Metabolix, Inc.

Yield10 Bioscience, Inc.

Massachusetts Institute of Technology

Purdue Research Foundation

Nuseed Nutritional US Inc.

Kristi D Snell

Oliver P Peoples

Jihong Tang

Meghna Malik

Frank Anthony Skraly

Nii Patterson

Venkatesh Bollina

Lauralynn Kourtz

Karen Bohmert-Tatarev

Nirmala Sharma

Anthony J Sinskey

John Wesley Frost

Maurice M Moloney

ChoKyun Rha

Thomas Martin Ramseier

Sang Jun Sim

Karen M Frost

Mariya Somleva

Jixiang Han

Madana M R Ambavaram

Zhigang Zhang

Kieran Ryan

Julie Beaulieu

Bradley Garrison

Stephanie Aquin

Scott A Hogan

Claire Burkitt

Susan McAvoy

Venkata Tavva

Yuanyuan Ji

Andrew Hertig

Matthew Martino

A transgenic land plant that expresses a polyhydroxy alkanoate synthase seed specifically, with cytosolic localization is disclosed. The plant includes a nucleic acid encoding the polyhydroxy alkanoate synthase and a seed-specific promoter operably linked to the nucleic acid. The seed-specific promoter drives expression of the polyhydroxy alkanoate synthase in cytosol of cells of seeds of the plant. The polyhydroxy alkanoate synthase includes a catalytic domain. The polyhydroxy alkanoate synthase does not include any sequence positioned to mediate translocation of the catalytic domain across any membrane of the cells, thereby resulting in the polyhydroxy alkanoate synthase being expressed seed specifically, with cytosolic localization.

Transgenic land plants that express a polyhydroxyalkanoate synthase seed specifically with cytosolic localization

A modified plant is provided. The modified plant includes a polynucleotide including a promoter operably linked to a coding sequence. The coding sequence encodes a transcription factor that has at least 30% sequence identity to one or more of SEQ ID NOs: 27, 204, 10, 19, 1-9, 11-18, 20-26, 28-203, 205, 237, or 242-310. The promoter is non-cognate with respect to the coding sequence. Also provided is a method for producing a modified plant having increased seed yield and/or oil content in comparison to a reference plant by transformation. Also provided is a method for producing a modified plant having increased seed yield and/or oil content in comparison to a reference plant by editing.

Modified plants comprising a polynucleotide comprising a non-cognate promoter operably linked to a coding sequence that encodes a transcription factor

A genetically engineered land plant that expresses a protein that has homology to a plant invertase inhibitor and/or a pectin methylesterase inhibitor and that increases seed yield with increased expression ('an ISY protein') is disclosed. The plant comprises a modified gene for the ISY protein. The ISY protein comprises one or more of an ISY protein ofcomprising SEQ ID NO: 2 or a fragment,homolog, or ortholog thereof. The modified gene comprises a promoter and a nucleic acid sequence encoding the ISY protein. The promoter is non-cognate with respect to the nucleic acid sequence encoding the ISY protein. The modified gene is configured such that transcription of the nucleic acid sequence is initiated from the promoter and results in expression of the ISY protein. A genetically engineered land plant that expresses an RNA that increases seed yield with increased expression also is disclosed.

Genetically engineered land plants that express an increased seed yield protein and/or an increased seed yield RNA

Transgenic plants having enhanced yield and having enhanced seed yield are disclosed. The transgenic plants are transformed with a transgenic polynucleotide encoding one or more metabolic enzymes. The metabolic enzymes can be from any biological source. The transgenic polynucleotide(s) comprises a nucleic acid sequences encoding the metabolic enzymes under the control of functional plant promoters, the one or more metabolic enzymes are targeted to the plastids by the addition of plastid targeting signals. Optionally the functional plant promoters are seed specific promoters and the metabolic enzymes are targeted to the plastids by the addition of plastid targeting peptide heterologous to the metabolic enzymes. Methods of making the transgenic plants and transgenic polynucleotides are disclosed. The magnitude of the increases in seed yield achieved with these transgenic plants are unprecedented.

Plants with enhanced yield and methods of construction

A genetically engineered land plant that expresses a plant CCP1-like mitochondrial transporter protein is provided. The genetically engineered land plant comprises a modified gene for the plant CCP1-like mitochondrial transporter protein. The plant CCP1-like mitochondrial transporter protein is an ortholog of CCP1 ofof SEQ ID NO: 1 derived from a source land plant. The plant CCP1-like mitochondrial transporter protein is localized to mitochondria of the genetically engineered land plant based on a mitochondrial targeting signal intrinsic to the plant CCP1-like mitochondrial transporter protein. The modified gene comprises (i) a promoter and (ii) a nucleic acid sequence encoding the plant CCP1-like mitochondrial transporter protein. The promoter is non-cognate with respect to the nucleic acid sequence. The modified gene is configured such that transcription of the nucleic acid sequence is initiated from the promoter and results in expression of the plant CCP1-like mitochondrial transporter protein.

Genetically engineered land plants that express a plant CCP1-like mitochondrial transporter protein

Transgenic plants having enhanced yield and having enhanced seed yield are disclosed. The transgenic plants are transformed with a transgenic polynucleotide encoding one or more metabolic enzymes. The metabolic enzymes can be from any biological source. The transgenic polynucleotide(s) comprises a nucleic acid sequences encoding the metabolic enzymes under the control of functional plant promoters, the one or more metabolic enzymes are targeted to the plastids by the addition of plastid targeting signals. Optionally the functional plant promoters are seed specific promoters and the metabolic enzymes are targeted to the plastids by the addition of plastid targeting peptide heterologous to the metabolic enzymes. Methods of making the transgenic plants and transgenic polynucleotides are disclosed. The magnitude of the increases in seed yield achieved with these transgenic plants are simply unprecedented.

Transgenic plants, plant material, plant cells, and genetic constructs for synthesis of biopolymers, for example polyhydroxyalkanoates ('PHA') are provided. In one embodiment, the transgenic plants synthesize polyhydroxybutyrate ('PHB'). In one embodiment the transgenic plant encodes siRNA for one or more of the genes encoding enzymes for producing PHA. In a more preferred embodiment, the siRNA expression is under the control of an inducible regulatory element. In another embodiment, the transgenic plant contains transgenes that encode expression enzymes that will degrade the polymer. In a preferred embodiment, the expression of these enzymes is under the control of a germination specific, inducible, or minimal promoter. In another embodiment, the transgenic plant contains transgenes encoding enzymes that increase carbon flow for polymer synthesis. In a preferred embodiment, these transgenes encode enzymes that increase carbon flow in the Calvin Cycle.

Generation of high polyhydroxybutyrate producing oilseeds

Transgenic plants that produce high levels of polyhydroxybutyrate and methods of producing them are provided. In a preferred embodiment the transgenic plants are produced using plastid transformation technologies and utilize genes which are codon optimized. Stably transformed plants able to produce greater than 10% dwt PHS in tissues are also provided.

Stable, fertile, high polyhydroxyalkanoate producing plants and methods of producing them

Methods and constructs for the introduction of multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway are provided. In one embodiment, the constructs contain two or more enzyme-encoding genes, each under the control of an inducible promoter and each with a polyadenylation signal. The constructs are used to produce transgenic plants, in which the expression of the enzymes are increased when a chemical inducing agent is applied, and a biosynthetic product of the series of enzymes encoded by the transgenes is produced. Constructs may be used which contain two or more enzyme-encoding genes under the control of one or more promoters activated by activator molecules or complexes expressed from a transgene or transgenes, which are themselves under the control of one or more inducible promoters and switched on following the external application of a chemical. The transgene or transgenes expressing the activator molecules or complexes may be included in the same construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. Alternatively, the transgene or transgenes expressing the activator molecules or complexes may be on a different construct from the construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. The activator molecule can be expressed using a constitutive promoter in an inactive form which is converted to the active form following application of the chemical inducing agent.

Chemically Inducible Expression of Biosynthetic Pathways

Transgenic plants, plant material, and plant cells for synthesis of polyhydroxyalkanoates, preferably poly(3-hydroxybutyrate) (also referred to a as PHB) are provided. Preferred plants that can be genetically engineered to produce PHB include plants that do not normally produce storage products such as oils and carbohydrates, and plants that have a CNAD-malic enzyme photosynthetic pathway. Such plants also advantageously produce lignocellulosic biomass that can be converted into biofuels. An exemplary plant that can be genetically engineered to produce PHB and produce lignocellulosic biomass is switchgrass,L. A preferred cultivar of switchgrass is Alamo. Other suitable cultivars of switchgrass include but are not limited to Blackwell, Kanlow, Nebraska 28, Pathfinder, Cave-in-Rock, Shelter and Trailblazer.

Production of polyhydroxybutyrate in switchgrass

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5' promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all nonessential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. A single 3&#x2032; termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and/or value added products, illustrated by the production of polyhydroxyalkanoates in plants.

Multi-gene expression constructs containing modified inteins

Methods for producing polyhydroxyalkanoates (PHAs) from fatty acid biosynthetic pathways using a 3-hydroxy acyl ACP thioesterase, a PHA synthase, and an acyl CoA synthetase, have been developed. Methodology for enabling PHA production from fatty acid biosynthetic pathways in non-native bacterial PHA producers and plants using an enzyme having the catalytic activity of 3-hydroxy acyl ACP thioesterase, an acyl CoA synthetase with substrate specificity for medium chain length 3-hydroxy fatty acids, and a medium chain length PHA synthase, has been developed. Acyl CoA synthetase activity can be supplied either by the endogenous acyl CoA synthetase of the host organism, when sufficiently expressed, or the host organism's activity can be supplemented by the expression of a recombinant acyl CoA synthetase gene. New strategies are described for plant based PHA production in the chloroplasts, cytosol, and peroxisomes of biomass crops as well as the plastids, cytosol, and peroxisomes of oil seed crops.

Production of medium chain length polyhydroxyalkanoates from fatty acid biosynthetic pathways

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5' promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5&#x2032;-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3&#x2032; termination sequence, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits and/or value added products.

Chemically inducible expression of biosynthetic pathways

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5' promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5&#x2032;-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3&#x2032; termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and/or value added products, illustrated by the production of polyhydroxyalkanoates in plants.

Growing community of inventors

Belmont, MA, United States of America

Kristi D Snell