title: Production of high levels of poly-3-hydroxybutyrate in plastids of Camelina sativa seeds.
abstract: Poly-3-hydroxybutyrate (PHB) production in plastids of Camelina sativa seeds was investigated by comparing levels of polymer produced upon transformation of plants with five different binary vectors containing combinations of five seed-specific promoters for expression of transgenes. Genes encoding PHB biosynthetic enzymes were modified at the N-terminus to encode a plastid targeting signal. PHB levels of up to 15% of the mature seed weight were measured in single sacrificed T1 seeds with a genetic construct containing the oleosin and glycinin promoters. A more detailed analysis of the PHB production potential of two of the best performing binary vectors in a Camelina line bred for larger seed size yielded lines containing up to 15% polymer in mature T2 seeds. Transmission electron microscopy showed the presence of distinct granules of PHB in the seeds. PHB production had varying effects on germination, emergence and survival of seedlings. Once true leaves formed, plants grew normally and were able to set seeds. PHB synthesis lowered the total oil but not the protein content of engineered seeds. A change in the oil fatty acid profile was also observed. High molecular weight polymer was produced with weight-averaged molecular weights varying between 600 000 and 1 500 000, depending on the line. Select lines were advanced to later generations yielding a line with 13.7% PHB in T4 seeds. The levels of polymer produced in this study are the highest reported to date in a seed and are an important step forward for commercializing an oilseed-based platform for PHB production.
authors.affiliation: Metabolix Oilseeds Inc, Saskatoon, SK, Canada.
authors.forename: Meghna R
authors.initials: MR
authors.lastname: Malik
authors.affiliation: Donald Danforth Plant Science Center, Saint Louis, MO, USA.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Yang
authors.affiliation: Metabolix Inc, Cambridge, MA, USA.
authors.forename: Nii
authors.initials: N
authors.lastname: Patterson
authors.affiliation: Metabolix Inc, Cambridge, MA, USA.
authors.forename: Jihong
authors.initials: J
authors.lastname: Tang
authors.affiliation: Donald Danforth Plant Science Center, Saint Louis, MO, USA.
authors.forename: Rachel L
authors.initials: RL
authors.lastname: Wellinghoff
authors.affiliation: Donald Danforth Plant Science Center, Saint Louis, MO, USA.|Department of Biological Sciences, Webster University, Saint Louis, MO, USA.
authors.forename: Mary L
authors.initials: ML
authors.lastname: Preuss
authors.affiliation: Metabolix Oilseeds Inc, Saskatoon, SK, Canada.
authors.forename: Claire
authors.initials: C
authors.lastname: Burkitt
authors.affiliation: Metabolix Oilseeds Inc, Saskatoon, SK, Canada.
authors.forename: Nirmala
authors.initials: N
authors.lastname: Sharma
authors.affiliation: Metabolix Oilseeds Inc, Saskatoon, SK, Canada.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ji
authors.affiliation: Donald Danforth Plant Science Center, Saint Louis, MO, USA.|Department of Biology, Washington University, Saint Louis, MO, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Jez
authors.affiliation: Metabolix Inc, Cambridge, MA, USA.
authors.forename: Oliver P
authors.initials: OP
authors.lastname: Peoples
authors.affiliation: Donald Danforth Plant Science Center, Saint Louis, MO, USA.
authors.forename: Jan G
authors.initials: JG
authors.lastname: Jaworski
authors.affiliation: Donald Danforth Plant Science Center, Saint Louis, MO, USA.|Center for Plant Science Innovation and Department of Biochemistry, University of Nebraska, Lincoln, NE, USA.
authors.forename: Edgar B
authors.initials: EB
authors.lastname: Cahoon
authors.affiliation: Metabolix Oilseeds Inc, Saskatoon, SK, Canada.|Metabolix Inc, Cambridge, MA, USA.
authors.forename: Kristi D
authors.initials: KD
authors.lastname: Snell
mesh_terms: D019607:Brassicaceae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D018525:Germination; D006885:Hydroxybutyrates / Q000737:chemistry / Q000378:metabolism*; D009928:Organ Specificity; D018515:Plant Leaves / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D018087:Plastids / Q000378:metabolism; D011091:Polyesters / Q000737:chemistry / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D036226:Seedlings / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes; D000094122:Polyhydroxybutyrates
chemical_list: D005227:Fatty Acids; D006885:Hydroxybutyrates; D011091:Polyesters; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates
keywords: Camelina sativa; bioplastics; oilseeds; poly-3-hydroxybutyrate; polyhydroxyalkanoate; polyhydroxybutyrate