. The predominance of Cluster I diazotrophs (100%) and Actinobacteria (60.1%) in CRP indicated that these microbial groups might be key players in ecosystem restoration. Impact of management practices soil enzyme activities. Following over 30 years of cropping management practices and evaluation of 60 soils with respect to activities of 14 enzymes involved in cycling carbon, nitrogen, phosphorus, and sulfur, data clearly suggested that returning crop residues to soil led to preserving soil biology and metabolic capacity; while grazing negatively impacted soil's capacity to cycle nutrients. Understanding the impact of long-term cropping systems on these enzyme activities in the soil can guide the development of management strategies that promote soil productivity while maintaining the sustainability of the system. Enzyme assay methods - standardization and development: Continued the effort to revise and standardize soil enzyme assay methods and developed two new soil enzyme assay methods (publications are in progress). Enzyme methods developed will support our effort to a systematic evaluation of ecosystem health components, focusing on microbial indicators to provide insight into the significance of changes in these key indicators. Highlights of key findings: Enzymes activities were significantly affected by incubation methods. Activities obtained by water-bath incubation were 134% to 145% of those by the air-mode incubation. Shaking during incubation in enzyme assays has a limited effect on activities determined. The detected differences between water-bath and air-mode incubations were attributed to temperature variations in the reaction mixtures of the assay. Although temperatures in all incubators were maintained at 37oC during the 1-hr incubation period, temperatures in the reactions mixtures in samples placed in the air-mode incubator never reached 37oC during the 1-hr incubation. Temperatures in the water bath and the sample reaction mixtures placed in the water-bath were stable and remained at the desired temperature throughout the incubation period, which would result in data to be more reproducible and accurate for meaningful comparison and interpretation. Presented an invited talk on the subject at an international conference in July 2016 in Bangor, United Kingdom and subsequently published a peer-reviewed article (lead author). Enhanced understanding on the capacity and mechanism of soil in adsorbing antibiotics. Highlights of key findings: Soil was effective in restraining environmental mobility of ciprofloxacin through adsorption. Sorption capacity varied among the tested soils, with >140,000 mg ciprofloxacin kg-1 soil detected. Sorption capacity and portioning coefficient of ciprofloxacin under acidic conditions were strongly and significantly correlated with soil clay content. Significant relationships with soil organic carbon content were also detected. Multiple soil properties governed their interactions and sorption behavior, signifying the importance of developing soil-based wastewater treatment facilities and site-specific management strategies to mitigate potential negative impacts. Implications of the result on biothreat: Centers for Disease Control and Prevention (CDC) Strategic National Stockpile (SNS) of antibiotics effective against anthrax (ciprofloxacin and doxycycline) has been prepared to respond to a large-scale anthrax attack in the United States. For an individual >67 lbs, the CDC prescribes 500 mg of ciprofloxacin twice a day for 10 consecutive days to treat anthrax. The human body metabolizes only about 10% of orally administered antibiotics, indicating that about 90% would be released into the environment. (i.e. an area of 2 million people would have about approximately 1,800 kg of ciprofloxacin released each day). Understanding sorption capacity and factors affecting adsorption to the soil is critical in protecting wastewater treatment infrastructure and alleviating threats to human and animal health. Microbial phosphorus in soil. I have teamed up with a faculty in environmental engineering to initiate a research project on the subject. Our topical submission was selected by National Science Foundation for further consideration in the 2019 funding cycle. In March 2019, I was invited to NSF to give a presentation to NSF program directors.

Publications

  • Type: Book Chapters Status: Published Year Published: 2018 Citation: Shiping Deng. 2018. Port Silt Loam: Oklahoma State Soil. In the state soil booklet developed under the auspices of the Soil Science Society of Americas K-12 Committee, Soil Science Society of America, Madison, Wisconsin, USA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Shiping Deng, Yan Wang, Mark Krzmarzick, and John Gustafson. 2019. Adsorption of Ciprofloxacin in Soil. Meeting Abstract, 110th Annual SSSA meeting, San Diego, Jan 6  9, 2019.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Yan Wang and Shiping Deng. 2019. Incubation Methods for Enzyme Assays. Meeting Abstract, 110th Annual SSSA meeting, San Diego, Jan 6  9, 2019.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Tang, Zhenya; Fangling Fan; Xinyue Wang; Xiaojun Shi; and Shiping Deng. 2017. Mercury in rice (Oryza sativa L.) and rice-paddy soils under long-term fertilizer and organic amendment. Ecotoxicol. Environ. Safety
  • Type: Theses/Dissertations Status: Other Year Published: 2019 Citation: Pebeck (Gentry), Kaylee. Microbial Influence on Switch grass Biomass Production. Master of Science in Plant and Soil Sciences, Oklahoma State University. Co-advise with Dr. Beatrix Haggard. (in progress)
  • Type: Theses/Dissertations Status: Other Year Published: 2019 Citation: Pasket, Amber. The Fate of Amended Phosphorus and Sorption of Ciprofloxacin in Soils. Master of Science in Plant and Soil Sciences Department of Plant and Soil Sciences, Oklahoma State University. Co-advise with Dr. Hailin Zhang. (in progress)
  • Type: Theses/Dissertations Status: Other Year Published: 2019 Citation: Alserae, Hussein. Cellulose-1-4-?- cellobiosidase activity in soils. Ph.D. in Soil Science. Department of Plant and Soil Sciences, Oklahoma State University. (in progress)


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Scientific communities, agricultural producers, stakeholders, and state/federal government agencies are the target audiences. Changes/Problems:There is a global phosphorus (P) crisis. The threat is two-fold, with P being deficient in nearly 67% of crop production land, while P pollution is generating widespread environmental concerns, especially related to water quality. Unfortunately, the global P supply is diminishing over time, and P reserves are finite, non-renewable, and non-substitutable. The only viable solution to the crisis is to improve P use efficiency through effective management practices. P use efficiency, crop deficiency, and pollution are interrelated and directly linked to the mobility of P in the terrestrial environment. Decades of research demonstrated that inorganic P has limited mobility, but organophosphates (organo-P) are relatively mobile in the environment. Further, the mobility of organo-P may be governed by microbial activities. There is a critical gap, however, in understanding what predominant mobile organo-P molecules are present, and what microbial members and processes produce them. Based on preliminary data from a century-old field experiment, the central hypothesis is that unique microbial members promote the formation of mobile organo-P compounds. The goal of this research is to identify predominant mobile organic-P compounds and the microbial members involved in their development in soils using a century-long experimental site. Specific objectives are (1) Identify predominant organo-P compounds in a soil that exhibited high P mobility following a century-long field experiment; (2) Characterize microbial communities involved in developing mobile organo-P molecules and identify biomarkers for these taxa and functional genes; and (3) Determine the distribution of these taxa throughout benchmark Oklahoma soils and their correlation with P and geochemical characteristics. What opportunities for training and professional development has the project provided?Training and advising graduate students andUndergraduate students. Training and supervising visiting scientists How have the results been disseminated to communities of interest? Attending and presenting at national and international conferences Publishing in peer-reviewed scientific journals Classroom teaching and discussions What do you plan to do during the next reporting period to accomplish the goals?We will continue current research effort and interests to develop and standardize enzyme assay methods for soil and to further our effort in understanding and revealing microbial diversity and activities that are important in agricultural production as well as preserving and maintaining soil and water quality in the environment. We also continue our effort in evaluating the fate and stability of antibiotics in soil and sludge. Recently, we have been in communication with NSF to initiate a project on microbial phosphorus in soil.

Impacts
What was accomplished under these goals? On enzyme assay methods: Enzymes activities were significantly affected by incubation methods. Activities obtained by water-bath incubation were 134% to 145% of those by the air-mode incubation. Shaking during incubation in enzyme assays has limited effect on activities determined. The detected differences between water-bath and air-mode incubations were attributed to temperature variations in the reaction mixtures of the assay. Although temperatures in all incubators were maintained at 37oC during the 1-hr incubation period, temperatures in the reactions mixtures in samples placed in the air-mode incubator never reached 37oC during the 1-hr incubation. The above-detected differences would vary depending on the size and capacity of the air-mode incubator as well as the number of samples assayed in each batch. Temperatures in the water bath and the sample reaction mixtures placed in the water-bath were stable and remained at the desired temperature throughout the incubation period, which would result in data obtained be more reproducible and accurate for meaningful comparison and interpretation. Continue leading an international effort to standardize and improve soil enzyme assay methods. Enzyme assay methods developed will support the systematic evaluation of ecosystem health components, focusing on soil biological indicators and the significance of changes in these indicators induced by changes in soil management practices. On the capacity and mechanism of soil in adsorbing antibiotics The soil was effective in restraining environmental mobility of ciprofloxacin through adsorption. Sorption capacity varied among the tested soils, with >140,000 mg ciprofloxacin kg-1 soil detected. Sorption capacity and portioning coefficient of ciprofloxacin under acidic condition were strongly and significantly correlated with soil clay content. Significant relationships with soil organic carbon content were also detected. Multiple soil properties governed their interactions and sorption behavior, signifying the importance of developing soil-based wastewater treatment facilities and site-specific management strategies to mitigate potential negative impacts. Implications of the result on biothreat: Centers for Disease Control and Prevention (CDC) Strategic National Stockpile (SNS) of antibiotics effective against anthrax (ciprofloxacin and doxycycline) has been prepared to respond to a large-scale anthrax attack in the United States. For an individual >67 lbs, the CDC prescribes 500 mg of ciprofloxacin twice a day for 10 consecutive days to treat anthrax. The human body metabolizes only about 10% of orally administered antibiotics, indicating that about 90% would be released into the environment. (i.e. an area of 2 million people would have about approximately 1,800 kg of ciprofloxacin released each day). Understanding sorption capacity and factors affecting adsorption to the soil is critical in protecting wastewater treatment infrastructure and alleviating threats to human and animal health. On soils under crop production Grazing negatively impacted soil's capacity to cycle nutrients, while returning crop residues to soil led to preserving soil biology and metabolic capacity. This is based on an evaluation of 14 soil enzymes in 60 soils that were under over 30 years of cropping management practices. On microbial phosphorus in soil I have always had an interest in phosphorus in soil. Last two years, I have teamed up with a faculty in environmental engineer to initiate a research project for NSF funding. Our proposal in the last round submission was selected for further consideration in 2019 funding cycle. An MS student under my supervision started to work on the subject for additional preliminary data.

Publications

  • Type: Book Chapters Status: Published Year Published: 2018 Citation: Shiping Deng. 2018. Port Silt Loam: Oklahoma State Soil. In the state soil booklet developed under the auspices of the Soil Science Society of America⿿s K-12 Committee, Soil Science Society of America, Madison, Wisconsin, USA.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Shiping Deng, Yan Wang, Mark Krzmarzick, and John Gustafson. 2019. Adsorption of Ciprofloxacin in Soil. Meeting Abstract, 110th Annual SSSA meeting, San Diego, Jan 6 ⿿ 9, 2019.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Yan Wang and Shiping Deng. 2019. Incubation Methods for Enzyme Assays. Meeting Abstract, 110th Annual SSSA meeting, San Diego, Jan 6 ⿿ 9, 2019.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Tang, Zhenya; Fangling Fan; Xinyue Wang; Xiaojun Shi; Shiping Deng, and Dingyong Wang. 2018. Mercury in rice (Oryza sativa L.) and rice-paddy soils under long-term fertilizer and organic amendment. Ecotoxicol. Environ. Safety 150: 116-122.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Richard P. Dick, Linda K. Dick; Shiping Deng; Xiufen Li, Ellen Kandeler; Christian Poll; Christopher Freeman; Timothy Graham Jones; Michael N. Weintraub; Kawthar A. Esseili; and Jyotisna Saxena. 2018. Cross-laboratory Comparison of Fluorimetric Microplate and Colorimetric Bench-scale Soil Enzyme Assays. Soil Biology and Biochemistry 121:240-248.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: McGowen, Eric B., Sumit Sharma, Shiping Deng, Hailin Zhang, and Jason G. Warren. 2018. An Automated Laboratory Method for Measuring CO2 Emissions from Soils. doi:10.2134/ael2018.02.0008. Agricultural & Environmental Letters 2018 3:180008
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Warren, Jason, Hailin Zhang, Shiping Deng, Eric McGowen and Sumit Sharma. 2019. Reply to ⿿Basis for Comparisons of Soil CO 2 Respiration Test Procedures⿝ doi:10.2134/ael2018.11.0064. Agricultural & Environmental Letters 4:180064.


Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Scientific communities, agricultural producer, stake holders, and state/federal government agencies are our target audiences. Changes/Problems:The long-term goal is to better understand the underlying basis of the importance and significance of biological and microbial activities in soils in relation to sustaining agricultural production, and maintaining and improving soil and environmental quality. Understanding the complex microbial community in the soil environment has proven to be a challenging task because of the vast diversity and enormously large population inhabiting the environment. Recent development of culture-independent molecular techniques has brought us hope for in-depth understanding of the soil black box. Unfortunately, changes in microbial community structure and diversity due to seasonal and temporal variations in nutrient or physical conditions are slow and gradual, making it difficult to interpret the data and obtain conclusive results. Therefore, in addition to continue our effort in developing and perfecting enzyme assay protocols and methods for evaluating microbial diversity and activities, we also obtained some bacterial isolates that are able to grow without nitrogen supplement in the growth media. Bacteria that confirmed with nitrogen fixation traits will also be evaluated for their potentials in promoting agricultural production. However, progress on this research effort is slow because of limited research funding. A new research project was initiated last year to strengthen the understanding of fate and stability of an antibiotic, ciprofloxacin, in soil and sludge. Preliminary study of this project was funded by the Vice President's Office at OSU. A research team has been established with the goal of seeking funding from the Department of Defense to strengthen biosecurity against bioterrorism. What opportunities for training and professional development has the project provided?Opportunity to train three visiting scientists as well as multiple undergrad and graduate students received hands-on experience in this research project. How have the results been disseminated to communities of interest? Attending and presenting at national and international conferences Publishing in peer-reviewed scientific journals Classroom teaching and discussions As an invited speaker to present at national/international conferences, i.e. the Tri-society meeting in Tampa, FL. What do you plan to do during the next reporting period to accomplish the goals?We will continue current research effort and interests to develop and standardize enzyme assay methods for soil; and to further our effort in understanding and revealing microbial diversity and activities that are important in agricultural production as well as preserving and maintaining soil and water quality in the environment. Recently, we are also involved in a project to evaluate fate and stability of antibiotics in soil and sludge, which is key to biosecurity against bioterrorism.

Impacts
What was accomplished under these goals? Cropping systems had significant impact on soil's capacity to cycle carbon, nitrogen, phosphorus, and sulfur through influencing on activities of soil enzymes. The effects of cropping systems on nitrogen- transforming enzyme activities were more significant than C-transforming enzyme activities. Following over 30 years of cropping management practices and evaluation of 60 soils with respect to activities of 14 enzymes involved in cycling carbon, nitrogen, phosphorus, and sulfur, data clearly suggested that returning crop residues to soil led to preserving soil biology and metabolic capacity; while grazing negatively impacted soil's capacity to cycle nutrients. Understanding the impact of long-term cropping systems on these enzyme activities in soil can guide the development of management strategies that promote soil productivity while maintain sustainability of the system. Continue leading an international effort to standardize soil enzyme assay methods. Enzyme assay methods developed will support systematic evaluation of ecosystem health components, focusing on soil biological indicators and the significance of changes in these indicators induced by changes in soil management practices.

Publications

  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2017 Citation: Du, Mengya, S.P. Deng, F.L. Fan, and H.L. Zhang. 2017. Grazing Lead to Lower Activities of Carbon-Transforming Enzymes in Soils. Meeting Abstract, 109th Annual ASA-SSSA-CSA Meeting, Tampa FL, Oct 22  25, 2017.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2017 Citation: Fan, F.L., M.Y. Du, Z.Y. Tang, S.P. Deng, and H.L. Zhang. 2017. Effect of Cropping Systems on Activities of Phosphorus- and Sulfur-Transforming Enzymes in Soils. Meeting Abstract, 109th Annual ASA-SSSA-CSA Meeting, Tampa FL, Oct 22  25, 2017.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2017 Citation: Du, Mengya, S.P. Deng, F.L. Fan, and H.L. Zhang. 2017. Negative Effect of Grazing on Activities of Nitrogen-Transforming Enzymes in Soils. Meeting Abstract, 109th Annual ASA-SSSA-CSA Meeting, Tampa FL, Oct 22  25, 2017.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2017 Citation: Dick, R.P., Lorenz, N. and S.P. Deng. 2017. Soil Enzyme Activity Assays: Protocols and Interpretations. Meeting Abstract, 109th Annual ASA-SSSA-CSA Meeting, Tampa FL, Oct 22  25, 2017.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2017 Citation: Deng, S.P., and R.P. Dick. 2017. Microplate Format Assay of Soil Enzymes and Method Standardization. Meeting Abstract, 109th Annual ASA-SSSA-CSA Meeting, Tampa FL, Oct 22  25, 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Deng, S., R. Dick, C. Freeman, E. Kandeler, M. N.Weintraub. 2017. Comparison and standardization of soil enzyme assay for meaningful data interpretation. J. Microbiol. Meth. 133:3234.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Xiaowei Pan, Michael D. Richardson, Shiping Deng, Robert J. Kremer, James T. English, Jeanne D. Mihail, Carl E. Sams, Peter C. Scharf, Kristen S. Veum, and Xi Xiong. 2017. Effect of Organic Amendment and Cultural Practice on Large Patch Occurrence and Soil Microbial Community. Crop Sci. 57:22632272.


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Scientific communities, agricultural producers, stake holders, and state/federal government agencies are our targeted audiences. Changes/Problems:The long-term goal is to better understand the underlying basis of the importance and significance of biological and microbial activities in soils in relation to sustaining agricultural production, and maintaining and improving soil and environmental quality. Understanding the complex microbial community in the soil environment has proven to be a challenging task because of the vast diversity and enormously large population inhabiting the environment. Recent development of culture-independent molecular techniques has brought us hope for in-depth understanding of the soil black box. Unfortunately, changes in microbial community structure and diversity due to seasonal and temporal variations in nutrient or physical conditions are slow and gradual, making it difficult to interpret the data and obtain conclusive results. Therefore, in addition to continue our effort in developing and perfecting enzyme assay protocols and methods for evaluating microbial diversity and activities, we also obtained some bacterial isolates that are able to grow without nitrogen supplement in the growth media. These bacteria are currently being characterized. The ones confirmed with nitrogen fixation traits will be further evaluated. It is our hope that these isolates could potentially strengthen our effort in promoting agricultural production and maintaining soil and environmental quality. However, growth conditions could influence nitrogen fixation traits. Characterization of the microbes is merely the first step in understanding these microbes. What opportunities for training and professional development has the project provided? Training and supervising graduate students as a major adviser on my project: 1 Training and advising graduate students on their projects as a member of graduate committees: 10. Training and advising undergraduate students conducting research on their projects: 1 Teaching and sharing knowledge with graduate/undergraduate students in courses taught or in the laboratory (16 students in SOIL 4483, 7 students in SOIL5383, 15 students in SOIL5483, 2 graduate students in the lab, 3 freshman visited the lab) Training and supervising visiting scientists on their projects: 3 How have the results been disseminated to communities of interest? Attending and presenting at national and international conferences Publishing in peer-reviewed scientific journals Classroom teaching and discussions As an invited speaker to present at an international conference in the United Kingdom What do you plan to do during the next reporting period to accomplish the goals?We will continue current research effort and interests to develop and standardize enzyme assay methods for soil; and to further our effort in understanding and revealing microbial diversity and activities that are important in agricultural production as well as preserving and maintaining soil and water quality in the environment. Special attention will be paid to characterize bacterial isolates with potential to fix atmospheric dinitrogen.

Impacts
What was accomplished under these goals? Based on evaluation of 60,797 16S rRNA and 42,931 nifH gene sequences, richness and diversity of microbial communities was enhanced by Manure but reduced by NPK, with the impact more prominent at the genus and species than phylum to family levels. The community abundance was predominantly influenced by carbon availability, while the richness and diversity of a community was governed by pH. A total of 25 or 16 microibal phyla were detected in soils at each of the two locations. Acidobacteria and Proteobacteria were the most abundant phyla detected at location 1; while Firmicutes, Actinobacteria and Chloroflexi were the most predomiant phyla detected at location 2. Abundance ratios of selected phyla may be of ecological significance and functions to indicate nutritional status, stress resistance, and metabolic capacity of a community. Of microbes habitating in a community, diazotrophs were especially competitive and thrived in soils receiving limited fertilization. Following century-long cultivation of winter wheat, Cyanobacteria comprised 2.6% of the microbial community in Check (without fertilization), whereas they were 0.19% and 0.05% in Manure and NPK, respectively. The observations were further supported by data obtained at location 2, where the abundance of Cyanobacteria in Native was 3.75%, in CRP was 0.01%, and in Wheat was 1.95%. The detected diazotrophic communities were constituted by distinct proportion of nifH clusters I and III. The predominance of Cluster I diazotrophs (100%) and Actinobacteria (60.1%) in CRP indicated that these microbial groups might be key players in ecosystem restoration. Help leading an international effort to standardize soil enzyme assay methods and presented an invited talk on the subject at an international conference in July 2016 in Bangor, United Kingdom. Enzyme methods developed will support our effort to systematic evaluation of ecosystem health components, focusing on microbial indicators to provide insight into the significance of changes in these key indicators.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: McGowen, Blake, Hailin Zhang, Jason G. Warren, Shiping Deng. 2016. An Alternative Method for Soil Microbial Activity Analysis. Meeting Abstract, 108th Annual ASA-SSSA-CSA Meeting, Phoenix AZ, Nov 6  9, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Pan, Xiaowei, Xi Xiong, Robert Kremer, James T. English, Michael D. Richardson, and Shiping Deng. 2016. Evaluation of Organic Amendments and Delivery Technologies for Control of Large Patch Disease (Rhizoctonia solani) on Zoysiagrass (Zoysia japonica) Fairways. Meeting Abstract, 108th Annual ASA-SSSA-CSA Meeting, Phoenix AZ, Nov 6  9, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Pan, Xiaowei, Michael Richardson, Shiping Deng, Robert Kremer, James English, and Xi Xiong. 2016. Soil Amendment for Control of Large Patch. Meeting Abstract, 108th Annual ASA-SSSA-CSA Meeting, Phoenix AZ, Nov 6  9, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Deng, S.P. and R.P. Dick. 2016. Comparison and standardization of soil enzyme assay for meaningful data interpretation. The 5th International Conference, Enzymes in the Environment: Activity, Ecology and Applications, Bangor, Wales, United Kingdom, July 24-28, 2016.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Konemann, C.E.; B.M. Kard; M.E. Payton; S.P. Deng; J.G. Warren; and T.M. Wilson. 2016. CO2 emissions from soil on Oklahomas Tallgrass Prairie Preserve in the Presence or Absence of Termites (Isoptera: Rhinotermitidae). Am. Midl. Nat. 176:60-71.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Katsalirou*, E., S. P. Deng, A. Gerakis; and D. L. Nofziger. 2016. Long-term management effects on soil P, microbial biomass P, and phosphatase activities in prairie soils. Eur. J. Soil Biol. 76: 61-69.
  • Type: Theses/Dissertations Status: Submitted Year Published: 2016 Citation: Li, X.F. 2016. Microbial Communities in Soil Ecosystems under Different Land Uses. Ph. D. Dissertation. Department of Plant and Soil Sciences, Oklahoma State University.
  • Type: Other Status: Published Year Published: 2016 Citation: Ascher, J., Deng, S., Dick, R., Insam, H., and van Gestel, C.A.M. 2016. Editorial on Tripmarks. Applied Soil Ecology vol. 100.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Scientific communities, agricultural producers, stake holders, and state/federal government agencies are our primary audiences. Changes/Problems:The long-term goal is to better understand the underlying basis of the importance and significance of biological and microbial activities in soils in relation to sustaining agricultural production, and maintaining and improving soil and environmental quality. Understanding the complex microbial community in the soil environment has proven to be a challenging task because of the vast diversity and enormously large population inhabiting the environment. Recent development of culture-independent molecular techniques has brought us hope for in-depth understanding of the soil black box. Unfortunately, changes in microbial community structure and diversity due to seasonal and temporal variations in nutrient or physical conditions are slow and gradual, making it difficult to interpret the data and obtain conclusive results. Therefore, in addition to continue our effort in developing and perfecting enzyme assay protocols and methods for evaluating microbial diversity and activities, we also started to isolate microbes that could potentially strengthen our effort in promoting agricultural production and maintaining soil and environmental quality. In particular, we are interested in free-living nitrogen fixers (bacteria that have the ability to obtained atmospheric nitrogen without partnering with other organisms). However, growth conditions are important influencing their nitrogen fixation traits. Thus, characterization of the microbes is the initial step of this research effort. What opportunities for training and professional development has the project provided? Training and supervising 1 graduate student as a major adviser Training and advising 11 graduate students as a member of graduate committees Training and advising 1 undergraduate student conducting research projects Teaching and sharing knowledge with 35 graduate/undergraduate students in courses taught or in the laboratory Training and supervising 2 visiting scientists How have the results been disseminated to communities of interest? Attending and presenting at national and international conferences Publishing in peer-reviewed scientific journals Classroom teaching and discussions As an invited speaker to present at higher education institutions What do you plan to do during the next reporting period to accomplish the goals?We will continue our current research effort and expand our interests to develop more enzyme assay methods, further our effort in understanding and revealing microbial diversity and activities that are important in agricultural production as well as preserving and maintaining soil and water quality in the environment. In the next funding cycle, special effort will be made to characterize bacterial isolates with potential fix atmospheric nitrogen.

Impacts
What was accomplished under these goals? Advanced understanding on the impact of management practices on soil biology, and ecosystem health and function, and diversity and community structure of diazotrophic microorganisms in agroecosystems. Based on evaluation of different ecosystems, such as Native, CRP (abandoned cropland currently under Conservation Reserve Programs), and Wheat as well as wheat field of different length of cultivation period, we found that: Bacterial community was more diverse in soils under continuous wheat cultivation than Native or CRP. If cultivation leads to soil degradation, bacterial diversity may not be a sensitive indicator of soil degradation. Firmicutes and Actinobacteria were the most dominant of phyla detected in Native and CRP, comprising 66.7~89.9% of the tested communities. In the soil under continuous wheat, Firmicutes and Chloroflexi were the most dominant, comprising 72.6%. CRP led to predominance of Actinobacteria, while continuous wheat cultivation promoted Firmicutes and Chloroflex. Proteobacteria and Acidobacteria, two of the most dominant groups reported in the literature, were only 8.1~17.5% of the tested communities. Ratios of Proteobacteria/Acidobacteira, presumably provide indication of soil nutritional status, was the highest in CRP (12.8), and similar in Native (2.73) and Wheat (2.24). Ratio of Firmicutes/Bacteroidetes was the highest in Native (95.2), and similar in CRP (60.9) and Wheat (60.4), which may reflect influence of cultivation. Cyanobacteria, was the highest in Native, followed by Wheat, and the least in CRP, suggesting the nutritional resilience of natural ecosystem in nitrogen acquisition. Sequences of nifH genes belonged mostly to cluster III for Native and cluster I for CRP or Wheat, with none belonged to cluster II or IV. Cluster I has been reported to be dominant in arctic tundra soil (65.3%) and marine systems (80%); while cluster III was dominant in tropical forest soil (46.9%) and termite guts (57%). These results suggest that different diazotrophic groups are important for soil N nutrition and acquisition in different environments. Bacterial composition was more similar in soils under 14-46 years of wheat cultivation than that under >100 years of cultivation. Chloroflexi was a predominant group in the soil under century-long wheat cultivation, but not in the soils that were cultivated with wheat for <46 years. However, Acidobacteria and Proteobacteria were the two most dominant phyla in all soils and ecosystems tested. The ratios of selected phylum, including ratios of Proteobacteria to Acidobacteria, Actinobacteria to Firmicuteswere, and Firmicutes to Bacteroidetes, were potential indicators of nutritional status, stress resilience, and metabolic capacity in soil. Continued the effort to revise and standardize soil enzyme assay methods. Enzyme methods developed will support our effort to systematic evaluation of ecosystem health components, focusing on microbial indicators to provide insight into the significance of changes in these key indicators. Thousands of bacterial isolates with potential to fix atmospheric nitrogen were obtained. Information generated from this project may be used to develop preventive strategies to guide future land use for enhanced sustainability and restoration of degraded soil ecosystems.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Pan, X.W., Xi Xiong, M.D. Richardson, J.T. English, R.J. Kremer and S.P. Deng. 2015. Effects of Organic Amendments and Cultural Practices for Control of Large Patch (Rhizoctonia solani) on Zoysiagrass (Zoysia japonica) Fairway. Meeting Abstract and oral presentation, 107th Annual ASA-SSSA-CSA Meeting, Minneapolis MN, Nov 15  18, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Pan, X.W., Xi Xiong, M.D. Richardson, J.T. English, R.J. Kremer and S.P. Deng. 2015. Effects of Organic Amendments and Cultural Practices on Soil Physical and Chemical Properties on Zoysiagrass (Zoysia japonica) Fairway. Meeting Abstract and poster presentation, 107th Annual ASA-SSSA-CSA Meeting, Minneapolis MN, Nov 15  18, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Xiufen Li, Shiping Deng, William R. Raun, Ying Teng and Yan Wang. 2015. Dominant Bacterial Phyla in Soils Under 18-114 Years of Wheat Cultivation. Meeting Abstract and poster presentation, 107th Annual ASA-SSSA-CSA Meeting, Minneapolis MN, Nov 15  18, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Xiaomin Ge, Shiping Deng and Luozhong Tang. 2015. Effects of Plant Litterfalls on Nitrogen Mineralization, Microbial Biomass and Enzyme Activities in Soils of a Poplar Ecosystem. ?. Meeting Abstract and poster presentation, 107th Annual ASA-SSSA-CSA Meeting, Minneapolis MN, Nov 15  18, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Natalie Doak and Shiping Deng. 2015. Are Phosphatase Activities Related to Microbial Abundance and Activity in Soil?. Meeting Abstract and poster presentation, 107th Annual ASA-SSSA-CSA Meeting, Minneapolis MN, Nov 15  18, 2015.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Yan, Y.F., S.Z. Fang, Y. Tian, S.P. Deng, L.Z. Tang and D.N. Chuong. 2015. Influence of Tree Spacing on Soil Nitrogen Mineralization and Availability in Hybrid Poplar Plantations. Open Access. Forests 6, 636-649. doi:10.3390/f6030636