Historic, Archive Document
Do not assume content reflects current
scientific knowledge, policies, or practices.
Reserve
aQRlll
.S64
1994
In-Depth Laboratory Review
October 19-21, 1 994
SYSTEMS
Soil Quality
Sustainable Agriculture
Composted Waste
Arbuscular Mycorrhizae
Pesticide Metabolism
Bioremediation
Biocontrol Management System
United States
Department of
Agriculture
National Agricultural Library
AGENDA
October 19, 1994
Theater, Log Lodge, BARC-East
8:15
8:30
8:40
9:20
10:20
10:30
11:30
12:00
Lincoln
PRE REVIEW "BREAK"
P. KEARNEY
P. MILLNER
Room, Log Lodge, BARC-East
GET ACQUAINTED SESSION AND SNACKS
INTRODUCTORY REMARKS
LAB OVERVIEW AND ISSUES
EXECUTIVE SESSION
BREAK
PATRICIA MILLNER
CHLOE RINGER
LUNCH
U.S.D.A., NAL
OCT 2 3 1998
•••••• Received
1:00
2:00
3:00
3:15
4:00
LAWRENCE SIKORA
DONALD KAUFMAN
BREAK
LAURA LENGNICK
REVIEW TEAM DISCUSSION
October 20, 1994
Lincoln Room Log Lodge, BARC-East
8:00
JEFFREY BUYER
9:00
WALTER MULBRY
10:00
BREAK
10:15
JEFFREY KARNS
11:15
SARA WRIGHT
12:15
LUNCH
1:15
TOUR FACILITIES
2:30
MEETING WITH SUPPORT STAFF
3:15
REVIEW TEAM DISCUSSION AND REPORT DRAFTING
October 21, 1994
Lincoln Room, Log Lodge, BARC-East
8:30 EXECUTIVE SESSION
10:30 END OF REVIEW
1
TABLE OF CONTENTS
AGENDA . 1
REVIEW TEAM . 3
USDA-ARS Organizational Background . 4
Definitions of Terms, Abbreviations and Acronyms used in ARS . 5
USDA Organizational Chart . 6
Beltsviile Area Organization Chart . 8
SOIL MICROBIAL SYSTEMS LABORATORY . 9
BACKGROUND . 9
PROBLEM . 9
MISSION . 9
RESEARCH ACCOMPLISHMENTS . 10
VISITING SCIENTISTS SINCE 1990 . 13
COOPERATIVE RESEARCH AGREEMENTS . 13
SUMMARY . 14
CRIS PROJECTS . 14
OLD CR1S PROJECTS . 14
DISTRIBUTION OF SCIENTIFIC PERSONNEL
AMONG CRIS PROJECTS . 15
SCIENTIFIC PERSONNEL AND EXPERTISE . 15
SUMMARY OF SOIL-MICROBIAL SYSTEMS LABORATORY
FINANCIAL RESOURCES . 16
1991 - 1994 PRODUCTIVITY SUMMARY . 17
SUPPORT STAFF . 18
SAFETY AND HEALTH REPORT . 18
RESPONSE TO RECOMMENDATIONS FROM PREVIOUS REVIEW (1993) . 19
CONTRIBUTIONS FROM SCIENTISTS . 20
Patricia D. Millner . 20
Chloe E. Ringer . 24
_ Lawrence J. Sikora . 26
Donald D. Kaufman . 30
Laura Lengnick . 33
Jeffrey S. Buyer . 35
2
Walter W. Mulbry . 38
Jeffrey S. Karns . 41
Sara F. Wright . 44
REVIEW TEAM
Dr. R. Ford Dennison, Chairman
Agronomy and Range Science
University of California
Davis, CA 35616
Phone: 916-752-4361
Dr. Ann Kennedy
USDA-ARS-PWA
Land Management & Water Conservation
Johnson Hall, Rm 215
Washington State Univ.
Pullman WA 99164-6421
Phone 509-335-1554
Dr. Allan Felsot
Food and Environmental Quality Lab.
Washington State University
Richland, WA 99164-1643
Phone: 509-375-9365
Dr. William Brinton, Jr.
Director, Woods End Research Laboratory, Inc.
1860 Rome Road
Mr. Vernon, ME 04352
Phone: 207-293-2488
3
■
DEFINITION OF SOME TERMS, ABBREVIATIONS AND ACRONYMS USED IN ARS
ARS: Agricultural Research Service. An agency in the Science and Education branch of USDA. ARS
has about 8,000 employees, including about 2,500 senior scientists. The Agency conducts research at
127 locations in the U.S. ARS is led by an Administrator and is divided geographically into eight
Areas, which are led by Area Directors.
BA: The Beltsviile Area includes the Beltsville Agricultural Research Center, the Beltsville Human
Nutrition Research Center, the U.S. National Arboretum, and the Glenn Dale Plant Distribution Station.
The Beltsville Area, at 6,600 acres, is the smallest Area geographically, but the largest in terms of
personnel and budget. About 1,450 employees, including about 440 scientists, work in the BA.
NPS: National Program Staff. Members are called National Program Leaders and each is a subject
matter specialist. NPS serves the Administrator of ARS in developing and coordinating research plans
and strategies on a national basis. NPS sets National program-directions, establishes priorities,
allocates resources, including this review, and acts as a clearing house for decision making.
Considerable interaction between Area managers and NPS is required to fulfill our respective roles.
INSTITUTES: The Beltsville Agricultural Research Center is composed of four Institutes (see
Appendix): the Plant Sciences Institute, the Livestock and Poultry Sciences Institute, the Natural
Resources Institute, and the Product Quality and Development Institute.
LABORATORIES: Laboratories are units located in the Institutes. Laboratories are led, both
scientifically and administratively, by Research Leaders. Typically, a Laboratory is comprised of 8-10
scientists, a scientific and clerical support staff and several temporary student and postdoctoral
employees. The program and mission of a Laboratory of this size must obviously be limited. In
reviewing a Laboratory, bear in mind that what appear to be discipline or program gaps are often filled
by collaboration with other Laboratories in the BA or elsewhere.
CRIS: Current Research Information System. This is an electronic system for the filing and retrieval of
information about individual agricultural research projects. In ARS, the terms "CRIS Work Unit" or the
acronym "CRIS" are used synonymously with "research project" or "project." New projects are planned
in coordination with NPS and are subjected to peer-review. The normal life of a project in ARS is 3 to 5
years.
SY: Scientist Year. This is the effort of a research scientist for 1 year. Fractional efforts (e.g., 0.5 SY)
in a given project are possible when a scientist works in more than one project during the course of a
fiscal year. The term is also used in ARS as a synonym for a research scientist [e.g., "I have six SYs
(research scientists). in my Laboratory'].
OTHER KINDS OF SCIENTIFIC PERSONNEL: Research scientists are responsible for all phases of
research. ARS also employs research associates ("postdocs"), support scientists (who have
responsibility for some portion of a project), technicians, students, and in some operations nonresearch
scientific personnel who perform work involving service to the public or to other government agencies.
AM: Administrative Management. This branch of ARS manages support activities, such as
procurement, facilities, fiscal allocations and personnel operations at all levels in ARS.
NOTE: The organizational scheme described above is presented graphically on the following pages.
4
ARS
Administrator
Associate Administrator
Deputy Administrator
National Program
Staff
Deputy Administrator
Administrative
Management
~i i r i i i
Pacific Northern Southern Beltsville Midwest North South Mid
West Plains Plains Area* Area* Atlantic Atlantic South
Area* Area* Area* Area* Area* Area*
[*Each Area is led by an Area Director and an Associate Area Director]
5
USDA Organizational Chart
6
Agricultural Research Service - Area Organization
Oeceobcr 1993
USOA, AGRICULTURAL RESEARCH SERVICE
6ELTSVILLE AREA
AREA OIRECTOR
K. 0. KURRELL
ASSOCIATE AREA DIRECTOR
G. C. HARTER
U.S. NATIONAL
ARBORETUM
BELTSVILLE HUMAN NUTRITION
RESEARCH CENTER
DIRECTOR
T. E. Elias
OIRECTOR
J. Spence
ADMINISTRATIVE
t FACILITIES MGR.
C. Kooberger
CARBOHYDRATE NUTRITION
J. G. Hallfrisch
ENERGY t PROTEIN NUTRITION
P. U. Hoe
EDUCATION UNIT
E. A. Neuaam
GARDENS UNIT
J. T. Judd
GERKPLASH UNIT
E. J. Garvey
G. R. Beecher
FLORAL t NURSERY
PLANTS RES. UNIT
J. C. Smith
R. H. Lawson
r
NATURAL RECOURCES
INSTITUTE
DIRECTOR
P. C. Kearney
a I KATE STRESS
S. Britz
ELECTRON H ICR OS COPE
U. P. Uergin
ENVIRONMENTAL CHEMISTRY
R. J. Wright
HYDROLOGY
U. J. Rauls (Acting)
REMOTE SENSING RESEARCH
G. F. Hart
SOIL MICROBIAL SYSTEMS
P. 0. Klllner
SYSTEMS RESEARCH
J. C. Ritchie (Acting)
LIVESTOCK AND POULTRY
SCIENCES INSTITUTE
DIRECTOR
T. J. Sexton
ANIMAL IMPROVEMENT PROGRAMS
H. 0. Noroan
BIOSYSTEKATIC PARASITOLOGY
J. R. LIchtenfels
GENE EVALUATION AND MAPPING
C. E. Rex road
GERKPLASH t GAMETE PHYSIOLOGY
L. A. Johnson
HELMINTHIC DISEASES
J. f. Urban
LIVESTOCK INSECTS
0. K. Kayes
MILK SECRETION t MASTITIS
R. H. Miller
HONRUHIHAHT ANIMAL NUTRITION
N. C. Steele
PROTOZOAN DISEASES
M. 0. Ruff
RESEARCH ANIMAL SERVICES
K. Hummel
RUHINANT NUTRITION
T. S. Ruasey
VETERINARY SERVICES
8. Stroud
ZOONOTIC DISEASES
R. fever
FACILITIES MANAGEMENT AND
PRODUCT QUALITY AND
OPERATIONS DIVISION
DEVELOPMENT INSTITUTE
DIRECTOR
OIRECTOR
W. G. Homer
0. 0. Bills
ADMINISTRATIVE BRANCH
FAMILY ECONOMICS
C. Bouie (Acting)
RESEARCH GROUP
F. Schwenk
FACILITY ENGINEERING
BRANCH
HORTICULTURAL CROPS
R. 0. Little
QUALITY
A. E. Uatada
FARM' OPERATIONS BRANCH
H. T. Badger
INSTRUMENTATION AND
SENSING
Y. R. Chen
MEAT SCIENCE RESEARCH
H. 8. Solomon
PESTICIDE RESEARCH
STAFF
0. 0. Bills
PLANT SCIENCES INSTITUTE
OIRECTOR
J. B. St. John
ASSOCIATE DIRECTOR
J. J. Hem
BEE RESEARCH
H . Shi manuk i
BIOCOWTROL OF PLANT OISEASES
R . 0. Luaxsden
FRUIT
R. F. Korcak
INSECT BIOCONTROL
J. L. Vaughn
INSECT CHEMICAL ECOLOGY
B. Leonhardt
INSECT NEUROBIOLOGY i HORMONE
J. A. Svoboda
MOLECULAR PLANT PATHOLOGY
R. E. Davis
NATIONAL GERKPLASH RESOURCES
A. K. Stoner
NEKATOLOGY
D. J. Chitwood
PLANT MOLECULAR BIOLOGY
A. K. Hattoo
SOYBEAN AND ALFALFA RESEARCH
0. L. Keister
SYSTEMATIC BOTAKY t MYCOLOGY
A. Y. Rossman
SYSTEMATIC ENTOMOLOGY
M. 6. Stoetzel (Acting)
VEGETABLE
S. L. Sinden
WEED SCIENCE
J. 0. Anderson
8
USDA-ARS SOIL MICROBIAL SYSTEMS LABORATORY
BACKGROUND
The Soil Microbial Systems Laboratory was established in 1984; it was previously
known as the Biological Waste Management and Organic Resources Laboratory. The
change in name was accompanied by a change in research direction away from waste
utilization in agriculture to very basic soil microbiology - plant - rhizosphere
investigations. In 1993, the Laboratory acquired staff, space, equipment, funds and
projects from the former Pesticide Degradation Laboratory. The molecular biology
experience of these new colleagues has been as welcome and productive addition.
PROBLEM
Public and private sectors demand that agricultural and other systems operate
sustainably, i.e., so they meet the needs and aspirations of the present without
compromising the ability of future generations to meet their own needs. Development of
alternative agricultural practices that balance the needs to conserve soil and water
quality, profitability, food safety and quality will address concerns about the impacts of
traditional and modern farming practices on sustainability and negative environmental
impacts. Such new approaches require more internalized nutrient cycling and fewer off-
farm inputs to increase system efficiency. Soil quality conservation and improvement
and use of organic matter will involve microbial processes. Improved agricultural system
functioning will require a better understanding of the critical biological components,
interactions, and nutrient transformations involved. How can crop/soil/nutrient plans be
managed so that optimal benefits of microbial activities are realized? Which microbial
processes are critical to the transition from conventional to alternative agricultural
practices and how and which practices need to be used to ease this transition.
MISSION
The mission of the laboratory is to conduct research on soil microbiological
processes that influence soil quality, especially the biological activity of soil.
Emphasis is placed on the development and application of basic principles and
technology of organic matter recycling, including composting, environmental,
biochemical, and genetic factors that influence interactions of soil-plant microbes
and microbiological processes critical in nutrient availability, agroecosystem
sustainability, and biodegradation of complex chemical compounds. The resultant
improvements in microbial cycling of plant nutrients which are derived from soil,
soil organic matter, crop residues, and animal manures will enhance soil quality,
conservation, and productivity by minimizing agricultural chemical inputs as well as
losses from erosion, nutrient runoff/leaching, and pest damage.
9
,
RESEARCH ACCOMPLISHMENTS
SOIL QUALITY AND MICROBIAL DIVERSITY
An assay was developed for pseudobactin, the siderophore of plant growth-promoting
Pseudomonas putida BIO, using monoclonal antibodies. This is the most sensitive assay for any
siderophore yet reported. A method was also developed to extract pseudobactin from soil
quantitatively; this is the first report of such for siderophores from soil. These two methods were
combined to measure pseudobactin in the rhizosphere of barley colonized with P. putida BIO.
This is the first directly measured concentration of a siderophore produced by an introduced
organism in a natural ecosystem. For the first time monoclonal antibodies to phytosiderophores
(iron chelators) secreted by graminaceous plants were produced.
A protein from the membrane that surrounds poly-p-hydroxybutyrate (PHB) granules was isolated.
This protein is missing in the bacteroid of a Fix' mutant of Bradyrhizobium japonicum. This
research suggests for the first time that PHB formation is important in N2 fixation conducted by
Bradyrhizobia.
A new solid medium was developed for enumeration and isolation of soil and rhizosphere
microorganisms without interference from the rapidly spreading bacterium Bacillus mycoides This
medium was used successfully in soil/rhizosphere studies to suppress the growth of B. mycoides
while enumerating and isolating other gram-positive bacteria.
Preliminary studies of the effects of the cropping systems on soil microbial community diversity
which was determined for bacteria by fatty acid methyl ester analysis, with calculations of
evenness and diversity, showed that total counts were not significantly different for the three
cropping systems studied (Rodale Institute Research Center's Farming Systems Trial). The
bacterial population isolated from the low-input animal system had the fewest genera and
species, and the lowest diversity and evenness. The bacterial populations from the conventional
and low-input legume systems had similar diversity and evenness. The fungal population from the
animal manure system had lower diversity and evenness than the other two systems. These
preliminary results suggest that conventional agricultural practices may maintain high indices of
microbial diversity in the rhizosphere. The functional significance of this needs to be investigated.
Arbuscular-Mycorrhizae
A significant accomplishment was achieved when the site of reaction of a monoclonal antibody on
vesicular-arbuscular mycorrhizal fungal hyphae was identified as a transient protein. All tests to
date indicate that the protein is present on actively growing, young hyphae and absent on older,
lysing, or melanized hyphae and after pot cultures have been allowed to dry at room temperature.
Thus, the presence of the protein is an indicator of activity of hyphae. Because the probe for the
protein is a monoclonal antibody, it is predicted that rapid, easily performed assays can be
developed to quantify the protein in soils. Also, preliminary tests have indicated that the protein
may be important in stabilization of soil aggregates which would be another breakthrough in
defining elements that contribute to soil quality.
Farming Systems and Compost Utilization Trials
Relevant historical data from RIRC's Farming systems Trial (FST) was reviewed; determined that
10
.
new statistical approaches were needed to evaluate the data because of variability in soil and
yield factors. A substantial library of aerial photographs of the RIRC area which covers the period
from 1948-1981 at irregular intervals of 4-7 years, and at least annual coverage of the years
1981-1989 was compiled. Such photographs have been essential in developing more accurate
soil maps of FST. Through computer digitization of soil reflectance, as recorded by aerial
photography, a new, more accurate soil map of RIRC Farming Systems Trial (FST) was generated
in cooperation with Dr. Eileen Perry. This map is currently used to accurately define and select
sampling sites for on-going data collection. Research was initiated to collect data needed to
characterize measurable variables such as soil reflectance, rooting depths, chlorophyll content,
plant height, rock fragment content, soil moisture, bulk density and crop yield. These data will be
used to develop a multivariate statistical procedure needed to analyze treatment effects in FST.
These procedures and results are being used by agronomists, soil scientists, extension agents
and farmers who are concerned with use, yield, and systems responses of sustainable agriculture
practices. Procedures developed in this investigation are being used successfully to delineate the
design of new, long-term experiments in such a way as to reduce variability due to soil
chemical/physical characteristics.
Cooperative investigations were conducted by Dr. M. Wander, Ohio State University, (presently at
University of Illinois, Urbana, Illinois) Dr, Bob Dudley, Eastern regional Research Center, and Dr.
Kaufman, on metabolism of C13-labeled acetate, benzoate, and catechol in FST conventional and
sustainable cropped soils. Treatment difference were apparent among treatments in C1302
evolution, which indicates a potentially different microbial utilization of these substrates by
microbes in the different cropping systems. NMR analysis of the treated/incubated soils is being
used to characterize the distribution of C13 products in soils.
A major collaborative long-term investigation was designed and initiated in 1992 by USDA-SMSL
and Rodale Institute Research Center scientists which involves composting of various
combinations of urban wastes (leaves, brush, newspapers, cafeteria wastes) and rural wastes
(animal manures), and subsequent utilization of the composted wastes in rotational field trials
which include both field and vegetable crops. The experiment is designed to last 15-20 years and
active research components of the project include formation, maintenance, and analysis of the
compost pile and process, utilization of the compost in field trials, effects of composts on soil
quality, crop yield and quality, nutrient cycling, leaching, and other environmental parameters.
Biodegradation of several starch-based plastic materials (golf tees, votive candle cups, film, and
pellets) were examined with microbial cultures in the laboratory and sequentially in compost piles
and field soils. Although most materials were still readily recognizable by shape, substantial loss
of composition did occur. Carbon-13 NMR spectra of the biodegradable materials showed
significant losses in starch contents, but few changes in composition of the petroleum-based
plasticizer in each item. Despite extensive degradation of the starch component, the votive
candle cups, and pellets were still readily identifiable in field soils after 11 weeks of composting
and 52 weeks in cropped field soil.
Working collaboratively with Penn state and RIRC, SMSL scientists have participated in
development of efficient methods to evaluate root architecture and dynamics in field and
vegetable cropping systems and to obtain data on interaction of sustainable agricultural practices
on root architecture and dynamics. Corn and bell pepper roots were excavated from several
depths in field plots where different cropping practices (permanent bed, moldboard plowed, or
minimum till time-fixed or variable traffic patterns) were used. Observed that a!! root forms were
dynamic within each treatment over time.
11
Bioremediation and Pesticide Metabolism
A total management system for coumaphos-containing cattle-dipping vats used in APHIS-VS's
Tick eradication Program was field tested at CFTRL in Mission, TX. Acidification of the vat
contents prevented accumulation of a toxic metabolite, formed in untreated vats due to the action
of anaerobic microbes, without using a biocide. The contents of the vat were then successfully
biodegraded by a consortium of bacteria. This allows maximum use of the coumaphos in the vat
accomplishing waste minimization, and allows for environmentally-sound disposal of vat contents.
Biodegradation studies were conducted using soils from cattle dipping waste pits that were
contaminated with high levels of coumaphos. Results showed that the coumaphos in all of the
soils could be rapidly biodegraded in soil slurries using indigenous soil microorganisms.
A cloning strategy based on DNA amplification to generate gene specific probes was used to
isolate, sequence and over-express the parathion hydrolase gene adpB from a Nocardia strain in
E. coli. This strategy was also used to isolate and characterize an s-triazine dechlorination gene
( trzA ) from the Rhodococcus corallinus. A collaborator has used the trzA gene to construct a
recombinant bacterial strain capable of dealkylating and dechlorinating atrazine and simazine.
Several genes were cloned from bacterial strains that degrade simpler s-triazine compounds likely
to be intermediates in the degradation of the major herbicides, atrazine, simazine, and cyanazine.
These studies demonstrated the importance of transmissible DNA elements and DNA/DNA
recombination in the evolution and spread of genes encoding the degradation of xenobiotic
compounds. The cloned genes may be of use in future construction of bacterial strains that
completely degrade triazine herbicides, which are typically relatively resistant to microbial
degradation.
A special 2-year project of a headquarters supported post doc resulted in production of a
monoclonal antibody which reacted with alachlor and diethyl analine, the simplest residue of this
herbicide. The antibody is being used in a flow immunosensor by the Environmental Chemistry
Laboratory to detect metabolites of alachlor in meat.
Sustainable Agriculture
Collected baseline measurements and uniformity trial data in preparation for designing a long-term
field crop exeriment on alternative agricultural practices. Established a pilot field study of
utilization of urban and rural wastes in sustainable agriculture. The study compares six
composts, two manures, and five inorganic wastes to each other and to fertilizer.
Demonstrated that one third compost N and two thirds fertilizer N equaled 100% fertilizer N in the
.growth chamber. These data suggest that farmers could reduce fertilizer costs by using
composts. Determined that addition of compost to soil did not stimulate the mineralization of soil
organic matter. Developed a new microbial biomass determination based on rehydration of dried
soils. Results were similar to the fumigation technique but used chemicals that are less
hazardous than chloroform. Proposed an indicator of soil quality based on the use efficiency of N
from organic and inorganic sources.
Alternatives to Methyl Bromide Soil Fumigation
New CRIS proposal was written and sent out for review.
12
SMSL VISITING SCIENTISTS SINCE 1990
Suichi Mihashi, University of Tokyo
Vladimir Yakovchenko, Kiev State University, Ukraine
Paulette Royt, George Mason University
Sergei Chernikov
Vitaly Nebesny, Ukranian Academy of Science
Wolfgang Hordt, Univ. Hohenbeim, Stuttgart, Germany
Thierry Heulin, C.N.R.S., Nancy, France
Steve Bentivenga, University of West Virginia
Chang Lu, Peoples Republic of China
Keng-Yeang Lum, Malaysian Agricultural Research and Development Institute, Kuala Lumpor,
Malaysia
COOPERATIVE RESEARCH AGREEMENTS
Soil and Agronomic Factors in Sustainable Agriculture - Specific Cooperative Agreement
University of Maryland Agronomy Department (1270-12000-015-01S)
Root Biology in Sustainable Agriculture - Specific Cooperative Agreement - Rodale Institute
Research Center (1270-12000-014-01S) u.i! cjki
Root Agriculture and Dynamics in Sustainable Agriculture - Specific Cooperative Agreement -
Pennsylvania State University (1270-12000-014-02S) ojo^K. Q fcocLaslt,
ft
nn
13
■
SUMMARY
SOIL-MICROBIAL SYSTEMS LABORATORY
CRIS PROJECTS
CWU NO.
TITLE
NET TO CRIS
DURATION
1270-12000-01 3-00D
Role of Vesicular Arbuscular
Mycorrhizae (VAM in Low Input
Agricultural Systems
155,157
2/91 - 2/96
1270-1 2000-0 14-00D
Production and Use of Rural/Urban
Waste Co-Compost: Microbial
Processes
265,211
6/94 - 6/99
1270-1 2000-01 5-QOD
Integrated Soil-Nutrient-Crop-
Microbial-Pest-Waste Management
Strategies for Sustainable Agriculture
296,095
8/94 - 8/99
1 270-1 2000-01 6-00D
Soil Quality and Soil Ecology in
Sustainable Agriculture
351,108
8/94 - 8/99
1270-1 2220-001 -00D
Integrated Soil/Crop Biocontrol
Management System: Sustainable
Alternatives to Methyl Bromide
232,162
8/94 - 8/99
1270-1 21 30-005-00D
Biochemistry and Molecular Biology of
Pesticide Metabolism in Soil and
Water
269,414
89 - 12/94
0500-00026-034-00D
Bioremediation of Contaminated Sites
to Protect Water Quality
60,000
10/94 - 9/95
TOTAL 1,629,147
OLD CRIS PROJECTS
CWU NO.
TITLE
DURATION
1 270-1 2000-009-00D
Analysis of Soil Microbiological Processes
Which Affect Low-Input Sustainable
Crop/Livestock System
3/89 - 2/94
1270- 12000-01 0-00D
Fertilizer and Cultural Management Effects on
Crop Nutrient Quality and Bioavailability
4/89 - 4/94
1270-12000-01 1-00D
Management of Beneficial Plant Microbe Soil
Interactions for Low Input Sustainable
Agriculture
4/89 - 4/94
1 270-1 2000-01 2-00D
Effects of Residue Decomposition in Low Input
Sustainable Agricultural Systems
5/89 - 5/94
14
'
DISTRIBUTION OF SCIENTIFIC PERSONNEL
AMONG CRIS PROJECTS
Scientist
1270-
12000-
013-00D
1270-
12000-
014-00D
1270-
12000-
015-00D
1270-
12000-
016-00D
1270-
12220-
001-00D
1270-
12130-
005-00D
0500-
00026-
034-00D
Buyer
.20
.20
.50
.10
Karns
.25
.75
Kaufman
.65
.20
.15
Lengnick
.70
.30
Millner
.35
.10
.10
.10
.35
Mulbry
.10
.10
.10
.70
Ringer
.50
.50
Sikora
.10
.50
.30
.10
Wright
.55
.10
.05
.30
SOIL MICROBIAL SYSTEMS LABORATORY
SCIENTIFIC PERSONNEL AND EXPERTISE
Dr. Jeffrey Buyer
Research Chemist
biochemistry, siderophore-metal
interactions, microbial ecology
Dr. Jeffrey Karns
Research Microbiologist
molecular biology, bioremediation,
biochemistry, microbial ecology
Dr. Donald Kaufman
Research Microbiologist
sustainable agriculture, composting,
pesticide metabolism
Dr. Patricia Millner
Research Microbiologist
& Research Leader
soil microbiology, composting,
bioaerosols, immunology,
mycology/mycorrhizae, molecular
systematics
Dr. Walter Mulbry
Research Microbiologist
molecular biology, bioremediation,
biochemistry
Dr. Lawrence Sikora
Research Microbiologist
immunology, compost, science, soil
science, nutrient cycling, soil quality
Dr. Sara Wright
Research Microbiologist
immunology, mycorrhizae
15
SUMMARY OF SOIL-MICROBIAL SYSTEMS LABORATORY
FINANCIAL RESOURCES
FY '93
FY '94
FY '95
Net to Location
1,401,531
1,927,508
1,959,864
Indirect Research Cost
275,135
382,474
392,322
Adjustments
Post Doc
40,690
50,650
9,550
Water Quality
60,000
60,000
BARD
4^51
4^61
Net to MU
1,126,396
1,545,034
1,567,542
Salary
811,737
762,106
1,024,618
All-Other
311,380
595,919
604,530
Ratio of Salary to All-Other
2.60
1.27
1.69
Percent Salaries
58.05
37.97
50.69
Total dollar per SY
199,750
286,734
288,781
Discretionary Funds per SY
39,626
103,677
62,219
Percent Discretionary
19.84
36.16
21.55
16
SOIL-MICROBIAL SYSTEMS LABORATORY
1991 - 1994 PRODUCTIVITY SUMMARY
Scientist
Peer Reviewed Pub.
Other
Abstract/Sci.
1st Author
Co-Author
Publications
Presentations
Buyer
3
2
4
2
Karns
2
7
23
Kaufman
1
3
3
12
Lengnick
2
1
1
3
Millner
4
3
2
15
Mulbry
5
1
10
Ringer
1
Sikora
5
4
2
10
Wright
4
2
3
Hornick*
1
2
4
4
‘Resigned 2/94
PEER REVIEWED PAPERS: 53
TOTAL PUBLICATIONS: 69
PUBLICATIONS PER SY PER YEAR
PEER REVIEWED
ALL PUBLICATIONS
1991-1994
5.3
6.9
17
■
SUPPORT STAFF
Ms. Dorothy Talmud
Secretary, Office Automation
Ms. Claire Reese
Office Automation Assistant
Ms. Nancy Enkiri
Microbiologist
Ms. Sara Reynolds
Microbiologist
Mr. Tim Prickett
Biological Laboratory Technician
Mr. Stanley Tesch
Biological Laboratory Technician
Dr. Cecil Tester
Chemist
Ms. Heidi Hartman
Cooperative Student - University of Maryland
Ms. Tracy Butler
Cooperative Student - University of Maryland
Mr. Pedro Del Valle
University of Maryland - Biological Lab. Technician
Dr. Laura Lengnick
Agronomist - Research Associate
Dr. Chloe Ringer
Plant Pathologist - Research Associate
Ms. Mary Jane Letaw
University of Maryland -Agricultural Technician
Ms. Leanne Teerlinck
RIRC- Research Assistant
Ms. Elizabeth Lyman
RIRC - Research Assistant
SAFETY AND HEALTH REPORT
All items identified by the Safety and Health Office have been corrected except for installation of
new stairs and railing along the path leading to the Chemical Storage bunker, Building 318 B.
This has been noted on every inspection since February 1990 and each time a work request was
submitted to FMOD for action; none has yet been taken. The lab safety program is ongoing with
each problem identified and corrective action taken by the Lab safety officer with cooperation from
every lab member.
18
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RESPONSE TO RECOMMENDATIONS FROM PREVIOUS REVIEW (1993)
Concern 1: It is recommended that all present work on the decomposition of inoculated corn
residues be terminated when current experiments are completed.
Response: All work was completed and published; no new work undertaken.
Concern 2: it is recommended that Drs. Kaufman, Payne, Perry and other cooperators publish
peer-reviewed manuscripts on the large amount of research conducted by ARS scientists at the
Rodale Research Center.
Response: To date, 3 papers have been published on completed work; analysis of data
has been hampered by the termination of temporary ARS employees, Drs. Payne and Perry, and
resignation of the participating Research Director at RIRC, and change in assignments of other
cooperators. Dr. Kaufman has been negotiating with ARS and RIRC staff involved in the work for
an agreeable plan for publication. He has determined that the data will need specialized statistical
handling, with modern georeferencing and mapping to account for spatial variability. Dr. Millner
has recommended and Dr. Kaufman has welcomed the suggestion that Dr. Lengnick become
involved in this project. It is the lab's intention to provide Dr. Lengnick with a full-time field
assistant to manage the daily matters at the BARC Sustainable Agriculture site so that she will
have time to undertake the statistical analysis of the RIRC/ARS data necessary for publication.
Concern 3: It is recommended that Dr. Laura Lengnick be asked to assume the coordinating
role in SMSL for the sustainable project.
Response: Dr. Lengnick was hired as a Research Associate in 1993 and has
coordinated the field crops research project of the BARC Sustainable Agriculture Program.
Concern 4: It is recommended that Dr. Millner continues to work with Dr. Kemper and Dr.
Walker on establishing a Federal Center on Sustained Agricultural Use of Urban and Rural
Wastes.
Response: In 1993, Dr. Millner conducted a special briefing session for the Beltsville
Area Director and staff regarding the research program, proposed operational plan, and funding
streams for the Center. NPS's requests for information and statement of proposed work were
promptly supplied. In addition, Dr. Millner provided information to The Composting Council to
assist them in establishing a working coalition of interested stakeholders. Thus far, no additional
funding has been received to support a Center. Dr. Millner has recently submitted a multi-agency
grant proposal to USEPA to conduct research on establishing sampling and analytical standards
for compost in the US. This project will be the first one in which a research unit at BARC would
take the major coordinating lead as envisioned in the plans for the Center.
Concern 5: It is recommended that Dr. Sikora continue research to more clearly define what is a
measure of soil quality. He is making good progress on his nitrogen flux concepts.
Response: Dr. Sikora has continued and expanded his research on measuring soil quality
through the help of Dr. Vladamir Yakovchenko, Visiting Scientist from University of Kiev, and an
University of Maryland cooperative student from agronomy. In addition, the expiring CRIS project
covering this research has been rewritten to include development of soil biological quality
indicators.
19
'
CONTRIBUTIONS FROM SCIENTISTS
I. Patricia D. Millner, Microbiologist
II. CRIS Project:
CRIS Project 1270-1 2000-01 3D: Role of Vesicular Arbuscular Mycorrhizae (VAM) in Low
Input Agricultural Systems
Objective: Determine relatedness of arbuscular mycorrhizal fungi (AMF) at the molecular level,
and use this to develop molecular probes useful for identification of taxa that have colonized field
samples of crop roots. Apply the technique to farming system field trials to determine changes in
AMF root colonization and soil networking by plant growth effectiveness taxa in comparison with
"weedy" non-beneficial taxa. Determine the relative effects of various alternative low-input
treatments on AMF crop/soil colonization such that taxa which facilitate transition from
conventional to alternative farming practices can be recognized and managed through cropping
practices rather than inoculation.
Progress: Methods have been developed to cleanly produce spores in soilless condition and to
reliably extract, amplify, and finally sequence rDNA from as few as one to ten spores of identified
AMF isolates. Using areas of the variable regions of the internal transcribed spacers and the
conserved 5.8s regions, it has been possible to confirm some species complexes and generic
groupings relatedness. This adds support to the previously proposed concepts based on
morphological, ontological, and fatty acid methyl ester patterns of our collaborators. It also
provides the basic data needed to proceed directly to construction and development of the taxon
specific probes.
Plans: Refine PCR primers to be selective for AMF DNA by generic and/or species groupings;
adapt the extraction and PCR technique to colonized roots of corn. Construct taxon-specific
oligonucleotide probes (TSOPs)for Glomus mosseae, G. etunicatum, G. occultum, Gigaspora
rosea, Gi. gigantea, Gi. albida, Acaulospora mellea, Entrophospora (species to be determined),
and other dominant AMF taxa present in the BARC long-term sustainable field crops research
study site (as determined in the baseline species survey). Verify probe specificities with standard
operating conditions. Use TSOPs to determine AMF colonization by crop and field location over
time and relate to cropping practices, and changes in biotic and abiotic soil factors.
Old CRIS Project 1270-12000-009-003: Analysis of Soil Microbiological Processes Which
Affect Low-Input Sustainable Crop/Livestock System
New CRIS Project 1 270-1 2000-014-00D: Production and Use of Rural/Urban Waste Co¬
compost: Microbial Processes
Objective: Develop microbial consortia that can be used to reliably produce disease suppressive
composts for control of certain soilborne plant diseases.
Progress: Worked with Dr. Ringer in planning preliminary growth chamber studies in which she
has found 6 rural/urban waste co-composts suppress Pvthium, which causes damping-off disease
of cucumber seedlings.
20
.
Plans: These composts and others will be further tested (see p.24). The physical, chemical, and
biological quality status of compost that is found suppressive will be characterized to increase our
understanding of the substrate factors that favor development and expression of suppressiveness
in compost. Effective microorganisms from these composts eventually will be used along with
other biocontrol microbes to develop consortia of plant beneficial microorganisms for production
of disease-suppressive compost.
New CRIS 12700-12222-001-00D Integrated Soil/Crop Biocontrol Management System:
Sustainable Alternatives to Methyl Bromide
Old CRIS 1270-1 2000-01 0-00D Fertilizer and Cultural Management Effects on Crop Nutrient
Quality and Bioavailability (Drs. Hornick and Chaney, previous members of SMSL staff worked
on this CRIS exclusively)
Objectives: Develop an integrated group of practices for conserving and improving soil and crop
health and productivity by combining the use of biocontrol agents, organic matter amendments,
microbial competition, alternate cover crops, low toxicity/residual pesticides/fungicides, and low-
cost field bed preparation techniques for high-value crops such as strawberries, asparagus, and
tomatoes.
Progress and Plans: This project is designed to work in concert with and to move the results of
other projects to the field by working toward the solution of a very urgent need. The project
proposal is currently under review. Meanwhile, compost quality assessments and disease
suppressive studies on CRIS 1270-12222-014-00D are proceeding so that we can move directly
along. Collaborators who have good linkages with commercial enterprises that are willing to
participate in field trials of promising alternatives to methyl bromide have been identified and are
ready to help with field trials.
III. Cooperators:
ARS:
All staff members of Soil Microbial Systems Laboratory, BARC
Dr. Jim Locke, Floral Crops Research Unit, National Arboretum, BARC
Mr. T. Badger, Farm Manager, BARC
Dr. Stan Nemec, Citrus & Subtropical Products Lab, Winter Haven, FL
Dr. David Douds, Plant & Soil Biophysics Research, Eastern Regional Res. Ctr., Philadelphia, PA
Other:
Dr. Joseph Morton, Dept, of Soil and Plant Sciences, West Virginia University, Morgantown
Dr. Raymond Weil, Department of Agronomy, University of Maryland, College Park, MD
Dr. F.R. Gouin, Department of Horticulture, University of Maryland, College Park, MD
Dr. John Haines, New York State Museum, Albany, NY
Dr. John Walker, Office of Solid Waste, USEPA, Washington, D.C.
The Composting Council, Alexandria, VA.
Dr. T. Heulin, CNRS, Nancy, France
21
m
IV. Curriculum Vitae:
Education:
1966-1970 Univ. of Maryland; major, Microbiology; minor, Chemistry; B.S. 1970.
1970-1975 Univ. of Maryland; major, Botany (Mycology); M.S. 1975.
1981-1984 Univ. of Maryland; major, Environmental Science; minors, Microbiology, Biochemistry;
Ph.D. 1984.
Professional Employment:
1969- 1970
1970- 1978
1977- 1978
1978- 1984
1985-1991
1991-
Biological Aid, Mycology Laboratory, Plant Protection Institute, USDA-ARS, Beltsville, MD.
Microbiologist, Mycology Laboratory, USDA, ARS, Beltsville, MD.
Microbiologist, Biological Waste Management and Soil Nitrogen Laboratory, Agricultural
Environmental Quality Institute, USDA. ARS. Beltsville, MD.
Microbiologist, Biological Waste Management and Organic Resources Laboratory, USDA,
ARS, Beltsville, MD.
Microbiologist, Soil-Microbial Systems Laboratory (formerly Biological Waste Management
Laboratory), ARS, USDA, Beltsville, MD.
Research Leader, Soil-Microbial Systems Laboratory, ARS, USDA, Beltsville, MD.
Membership in Professional Societies:
Mycological Society of America Soil Ecology Society
American Society for Microbiology Soil and Water Conservation Society
American Phytopathological Society
American Society of Agronomists
Offices and Committee Assignment Held in Professional Societies:
Member, Sustaining Membership Committee, 1980-1983, Mycological Society of America.
Member, Mycorrhizae Committee, 1990-1993, American Phytopathological Society.
Member, Advisory Committee, 1990-Present, International Collection of Vesicular and Arbuscular-Vesicular
Mycorrhizal Fungi, West Virginia University, Morgantown (NSF funded culture collection)
Honors and Awards:
1967 Alpha Lambda Delta, Elected Member, Freshman Woman's Honorary
1969 Sigma Alpha Omicron, Elected Member, Microbiology Honorary
1 969 Mortar Board Scholarship recipient
1970 B.S. Degree with honors
1977 USDA Superior Service Award, Team member of the Sewage Sludge Land Utilization Research
Group "in recognition for outstanding team effort and response to an urgent national need for
research information on safe and beneficial use of sewage sludge on agricultural land."
1984 Nominated to receive the Outstanding Dissertation Award from the University of Maryland (College
Park). Regents decided to forgo all awards that year for fiscal reasons.
1 991 Nominated for WISE (Women in Science Engineering) award
1992 ARS Kinney Post-Doctoral Research Associate for proposed work on "Microbial Enhancement of
Farm/Urban Waste Compost" for FY-93.
Referred Publications Since 1991:
Millner, P. D. Characterization and use of vesicular-arbuscular mycorrhizae in agricultural
production systems. Plant and Soil 32:335-342. 1991.
22
Lewis, J. A., lumsden, R. D., Millner, P. D. and Keinath, A. P. Suppression of damping-off of
peas and cotton in the field with composted sewage sludge. Crop Protection 11:260-266. 1992.
Millner, P. D. and Kitt, D. G. The Beltsville method for soilless production of V-A mycorrhizal
fungi. Mycorrhiza 2:9-15. 1992
Wright, S. F. and Millner, P. D. Dynamic processes of vesicular-arbuscular mycorrhizae: A
mycorrhizosystem within the agroecosystem, pp. 29-59. IN: J.L. Hatfield, et al. (eds.) Soil
Biology: Effects on Soil Quality. Advances in Soil Science. Lewis Publ., Boca Raton, FL. 1993
Syliva, D.M., D.O. Wilson, J. H. Graham, J.J. Maddox, P.D. Millner, J.B. Morton, H.D. Skipper, S.
F. Wright and A.G. Jarstfed. Evaluation of vesicular-arbuscular mycorrhizae in diverse plants and
soils. Soil Biol. Biochem. 25:705-713. 1993
Millner, P.D. and D. G. Kitt. Mycorrhizal colonization of corn co-inhabited by biopesticidal-
recombinant Clavibacter xyli subsp. cynodontis. Microb. Releases 2:81-84. 1993.
Millner, P. D. et al. Bioaerosols Associated with Composting. J. Regulatory Toxicol. Pharmacol.
(Accepted for Publication as a supplement). 69 pp.
Other:
Millner, P. D. "VA Mycorrhizae, Soil Fertility and Crop Production in Africa", World Bank,
Proceedings of Workshop on Managing The Fertility of Soils in Africa , Washington, D.C. 1992,
Millner, P. D. "Inoculum Production and Use of VA Mycorrhize in Soils" , Proceedings of OECD
Workshop on Establishment of Microbial Inocula in Agriculture, Maui, Hawaii (accepted for
publication in a section of J. Alternative Agriculture). 1994.
Millner, P.D. Book Review: "Mycorrhizal Fungi in Sustainable Agriculture". J. Alternative
Agriculture 8(4): 1993.
23
.
'
I. Chloe E. Ringer, Postdoctoral Research Associate
II. CRIS Project:
Old CRIS Project 1270-120Q0-009-00D: Analysis of Soil Microbiological Processes Which
Affect Low-Input Sustainable Crop/Livestock System
New CRIS Project 1270-12000-014-00D: Production and Use of Rural/Urban Waste Co¬
compost: Microbial Processes
Objective: Determine the potential usefulness of composts and compost extracts in controlling
specific soilborne and foliar plant pathogens.
Progress: Conducted preliminary growth chamber studies on the usefulness of various
rural/urban waste co-composts for suppression of Pvthium damping-off disease of cucumber
seedlings in soilless potting mixes composed of 1:1 peatiperlite and 10 or 30% concentrations of
the following waste co-composts:
broiler carcasses/broiler manure/sawdust/straw
broiler litter/leaves
dairy manure/leaves/straw/newspaper
horse manure/cow manure/straw/sawdust/clay
broiler litter/food waste/biodegradable plastic/leaves
leaves/lawn waste
All six composts were highly suppressive to Pvthium damping-off of cucumber. Severe disease
losses in sterile, inoculated controls provided evidence that the suppression was due to the
presence of beneficial microorganisms in the composts. Further, plant growth in most nonsterile
compost mixes was comparable to growth in a popular, commercial soilless mix. These results
will strengthen interest in on-farm composting of rural/urban wastes and contribute to the future
development of a marketable, value-added farm product, while reducing the use of peat and
chemical inputs.
Plans: These composts and others will be further tested to evaluate their effectiveness in
suppressing other soilborne plant pathogens, such as Phvtophthora spp. and Rhizoctonia solani.
on cucumber seedlings and other horticultural crops. Additionally, compost extracts will be
evaluated for suppression of foliar plant pathogens. Effective microorganisms from these
composts eventually will be used to develop consortia of plant beneficial microorganisms for the
commercial production of reliably high-quality, disease-suppressive compost.
III. Cooperators:
Dr. K.P. Hebbar, Biocontrol of Plant Diseases Lab, ARS, Beltsville, MD
Dr. R.L. Chaney, Environmental Chemistry Lab, ARS, Beltsville, MD
Dr. L.E. Carr, Agricultural Extension Service, University of Maryland, Princess Anne, MD
Dr. F.R. Gouin, Department of Horticulture, University of Maryland, College Park, MD
Rodale Institute Research Center, Kutztown, PA
Mr. G. Leidig, Autrusa Compost Consulting, Blue Bell, PA
Mr. P. Boop, Briar Patch Organic Farms, Mifflinburg, PA
24
.
IV. Curriculum Vitae:
Education:
1975 B.S. Botany, University of Maryland, College Park, MD
1985 M.S. Plant Pathology, University of Maryland, College Park, MD
1992 Ph.D. Plant Pathology, University of Maryland, College Park, MD
Professional Employment:
1976-1981
1981-1985
1985-1986
1987-1992
4/93-9/93
1 /94-
Plant Pathologist, Biocontrol of Plant Diseases Lab, USDA-ARS, Beltsville, MD
Graduate Research Assistant, Department of Botany, University of Maryland, College Park,
MD
Graduate Teaching Assistant, Department of Botany, University of Maryland, College Park,
MD
Graduate Student, Department of Botany, University of Maryland, College Park, MD
Acting Director, Plant Diagnostic Lab, Department of Botany, University of Maryland, College
Park, MD
Postdoctoral Research Associate, Soil Microbial Systems Lab, USDA-ARS, Beltsville, MD
Membership in Professional Societies:
American Phytopathological Society
Publications:
Ringer, C.E., and Grybauskas, A.P. 1994. Infection cycle components and disease progress of gray leaf
spot on field corn. Plant Disease (In press).
25
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Lawrence J. Sikora, Microbiologist
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I. CRIS Project:
Old CRIS Project 1270-1 2000-01 2-00D: Effects of residue d
sustainable agriculture
S\Ycr* - /ry/J Ck
lecomposition in low-inpur
New CRIS Project 1270-1 2000-015-00D: Integrated Soil-Nutrient-crop-microbial-Pest-Waste
Management Strategies for Sustainable Agriculture. 0r*\p <5^ U$ £ J
Objective: Determine the effects of compost-fertilizer combinations on the yielca and nytrient
content of crops.
Progress: Adding a mixture of compost and fertilizer equalling to the N requirement for wheat
showed that compost N can substitute for one third of the inorganic N requirement. These data
suggest that farmers can save on their fertilizer costs when using composts in this manner and
benefit further from the additional organic matter in compost. Similar studies using fescue showed
that a refuse-sewage sludge co-compost could substitute for one third of the inorganic N
requirement but sewage sludge compost did not demonstrate the same benefit. The use of
ammonium nitrate or urea as the fertilizer source did not effect these results. Addition of compost
to soil did not increase the mineralization of soil organic matter.
Plans: Continue to evaluate mixtures or blends of composts with fertilizers to maximize the
benefits of composts while minimizing the additions of non-nutrients such as heavy metals from
composts made from urban wastes. Develop control strategies for composting that will maximize
benefits and remove hindrances of composts and composting such as product instability and odor
generation.
Old CRIS Project 1270-12000-009-00D: Analysis of Soil Microbiological Processed which
Affect Low-Input Sustainable Crop/Livestock System
New CRIS Project 1270-1 2000-01 4-00D:
compost;
Production and Use of Rural/Urban Waste Co-
Objective: Determine the effects of low-input agricultural practices on soil microbial C and N
components and transformations.
Progress: The Farming systems Trial at the Rodale Institute Research Center has been studied
extensively to determine the effect of manures or legumes as compared to inorganic fertilizer as
the source of nitrogen for crop growth . Decomposition studies indicated a higher level of
respiration for both legume and manure systems. Other components that were higher in the
manure or legume low-input systems were microbial biomass, N mineralization and N flux. Yield
data for the 13 year study did not show yield responses (decreases or increases) to the low-input
treatments possible due to the organic matter variation of greater than 100% across the field .
Efforts to map the area by sampling and by aerial photography indicated that soil brightness
measurements of aerial photos could predict soil organic matter content with reasonable
accuracy. Normalizing the yield data for organic matter levels is being used to determine
treatment effects. A soil quality indicator based on nitrogen efficienpy was proposed /
_ — ksfe — SrA 7 J cA~
26
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Plans: Establish the principles of co-composting using laboratory scale composters. Determine
through field utilization trials the environmental safety and agronomic/soil quality benefits of
composted rural/urban waste materials on crop lands. Continue pilot field project evaluating the
effects of urban and rural composts on field crops. Test the effects and survival of microbial
inoculant for composting to enhance the compost as a soil biofertilizer.
Old CRIS; 1270-12000-011-00D: Management of beneficial plant microbe soil interactions
for low input sustainable agriculture.
NEW CRIS 1270-12000-016-00D: Soil Quality and Soil Ecology in Sustainable Agriculture.
Objective: Analyze Bradyrhizobium japonicum-soybean interaction to improve the N2 fixation
capacity.
Progress: A mutant of Bradyrhizobium japonicum which does not fix N2 was found to have low
levels of poly-p-hydroxybutyrate (PHB) in the bacteroid form. A monoclonal antibody was raised
which reacts with a 14 kDa component that resides on the membrane that surrounds PHB and is
missing in the mutant. Other scientists hypothesize that this component is a membrane stabilizer
which separates the hydrophilic cytoplasm from the hydrophobic granule. Without PHB, N2
fixation in B. japonicum may be reduced or eliminated especially during seed filling. This research
demonstrates the possible importance of PHB in N2 fixation in Bradyrhizobia.
Plans: Extensive sampling of the FST plots at Rodale allowed preliminary evaluation of soil
factors which may be used to determine soil quality. Studies will be continued at Rodale and at
two other long term sites in Maryland to test a biological indicators of soil quality such as
respiration, biomass content and biomass turnover of nitrogen. Guidelines will be developed for
uses of composts with fertilizer to reduce the total amount of costly external nutrient inputs.
Determine the effect of composts on the microbial diversity of soils and rhizosphere.
il. Cooperators
ARS:
Dr. Dave Kuykendall, Beltsville, MD
Dr. Ron Korcak, FL, Beltsville, MD
Dr. Jeff Buyer, SMSL, Beltsville, MD
Dr. Pat Millner, SMSL, Beltsville, MD
Dr. Don Kaufman, SMSL, Beltsville, MD
IV. Curriculum Vitae:
Educational Background:
1962-1966 Wayne State University; B.S. Biology 1966
1969-1971 North Dakota State University: M.S. Bacteriology; Biochemistry minor 1971
1971-1973 University of Idaho; Ph.D. Bacteriology 1973
Research Experience:
1966-1969 Research Assistant, Worcester Foundation for Experimental Biology, Shrewsbury, MA
1969-1971 Graduate Research Assistant in Bacteriology, North Dakota State University, Fargo, ND
1971-1973 Graduate Research Assistant in Bacteriology, University of Idaho, Moscow. ID
Other:
Dr. Laurie Drinkwater, Rodale Institute
Research Center, Kutztown, PA
Dr. Vladimir Yakovchenko, Kiev, Ukraine
Mr. Jerry Pearson, Aurelia Corp.
27
1973-1975 Post Doctoral Research Assoc., Soil Science Dept., University of Wisconsin, Madison, Wl
1976-1978 Research Chemist, Biological Waste Managements Nitrogen Lab, USDA-ARS, Beltsville MD
1978-1994 Microbiologist, Soil-Microbial Systems Lab., USDA-ARS, Beltsville, MD
Professional Society Activities:
American Society of Agronomy, Membership Committee, 1984- present
American Society of Microbiology
Sigma Xi
Soil Science Society of America representative to Planning Committee for the Third National Symposium on
Individual and Small Community Sewage Treatment sponsored by American Society of Agricultural
Engineering, held Dec. 14-15, 1981
Served on committee that organized research plot field trips at 1 983 Northeast regional meeting of American
Society of Agronomy
Honors and Awards:
USDA Superior Service Award, 1977 - Team member of the Sewage Sludge Land Utilization Research
Group, "In recognition for outstanding team effort and response to urgent national need for research
information on safe and beneficial uses of sewage e sludge on agricultural land."
USDA Certificate of Merit, 1990 - Recognition in assuming responsibility for maintaining and further
developing the monoclonal antibody technology in SMSL.
USDA Certificate of Appreciation, 1990 - Recognition of committee participation for 1st BARC Orientation
and Poster Day.
USDA Certificate of Appreciation, 1 992 - Service on 1 992 BARC Poster Day planning Committee.
USDA Certificate of Appreciation, 1994 - Serve as Mentor in Partners in Education Program.
Referred Publications Since 1991:
Buyer, J. S., L. J. Sikora and M. G. Kratzke. 1991. Development of a detection system for ferric
pseudobactin using monoclonal antibodies. Plant and Soil 130:243-247.
Mihashi, S, L. J. Sikora, J. S. Buyer, S. Fushiya and S. Mori. 1992 Development and
characterization of a monoclonal antibody to phytosiderophres. Plant Cell Physiol. 33:151-156.
Buyer, J. S., M. G. Kratzke and L. J. Sikora. 1993. A method for detection of pseudobactin, the
siderophore produced by a plant-growth-promoting Pseudomonas strain, in the barley
rhizosphere. Appl. Environ. Microb. 59:677-681.
Sikora, L. J. 1993. Effect of recombinant endophyte containing Bacillus thuringiensis delta
endotoxin on some rhizosphere populations. Microb. Releases 2:109-112.
Buyer, J. S., M. G. Kratzke and L. J. Sikora. 1994. Microbial siderophores and rhizosphere
ecology. IN Biochemistry of Metal Micronutrient in the Rhizosphere. Ed. J. A. Mathey, D. E.
Crowley and D. G. Luster. Lewis Publishers, Boca Raton. LA, pp. 67-80.
Sikora, L. J. and M. I. Azad. 1993. Effect of compost-fertilizer combinations on wheat yields.
1993 Comp. Sci. & Util. 1:93-96.
Sikora., L. J., L. D. Kuykendall, R. S. Dwivedi, E. M. Herman and N. K. Enkiri. 1994.
Characterization of a 14 kDa component with low expression in a unique Nod+Fix Bradyrhizobium
japonicum. Microbiol. 140:000-000.
28
'
Sikora, L. J., V. Yakovchenko, and D. D. Kaufman. 1994. A proposed soil quality indicator. Proc.
The 3rd Wye International Conf. on Sustainaable Agriculuture. Ed. H. Cook and H. Lee.
Sikora, L. J., V. Yakovchenko, and D. D. Kaufman. 1994. Comparison of the rehydration method
for biomass determination to fumigation-incubation and substrate-induced respiration method.
Soil Biol. Biochem. 26:000-000.
I. Donald D. Kaufman, Microbiologist
II. CRIS Project:
Old CRIS Project 1 270-1 2000-009-00D: 1 270-1 2000-009-00D: Analysis of Soil
Microbiological Processes Which Affect Low-Input Sustainable Crop/Livestock System
New CRIS Project 1 270-1 2000-014-00D - Production and Use of Rural/Urban Waste Co-
Compost: Microbial Processes.
Objective: (a) To develop improved methods for assessing, predicting, and monitoring compost
quality based on changes in chemical, physical and microbiological properties; (b) to assess the
effects of specific microbial inoculants during the composting process in facilitating development
of a more consistently uniform compost product, and enhancing compost performance as a soil
biofertilizer; and (c) to determine through field utilization trials the environmental safety and
agronomic/soil quality benefits of composting rural/urban waste materials on croplands, or as a
farm marketable product.
Progress: (a) Methods for assessing compost pH, bulk density, cation exchange capacity, N02,
NH4/NH3 determinations, C:N ratio, 02 and C02 respiration, water holding capacity, phytotoxicity,
and weed seed population were examined on samples of several rural/urban waste mixtures
throughout the 1992-1993 and 1993-1994 composting seasons. These methods, and others
being developed are important for development of in-field assessments of compost quality for
consistent production of high quality composts for specific agricultural production systems; (b)
Experiments have been conducted with specific compost inoculants on specific substrates to
assess their efficacy in degrading such components in composting materials. Development of
more effective inoculants will facilitate more consistent production of high quality composts; (c) A
long term compost utilization trial was initiated in collaboration with Rodale Institute Research
Center Scientists, to assess the impact of rural/urban composts on environmental safety and
agronomic benefits of these materials in rotational cropping systems. Cropping systems selected
include both field and vegetable crops.
Plans: New and improved tests will be developed and applied to characterization of compost
quality. Results from existing and new improved tests will be correlated with results from field
tests where compost utilization occurs. By matching compost production with utilization it will
facilitate development of a more consistent uniform compost product which when used in field will
further enhance compost performance as a soil biofertilizer.
III. Cooperators:
ARS:
Dr. John Doran, Soil and Water Conservation Research Lab., Lincoln, NE
Dr. Robert Dudley, Core Research Applications, Western Regional Res. Ctr., Philadelphia, PA
Dr. David Douds, Plant & Soil Biophysics Research, Eastern Regional Res. Ctr., Philadelphia, PA
Dr. Jeffrey Buyer, Soil-Microbial Systems Lab., BA, Beltsville, MD
Dr. Lawrence Sikora, Soil-Microbial Systems Lab., BA, Beltsville, MD
Dr. Vladimir Yakovchenko, Soil-Microbial Systems Lab., BA, Beltsville, MD
Dr. Marion Simpson, Soil-Microbial Systems Lab., BA, Beltsville, MD
Dr. Chloe Ringer, Soil-Microbial Systems Lab., BA, Beltsville, MD
Dr. Patricia Millner, Soil-Microbial Systems Lab., BA, Beltsville, MD
30
.
Dr. Richard Zobei, LJ.S. Plant, Soil and Nutrition Research Lab., Ithaca, N.Y.
Dr. Jim Power, Soil and Water Conservation Research Lab., Lincoln, NE
Dr. Sam Smith, Water Quality Research Lab., Durant, OK
Dr. Dora! Kemper, National Program Staff, USDA, Beltsville, MD
Dr. Ben Coffman, Weed Science Lab., BA, Beltsville, MD
Other:
Dr. Eileen Perry, Batelle N.W., Richland, Washington
Dr. Michelle Wander, Dept. Agronomy, Univ. Illinois, Urbana, IL
Dr. Laurie Drinkwater, Rodale Institute Research Center, Kutztown, PA
Dr. Carolyn Reider, Rodale Institute Research Center, Kutztown, PA
Dr. Peggy Wagoner, Rodale Institute Research Center, Kutztown, PA
Dr. Steve Peters, Rodale Institute Research Center, Kutztown, PA
Dr. Kim Kroll, Rodale Institute Research Center, Kutztown, PA
IV. Curriculum Vitae:
Education:
1951-52 Kent State Univ. Kent, OH; Biology Department (Biology, Chemistry, B.Sc.)
1955-58 Kent State Univ., Kent, OH; Biology Department (Microbiology); M.Sc.
1958-62 Ohio State Univ., Columbus, OH; Plant Pathology; Ph.D. 1962.
Professional Employment:
1952-1955
1955-1956
1957- 1958
1958- 1962
1962- 1963
1963- 1965
1965-1970
1967-1968
1970-1977
1978-1984
1984-1991
1991-
Laboratory Assistant, Kent State Univ., Kent, OH
Graduate Assistant, Kent State Univ., Kent, OH
Graduate Assistant, Kent State Univ., Kent, OH
Research Assistant, Ohio State Univ., Columbus, OH, and Ohio Agricultural Experiment
Station, Wooster, OH
Research Microbiologist, Pesticide Degradation Laboratory, USDA, ARS, Beltsville, MD
Research Plant Pathologist, Pesticide Degradation Laboratory, USDA, ARS, Beltsville, MD
Research Microbiologist, Pesticide Investigations - Behavior in soils, Crops Research
Division, USDA, ARS, Beltsville, MD
Fuibright-Hays Lecturer in Soil Microbiology, Plant Pathology and Environmental
Microbiology, Khon Kaen Univ., Khon Kaen, Thailand (leave of absence)
Research Soil Microbiologist, Pesticide Degradation Laboratory, USDA, ARS, Beltsville, MD
Research Soil Microbiologist, Pesticide Degradation Laboratory, USDA, ARS, Beltsviile, MD
Research Leader, Soil-Microbial Systems Laboratory, USDA, ARS, Beltsville, MD
Senior Scientist, ARS, Soil-Microbial Systems Laboratory, Rodale Institute Research
Center, Kutztown, PA
Membership in Professionai Societies:
Soil Ecological Society of America
American Society of Microbiology
International Society of Plant Pathology
American Chemical Society: Pesticide Chemistry Division
Sigma Xi
American Chemical Society: Pesticide Chemistry Division
Offices and Committee Assignments held in Professional Societies:
Member of Plant Science Seminar Committee, Piant Industry Station, USDA, Beltsville, MD, 1965-67
Chairman, Pesticide Degradation Laboratory Seminar Committee, 1972-74
Chairman, Pesticide Degradation Laboratory, Manuscript Review Committee, 1972-75
Chairman, Agricultura! Environmental Quality Institute Seminar Committee, 1973-74
Member, Biological Controi Committee, American Phytopathoiogical Society, 1975-78
31
Member, Committee on Chemical Control, International Society for Plant Pathology, 1975-present
Founding and Elected Board Member, Society of Environmental Toxicology & Chemistry (SETAC), 1978-83
Member, Publications Committee of SETAC; Member, Editorial Board of Environmental Toxicology and
Chemistry, SETAC Journal; and Founding Editor, SETAC News, 1980-82
Honors and Awards
Gamma Sigma Delta (Nat. Hon. Soc. Agric.), 1960
Sigma Xi, Member, 1961
Fulbright-Hays Lectureship in Soil Microbiology and Plant Pathology, Khonkaen University, Khon Kaen
Thialand, 1967-68
American Chemical Society, nominated for and awarded full membership under Bylaw I, Section 3(a)(7) for
"significant achievement in chemistry", 1977
Distinguished Alumni Award, The Ohio State University, "for recognition of his significant contributions to soil
microbiology", 1980
Certificate of Merit: for "Outstanding coordination of ARS scientists with researchers and their collaborators
at Rodale Institute Research Center on Sustainable Agriculture projects. $1000, December 1992.
Publications Since 1991:
Reichel, H., H. D. Sislerand D. D. Kaufman. Inducers, substrates, and inhibitors of a propanol-
degrading amidase of Fusarium oxvsporum. Pesticide Biochem. and Physiol. 39:240-250. 1991.
Perry, E. M., Kaufman, D. D., Hart, G. F,, and Payne, B. Spatial analysis of remotely-sensed and
groundbased data to explain location effects in crop yields. 1993 Proceedings ASPRS/ACSM
Annual Convention: Agricultural Remote Sensing Applications. Vol II., New Orleans, LA. pp. 1 1Q-
114. 1993.
Young, A. L., S. A. Tolin, J. N. Rutger, D. D. Kaufman and M. B. Steinbock. 1994. Plant
Biotechnology in China: Trip Report, Sept. 16-30, 1993. Prepared for: Research and Scientific
Exchange Division, Office of International Cooperation and Development, USDA, Washington,
D.C. 34 pp.
Parr, J. F., S. B. Hornick, and D. D. Kaufman. 1994. Use of microbial inoculants and organic
fertilizers in agricultural production. Proc. Inti. Conf. on Use of Microbial and Organic Fertilizers in
Agriculture. June 13-14, Suweon, Korea. In Press.
Kaufman, D. D., R. L. Dudley, C. R. Reider, L. M. Teerlinck, and B. W. Lyman. 1994.
Biodegradation of several starch-based plastic materials in microbial cultures, compost pile, and
field soil. Journal of Biodegradation. Approved for publication August 5, 1994.
Wander, M. M., D. S. Hedrick, D. Kaufman, S. J. Traina, B. R. Stinner, S. Kehermeyer and D. C.
White. 1994. The functional significance of the microbial biomass and biologically-active soil
organic matter in organic and conventionally managed soils. Plant and Soil, Special Issue on Soil
Biodiversity (In Press).
Wander, M. M., D. S. Hedrick, D. Kaufman, S. J. Traina, B. R. Stinner, S. Kehermeyer and D. C.
White. 1994. The functional significance of the microbial biomass and biologically-active soil
organic matter in organic and conventionally managed soils. Soil Ecological Society Special Issue
on Soil Biodiversity (In Press).
32
.
'
I. Laura Lengnick, Research Agronomist
II. CRIS Project:
Old CRIS Project 1270-12000-012-00D: Effect of Residue Decomposition in Low Input
Sustainable Agricultural Systems
New CRIS Project 1270-1 2000-015-00D: Integrated Soil-Nutrient-Crop-Microbial-Pest-Waste
Management Strategies in Sustainable Agriculture
Objective: The field crops research project will study the short- and long-term effects of
sustainable and conventional production practices on biological, environmental, and economic
aspects of field crop production in the Middle Atlantic states. Experimental plots are scheduled to
be established in the spring of 1995 following a two year period of site characterization and
experimental design.
Progress: Based on the geostatistical analysis of an initial soil survey conducted at the start of
the project in May 1993, an intensive soil survey of the site on a 25 m square grid (300 points)
was planned and completed this past spring. The first two of three planned uniformity crops
(corn/winter wheat/soybean) were unsuccessful as a result of severe climatic conditions in the
summer of 1993 and late winter of 1994. No-till corn was planted in May 1994 and conditions
during the growing season have been very favorable for corn production. Data on crop growth
and development have been collected on a 25 m square grid (300 points) throughout the growing
season and biomass and grain yield data will be collected on this grid at crop maturity. Other data
have also been acquired during the characterization period: color infrared aerial photography of
the crop canopy during the 1993 and 1994 growing seasons; a digital elevation model of the site;
and, black and white aerial photography of previous land use on the site during the period of 1930
to 1960. Development of a geographic information systems (GIS) data base to support
comprehensive geostatistical analyses of spatial variation in soil characteristics and crop
productivity is underway. All photography has been scanned into the GIS data base. Soils point
data have been entered into the GIS data base and geostatistical analyses are currently being
conducted on these data. Planning of the specific experimental treatments got underway with the
meeting of a Farmer/Extension planning group in August 1994. This group proposed 9 potential
farming systems to be considered as treatments for the project.
Plans: Additional soil physical and microbiological characterization will be completed by soil
series for N mineralization, soil moisture relations, hydraulic conductivity, infiltration, aggregation,
particulate organic matter, total microbial biomass C and N, nematode diversity and populations,
and microbial biomass diversity (major groups). Additional uniformity crop data will be collected
on a no-till hairy vetch cover crop that will be planted following corn harvest. All data collected at
the site will be compiled in an ARC/INFO GIS data base and analysis of spatial characteristics will
be used to aid in the design of homogeneous blocks across the 16 ha site. A BARC research
planning group will meet in November 1994 to refine the systems proposed by the Farmer/
Extension planning group. Experimental treatments will be finalized in January 1995, following
final review by the Farmer/Extension planning group. In early 1995, a field manager will be hired
to manage the final phase of site preparation and all field operations once the experiment is
underway.
33
.
III. Cooperators:
ARS:
Mr. G. Hart and Mr. W. Dulaney, Remote Sensing Research Laboratory
Dr. J. Teasdale, Weed Science Laboratory
Dr. D. Fravel and Dr. R. Lumsden, Biocontrol of Plant Diseases Laboratory
Dr. Z. Handoo, Nematology Laboratory
Mr. R. Hoover, Farm Branch-West
Other:
Dr. R. Weil, University of Maryland, College Park, MD
Dr. R. Forney, Remington Farms Project, Chestertown, MD
IV. Curriculum Vitae:
Education:
1992 Ph.D. Agronomy, The Pennsylvania State University, University Park
1985 M.S. Soil Science, North Carolina State University, Raleigh
1981 B.S. Agronomy, University of Maryland, College Park
Professional Employment:
1993 -
1992
1988 -1991
1987
1985 -1987
1985
1982 -1984
Project Coordinator, Sustainable Agriculture-Field Crops, USDA-ARS, Beltsville, MD
Congressional Science Fellow, American Society of Agronomy, Washington, DC
Graduate Research Asst., Dept, of Agronomy, Pennsylvania State Univ., University Pk., PA
Statistical Analyst, Dept, of Forestry, Pennsylvania State University, University Park, PA
Agricultural Research Technician I, Crop Science Dept., NC State University, Raleigh, NC
Graduate Teaching Assistant, Soil Science Dept., NC State University, Raleigh, NC
Graduate Research Assistant, Soil Science Dept., NC State University, Raleigh, NC
Professional Activities:
Contributor, Agronomy News, American Society of Agronomy, Madison, Wl, 1992.
Review Panel Member. National Academy of Sciences, 1 992.
Graduate Representative to Faculty, Dept of Agronomy, Pennsylvania State Univ, University Park 1989-90
Contributing Editor, Stewardship News. Carolina Farm Stewardship Assoc., Carrboro, NC 1984 to 1987.
Board Member, Carolina Farm Stewardship Association. 1985, 1986.
NC State Univ Repr, Coordinating Committee, 1st Ann. No. Carolina Alternative Farming Field Days, 1985.
Coach. NC State University Soil Judging Team. 1983.
Refereed Publications Since 1991:
Lengnick, L.L. and R.H. Fox. Simulation by NCSWAP of seasonal nitrogen dynamics in corn: I.
Soil Nitrate. Agronomy Journal 86 (1): 167-175, 1994.
Lengnick, L.L. and R.H. Fox. Simulation by NCSWAP of seasonal nitrogen dynamics in corn: II.
Corn growth and yield. Agronomy Journal 86 (1): 176-181, 1994.
Jabro, J.D., J.M. Jemison,Jr., L.L. Lengnick, R.H. Fox and D.D. Fritton. Field validation and
comparison of LEACHM and NCSWAP models for predicting nitrate leaching. Trans American
Soc Agri Eng 36(6): 1651-1657, 1993.
34
I. Jeffrey S. Buyer, Research Chemist
II. CRIS Project:
Old CRIS Project 1270-12000-01 1-00D: MANAGEMENT OF BENEFICIAL PLANT MICROBE
SOIL INTERACTIONS FOR LOW INPUT SUSTAINABLE AGRICULTURE
New CRIS Project 1270-12000-016-00D: Soil Quality and Soil Ecology in Sustainable
Agriculture
Objective: Determine the occurrence of microbial and plant siderophores in the rhizosphere and
the role of regulating iron in the rhizosphere as a management tool for controlling harmful
microbial populations.
Progress: We have developed immunoassays to pseudobactin, the siderophore produced by
plant growth promoting Pseudomonas B10, and to the plant siderophore mugineic acid, and
measured pseudobactin in the rhizosphere of barley inoculated with B10. We have developed
monoclonal antibodies to chrysobactin, the siderophore produced by plant pathogenic Erwinia
chrysanthemi , and are currently using these antibodies to develop a quantitative assay.
Plans: In order to study the impact of siderophores on microbial populations we are planning to
study the effect of inoculated B10 on microbial diversity. Diversity will be measured by fatty acid
methyl ester (FAME) analysis of isolates. Community-level diversity will be analyzed by
phospholipid analysis or DNA analysis of soil extracts. Isolates will be screened for their ability to
use pseudobactin and identified by FAME analysis. This experiment will allow us to determine if
inoculation with a biocontrol organism affects microbial diversity and if any impact on diversity is
due to siderophore production.
Objective: Evaluate the impact of soil microbial diversity on soil quality and sustainable
agriculture.
Progress: Rhizosphere bacteria, actinomycetes, and fungi were enumerated and isolated from
the Farming Systems Trial, Rodale Institute Research Center. Isolates are being identified by
fatty acid methyl ester analysis. Preliminary results from 1993 indicate that diversity of culturable
bacteria and fungi is lowest in the rhizosphere of plants treated with animal manure as the only
nitrogen input.
Plans: Isolates will be examined for particular biological functions important in sustainable
agriculture, such as cellulose decomposition, phosphate solubilization, and organic nitrogen
mineralization. Community structure will be determined by phospholipid analysis, DNA analysis,
and carbon source utilization patterns. A similar study will be carried out at the Sustainable
Agriculture Field Site at Beltsville in order to follow diversity during the transition from conventional
to low-input methods.
Objective: Determine relationships among selected VAM fungal isolates.
Progress: A method for fatty acid analysis of spores, at 100 times the sensitivity of previous
methods, was developed.
35
.
Plans: Selected VAM fungal isolates will be grown and spores collected for fatty acid analysis.
Numerical taxonomy of fatty acid composition of spores will be used to study VAM taxonomy.
The great sensitivity of our method may allow us to analyze single spores of certain genera, thus
allowing us to study clonal diversity.
III. Cooperators:
ARS:
Dr. L.J. Sikora, Soil Microbial Systems Laboratory, Beltsville, MD
Dr. D.R. Kaufman, Soil Microbial Systems Laboratory, Beltsville, MD
Dr. J.S. Karns, Soil Microbial Systems Laboratory, Beltsville, MD
Dr. P.D. Millner, Soil Microbial Systems Laboratory, Beltsville, MD
Dr. S.J. Wright, Soil Microbial Systems Laboratory, Beltsville, MD
Other:
Dr. M.J. Miller, University of Notre Dame, Notre Dame, IN
Dr. Y. Chen, Hebrew University of Jerusalem, Rehovot, Israel
Dr. Y. Hadar, Hebrew University of Jerusalem, Rehovot, Israel
Mr. Wolfgang Hordt, Universitat Hohenheim, Stuttgart, Germany
Dr. T. Heulin, CNRS, Nancy, France
IV. Curriculum Vitae:
Education and Experience:
1973 to 1977
1977- 1978
1978- 1980
1980-1985
1985-1987
1987-1990
1990-
State University of New York, Stony Brook, NY; Biochemistry Department, B.S. 1977
Research Assistant, New York Ocean Sciences Laboratory, Montauk, N.Y.
Technical Specialist, Marine Sciences Research Center, S.U.N.Y. at Stony Brook, N.Y.
University of California, San Diego, CA; Chemistry Department, Ph.D. 1985. Title of thesis
Siderophores of Plant Growth Promoting and Plant Deleterious Pseudomonads
Visiting scholar, Laboratory of J. Neilands, Department of Biochemistry, U C Berkeley, CA
Research Chemist (Postdoctoral), Soil-Microbial Systems Laboratory, USDA, ARS, .
Research Chemist, Soil Microbial Systems Laboratory, USDA, ARS, Beltsville, MD.
Professional Societies
American Association for the Advancement of Science
American Chemical Society
American Society for Microbiology
Invited Seminars
"Iron and Ecology", University of Maryland, College Park, 1991.
"Siderophores and Rhizosphere Ecology", University of Delaware, Newark, 1991.
"Microbial Siderophores and Rhizosphere Ecology", George Mason University, 1992.
"Microbial Siderophores and Rhizosphere Ecology", American Chemical Society National Meeting, 1992.
"Rhizosphere Siderophores, Microbial Ecology and Plant Uptake of Iron", Amer Soc Agron Meeting, 1 993.
"Siderophore Production by Pseudomonas BIO in the Rhizosphere", Amer Soc Microbiology 1994.
Honors and Awards
Individual National Research Service Award, 1985-87.
36
'
Peer Reviewed Publications since 1991
Buyer, J. S., de Lorenzo, V., and J. B. Neilands (1991) Production of the siderophore aerobactin
by a halophilic pseudomonad. Appl. Environ. Microbiol. 57:2246-2250.
Buyer, J. S., Sikora, L. J., and M. G. Kratzke (1991) Development of a detection system for ferric
pseudobactin using monoclonal antibodies. Plant and Soil 130:243-247. (republished after peer
review from invited book chapter)
Mihashi, S., Sikora, L. J., Buyer, J. S., Fushiya, S., and S. Mori (1992) Development and
characterization of a monoclonal antibody to phytosiderophores. Plant Cell Physiol. 33:151-156.
Buyer, J. S., Kratzke, M. G., and L. J. Sikora (1993) A method for detection of pseudobactin, the
siderophore produced by a plant growth Pseudomonas, in the barley rhizosphere. Appl. Environ.
Microbiol. 59:677-681.
Persmark, M., Pittman, P., Buyer, J. S., Schwyn, B., Gill, P. R., and J. B. Neilands (1993)
Isolation and structure of Rhizobactin 1021, a siderophore from the alfalfa symbiont Rhizobium
meliloti 1021. J. Am. Chem. Soc. 115:3950
Invited Book Chapters
Buyer, J. S., Sikora, L. J., and M. G. Kratzke (1991) Development of a detection system for ferric
pseudobactin using monoclonal antibodies. IN Y. Chen and Y. Hadar (eds), Iron Nutrition and
Interactions in Plants, Kluwer Academic Publishers, The Netherlands, pp. 283-287.
Buyer, J. S. and L. J. Sikora (1991) Rhizosphere interactions and siderophores. IN D. L. Keister
and P. B. Cregan (eds), The rhizosphere and plant growth, Kluwer Academic Publishers, The
Netherlands, pp. 263-269.
Buyer, J. S., Kratzke, M. G., and L. J. Sikora (1994) Microbial siderophores and rhizosphere
ecology. IN D. Crowley, D. Luster, and J. Manthey (eds), The biochemistry of metal
micronutrients in the rhizosphere, Lewis Publishers, Chelsea, Michigan, pp. 67-80.
Reviews
Loper, J. E. and J. S. Buyer (1991) Siderophores in microbial interactions on plant surfaces.
Molecular Plant-Microbe Interactions 4:5-13.
37
'
I. Walter W. Mulbry, Microbiologist
II. CRIS Project:
1270-1 21 30-005-00D: Biochemistry and Molecular Biology of Pesticide Metabolism in Soil
and Water
Objective: To isolate and characterize microorganisms that metabolize agricultural chemicals of
environmental concern and to determine the biochemical and genetic basis of pesticide
metabolism in microorganisms; to isolate and characterize pesticide degradation genes; to
develop microbially based waste disposal technologies that protect the environment and
groundwater, and; to determine the effects of environmental factors on rates of pesticide
metabolism to develop predictive models of in situ metabolism.
Progress: A novel cloning strategy was developed to isolate biodegradation genes from Gram-
positive bacteria using DNA amplification to generate gene specific probes. This strategy has
broad applicability for isolating previously "unclonable" genes from diverse organisms. This
strategy was used to clone, sequence and over-express the new parathion hydrolase gene adpB
from a Nocardia strain in E. coli. More recently, this strategy was used to isolate and characterize
an s-triazine dechlorination gene (trzA) from the organism Rhodococcus corallinus. A
collaborating Canadian scientist has used this gene to construct a recombinant bacterial strain
that is capable of dealkylating and dechlorinating atrazine and simazine.
DNA fragments from the cloned organophosphate degradation gene adpB have been used to
characterize spontaneous mutants of Nocardia that have lost the ability to produce the adpB
hydrolase. The goal of this research is to gain insight into the general mechanisms by which
environmental organisms quickly change their metabolic characteristics.
Plans: Use the s-triazine degradation gene trzA as a gene probe to isolate other s-triazine
degradation genes. Introduce this gene into other organisms having N-dealkylation activities
(such as Streptomyces sp PS1/5) in order to construct microorganisms capable of constituitively
degrading s-triazines in contaminated soils. Use the organophosphate degradation gene acfpB to
monitor the occurrence and spread of this gene in the microbial community associated with
organophosphate contaminated soils. Further characterize and manipulate the genetic elements
responsible for the loss of the adpB gene in Nocardia to construct bacterial suicide vectors.
0500-00026-034-00D: Bioremediation of Contaminated Sites to Protect Water Quality
Objective: To develop safe, effective and economical methods to eliminate potential point
sources of ground and surface water contamination resulting from sites that have been
contaminated by agrochemicals. More specifically, to refine the use of specific microbial isolates
and consortia to bioremediate soils and waters from pesticide contaminated sites.
Progress: Biodegradation studies were conducted using soils from eight cattle dipping waste pits
that were contaminated with high concentrations of the organophosphate insecticide coumaphos.
The results showed that the coumaphos in all of the soils could be biodegraded in soil slurries
using indigenous soil microorganisms.
38
,
Plans: Determine coumaphos degradation rates in moist buffered soils and evaluate whether this
strategy would be practical for the remediation of contaminated soils. Determine the extent and
rate of coumaphos degradation in dip waste applied to trickling gravel filters. Evaluate different
filter matrices with regard to effectiveness and cost. Determine optimal operating parameters and
scale-up factors necessary for demonstration scale experiments using trickling gravel filters.
III. Cooperators:
ARS:
Mr. E.H. Ahrens, Cattle Fever Tick Research Lab, Mission, TX.
Dr. R.B. Davey, Cattle Fever Tick Research Lab, Mission, TX.
Dr. Daniel Shelton, Environmental Chemistry Lab, Beltsville, MD
Dr. Larry Sikora. Soil Microbial Systems Lab, Beltsville, MD
Dr. Patricia Milner, Soil Microbial Systems Lab, Beltsville, MD
Other:
Dr. Ram Behki, Agriculture Canada, Ottawa, Canada
Dr. B.M. Pogell, Center for Agricultural Biotechnology, Univ. of MD, College Park, MD
IV. CURRICULUM VITAE
Education
1975-79 Duke University: Major, Zoology; B.S. 1979.
1980-82 University of Utah; Major, Biology.
1984-87 University of Maryland; Marine, Environmental, and Estuarine SciencesProgram; Ph.D. 1988.
Professional Employment
1989- Microbiologist; USDA/ARS Agricultural Research Center, Natural Resources Institute, Pesticide
Degradation Laboratory, Beltsville, MD. Promoted to GS-13 8/92. Transferred to Soil Microbial
Systems Laboratory 10/93.
1988-89 Research Associate, Dept, of Biology, University of Maryland Baltimore County.
1987-88 Research Affiliate, GS-9; USDA/ARS Agricultural Research Center, Natural Resources Institute,
Pesticide Degradation Laboratory, Beltsville, MD.
Honors and Awards
1 992 Appointed to the Interational Union of Pure and Applied Chemistry (IUPAC) Task Force on Scientific
Aspects of the Destruction of Chemical Warfare Agents.
1 990 Research grant from Ciba Geigy Corporation for project "Isolation and characterization of s-triazine
dechlorination genes from Rhodococcus corallinus."
1 987 Research grant from Maryland Center for Agricultural Biotechnology for project "Genetic Analysis of
Parathion Hydrolase Genes."
Membership in Professional Societies
American Chemical Society
American Society for Microbiology
Air and Waste Management Association
Union of Concerned Scientists
39
,
Refereed Publications since 1991:
Lovett, P. S., Ambulos, N. P., Mulbry, W. W., Noguchi, N., and Rogers, E. J. UGA can be
decoded as tryptophan at low efficiency in Bacillus subtilis. J. Bacteriol. 173:1810-1812. 1991.
Mulbry, W. W. and Kearney, P. C. Degradation of pesticides by micro-organisms and the
potential for genetic manipulation. Crop Protection 10:334-346. 1991.
Mulbry, W. W. and Eaton, R. W. Purification and characterization of the N-methylcarbamate
hydrolase from Pseudomonas strain CRL-OK. App. Environ. Microbiol. 57:3679-3682. 1991.
Mulbry, W. W. The aryldialkylphosphatase-encoding gene adpB from Nocardia sp. strain B-1:
cloning, sequencing and expression in Eschenchia coli. Gene 121:149-153. 1992.
Mulbry, W. W. Purification and characterization of an inducible s-triazine hydrolase from
Rhodococcus corallinus NRRL B-15444R. App. Environ. Microbiol. 59:3533-3538. 1993.
Mulbry, W. W., Del Valle, P., Karns, J. S., and Shelton, D. R. Biodegradation of the
organophosphate insecticide coumaphos in contaminated soils. Proceedings of the 1994 Annual
Meeting of the Air and Waste Management Assocation. In press.
Muldoon, M.T., Nelson, J.O. Pesticide waste treatment monitoring of s-triazines uising
immunoassy. J. Agric. Food Chem. 42:1686-1692. 1994.**
Muldoon, M.T., Nelson, J. O. Evlauation of the effects of selected agricultural materials on an
ELISA for s-triazines. Food Agric. Immunol. 1994. (In press).**
Muldoon, M.T., Nelson, J.O. Immunochemical approach for pesticide waste treatment monitoring
of s-triazines. ACS Sym. Ser. 1994 (In press).**
** Dr. M. T. Muldoon complete his doctoral research in Dr. Mulbry's laboratory in 1993-94.
40
I. Jeffrey S. Karns, Microbiologist
II. CRIS Project:
1270-12130-005-00D: Biochemistry and Molecular Biology of Pesticide Metabolism in Soil
and Water
Objective: To isolate and characterize microorganisms that metabolize agricultural chemicals of
environmental concern and to determine the biochemical and genetic basis of pesticide
metabolism in microorganisms; to isolate and characterize pesticide degradation genes; to
develop microbially based waste disposal technologies that protect the environment and
groundwater, and; to determine the effects of environmental factors on rates of pesticide
metabolism to develop predictive models of in situ metabolism.
Progress: The nucleotide sequence of a unique repeated element associated with s-triazine
degradation genes in Pseudomonas was determined. The sequence revealed that the element
has the structural characteristics of a bacterial insertion element including 27 base-pair inverted
repeats at the ends of the element. Comparison with sequences in DNA databanks revealed that
this element shared some degree of homology with known insertion elements but represents a
heretofore unreported element. The genes encoding the enzymes for the degradation of the s-
triazine compounds ammelide and cyanuric acid were shown to be plasmid encoded in a strain of
Klebsiella pneumoniae that used these compounds as a nitrogen source. Cyanuric
amidohydrolase, the enzyme responsible for cleavage of the s-triazine ring, was purified to
homogeneity and characterized. Barbituric acid was found to be a potent competitive inhibitor if
this enzyme reaction. A total-vat management system for the preservation and disposal of
coumaphos-containing cattle-dips generated by the APHIS-Veterinary Services tick eradication
program in Texas was field tested. It was shown that acidifying the vats with triple
superphosphate fertilizer prevented formation of toxic potasan allowing maximum use of the
coumaphos suspension in the vat. Neutralization of the vat contents and introduction of air
allowed extensive biodegradation of the coumaphos in the spent cattle-dip. A screening study
was conducted which showed that the organism Streptomyces sp. PS1/5 was able to metabolize
a wide spectrum of herbicides in broth culture.
Plans: Continue to work with ARS scientists at the Cattle Fever Tick Research Lab in Mission,
TX on monitoring the performance of the total vat management system in several working cattle¬
dipping vats. Aid them in research into chemical alternatives to coumaphos. To clone the gene
encoding the enzyme biuret amidohydrolase (the product of enzymatic ring-cleavage of s-
triazines) and to characterize the enzyme.
0500-00026-034-00D: Bioremediation of Contaminated Sites to Protect Water Quality
Objective: To develop safe, effective and economical methods to eliminate potential point
sources of ground and surface water contamination resulting from sites that have been
contaminated by agrochemicals. More specifically, to refine the use of specific microbial isolates
and consortia to bioremediate soils and waters from pesticide contaminated sites.
Progress: Collected soils from several coumaphos contaminated waste pits along the border
between Texas and Mexico. Initiated studies on the biodegradation of atrazine in soil by
Streptomyces sp. PS1/5.
41
.
Plans: Develop methods for the use of microorganisms of the genus Streptomyces for the
bioremediation of pesticide contaminated soils through solid-state fermentation. Optimize
parameters for the degradation of atrazine then look at metabolism of other pesticides.
Characterize the products formed from pesticides by Streptomyces and isolate microorganisms to
further degrade those products. Continue to support Dr. Mulbry's research into methods for the
practical biodegradation of coumaphos in contaminated soils.
III. Cooperators:
ARS:
Mr. E.H. Ahrens, Cattle Fever Tick Research Lab, Mission, TX.
Dr. R.B. Davey, Cattle Fever Tick Research Lab, Mission, TX.
Dr. Cathleen Hapeman, Environmental Chemistry Lab, Beltsville, MD
Dr. Daniel Shelton, Environmental Chemistry Lab, Beltsville, MD
Dr. Larry Sikora, Soil Microbial Systems Lab, Beltsville, MD
Dr. Jeffrey Buyer, Soil Microbial Systems Lab, Beltsville, MD
Other:
Dr. B.M. Pogell, Center for Agricultural Biotechnology, Univ. of Maryland, College Park, MD
Dr. Alan Sexstone, Dept, of Soil Microbiology, West Virginia University, Morgantown, WV
IV. CURRICULUM VITAE
Education:
1975 B.S. Medical Technology, The Pennsylvania State University, University Park, PA
1981 Ph.D. Microbiology, Virginia Commonwealth University, Richmond, VA
Professional Employment:
1 981 -1 983 Post-Doctoral Fellow, Dept, of Microbiology and Immunology, University of Illinois at Chicago,
Health Sciences Center, Chicago, IL.
1 983- 1 984 Postdoctoral Research Associate, Dept, of Entomology, Univ. of Maryland, College Park
1984- 1993 Microbiologist, Pesticide Degradation Lab, USDA, ARS, Beltsville, MD.
1993- Microbiologist, Soil Microbial Systems Lab, USDA, ARS, Beltsville, MD.
Membership in Professional Societies:
American Society for Microbiology (ASM)
American Chemical Society (ACS)
American Society for the Advancement of Science (AAAS)
Editorial Board Appointments:
Member of editorial board of Applied and Environmental Microbiology, 1988-1994
Refereed Publications since 1991:
Karns, J.S. and P. H. Tomasek. Carbofuran hydrolase- Purification and properties. J. Ag. Food
Chem. 39:1004-1008. 1991.
42
Eaton, R.W. and J.S. Karns. Cloning and Analysis of s-Triazine Catabolic Genes from
Pseudomonas NRRLB-12227. J. Bacteriol. 173:1215-1222. 1991.
Eaton, R. W. and J. S. Karns. Cloning and Comparison of the DNA Encoding Ammelide
Aminohydrolase and Cyanuric Acid Amidohydrolase from Three s-Triazine-Degrading Bacterial
Strains. J. Bacteriol. 173:1363-1366. 1991.
Smith, J. M., G. F. Payne, J. A. Lumpkin, and J. S. Karns. Enzyme Based Strategy for Toxic
Waste Treatment and Waste Minimization. Biotechnology and Bioengineering 39:741-752. 1992.
Karns, J.S. Biotechnology in Bioremediation of Pesticide Contaminated Sites: Past, Present, and
Future. hr Pesticide Waste Management. Bourke, et al. , (Eds.). ACS Symposium Series 510,
Chapter 12. 1992.
Shelton, D.R., J.S. Karns and C.J. Hapeman-Somich. Biological methods for the disposal of
cattle-dip waste, hr Pesticide Waste Management. Bourke, et al., (Eds.). ACS Symposium
Series 510, Chapter 12. 1992.
Davey, R.B., E.H. Ahrens, J.E. George, and J.S. Karns. Efficacy of freshly mixed coumaphos
suspensions adjusted to various pH levels for treatment of cattle infested with Boophilus
annulatus (Say) (Acari: Ixodidae). Manuscript accepted for publication in Preventative Veterinary
Medicine (8/8/94)
McClung, G., W. A. Dick, and J. S. Karns. Degradation of EPTC by Isolated soil microorganisms.
Manuscript accepted for publication in Jnl. Agric. Food Chem. (Sent to Journal)
Mulbry, W. W., Del Valle, P., Karns, J. S., and Shelton, D. R. Biodegradation of the
organophosphate insecticide coumaphos in contaminated soils. Proceedings of the 1994 Annual
Meeting of the Air and Waste Management Assocation. In press.
Additional work completed by cooperators working on this project as part of M.T. Muldoon's
dissertation research while this program was still affliated with Pesticide Degradation Lab.
Muldoon, M.T., Fries, G.F., Nelson, J.O. Evaluation of ELISA for the multianalyte analysis of s-
triazines in pesticide waste and rinsate. J. Agric. Food Chem. 41:322-328. 1993.
Muldoon, M.T., Huang, R.N. Hapeman, C.J., Fries, G.F., Ma, M., Nelson, J.O. Hapten synthesis
and immunoassay development for the analysis of chlorodiamino-s-triazine in treated pesticide
waste and rinsate. J. Agric. Food Chem. 42:747-755. 1994.
43
I. Sara F. Wright, Soil Scientist
II. CRIS Project:
127O-1200O-O13-0OD: Role of Vesicular Arbuscular Mycorrhizae (VAM) in Low Input
Agricultural Systems
Objective: Determine the effect of soil and crop management practices in model sustainable
agriculture systems on the activity of indigenous mycorrhizal fungi. Utilize fatty acid profiles to
identify fungal species contributing to the hyphal network.
Progress: An immunological test for activity of VAM fungal hyphae has been developed. A
monociona! antibody (MAb) was developed against a glycoprotein present on actively growing,
young hyphae but not on older, melanized, or lysing hyphae. This is the first potentially useful
tool to assess activity of VAM fungal hyphae, the most important VAM fungal contribution to an
agroecosystem.
Fatty acid profiles for identification of VAM fungal isolates are being tested by Jeff Buyer. This
method of identification is still in the preliminary stage of method development. We are currently
supplying Jeff with single and multiple spore samples of different isolates collected from
successive generations of pot cultures.
Plans: The nature of the protein will be further defined and an assay will be developed to utilize
the MAb to quantify the protein in field soils. The assay will be used to quantitatively assess
activity of VAM fungi in model sustainable agriculture systems at Beltsville at sites chosen for
coordination with other soil tests.
Fatty acid profiles or conventional morphology will be used for identification of spores from trap
cultures of different treatments in the sustainable agriculture plots at Beltsville.
Old CRIS Project 1270-1 2000-01 2-00D: Effects of Residue Decomposition in Low Input
Sustainable Agricultural Systems
New CRIS Project 1270-1 2000-01 5-00D: Integrated Soil-Nutrient-Crop-Microbial-Pest-Waste
Management Strategies for Sustainable Agriculture
Objective: Define the effects of isolated inputs to a sustainable system on VAM fungi as
measured by activity of hyphae.
Progress: The recently discovered antibody to measure activity of VAM fungal hyphae is being
used to develop an enzyme-linked immunosorbent assay (ELISA) to quantify the targeted protein
under field conditions.
Plans: Assay activity of VAM hyphae under controlled conditions in greenhouse conditions using
field soils to predict changes due to crops, fertilizer nutrients, and composed waste additions.
44
Old CRIS Project 1270-12000-011-00D: Management of Beneficial Plant Microbe Soil
Interactions for Low Input Sustainable Agriculture
New CRIS Project 1270-12000-010-00D: Soil Quality and Soil Ecology in Sustainable
Agriculture
Objective: To determine the role of VAM fungi in formation of water stable soil aggregates.
Progress: Preliminary tests have indicated that the protein found on active hyphae of VAM fungi
is on the surface of water stable aggregates. This is the first indication that these fungi contribute
a definable proteinaceous compound to aggregates, although there has been speculation on that
VAM fungi contribute compounds to aggregates.
Plans: Cooperative work with Mike Miller at Argonne National Labs will be initiated to further
define the presence of the immunologically reactive protein detected on water stable aggregates.
Time-course studies and comparisons among genera and species of mycorrhizae for their
contribution of the protein to soil aggregates will be made.
Old CRIS Project 1270-12000-010-00D: Fertilizer and Cultural Management Effects on Crop
Nutrient Quality and Bioavailability
New CRIS Project 1270-12220-001-00D: Integrated Soil/Crop Biocontrol Management
System: Sustainable Alternatives to Methyl Bromide
Objective: To determine the usefulness of VAM fungi in biocontrol of pathogens on selected
crops currently produced after methyl bromide fumigation.
Progress: Project Outline is undergoing review.
Plans: Assess the contribution of VAM fungal isolates to biocontrol of pathogens of strawberries
under greenhouse conditions using pasteurized soils. Test superior isolates under field
conditions.
III. Cooperators:
ARS:
Dr. D. D. Douds, Eastern Regional Research Center, Philadelphia, PA
Dr. J. S. Buyer, Soil Microbial Systems Lab., Beltsville, MD
Dr. S. J. Lehotay, Environmental Chemistry Lab., Beltsville, MD
Other:
Dr. Frank Dazzo, Michigan State University, East Lansing, Ml
Dr. R. M. Miller, Argonne National Laboratory, Argonne, IL
Dr. J. B. Morton, West Virginia University, Morgantown, WV
Members of the Mycorrhizae section of the S-226 Regional Project
45
.
IV. Curriculum Vitae:
Education:
1962 B.S. Medical Technology, University of Texas, Austin, TX
1980 Ph.D. Soil Science, Texas A&M University, College Station, TX
Professional Employment:
1981- 1982
1982- 1984
1982-1991
1991-Present
Assistant Professor, West Virginia University, Morgantown, WV
Research Scientist, Cooperative Agreement with West Virginia University,
Appalachian Soil & Water Conservation Research Lab., Beckley, WV
Research Scientist, Appalachian Soil & Water Research Lab., Beckley, WV
Research Scientist, Soil Microbial Systems Lab., Beltsville, MD
Membership in Professional Societies:
American Society for Microbiology
Soil Science Society of America
American Society of Agronomy
Offices and Committee Assignment Held in Professional Societies:
Soil Science Society of America, Program Session Chairperson, 1983
American Society of Agronomy, Fellows Selection Committee, 1985-1986
West Virginia Association of Professional Soil Scientists, President-elect and President, 1985- 1987
American Society for Microbiology, Councilor for the Allegheny Branch, 1987-1988
American Society for Microbiology, Allegheny Branch President-elect and President, 1 988-1 990
American Society for Microbiology, Allegheny Branch Newsletter editor, 1990
Southern Regional Project (S-226) Program chairperson, 1990
International Collection of Vesicular and Arbuscular-Vesicular Mycorrhizal Fungi, Advisory
Committee, 1990-Present
Honors and Awards:
USDA Certificate of Merit, 1989
West Virginia Celebrate Women Award for outstanding achievement in science, 1990
Refereed Publications Since 1991:
Wright, S. F., and S. K. Zeto. 1991. Effects of pH and Al3+ on survival of Rhizobium
leguminosarum bv. trifolii in a simple solution and on nodulation of red clover in acid soils, pp.
603-609, in R. J. Wright, V. C. Baligar, and R. P. Murrman (eds.) Plant-Soil Interactions at Low
pH. Kiuwer Academic Publishers, Boston.
Wright, S. F., 1992. Immunological techniques for detection, identification, and enumeration of
microorganisms in the environment, pp. 45-63. in M. A. Levin, R. J. Seidler, M. Rogul (eds.)
Microbial Ecology Principles, Methods and Applications. McGraw-Hill, Inc., New York.
Wright, S. F. and P. D. Millner. 1993. Dynamic processes of vesicular-arbuscular mycorrhizae: A
mycofrhizosystem within the agroecosystem, pp. 29-59. in J. L. Hatfield and B. A. Stewart (eds)
Advances in Soil Science, Soil Biology: Effects on Soil Quality. Lewis Publishers, Boca Raton,
FL.
46
■
Sylvia, D. M., D. O. Wilson, J. H. Graham, J. J. Maddox, P. Millner, J. B. Morton, H. D. Skipper, S.
F. Wright and A G. Jarstfer. 1993. Evaluation of vesicular-arbuscular mycorrhizal fungi in diverse
plants and soils. Soil Biology & Biochemistry 25:705-713.
Olsen, P., S. Wright, M. Collins, and W. Rice. 1994. Patterns of reactivity between a panel of
monoclonal antibodies and forage Rhizobium strains. Applied and Environmental Microbiology
60:654-661.
Wright, S. F. 1994. Serology and conjugation of antibodies, pp. 593-645. in R. W. Weaver, et al.
(eds.) Methods of Soil Analysis, Part 2. Microbiological and Biochemical Properties. Soil Science
Society of America, Inc., Madison, Wl.
47
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