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ACCESSION NO: 0172393 SUBFILE: CRIS
PROJ NO: NCZ04158 AGENCY: CSREES NC.Z
PROJ TYPE: MCINTIRE-STENNIS PROJ STATUS: EXTENDED
START: 01 SEP 1996 TERM: 30 SEP 2002 FY: 2000

INVESTIGATOR: Yamamoto, Y.

PERFORMING INSTITUTION:
FORESTRY
NORTH CAROLINA STATE UNIV
RALEIGH, NORTH CAROLINA 27695

PLANT IMPROVEMENT FOR WASTEWATER REMEDIATION AND METABOLIC ENGINEERING FOR
NEW PRODUCT PRODUCTION

OBJECTIVES: 1)Identify Paulowina genotypes with superior ability for
wastewater remediation and wood production. Develop silvicultural
management practices for remediation plantations. Continue genetic
engineering work with focus on heavy metal accumulation. 2) Develop
duckweed as ahigh efficiencey, gene expression system for protein
production. Continue development of gene transfer methods, enhanced gene
expression, and protein subcelluar localization. Develop production methods
for duckweed, field test species and genotypes for remediation abilities.

APPROACH: Paulowina. Conduct experiments on Agrobacterium rhizogenes
mediated transformation and production of hairy rooted plants. Continue
assessment of remediation field plantings for growth, wood production, and
analysis of tissue samples for wastewater contaminant uptake. Duckweed.
Continue experiments on Agrobacterium mediated transformation. Isolation of
DNA sequences that regulate gene expression levels and subcellular
localization of foreign gene-encoded proteins. Growth paramters controlling
duckweed production in air life columns and fermentors will be determined.
Determine yields and purification methods for foreign gene-encoded
proteins. Gene transfer of the Neurospora metallothionein gene into
duckweed and the subsequent sensitivity to and accumulation of heavy metal
ions will be undertaken.

PROGRESS: 2000/01 TO 2000/12
Genetic transformation of duckweed has been improved. Protocols with both
Agrobacterium-mediated transformation (publication accepted) and
microprojectile bombardment have been developed. Growth enhancement of wild
type and transgenic duckweed expressing a bacterial cellulase was achieved
using high carbon dioxide in collaboration with Dr. Tisserat at USDA-NCAUR.
No detrimental effects on the cellulase activity were detected (in
collaboration with Dr. Freer at USDA-NCAUR). Several duckweed geographic
isolates from southeastern US have been selected for growth in swine
wastewater. Genetic improvement and field testing of Paulownia elongata
clones for waste remediation continued during this reporting period.

IMPACT: 2000/01 TO 2000/12
Growth enhancement of transgenic duckweed using carbon dioxide is
significant because carbon dioxide appears to eliminate needs for a carbon
source in culture. Southeastern duckweed genotypes are adapted to the
regional climate and are expected to perform better in the field. These
isolates may be more desirable than introducing exotic genotypes from the
ecological standpoint.

PUBLICATIONS: 2000/01 TO 2000/12
1. Bergmann, B.A., Cheng, J., Classen, J. and Stomp, A.-M. (2000) In vitro
selection of duckweed geographical isolates for potential use in swine
lagoon effluent renovation. Bioresource Technology 73: 13-20
2. Bergmann, B.A., Cheng, J., Classen, J. and Stomp, A.-M. (2000) Nutrient
removal from swine lagoon effluent by duckweed. Transactions of the ASAE
43: 263-269
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