US 2011 0145948A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0145948 A1 McKersie et al. (43) Pub. Date: Jun. 16, 2011
(54) TRANSGENIC PLANTS COMPRISING AS Related U.S. Application Data TRANSGENE A PHOSPHATDATE (60) Provisional application No. 61/090,308, filed on Aug. CYTDYLYLTRANSFERASE 20, 2008, provisional application No. 61/090,625, filed on Aug. 21, 2008, provisional application No. (75) Inventors: Bryan McKersie, Raleigh, NC 61/090,669, filed on Aug. 21, 2008. (US); Wesley Bruce, Raleigh, NC (US) Publication Classification (51) Int. Cl. (73) Assignee: BASG Plant Science GmbH, CI2N 5/82 (2006.01) Ludwigshafen (DE) AOIH 5/00 (2006.01) C7H 2L/00 (2006.01) (21) Appl. No.: 13/058,865 (52) U.S. Cl...... 800/287: 800/278; 800/298; 536/23.2 (57) ABSTRACT (22) PCT Fled: Aug. 10, 2009 Polynucleotides are disclosed which are capable of enhanc ing yield of a plant transformed to contain such polynucle (86) PCT NO.: PCT/EP2009/060332 otides. Also provided are methods of using Such polynucle otides and transgenic plants and agricultural products, S371 (c)(1), including seeds, containing such polynucleotides as trans (2), (4) Date: Feb. 14, 2011 genes.
Patent Application Publication Jun. 16, 2011 Sheet 2 of 15 US 2011/O145948A1
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Figure 3
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Patent Application Publication Jun. 16, 2011 Sheet 5 of 15 US 2011/O145948A1
Figure 5
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Patent Application Publication Jun. 16, 2011 Sheet 9 of 15 US 2011/O145948A1
Figure 9
Riboflavin biosynthesis in wild-type plants
PLASTD
PLASTD PENTOSE PHOSPHATE PATHWAY G6P -> --> --> --> R5P V DBP
MEMBRANE TRANSPORT PROTEIN RIBOFLAVIN
AAPP DRPP N / DAPP A GTP TSL PENTOSE PHOSPHATEPATHWAY ENVELOPE MEMBRANE G6P --> --> --> --> R5P PLASMALEMMA MEMBRANE Patent Application Publication Jun. 16, 2011 Sheet 10 of 15 US 2011/O145948A1
Figure 10A
A - Riboflavin biosynthesis in transgenic plants
PLASTID
PLASTID PENTOSE PHOSPHATEPATHWAY G6P -> --> --> --> R5P
DBP
WII DR X RIBOFLAVIN
CYTOSOLIC i PENTOSEPHOSPHATE PATHWAY ENVELOPE MEMBRANE G6P -> --> --> --> R5P PLASMALEMMA MEMBRANE Patent Application Publication Jun. 16, 2011 Sheet 11 of 15 US 2011/O145948A1
Figure 10B B - Riboflavin biosynthesis in transgenic plants
PLASTI
PTASTIT PENTOSE PHOSPHATE
EMBRANE RIBOFLAVIN RANSPORT RIBOFLAVIN
AAPP DRPP
CYT(SLIC PENTOSE PHOSPHATE ENVELOPE MEMBRANE G6P -> --> --> --> R5P PLASMALEMMA MEMBRANE Patent Application Publication Jun. 16, 2011 Sheet 12 of 15 US 2011/O145948A1
Figure 10C C-Riboflavin biosynthesis in transgenic plants
PLASTD
PLASTI IENTOSEPH) SHATE. PATHWAY G6P -> --> --> --> R5P V. DBP
PENTOSEPHOSPEHTEPATHWAYCYTOSOLIC i ENVELOPE MEMBRANE G6P -> --> --> --> R5P PLASMALEMMA MEMBRANE Patent Application Publication Jun. 16, 2011 Sheet 13 of 15 US 2011/O145948A1
Figure 11
Vitamin B6 biosynthesis
R5P G3PIDAP 4PHT D5P
Poly 1 PCXA
POx2 PCXJ
PLP OxH PNP PMP
OxY POXK POXK
PL PM PN Patent Application Publication Jun. 16, 2011 Sheet 14 of 15 US 2011/O145948A1
Figure 12
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TRANSGENC PLANTS COMPRISING AS synthesis. Since crop yield is dependent on the fixation of TRANSGENE A PHOSPHATDATE carbon dioxide in photosynthesis, water uptake and transpi CYTDYLYLTRANSFERASE ration are contributing factors to crop yield. Plants which are able to use less water to fix the same amount of carbon dioxide or which are able to function normally at a lower water 0001. This application claims the priority benefit of U.S. potential have the potential to conduct more photosynthesis provisional patent application Ser. No. 61/090.308, filed Aug. and thereby to produce more biomass and economic yield in 20, 2008; U.S. provisional patent application Ser. No. 61/090, many agricultural systems. 625, filed Aug. 21, 2008; and U.S. provisional patent appli 0006 Agricultural biotechnologists have used assays in cation Ser. No. 61/090,669, filed Aug. 21, 2008. The entire model plant systems, greenhouse studies of crop plants, and contents of each of the above-referenced applications is field trials in their efforts to develop transgenic plants that incorporated herein by reference. exhibit increased yield, either through increases in abiotic stress tolerance or through increased biomass. For example, FIELD OF THE INVENTION water use efficiency (WUE), is a parameter often correlated with drought tolerance. Studies of a plant's response to des 0002 This invention relates generally to transgenic plants iccation, osmotic shock, and temperature extremes are also which overexpress isolated polynucleotides that encode employed to determine the plants tolerance or resistance to polypeptides active in lipid metabolism, in specific plant tis abiotic stresses. Sues and organelles, thereby improving yield of said plants. 0007 An increase in biomass at low water availability may be due to relatively improved efficiency of growth or reduced BACKGROUND OF THE INVENTION water consumption. In selecting traits for improving crops, a 0003 Population increases and climate change have decrease in water use, without a change in growth would have brought the possibility of global food, feed, and fuel short particular merit in an irrigated agricultural system where the ages into sharp focus in recent years. Agriculture consumes water input costs were high. An increase in growth without a 70% of water used by people, at a time when rainfall in many corresponding jump in water use would have applicability to parts of the world is declining. In addition, as land use shifts all agricultural systems. In many agricultural systems where from farms to cities and suburbs, fewer hectares of arable land water Supply is not limiting, an increase in growth, even if it are available to grow agricultural crops. Agricultural biotech came at the expense of an increase in water use also increases nology has attempted to meet humanity's growing needs yield. through genetic modifications of plants that could increase 0008. Agricultural biotechnologists also use measure crop yield, for example, by conferring better tolerance to ments of other parameters that indicate the potential impact of abiotic stress responses or by increasing biomass. a transgene on crop yield. For forage crops like alfalfa, silage 0004 Crop yield is defined herein as the number of bush corn, and hay, the plant biomass correlates with the total els of relevant agricultural product (such as grain, forage, or yield. For grain crops, however, other parameters have been seed) harvested per acre. Crop yield is impacted by abiotic used to estimate yield, such as plant size, as measured by total stresses, such as drought, heat, Salinity, and cold stress, and by plant dry weight, above-ground dry weight, above-ground the size (biomass) of the plant. Traditional plant breeding fresh weight, leaf area, stem Volume, plant height, rosette strategies are relatively slow and have in general not been diameter, leaf length, root length, root mass, tiller number, Successful in conferring increased tolerance to abiotic and leaf number. Plant size at an early developmental stage stresses. Grainyield improvements by conventional breeding will typically correlate with plant size later in development. A have nearly reached a plateau in maize. The harvest index, larger plant with a greater leafarea can typically absorb more i.e., the ratio of yield biomass to the total cumulative biomass light and carbon dioxide than a smaller plant and therefore at harvest, in maize has remained essentially unchanged dur will likely gain a greaterweight during the same period. There ing selective breeding for grain yield over the last hundred is a strong genetic component to plant size and growth rate, years. Accordingly, recent yield improvements that have and so for a range of diverse genotypes plant size under one occurred in maize are the result of the increased total biomass environmental condition is likely to correlate with size under production per unit land area. This increased total biomass another. In this way a standard environment is used to has been achieved by increasing planting density, which has approximate the diverse and dynamic environments encoun led to adaptive phenotypic alterations, such as a reduction in tered at different locations and times by crops in the field leaf angle, which may reduce shading of lower leaves, and 0009 Harvest index is relatively stable under many envi tassel size, which may increase harvest index. ronmental conditions, and so a robust correlation between 0005. When soil water is depleted or if water is not avail plant size and grain yield is possible. Plant size and grain able during periods of drought, crop yields are restricted. yield are intrinsically linked, because the majority of grain Plant water deficit develops if transpiration from leaves biomass is dependent on current or stored photosynthetic exceeds the supply of water from the roots. The available productivity by the leaves and stem of the plant. As with water supply is related to the amount of water held in the soil abiotic stress tolerance, measurements of plant size in early and the ability of the plant to reach that water with its root development, under standardized conditions in a growth system. Transpiration of water from leaves is linked to the chamber or greenhouse, are standard practices to measure fixation of carbon dioxide by photosynthesis through the potential yield advantages conferred by the presence of a stomata. The two processes are positively correlated so that transgene. high carbon dioxide influx through photosynthesis is closely 0010 Plant membranes contain diverse molecular species linked to water loss by transpiration. As water transpires from and the composition of these membranes change in response the leaf, leaf water potential is reduced and the stomata tend to environmental cues during acclimation processes and as a to close in a hydraulic process limiting the amount of photo consequence of cellular injury from environmental stress. US 2011/O 145948 A1 Jun. 16, 2011
Plant membranes are generally considered to be a primary conditions. Biosynthesis of riboflavin requires GTP and ribu site of injury following exposure to low temperature stress lose-5-phosphate (R5P) as precursors. The microbial ribofla and various forms of oxidative stress such as occur during vin biosynthesis genes RibA-RibF have been cloned and bio water deprivation. This degradation may involve the action of chemically characterized. specific hydrolytic enzymes or may be the consequence of (0015 Homologs of RibA, RibB and RibE have been oxidative reactions mediated by free radicals. In the case of cloned from plants, and based on sequence analysis, the Sub oxidative free radical reactions, degradation may occurat the cellular localization of the plant proteins has been deduced. unsaturated double bonds of the fatty acid acyl chain or at the Proteins that function in the plastid have a typical amino acid ester bond linking the fatty acid acyl chain to the glycerol domain at the N terminus of the protein that acts as a targeting backbone of the phospholipid. This degradation if sufficiently sequence to direct the protein into the plastid. The plant severe will promote cell death, but in more moderate circum enzymes involved in riboflavin synthesis contain this plastid stances, degradation products are components of cell signal targeting sequence, whereas those that encode proteins ing mechanisms that promote an acclimation response. Dur involved in the biosynthesis of FAD from riboflavin do not. ing the acclimation process, plants adapt and develop greater Therefore, the synthesis of the coenzymes FMN and FAD in tolerance of environmental stress. Coincidentally, plants gen plants is believed to occur sequentially in the plastid and the erally alter both the quantity of membrane components in cytosol compartments. In wild type plants, the conversion of each cell and the composition of those membranes. These GTP to 5-amino-6-ribitylamino-2,4(1H.3H)-pyrimidinedi alterations in composition are coincident with increased tol one (ARP) occurs by several enzymatic reactions that are erance of the whole plant. Common changes include changes localized in plastids. The plastid compartment contains a in fatty acid unsaturation and phospholipid head groups. In pentose phosphate pathway which forms R5P. Within the addition, genetic differences among plants contribute to both plastid, R5P is metabolized to 3,4-dihydroxy-2-butanone differences in membrane composition, stress signaling 4-phosphate (DBP) which is combined by the enzymatic mechanisms and the whole plant's ability to tolerate stress. action of 6,7-dimethyl-8-ribityllumazine synthase (RibH) to Thus plant membranes are considered to be a central site for form 6,7-dimethyl-8-ribityllumazine (DR). DR is converted perceiving, tolerating and responding to environmental to riboflavin by the enzyme riboflavin synthase in the plastid StreSS. and then transported to the cytosol via an unknown mecha 0011 Phospholipids are the major structural components nism. In the cytosol, riboflavin is phosphorylated to FMN and of biological membranes and also serve as important-signal converted to FAD. FIG. 9 depicts the compartmentalized ing molecules. Phospholipids are commonly synthesized in riboflavin and FAD biosynthetic pathway in wild type plants. the endoplasmic reticulum and transported to other mem (0016 Enzyme I of FIG. 9 is GTP cyclohydrolase II (EC branes, but phospholipids can be synthesized in other cell 3.4.5.25), or RibA, which catalyzes the first step in riboflavin compartments, including the mitochondria and chloroplast. synthesis. RibA is sometimes found as a bifunctional enzyme Phospholipases hydrolyze phospholipids, and in plants, three with 3,4-dihydroxy-2-butanone 4-phosphate synthase classes of phospholipases have been reported including phos (DHBP synthase) activity in addition to GTP cyclohydrolase pholipase A (PLA), phospholipase C (PLC), and phospholi II activity. pase D (PLD). PLCs hydrolyze phosphotidylinositol 4.5- (0017 Enzyme VIII of FIG.9 is RibH (also known as RibE, bisphosphate (PIP2) to inositol 1,4,5-triphosphate and riboflavin synthase subunit beta, and lumazine synthase). diacylglycerol, which are components of the inositol signal U.S. Pat. Nos. 6,146,866 and 6.323,013 disclose the cloning ing pathway. of lumazine synthase from spinach, tobacco, Arabidopsis, 0012. As set forth above, plants can acclimate to environ and Magnaporthe grisea. The chloroplast targeting mental stress through moderate increases in the activity of sequences of the spinach, tobacco, and Arabidopsis RibH enzymes that alter fatty acid oxidation. Most eukaryotic cells polypeptides are identified in U.S. Pat. Nos. 6,146,866 and have two fatty-acid beta-Oxidation systems, one in mitochon 6,323,013. dria and the other in peroxisomes. The Escherichia coli gene 0018. A second class of cofactors is the vitamin group B2341 encodes a bifunctional anaerobic fatty acid oxidation known as vitamin B6. Three compounds belong to the vita complex protein associated with both enoyl-CoA hydratase min group: pyridoxal, pyridoxine, and pyridoxamine, all of and 3-hydroxyacyl-CoA epimerase activity. WO 2006/ which are widely distributed in animals and plants, especially 069610 and WO 2007/087815 disclose metabolic changes in in cereal grains. Pyridoxal and pyridoxamine also occur in plants transformed with the E. coli gene B2341 (SEQ ID nature as their phosphate derivatives, pyridoxal 5'-phosphate NO:21). (PLP) and pyridoxamine 5'-phosphate (PMP), which are the 0013 Many enzymes exist as proenzymes or Zymogens coenzyme forms of the vitamin. PLP participates in catalysis that require activation by a non-protein molecule, or cofactor, of several important reactions of amino acid metabolism, in order to exhibit full activity. Cofactors may be loosely Such as transamination, decarboxylation, and racemization. categorized as coenzymes, prosthetic groups, or metal acti 0019 De novo synthesis of PLP occurs only in bacteria, vators. A coenzyme is a small, heat-stable organic molecule fungi, and plants. In Escherichia coli, de novo synthesis that readily dissociates from a proenzyme that functions as a occurs through condensation of 4-phosphohydroxy-L-threo carrier of chemical groups between enzymes. Prosthetic nine and deoxyXyulose 5-phosphate to form pyridoxine groups are firmly bound to the proenzyme and form a perma 5'-phosphate (PNP). The condensation reaction is catalyzed nent part of the protein structure. by the concerted action of the PdxA and Pdx-Jenzymes. In the 0014. The vitamin riboflavin is a component of the coen E. coli de novo pathway, PNP is then oxidized by the PdxH Zymes flavin adenine dinucleotide (FAD) and flavin mono oxidase to form PLP. Recently a different de novo PLP bio nucleotide (FMN), which are required in the enzymatic oxi synthetic pathway has been identified which is independent dation of carbohydrates and other electron transport reactions of deoxyxyulose 5-phosphate. In this pathway, PLP is syn critical for plants in their response to environmental stress thesized from ribose 5-phosphate or R5P and either glycer US 2011/O 145948 A1 Jun. 16, 2011 aldehyde 3-phosphate or dihydroxyacetone phosphate, via the products of two genes designated PDX1 and PDX2, TABLE 1-continued which show no homology to any of the E. coli PLP synthetic Amino genes. The PDX1 and PDX2 gene products are predicted to acid function as a glutamine amidotransferase, with PDX2 as the Polynucleotide SEQ glutaminase domain and PDX1 as the acceptor/synthase Gene Name Organism SEQID NO: ID NO: domain. The pdx1 gene product of the filamentous fungus BO452 E. coi 27 28 Cercospora nicotianae is highly homologous to a conserved BN42634969 B. naptis 29 30 gene family designated SOR1, which is widespread in BNP53O2 30 B. naptis 31 32 archeabacteria, eubacteria, plants, and fungi. The two path GMSae.90f11 G. max 33 34 YNL2O2W S. cerevisiae 35 36 ways of PLP de novo synthesis are autoexclusive, that is, HA66688442 Helianthus annuit is 37 38 organisms have the genes for one or the other pathway, but not YKL14OW S. cerevisiae 39 40 both. SLL1023 Synechocystis sp. 41 42 0020 PLP may also be synthesized by a second pathway, PCC 6803 designated a salvage pathway, through which pyridoxal (PL), SLRO2S2 Synechocystis sp. 43 44 b3803 E. coi 45 46 pyridoxine (PN), and pyridoxamine (PM) taken up from the BNS1286476 B. naptis 47 48 cell's growth medium. The salvage pathway is present in GMS9791864 G. max 49 50 addition to the de novo synthetic pathway in E. coli, and is the ZMBFbO243O4 Z. mays 51 52 b3209 E. coi 53 S4 only means by which mammalian cells can make PLP. In the GMSS34d01 G. max 55 56 E. coli salvage pathway, PL, PN, and PM are first phospho HAO3MC1392 H. anniitis 57 58 rylated by kinases to form PLP pyridoxine 5'-phosphate b2578 E. coi 59 60 (PNP), and PMP respectively. PNP and PMP are oxidized by b2682 E. coi 61 62 b328S E. coi 63 64 the PdxH oxidase referenced above. The pdxK gene encodes b1938 E. coi 65 66 a PN/PL/PM kinase, and the pdxY gene encodes a PL kinase, SLL1894 S. sp. PCC 6803 67 68 and the products of both genes share a number of conserved GMO8OOOO37 G. max 69 70 motifs with the PfkB superfamily of carbohydrate kinases. SLL1282 S. sp. PCC 6803 71 72 GMO6MC29296 G. max 73 74 Homologs of the PdxK and PdxY kinases have been identi ZMO7MCOO430 Z. mays 75 76 fied from humans, Trypanosoma brucei, Haemophilus influ ZMO7MC231.87 Z. mays 77 78 enzae, Caenorhabditis elegans, Rattus norvegicus, Saccha b1636 E. coi 79 8O romyces cerevisiae, and Salmonella typhimurium. pdxH E. coi 81 82 ECpdxK E. coi 83 84 0021 Although some genes that are involved in stress TBpdxK Trypanosoma brucei 85 86 responses, water use, and/or biomass in plants have been CEpdxK Caenorhabditis 87 88 characterized, to date, success at developing transgenic crop elegans plants with improved yield has been limited, and no such STyfei Saimoneiia 89 90 typhimurium plants have been commercialized. There is a need, therefore, HIyfei Haemophilus. 91 92 to identify additional genes that have the capacity to increase influenzae yield of crop plants. Yn8fp S. cerevisiae 93 94 SLR1779 S. sp. PCC 6803 95 96 SUMMARY OF THE INVENTION bdx. E. coi 97 98 bdx1.1 A. thaliana 99 100 0022. The present inventors have discovered that there are bdx1.3 A. thaliana 101 102 three critical components that must be optimized to achieve CNpdx1 Cercospora 103 104 nicotianae improvement in plant yield through transgenic expression of SCpdx1 S. cerevisiae 105 106 certain polypeptides When targeted under the regulatory ele BSpdx1 Bacilius subtiis 107 108 ments as described herein, the polynucleotides and polypep OSpdx1 Oryza sativa 109 110 HBpdx1 Hevea brasiliensis 111 112 tides set forth in Table 1 are capable of improving yield of SLpdx1 Stellaria longipes 113 114 transgenic plants BNpdx1 B. naptis 115 116 PPpdx1 Physcomitrella patens 117 118 TABLE 1. SPpdx1 Schizosaccharomyces 119 120 pombe Amino acid Polynucleotide SEQ 0023. In one embodiment, the invention provides a trans Gene Name Organism SEQID NO: ID NO: genic plant transformed with an expression cassette compris YBRO29C S. cerevisiae 1 2 ing, in operative association, an isolated polynucleotide BNO4MC3O8OS Brassica naptis 3 4 encoding a promoter capable of enhancing gene expression in ZMO6MC30283 Zea mays 5 6 roots and shoots; an isolated polynucleotide encoding a Sub YKL192C S. cerevisiae 7 8 cellular targeting peptide; and an isolated polynucleotide BN1004MS43616414 B. napus 9 10 GMO6MCO7589 G. max 11 12 encoding a full-length phosphatidate cytidylyltransferase HA1004MS66693619 Heianihats annuus 13 14 polypeptide, wherein the transgenic plant demonstrates YDRO18C S. cerevisiae 15 16 increased yield as compared to a wild type plant of the same GMO6MC27072 G. max 17 18 variety which does not comprise the expression cassette. ZMO6MCO4863 Z. mays 19 2O B2341 E. coi 21 22 0024. In another embodiment, the invention provides a ZMO6MCO4303 Z. mays 23 24 transgenic plant transformed with an expression cassette ZMO6MC15742 Z. mays 25 26 comprising in operative association, an isolated polynucle otide encoding a promoter capable of enhancing expression US 2011/O 145948 A1 Jun. 16, 2011 in leaves; an isolated polynucleotide encoding a mitochon length Succinate-CoA ligase polypeptide; wherein the trans drial transit peptide; and an isolated polynucleotide encoding genic plant demonstrates increased yield as compared to a a full-length acyl-carrier protein, wherein the transgenic plant wild type plant of the same variety which does not comprise demonstrates increased yield as compared to a wildtype plant the expression cassette. of the same variety which does not comprise the expression 0031. In another embodiment, the invention provides a CaSSette. method of increasing yield of a plant by transforming a wild 0025. In another embodiment, the invention provides a type plant with an expression cassette comprising, in opera transgenic plant transformed with an expression cassette comprising, in operative association, isolated polynucleotide tive association, an isolated polynucleotide encoding a pro encoding a promoter; an isolated polynucleotide encoding a moter and an isolated polynucleotide encoding a full-length Subcellular targeting peptide; and an isolated polynucleotide cobalt-precorrin-6A reductase polypeptide; wherein the encoding a full-length acyltransferase polypeptide; wherein transgenic plant demonstrates increased yield as compared to the transgenic plant demonstrates increased yield as com a wild type plant of the same variety which does not comprise pared to a wild type plant of the same variety which does not the expression cassette. comprise the expression cassette. 0032. In another embodiment, the invention provides a 0026. In another embodiment, the invention provides a transgenic plant transformed with an expression cassette method of increasing yield of a plant by transforming a wild comprising, in operative association, an isolated polynucle type plant with an expression cassette comprising, in opera otide encoding a promoter and an isolated polynucleotide tive association, an isolated polynucleotide encoding a pro encoding a full-length polypeptide having uroporphyrin-III moter capable of enhancing gene expression in leaves; an C-methyltransferase activity and a HemX signature isolated polynucleotide encoding a mitochondrial transit pep sequence; wherein the transgenic plant demonstrates tide; and an isolated polynucleotide encoding a full-length increased yield as compared to a wild type plant of the same bifunctional anaerobic fatty acid oxidation complex polypep variety which does not comprise the expression cassette. tide, regenerating transgenic plants from the transformed 0033. In another embodiment, the invention provides a plant cell, and selecting higher-yielding plants from the trans transgenic plant transformed with an expression cassette genic plants. comprising in operative association, an isolated polynucle 0027. In another embodiment, the invention provides a otide encoding a promoter capable of enhancing gene expres method of increasing yield of a plant by transforming a wild Sionin roots and shoots and an isolated polynucleotide encod type plant with an expression cassette comprising, in opera ing a full-length polypeptide having isoprenoid biosynthesis tive association, an isolated polynucleotide encoding a pro activity and a DJ-1 Pfp1 signature sequence; wherein the moter capable of enhancing gene expression in leaves; an transgenic plant demonstrates increased yield as compared to isolated polynucleotide encoding a subcellular targeting pep a wild type plant of the same variety which does not comprise tide; and an isolated polynucleotide encoding a full-length the expression cassette. acyl-CoA thioesterase polypeptide; wherein the transgenic 0034. In another embodiment, the invention provides a plant demonstrates increased yield as compared to a wildtype transgenic plant transformed with an expression cassette plant of the same variety which does not comprise the expres comprising, in operative association, an isolated polynucle sion cassette. otide encoding a promoter capable of enhancing gene expres 0028. In another embodiment, the invention provides a sion in roots and shoots; and an isolated polynucleotide method of increasing yield of a plant by transforming a wild encoding a full-length polypeptide having LysE type translo type plant with an expression cassette comprising, in opera cator activity and a LysE signature sequence comprising tive association, an isolated polynucleotide encoding a pro amino acids 14 to 195 of SEQID NO:60; wherein the trans moter, an isolated polynucleotide encoding a Subcellular genic plant demonstrates increased yield as compared to a targeting peptide; and an isolated polynucleotide encoding a wild type plant of the same variety which does not comprise full-length sterol esterase polypeptide; wherein the trans the expression cassette. genic plant demonstrates increased yield as compared to a 0035. In another embodiment, the invention provides a wild type plant of the same variety which does not comprise transgenic plant transformed with an expression cassette the expression cassette. comprising, in operative association, an isolated polynucle 0029. In another embodiment, the invention provides a otide encoding a promoter capable of enhancing gene expres method of increasing yield of a plant by transforming a wild Sionin roots and shoots and an isolated polynucleotide encod type plant with an expression cassette comprising, in opera ing a full-length polypeptide having LIV-E family branched tive association, an isolated polynucleotide encoding a pro chain amino acid transport activity and an AZIC signature moter capable of enhancing gene expression in leaves; an sequence; wherein the transgenic plant demonstrates isolated polynucleotide encoding a mitochondrial targeting increased yield as compared to a wild type plant of the same peptide; and an isolated polynucleotide encoding a full variety which does not comprise the expression cassette. length 2,4-dienoyl-CoA reductase polypeptide; wherein the 0036. In another embodiment, the invention provides a transgenic plant demonstrates increased yield as compared to transgenic plant transformed with an expression cassette a wild type plant of the same variety which does not comprise comprising, in operative association, an isolated polynucle the expression cassette. otide encoding a promoter capable of enhancing gene expres 0030. In another embodiment, the invention provides a sion in roots and shoots; and an isolated polynucleotide method of increasing yield of a plant by transforming a wild encoding a truncated DNA-binding polypeptide having a type plant with an expression cassette comprising, in opera sequence comprising amino acids 1 to 102 of SEQID NO:64; tive association, an isolated polynucleotide encoding a pro wherein the transgenic plant demonstrates increased yield as moter, an isolated polynucleotide encoding a plastid transit compared to a wild type plant of the same variety which does peptide; and an isolated polynucleotide encoding a full not comprise the expression cassette. US 2011/O 145948 A1 Jun. 16, 2011
0037. In another embodiment, the invention provides a in increased tolerance to an environmental stress, and/or transgenic plant transformed with an expression cassette growth, and/or yield under normal and/or stress conditions as comprising, in operative association, an isolated polynucle compared to a wild type variety of the plant. otide encoding a promoter capable of enhancing gene expres 0045. In still another embodiment, the invention provides Sionin roots and shoots and an isolated polynucleotide encod a method of increasing a plant's tolerance to an environmental ing a full-length polypeptide having a first Ysc J. FliF stress, and/or growth, and/or yield. The method comprises the signature sequence and a second YscJ. FliF signature steps of transforming a plant cell with an expression cassette sequence; wherein the transgenic plant demonstrates comprising an isolated polynucleotide of Table 1, and gener increased yield as compared to a wild type plant of the same ating a transgenic plant from the plant cell, wherein the trans variety which does not comprise the expression cassette. genic plant comprises the polynucleotide. 0038. In another embodiment, the invention provides a transgenic plant transformed with an expression cassette BRIEF DESCRIPTION OF THE DRAWINGS comprising, in operative association, an isolated polynucle 0046 FIG. 1 shows an alignment of the amino acid otide encoding a promoter, and an isolated polynucleotide sequences of full-length phosphatidate cytidylyltransferase encoding a full length GTP cyclohydrolase II polypeptide polypeptide designated YBR029C (SEQ ID NO:2), which does not comprise a Subcellular targeting peptide; BN04MC30805 (SEQID NO:4) and ZMO6MC30283 (SEQ wherein the transgenic plant demonstrates increased yield as ID NO:6) The alignment was generated using Align X of compared to a wildtype plant of the same variety that does not Vector NTI Advance 10.3.0. comprise the expression cassette. 0047 FIG. 2 shows an alignment of the amino acid 0039. In another embodiment, the invention provides a sequences of the acyl-carrier proteins designated YKL 192O transgenic plant transformed with an expression cassette (SEQ ID NO:8), BN1004MS43616414 (SEQ ID NO:10), comprising, in operative association, an isolated polynucle GM06MC07589(SEQ ID NO:12), and otide encoding a promoter and an isolated polynucleotide HA1004MS66693619 (SEQID NO:14). The alignment was encoding a full length lumazine synthase polypeptide which generated using Align X of Vector NTI Advance 10.3.0. does not comprise a Subcellular targeting peptide; wherein 0048 FIG. 3 shows an alignment of the amino acid the transgenic plant demonstrates increased yield as com sequences of the acyltransferases designated: YDRO18C pared to a wild type plant of the same variety that does not (SEQ ID NO:16), GMO6MC27072 (SEQ ID NO:18) and comprise the expression cassette. ZMO6MC04863 (SEQ ID NO:20). The alignment was gen 0040. In another embodiment, the invention provides a erated using Align X of Vector NTI Advance 10.3.0. transgenic plant transformed with an expression cassette 0049 FIG. 4 shows an alignment of the amino acid comprising, in operative association, an isolated polynucle sequences encoding bifunctional anaerobic fatty acid oxida otide encoding a promoter and an isolated polynucleotide tion complex polypeptides designated b2341 (SEQ ID encoding a full length polypeptide capable of enhancing pyri NO:22), ZMO6MC04303 (SEQ ID NO:24) and doxal 5'-phosphate synthesis; wherein the transgenic plant ZMO6MC15742 (SEQ ID NO:26). The alignment was gen demonstrates increased yield as compared to a wildtype plant erated using Align X of Vector NTI Advance 10.3.0. of the same variety that does not comprise the expression 0050 FIG. 5 shows an alignment of the amino acid cassette. In this embodiment, the expression cassette may sequences encoding acyl-CoA thioesterase polypeptides des further comprise a polynucleotide encoding a mitochondrial ignated B0452 (SEQ ID NO:28), BN42634969 (SEQ ID or plastid transit peptide. NO:30), BNP5302 30 (SEQID NO:32), and GMsae90f11 0041. In a further embodiment, the invention provides a (SEQ ID NO:34). The alignment was generated using Align seed produced by the transgenic plants described above, X of Vector NTI Advance 10.3.0. wherein the seed is true breeding for a transgene comprising 0051 FIG. 6 shows an alignment of the amino acid the expression vectors described above. Plants derived from sequences encoding 2,4-dienoyl-CoA reductasepolypeptides the seed of the invention demonstrate increased tolerance to designated YNL202W (SEQ ID NO:36) and HA66688442 an environmental stress, and/or increased plant growth, and/ (SEQ ID NO:38). The alignment was generated using Align or increased yield, under normal or stress conditions as com X of Vector NTI Advance 10.3.0. pared to a wild type variety of the plant. 0.052 FIG. 7 shows an alignment of the amino acid 0042. In a still another aspect, the invention concerns sequences of uroporphyrin-III C-methyltransferases desig products produced by or from the transgenic plants of the nated b3803 (SEQ ID NO:46), BN51286476 (SEQ ID invention, their plant parts, or their seeds. Such as a foodstuff. NO:48), GM59791864 (SEQ ID NO:50), and feedstuff, food Supplement, feed Supplement, fiber, cosmetic ZMBFb0243J04 (SEQID NO:52). The alignment was gen or pharmaceutical. erated using Align X of Vector NTI Advance 10.3.0. 0043. The invention further provides certain isolated poly 0053 FIG. 8 shows an alignment of the amino acid nucleotides identified in Table 1, and certain isolated sequences of the isoprenoid biosynthesis proteins designated polypeptides identified in Table 1. The invention is also b3209 (SEQID NO:54), GMss34d01 (SEQID NO:56), and embodied in recombinant vector comprising an isolated poly HAO3MC1392 (SEQID NO:58). The alignment was gener nucleotide of the invention. ated using Align X of Vector NTI Advance 10.3.0. 0044. In yet another embodiment, the invention concerns a 0054 FIG. 9 shows a flow diagram of the riboflavin/FAD method of producing the aforesaid transgenic plant, wherein biosynthesis pathway in wild type plants. Enzyme designa the method comprises transforming a plant cell with an tions are as follows: Enzyme I is GTP cyclohydrolase II expression vector comprising an isolated polynucleotide of (RibA); Enzyme II is 2,5-diamino-6-ribosylamino-4(3H)-py the invention, and generating from the plant cell a transgenic rimidinone 50-phosphate deaminase; Enzyme III is 5-amino plant that expresses the polypeptide encoded by the poly 6-ribosylamino-2,4(1H.3H)-pyrimidinedione 50-phosphate nucleotide. Expression of the polypeptide in the plant results reductase: Enzyme IV is 2,5-diamino-6-ribosylamino-4 US 2011/O 145948 A1 Jun. 16, 2011
(3H)-pyrimidinone 50-phosphate reductase: Enzyme V is invention, changes in different phenotypic traits may improve 2,5-diamino-6-ribitylamino-4(3H)-pyrimidinedione yield. For example, and without limitation, parameters such 50-phosphate deaminase; Enzyme VI is a hypothetical phos as floral organ development, root initiation, root biomass, phatase: Enzyme VII is 3,4-dihydroxy-2-butanone-4-phos seed number, seed weight, harvest index, tolerance to abiotic phate synthase: Enzyme VIII is 6,7-dimethyl-8-ribityllu environmental stress, reduction of nutrient, e.g., nitrogen or mazine synthase (RibH); Enzyme IX is riboflavin synthase: phosphorus, input requirement, leaf formation, phototro Enzyme X is riboflavin kinase; and Enzyme XI is FAD syn pism, apical dominance, and fruit development, are Suitable thetase. Intermediates in the biosynthesis of riboflavin and measurements of improved yield. Any increase in yield is an FAD are GTP: DAPP (2,5-diamino-6-ribosylamino-4(3H)- improved yield in accordance with the invention. For pyrimidinone 50-phosphate); AAPP (5-amino-6-ribosy example, the improvement in yield can comprise a 0.1%, lamino-2,4(1H.3H)-pyrimidinedione 50-phosphate); DRPP 0.5%, 1%, 3%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, (2,5-diamino-6-ribitylamino-4(3H)-pyrimidinedione 70%, 80%, 90% or greater increase in any measured param 50-phosphate); ARPP (5-amino-6-ribitylamino-2,4(1H.3H)- eter. For example, an increase in the bu?acre yield of soybeans pyrimidinedione 50-phosphate); ARP; G6P (Glucose-6- or corn derived from a crop comprising plants which are phosphate); riboflavin; with RSP, DBP, DR, FMN, and FAD transgenic for the nucleotides and polypeptides of Table 1, as as set forth above. compared with the bu?acre yield from untreated soybeans or 0055 FIG. 10A shows a flow diagram of the proposed corn cultivated under the same conditions, is an improved riboflavin/FAD biosynthesis pathway in the transgenic plants yield in accordance with the invention. of the invention. Abbreviations areas set forth in FIG. 9. FIG. 0061. As defined herein, a “transgenic plant' is a plant that 10A shows the pathway with RibH targeted to the cytosol. has been altered using recombinant DNA technology to con FIG. 10B shows the pathway with RibA targeted to the cyto tain an isolated nucleic acid which would otherwise not be sol; FIG. 10C shows the pathway with both RibH and RibA present in the plant. As used herein, the term “plant' includes targeted to the cytosol. a whole plant, plant cells, and plant parts. Plant parts include, 0056 FIG. 11 shows a flow diagram of vitamin B6 syn but are not limited to, stems, roots, ovules, stamens, leaves, thesis as it relates to the present invention. Abbreviations: embryos, meristematic regions, callus tissue, gametophytes, DAP: dihydroxyacetone-P: G3P: glyceraldehyde-3P; with sporophytes, pollen, microspores, and the like. The trans PL, PLP, PM, PN, PNP, and R5P as set forth above. genic plant of the invention may be male sterile or male 0057 FIG. 12 shows an alignment of the amino acid fertile, and may further include transgenes other than those sequences of GTP cyclohydrolase II designated SLL 1894 that comprise the isolated polynucleotides described herein. (SEQ ID NO:68) and Gm018000037 (SEQID NO:70), The 0062. As used herein, the term “variety” refers to a group alignment was generated using Align X of Vector NTI of plants within a species that share constant characteristics Advance 10.3.0. Amino acids 1 to 49 of SEQ ID NO:70 that separate them from the typical form and from other correspond to the Subcellular targeting sequence. possible varieties within that species. While possessing at 0058 FIG. 13 shows an alignment of the amino acid least one distinctive trait, a variety is also characterized by sequences of the lumazine synthase polypeptides designated some variation between individuals within the variety, based SLL1282 (SEQ ID NO:72), GMO6MC29296 (SEQ ID primarily on the Mendelian segregation of traits among the NO:74), ZMO7MC00430 (SEQ ID NO:76) and progeny of Succeeding generations. A variety is considered ZM07MC23.187 (SEQ ID NO:78). The alignment was gen “true breeding for a particular trait if it is genetically erated using Align X of Vector NTI. Advance 10.3.0. Subcel homozygous for that trait to the extent that, when the true lular targeting sequences correspond to amino acids 1 to 53 of breeding variety is self-pollinated, a significant amount of SEQ ID NO:74; amino acids 1 to 56 of SEQID NO:76; and independent segregation of the trait among the progeny is not amino acids 1 to 80 of SEQID NO:78. observed. In the present invention, the trait arises from the transgenic expression of one or more isolated polynucle DETAILED DESCRIPTION OF THE PREFERRED otides introduced into a plant variety. As also used herein, the EMBODIMENTS term “wild type variety” refers to a group of plants that are 0059. Throughout this application, various publications analyzed for comparative purposes as a control plant, wherein are referenced. The disclosures of all of these publications the wild type variety plant is identical to the transgenic plant and those references cited within those publications in their (plant transformed with an isolated polynucleotide in accor entireties are hereby incorporated by reference into this appli dance with the invention) with the exception that the wildtype cation in order to more fully describe the state of the art to variety plant has not been transformed with an isolated poly which this invention pertains. The terminology used herein is nucleotide of the invention. The term “wild type' as used for the purpose of describing specific embodiments only and herein refers to a plant cell, seed, plant component, plant is not intended to be limiting. As used herein, “a” or “an can tissue, plant organ, or whole plant that has not been geneti mean one or more, depending upon the context in which it is cally modified with an isolated polynucleotide in accordance used. Thus, for example, reference to “a cell' can mean that at with the invention. least one cell can be used. 0063. The term “control plant’ as used herein refers to a 0060. In one embodiment, the invention provides a trans plant cell, an explant, seed, plant component, plant tissue, genic plant that overexpresses an isolated polynucleotide plant organ, or whole plant used to compare against trans identified in Table 1 in the subcellular compartment and tissue genic or genetically modified plant for the purpose of identi indicated herein. The transgenic plant of the invention dem fying an enhanced phenotype or a desirable trait in the trans onstrates an improved yield as compared to a wild type vari genic or genetically modified plant. A "control plant may in ety of the plant. As used herein, the term “improved yield' Some cases be a transgenic plant line that comprises an empty means any improvement in the yield of any measured plant vector or marker gene, but does not contain the recombinant product, such as grain, fruit or fiber. In accordance with the polynucleotide of interest that is present in the transgenic or US 2011/O 145948 A1 Jun. 16, 2011 genetically modified plant being evaluated. A control plant the like. Especially preferred are A. thaliana, Nicotiana may be a plant of the same line or variety as the transgenic or tabacum, rice, oilseed rape, canola, Soybean, corn (maize), genetically modified plant being tested, or it may be another cotton, and wheat. line or variety, Such as a plant known to have a specific phenotype, characteristic, or known genotype. A Suitable A. Phosphatidate Cytidylyltransferase control plant would include a genetically unaltered or non 0067. In one embodiment, the invention provides a pro transgenic plant of the parental line used to generate a trans vides a transgenic plant transformed with an expression cas genic plant herein. sette comprising, in operative association, an isolated poly 0064. As defined herein, the term “nucleic acid' and nucleotide encoding a promoter capable of enhancing gene "polynucleotide are interchangeable and refer to RNA or expression in roots and shoots; an isolated polynucleotide DNA that is linear or branched, single or double stranded, or encoding a subcellular targeting peptide; and an isolated a hybrid thereof. The term also encompasses RNA/DNA polynucleotide encoding a full-length phosphatidate cytidy hybrids. An "isolated nucleic acid molecule is one that is lyltransferase polypeptide, wherein the transgenic plant dem Substantially separated from other nucleic acid molecules onstrates increased yield as compared to a wild type plant of which are present in the natural Source of the nucleic acid (i.e., the same variety which does not comprise the expression sequences encoding other polypeptides). For example, a cassette. In accordance with the invention, when the promoter cloned nucleic acid is considered isolated. A nucleic acid is is capable of enhancing expression in roots or shoots, the also considered isolated if it has been altered by human inter Subcellular targeting peptide is a plastid transit peptide. 0068. As indicated in Table 2 below, when the S. cerevisiae vention, or placed in a locus or location that is not its natural gene product YBR029C (SEQ ID NO:2) is targeted to the site, or if it is introduced into a cell by transformation. More chloroplast and the gene's transcriptional expression is driven over, an isolated nucleic acid molecule. Such as a cDNA by the Super promoter, transgenic plants demonstrate molecule, can be free from some of the other cellular material improved response to water-limiting conditions. Moreover, with which it is naturally associated, or culture medium when Table 3 indicates that under well-watered conditions, plants produced by recombinant techniques, or chemical precursors expressing YBR029C which is targeted to the plastid or to or other chemicals when chemically synthesized. While it mitochondria were larger than control plants. Gene may optionally encompass untranslated sequence located at YBR029C encodes a phosphatidate cytidylyltransferase (EC both the 3' and 5' ends of the coding region of a gene, it may 2.7.7.41), also known as CDP-diacylglycerol synthase be preferable to remove the sequences which naturally flank (CDS), which catalyzes the synthesis of CDP-diacylglycerol the coding region in its naturally occurring replicon. from CTP and phosphatidate. CDS is a membrane-bound 0065. As used herein, the term “environmental stress’ protein, with eight predicted membrane spanning regions in refers to a sub-optimal condition associated with salinity, potato and Arabidopsis. Phosphatidate cytidylyltransferases drought, nitrogen, temperature, metal, chemical, pathogenic, are characterized, in part, by a distinctive signature sequence or oxidative stresses, or any combination thereof. As used of “S-X-LIVMF-K-R-X(4)-K-D-x-GSA-X(2)-LIF herein, the term “drought” refers to an environmental condi PGS-x-H-G-G-LIVMF-X-D-R-LIVMFT-D” where tion where the amount of water available to support plant amino acid positions within square brackets can be any of the growth or development is less than optimal. As used herein, designated residues, and unbracketed amino acid positions can only be that specific amino acid residue. Such conserved the term “fresh weight” refers to everything in the plant signature sequences are exemplified in the phosphatidate including water. As used herein, the term “dry weight” refers cytidylyltransferase proteins set forth in FIG. 1. to everything in the plant other than water, and includes, for 0069. The transgenic plant of this embodiment may com example, carbohydrates, proteins, oils, and mineral nutrients. prise any polynucleotide encoding phosphatidate cytidylyl 0066. Any plant species may be transformed to create a transferase. Preferably, the transgenic plant of this embodi transgenic plant in accordance with the invention. The trans ment comprises a polynucleotide encoding a polypeptide genic plant of the invention may be a dicotyledonous plant or having phosphatidate cytidylyltransferase activity, wherein a monocotyledonous plant. For example and without limita the polypeptide comprises a phosphatidate cytidylyltrans tion, transgenic plants of the invention may be derived from ferase signature sequence selected from the group consisting any of the following diclotyledonous plant families: Legumi of amino acids 351 to 377 of SEQID NO:2, amino acids 341 nosae, including plants such as pea, alfalfa and Soybean: to 367 of SEQID NO:4, or amino acids 340 to 366 of SEQID Umbelliferae, including plants such as carrot and celery; NO:6. More preferably, the transgenic plant of this embodi Solanaceae, including the plants such as tomato, potato, aub ment comprises a polynucleotide encoding phosphatidate ergine, tobacco, and pepper, Cruciferae, particularly the cytidylyltransferase domain having a sequence comprising genus Brassica, which includes plant such as oilseed rape, amino acids 65 to 377 of SEQIDNO:2, amino acids 54 to 367 beet, cabbage, cauliflower and broccoli); and A. thaliana; of SEQID NO:4, or amino acids 53 to 366 of SEQID NO:6. Compositae, which includes plants such as lettuce, Mal Most preferably, the transgenic plant of this embodiment vaceae, which includes cotton; Fabaceae, which includes comprises a polynucleotide encoding phosphatidate cytidy plants such as peanut, and the like. Transgenic plants of the lyltransferase polypeptide comprising amino acids 1 to 457 of invention may be derived from monocotyledonous plants, Such as, for example, wheat, barley, Sorghum, millet, rye, SEQ ID NO:2, amino acids 1 to 367 of SEQ ID NO:4, or triticale, maize, rice, oats and Sugarcane. Transgenic plants of amino acids 1 to 425 of SEQID NO:6. the invention are also embodied as trees such as apple, pear, quince, plum, cherry, peach, nectarine, apricot, papaya, B. Acyl Carrier Protein mango, and other woody species including coniferous and 0070. In another embodiment, the invention provides a deciduous trees such as poplar, pine, Sequoia, cedar, oak, and transgenic plant transformed with an expression cassette US 2011/O 145948 A1 Jun. 16, 2011 comprising in operative association, an isolated polynucle comprising, in operative association, isolated polynucleotide otide encoding a promoter capable of enhancing expression encoding a promoter; an isolated polynucleotide encoding a in leaves; an isolated polynucleotide encoding a mitochon Subcellular targeting peptide; and an isolated polynucleotide drial transit peptide; and an isolated polynucleotide encoding encoding an acyltransferase polypeptide; wherein the trans an acyl carrier protein; wherein the transgenic plant demon genic plant demonstrates increased yield as compared to a strates increased yield as compared to a wild type plant of the wild type plant of the same variety which does not comprise same variety which does not comprise the expression cas the expression cassette. Sette. 0075. As set forth in Table 5 below, when the S. cerevisiae 0071. As shown in Table 4 below, when the S. cerevisiae acyltransferase gene product YDR018C (SEQID NO:16) is gene product YKL192O (SEQ ID NO:8) is targeted to the targeted to the chloroplast or mitochondria under the control mitochondria under the control of the USP promoter (SEQID of the Super promoter or the USP promoter (SEQ ID NO:123), transgenic plants demonstrate improved response NO:123), transgenic plants demonstrate improved response to water-limiting conditions. Gene YKL 192O encodes an to water-limiting conditions. Moreover, Table 6 indicates that acyl-carrier protein (ACP), containing a phosphopantetheline under well-watered conditions, plants expressing YDR018C binding domain (PF00550). Acyl carrier proteins catalyze a under control of the USP promoter (SEQ ID NO:123), and condensation reaction to form peptide bonds in non-riboso targeted to the mitochondria, were larger than control plants. mal protein biosynthesis. Acyl carrier protein is a universal 0076. The transgenic plant of this embodiment may com and highly conserved carrier of acyl groups in fatty acid prise any polynucleotide encoding an acyltransferase biosynthesis. The amino-terminal region of the ACP proteins polypeptide. Preferably, the transgenic plant of this embodi is well defined and consists of alpha four helices arranged in ment comprises a polynucleotide encoding an acyltransferase a right-handed bundle held together by inter-helical hydro polypeptide comprising an acyltransferase domain selected phobic interactions. from the group consisting of amino acids 108 to 272 of SEQ 0072 Phosphopantetheline (or pantetheline 4' phosphate) is ID NO:16, amino acids 80 to 222 of SEQID NO:18, or amino the prosthetic group of ACP in some multienzyme complexes, acids 116 to 258 of SEQ ID NO:20. More preferably, the where it serves as a Swinging arm for the attachment of transgenic plant of this embodiment comprises a polynucle activated fatty acid and amino acid groups. Phosphopanteth otide encoding an acyltransferase having a sequence com eine binding domains are characterized, in part, by the pres prising amino acids 1 to 396 of SEQID NO:16, amino acids ence of the distinctive phosphopantetheline attachment site 1 to 384 of SEQID NO:18, or amino acids 1 to 332 of SEQID signature sequence, “DEQGSTALMKRH-LIVMFYS NO:2O. TAC-IGNQ-LIVMFYAG-DNEKHS-S-LIVMST D. Bifunctional anaerobic fatty acid oxidation complex {PCFY-ISTAGCPQLIVMF-LIVMATN-DENQG polypeptide TAKRHLM-LIVMWSTA-LIVGSTACR-LPIY}- {VY-LIVMFA” whereamino acid positions within square 0077. In another embodiment, the invention provides a brackets can be any of the designated residues, amino acid method of increasing yield of a plant species by transforming positions within curly brackets can be any amino acid residue a wild type cell of said species with an expression cassette except the one(s) listed and unbracketed amino acid positions comprising, in operative association, an isolated polynucle can only be that specific amino acid residue. The phospho otide encoding a promoter capable of enhancing expression pantetheline moiety is attached to serine residue indicated in in leaves; an isolated polynucleotide encoding a mitochon bold italic in the above signature sequence. This serine resi drial transit peptide; and an isolated polynucleotide encoding due is present in the amino terminus of helix II, a domain of a bifunctional anaerobic fatty acid oxidation complex the protein referred to as the recognition helix and which is polypeptide In a second step, transgenic plantlets are regen responsible for the interaction of ACPs with the enzymes of erated from the transformed plant cell. In a third step, the type II fatty acid synthesis. transgenic plantlets are subjected to a yield-related assay, and 0073 Preferably, the transgenic plant of this embodiment higher-yielding plants are selected from the regenerated comprises a polynucleotide encoding a full-length ACP com transgenic plants. prising a phosphopantetheline binding site signature sequence 0078. As shown in Tables 7 and 8 below, when transcrip selected from the group consisting of amino acids 77 to 92 of tion of the E. coli gene B2341 (SEQ ID NO:21) is under SEQID NO:8, amino acids 73 to 88 of SEQIDNO:10, amino control of the USP promoter (SEQID NO:123) and the gene acids 75 to 90 of SEQID NO:12, or amino acids 84 to 99 of product (SEQ ID NO:22) is targeted to the mitochondria, SEQID NO:14. More preferably, the transgenic plant of this transgenic plants tend to be larger and darker green than embodiment comprises a polynucleotide encoding an acyl control plants. Gene B2341 encodes a protein that comprises carrier protein comprising an acyl carrier protein domain three domains: a domain characteristic of the ECH or enoyl selected from the group consisting of amino acids 49 to 106 of CoA hydratase/isomerase family (PF00378); a C-terminal SEQ ID NO:8, amino acids 49 to 102 of SEQ ID NO:10, domain characteristic 3HCDH or 3-hydroxyacyl-CoA dehy amino acids 51 to 104 of SEQID NO:12, amino acids 60 to drogenase (PF00725); and a 3HCDH N,3-hydroxyacyl-CoA 113 of SEQID NO:14. Most preferably, the transgenic plant dehydrogenase, NAD binding domain (PF02737). The of this embodiment comprises a polynucleotide encoding an 3HCDH domain is characterized, in part, by the presence of acyl-carrier protein comprising amino acids 1 to 125 of SEQ the signature sequence, “IDNES-x(2)-GA-F-LIVM IDNO:8, amino acids 1 to 117 of SEQID NO:10, amino acids FYA-x-INTI-R-x(3)-PA-LIVMFY-LIVMFYST-x(5, 1 to 128 of SEQID NO:12, oramino acids 1 to 119 of SEQID 6)-LIVMFYCT-ILIVMFYEAH-x(2)-(GVEI”, where NO:14. amino acid positions within square brackets can be any of the designated residues, and unbracketed amino acid positions C. Acyltransferase can only be that specific amino acid residue. All the sequences 0074. In another embodiment, the invention provides a shown in FIG. 4 exhibit this characteristic signature transgenic plant transformed with an expression cassette sequence, the only deviation within this group is the presence US 2011/O 145948 A1 Jun. 16, 2011
of a leucine residue at position 498 in ZMO6MC15742 (SEQ high levels of activity on medium- and long chain acyl CoAS. ID NO:26) instead of the canonical arginine residue. Two families of ACHs have been identified in A. thaliana. 007.9 The method of this embodiment may employ any One family, consisting of AtACH1 and AtACH2, appears to polynucleotide encoding a bifunctional anaerobic fatty acid be peroxisomal, as they have type-1 peroxisomal targeting oxidation complex polypeptide comprising an ECH domain, sequences. The other family, consisting of AtACH4 and a 3HCDH-C terminal domain and a 3HCHD-NAD binding AtACH5, resides in the endoplasmic reticulum. domain. Preferably, the method of this embodiment employs I0083. The transgenic plant of this embodiment may com a polynucleotide encoding a bifunctional anaerobic fatty acid prise any polynucleotide encoding an acyl-CoA thioesterase. oxidation complex polypeptide comprising three domains: a) Preferably, the transgenic plant of this embodiment com an ECH domain selected from the group consisting of amino prises a polynucleotide encoding a full-length polypeptide acids 17 to 190 of SEQID NO:22; amino acids 17 to 187 SEQ having acyl-CoA thioesterase activity, wherein the polypep ID NO:24 and amino acids 18 to 187 SEQ ID NO:26; b) a tide comprises a domain selected from the group consisting of 3HCDH domain selected from the group consisting of amino amino acids 107 to 184 of SEQID NO:28, amino acids 54 to acids 489 to 513 of SEQID NO:22, amino acids 490 to 514 of 138 of SEQ ID NO:30, amino acids 127 to 211 of SEQ ID SEQ ID NO:24, amino acids 490 to 514 of SEQID NO:26: NO:32, and amino acids 3 to 87 of SEQ ID NO:34. More and c) a 3HCDH-N domain selected from the group consist preferably, the transgenic plant of this embodiment com ing of amino acids 310 to 490 of SEQID NO:22; amino acids prises a polynucleotide encoding an acyl-CoA thioesterase 312 to 491 of SEQID NO:24 and amino acids 312 to 491 SEQ having a sequence comprising amino acids 1 to 286 of SEQ ID NO:26 More preferably, the method of this embodiment ID NO:28, amino acids 1 to 248 of SEQ ID NO:30, amino employs a polynucleotide encoding a bifunctional anaerobic acids 1 to 212 of SEQID NO:32, or amino acids 1 to 197 of fatty acid oxidation complex polypeptide having a sequence SEQID NO:34. comprising amino acids 1 to 714 of SEQ ID NO:22, amino F. 2,4-dienoyl-CoA Reductase acids 1 to 723 of SEQID NO:24, or amino acids 1 to 727 of I0084. In another embodiment, the invention provides a SEQID NO:26. transgenic plant transformed with an expression cassette 0080. The invention is also embodied in a transgenic plant comprising, in operative association, an isolated polynucle comprising a polynucleotide encoding a bifunctional anaero otide encoding a promoter capable of enhancing expression bic fatty acid oxidation complex polypeptide comprising a) in leaves; an isolated polynucleotide encoding a mitochon an ECH domain selected from the group consisting of amino drial transit peptide; and an isolated polynucleotide encoding acids 17 to 190 of SEQID NO:22; amino acids 17 to 187 SEQ a full-length 2,4-dienoyl-CoA reductase polypeptide; ID NO:24 and amino acids 18 to 187 SEQ ID NO:26; b) a wherein the transgenic plant demonstrates increased yield as 3HCDH domain selected from the group consisting of amino compared to a wild type plant of the same variety which does acids 489 to 513 of SEQID NO:22, amino acids 490 to 514 of not comprise the expression cassette. SEQ ID NO:24, amino acids 490 to 514 of SEQID NO:26: I0085. As shown in Table 10 below, when transcription of and c) a 3HCDH-N domain selected from the group consist the S. cerevisiae geneyNL202W (SEQID NO:35) is targeted ing of amino acids 310 to 490 of SEQID NO:22; amino acids to mitochondria under control of the USP promoter, under 312 to 491 of SEQID NO:24 and amino acids 312 to 491 SEQ cycling drought conditions, transgenic plants were larger than ID NO:26 More preferably, the transgenic plant of this control plants. YNL202W is a 2,4-dienoyl-CoA reductase embodiment comprises a polynucleotide encoding a bifunc (EC 1.3.1.34, DECR), an auxiliary enzyme of beta-oxidation. tional anaerobic fatty acid oxidation complex polypeptide DECR participates in the degradation of unsaturated fatty having a sequence comprising amino acids 1 to 714 of SEQ enoyl-CoA esters having double bonds in both even- and ID NO:22, amino acids 1 to 723 of SEQID NO:24, or amino odd-numbered positions in peroxisome. It catalyzes the acids 1 to 727 of SEQID NO:26. NADP-dependent reduction of 2,4-dienoyl-CoA to yield E. Acyl-CoA Thioesterase trans-3-enoyl-CoA. I0086. The transgenic plant of this embodiment may com 0081. In another embodiment, the invention provides a prise any polynucleotide encoding a 2,4-dienoyl-CoA reduc transgenic plant transformed with an expression cassette tase. Preferably, the transgenic plant of this embodiment comprising, in operative association, an isolated polynucle comprises a polynucleotide encoding a full-length polypep otide encoding a promoter capable of enhancing gene expres tide having 2,4-dienoyl-CoA reductase activity, wherein the sion in leaves; an isolated polynucleotide encoding a mito polypeptide comprises a domain selected from the group chondrial transit peptide; and an isolated polynucleotide consisting of amino acids 107 to 270 of SEQID NO:36 and encoding a full-length acyl-CoA thioesterase polypeptide; amino acids 54 to 227 of SEQIDNO:38. More preferably, the wherein the transgenic plant demonstrates increased yield as transgenic plant of this embodiment comprises a polynucle compared to a wild type plant of the same variety which does otide encoding a 2,4-dienoyl-CoA reductase having a not comprise the expression cassette. sequence comprising amino acids 1 to 296 of SEQID NO:36 0082. As shown in Table 9 below, when transcription of or amino acids 1 to 264 of SEQID NO:38. the E. coli gene b0452 is targeted to mitochondria under the control of the USP promoter, transgenic plants were larger G. Sterol Esterase than control plants, both under well-watered and drought conditions. Gene B0452 (SEQ ID NO:27) encodes an acyl I0087. In another embodiment, the invention provides a CoA thioesterase (EC 3.1.2.2). Acyl-CoA thioesterases transgenic plant transformed with an expression cassette (ACH) are a group of enzymes that catalyze the hydrolysis of comprising, in operative association, an isolated polynucle acyl-CoAs to the free fatty acid and coenzyme A (CoASH), otide encoding a promoter, an isolated polynucleotide encod providing the potential to regulate intracellular levels of acyl ing a Subcellular transit peptide; and an isolated polynucle CoAs, free fatty acids and CoASH. This enzyme displays otide encoding a full-length sterol esterase; wherein the US 2011/O 145948 A1 Jun. 16, 2011
transgenic plant demonstrates increased yield as compared to corrin-6A reductase catalyzes the reduction of the macro a wild type plant of the same variety which does not comprise cycle of cobalt-precorrin-6X into cobalt-precorrin-6Y. the expression cassette. Cobalt-precorrin-6Y is a co-factor of many enzymes. 0088 As shown in Table 11 below, when transcription of 0.095 The transgenic plant of this embodiment may com the S. cerevisiae geneyKL140W (SEQID NO:39) is targeted prise any polynucleotide encoding a cobalt-precorrin-6A to plastids under control of the PCUbi promoter, transgenic reductase. Preferably, the transgenic plant of this embodi plants were larger than control plants under well-watered ment comprises a polynucleotide encoding a full-length conditions. Table 11 also shows that when YKL140W tran Scription was targeted to mitochondria under control of the polypeptide encoding a cobalt-precorrin-6A reductase hav USP promoter, transgenic plants were larger than control ing a sequence comprising amino acids 1 to 261 of SEQID plants under cycling drought conditions. YKL140W encodes NO:44. a sterol esterase (EC 3.1.1.13). In yeast, sterol esterase medi ates the hydrolysis of stearyl esters and is required for mobi J. Uroporphyrin-III C-methyltransferase lization of stearyl ester, thereby playing a central role in lipid 0096. In another embodiment, the invention provides a metabolism. This enzyme may have weak lipase activity transgenic plant transformed with an expression cassette toward triglycerides under some conditions. comprising, in operative association, an isolated polynucle 0089. The transgenic plant of this embodiment may com otide encoding a promoter capable of enhancing expression prise any polynucleotide encoding a sterol esterase. Prefer in leaves, and an isolated polynucleotide encoding a full ably, the transgenic plant of this embodiment comprises a length uroporphyrin-III C-methyltransferase; wherein the polynucleotide encoding a full-length polypeptide having transgenic plant demonstrates increased yield as compared to sterol esterase activity, wherein the polypeptide has a a wild type plant of the same variety which does not comprise sequence comprising amino acids 1 to 548 of SEQID NO:40. the expression cassette. Table 14 below shows that when the E. coli gene b3803 (SEQID NO:45) is expressed under con H. Succinate-CoA Ligase trol of the USP promoter, transgenic plants were larger than 0090. In another embodiment, the invention provides a control plants under well-watered growth conditions. The transgenic plant transformed with an expression cassette b3803 gene encodes a uroporphyrin-III C-methyltransferase, comprising, in operative association, an isolated polynucle also known as S-adenosyl-L-methionine-dependent Uropor otide encoding a promoter; an isolated polynucleotide encod phyrinogen-III C-methyltransferase (SUMT), an enzyme ing a chloroplast transit peptide; and an isolated polynucle involved in the biosynthesis of siroheme and cobalamin (vita otide encoding a full-length Succinate-CoA ligase min B12). SUMT (EC 2.1.1.107) is a branchpoint enzyme polypeptide; wherein the transgenic plant demonstrates that plays a key role in the biosynthesis of modified tetrapy increased yield as compared to a wild type plant of the same rroles. By catalyzing the transformation ofuroporphyrinogen variety which does not comprise the expression cassette. III into precorrin-2, SUMT controls the flux to compounds 0091. As shown in Table 12 below, when transcription of Such as vitamin B12 and Siroheme, an important co-factor of the Synechocystis gene SLL 1023 (SEQID NO:41) is targeted nitrate reductase and Sulfite reductase enzymes. In plants, to plastids under control of the PCUbi promoter, transgenic uroporphyrinogen III enters the pathway that leads to chlo plansts are larger than control plants, both under cycling rophyll synthesis. drought and well-watered conditions. SLL 1023 is a succi 0097. The transgenic plant of this embodiment may pref nate-CoA ligase (EC 6.2.1.5). Succinate-CoA ligase cata erably comprise any polynucleotide encoding a uroporphy lyzes the reaction of GTP+succinate--CoA=GDP+phos rin-III C-methyltransferase. More preferably, the transgenic phate--succinyl-CoA. This reaction is part of the TCA cycle plant of this embodiment comprises a polynucleotide encod for Sugar metabolism. ing a full-length polypeptide having uroporphyrin-III C-me 0092. The transgenic plant of this embodiment may com thyltransferase activity, wherein the polypeptide comprises a prise any polynucleotide encoding a Succinate-CoA ligase. domain comprising amino acids 101 to 356 of SEQ ID Preferably, the transgenic plant of this embodiment com NO:46:amino acids 97 to 353 of SEQID NO:48; amino acids prises a polynucleotide encoding a Succinate-CoA ligase hav 91 to 346 of SEQ ID NO:50; or amino acids 100 to 355 of ing a sequence comprising amino acids 1 to 401 of SEQID SEQID NO:52. Most preferably, the transgenic plant of this NO:42. embodiment comprises a polynucleotide encoding a uropor phyrin-III C-methyltransferase having a sequence compris I. Cobalt-precorrin-6A Reductase ing amino acids 1 to 393 of SEQID NO:46, amino acids 1 to 368 of SEQ ID NO:48; amino acids 1 to 363 of SEQ ID 0093. In another embodiment, the invention provides a transgenic plant transformed with an expression cassette NO:50, or amino acids 1 to 379 of SEQID NO:52. comprising, in operative association, an isolated polynucle otide encoding a promoter and an isolated polynucleotide K. Isoprenoid Biosynthesis Protein encoding a full-length cobalt-precorrin-6A reductase 0098. In another embodiment, the invention provides a polypeptide; wherein the transgenic plant demonstrates transgenic plant transformed with an expression cassette increased yield as compared to a wild type plant of the same comprising, in operative association, an isolated polynucle variety which does not comprise the expression cassette. otide encoding a promoter capable of enhancing gene expres 0094. As shown in Table 13 below, when the Synechocys Sionin roots and shoots and an isolated polynucleotide encod tis gene SLR0252 (SEQID NO:43) is expressed under con ing a full-length polypeptide having isoprenoid biosynthesis trol of the PcUbi promoter, transgenic plants were larger than activity; wherein the transgenic plant demonstrates increased control plants under cycling drought conditions. SLR0252 is yield as compared to a wild type plant of the same variety a cobalt-precorrin-6A reductase (EC 1.3.1.54). Cobalt-pre which does not comprise the expression cassette. US 2011/O 145948 A1 Jun. 16, 2011
0099 Table 15 below shows that when the E. coli gene comprising, in operative association, an isolated polynucle b3209 (SEQ ID NO:53) is expressed under control of the otide encoding a promoter capable of enhancing gene expres Super promoter, transgenic plants are larger than control sion in roots and shoots; and an isolated polynucleotide plants under well-watered growth conditions. While the spe encoding a full-length polypeptide having LIV-E family cific function of the b3209 protein is not known, it has been branched-chain amino acid transport activity; wherein the shown to increase lycopene production in E. coli when over transgenic plant demonstrates increased yield as compared to expressed. Lycopene is an important component of the pho a wild type plant of the same variety which does not comprise tosystem and a potent anti-oxidant that protects cells from the expression cassette. oxidative damage. In plant cells it is also converted to other 0105. As shown in Table 17 below, when the E. coli gene carotenoids and precursors for plant hormones. Isoprenoid b2682 (SEQ ID NO:61) is expressed under control of the biosynthesis proteins are characterized, in part, by the pres Super promoter, transgenic plants were larger than control ence of a DJ-1 Pfp1 signature sequence. Such signature plants under well-watered growth conditions. The b2682 sequences are exemplified in the isoprenoid biosynthesis pro gene encodes a LIV-E family branched-chain amino acid teins set forth in FIG. 8. transport protein, a membrane protein with five predicted 0100. The transgenic plant of this embodiment may com transmembrane domains involved in the export of L-valine. prise any polynucleotide encoding an isoprenoid biosynthesis These proteins are characterized, in part, by the presence of a protein. Preferably, the transgenic plant of this embodiment AZIC signature sequence. comprises a polynucleotide encoding a full-length polypep 0106 The transgenic plant of this embodiment may com tide having isoprenoid biosynthesis activity, wherein the prise any polynucleotide encoding a branched-chain amino polypeptide comprises a DJ-1 Pfp1 signature sequence acid transporter. Preferably, the transgenic plant of this selected from the group consisting of amino acids 46 to 208 of embodiment comprises a polynucleotide encoding a full SEQID NO:54: amino acids 32 to 189 of SEQID NO:56; and length polypeptide having LIV-E family branched-chain amino acids 25 to 151 of SEQID NO:58. Most preferably, the amino acid transport activity, wherein the polypeptide com transgenic plant of this embodiment comprises a polynucle prises a AZIC signature sequence comprising amino acids 23 otide encoding an isoprenoid biosynthesis protein having a to 167 of SEQ ID NO:62. Most preferably, the transgenic sequence comprising amino acids 1 to 220 of SEQID NO:54, plant of this embodiment comprises a polynucleotide encod amino acids 1 to 231 of SEQID NO:56, or amino acids 1 to ing a branched-chain amino acid transporter having a 161 of SEQID NO:58. sequence comprising amino acids 1 to 245 of SEQID NO:62. L. LysE Type Translocator N. DNA-Binding Protein 0101. In another embodiment, the invention provides a 0107. In another embodiment, the invention provides a transgenic plant transformed with an expression cassette transgenic plant transformed with an expression cassette comprising, in operative association, an isolated polynucle comprising, in operative association, an isolated polynucle otide encoding a promoter capable of enhancing gene expres otide encoding a promoter capable of enhancing gene expres Sionin roots and shoots and an isolated polynucleotide encod sion in roots and shoots; and an isolated polynucleotide ing a full-length polypeptide having LysE type translocator encoding a truncated DNA-binding polypeptide; wherein the activity; wherein the transgenic plant demonstrates increased transgenic plant demonstrates increased yield as compared to yield as compared to a wild type plant of the same variety a wild type plant of the same variety which does not comprise which does not comprise the expression cassette. the expression cassette. 0102. As shown in Table 16 below, when the E. coli gene 0108. As shown in Table 18 below, when the E. coli gene b2578 (SEQ ID NO:59) is expressed under control of the b3285 (SEQ ID NO:63) is expressed under control of the Super promoter, transgenic plants were larger than control Super promoter, transgenic plants are larger than control plants under well-watered conditions. The b2578 gene plants under well-watered growth conditions. The b3285 encodes a LysE type translocator protein, a membrane protein gene encodes a truncated DNA-binding protein comprising with six predicted transmembrane domains, which is the C-terminal portion of the E. coli SMF protein (public involved in maintaining intercellular levels of L-lysine by database accession number YP 026211), and may facilitate exporting excess L-lysine from the cell. These proteins are the access of DNA-modifying proteins to genomic DNA. characterized, in part, by the presence of a LysE signature Such DNA-binding proteins are characterized, in part, by the Sequence. presence of an SMF signature sequence. 0103) The transgenic plant of this embodiment may com 0109 The transgenic plant of this embodiment may com prise any polynucleotide encoding a LysE type translocator. prise any polynucleotide homologous to the polynucleotide Preferably, the transgenic plant of this embodiment com encoding the truncated DNA-binding protein of SEQ ID prises a polynucleotide encoding a full-length polypeptide NO:64. Preferably, the transgenic plant of this embodiment having LysE type translocator activity, wherein the polypep comprises a polynucleotide encoding a DNA-binding protein tide comprises a LysE signature sequence comprising amino having a sequence comprising amino acids 1 to 102 of SEQ acids 14 to 195 of SEQ ID NO:60. Most preferably, the ID NO:64. transgenic plant of this embodiment comprises a polynucle otide encoding a LySE type translocator having a sequence O.Y.Sc.J/FliF Protein comprising amino acids 1 to 195 of SEQID NO:60. 0110. In another embodiment, the invention provides a transgenic plant transformed with an expression cassette M. Branched-Chain Amino Acid Transporter comprising, in operative association, an isolated polynucle 0104. In another embodiment, the invention provides a otide encoding a promoter capable of enhancing gene expres transgenic plant transformed with an expression cassette sion in roots and shoots; and an isolated polynucleotide US 2011/O 145948 A1 Jun. 16, 2011
encoding a full-lengthysc/FliF family polypeptide; wherein 0116. In another embodiment, the invention provides a the transgenic plant demonstrates increased yield as com transgenic plant transformed with an expression cassette pared to a wild type plant of the same variety which does not comprising, in operative association, an isolated polynucle comprise the expression cassette. otide encoding a promoter and an isolated polynucleotide 0111. As shown in Table 19 below, when the E. coli gene encoding a lumazine synthase polypeptide which does not b1938 (SEQ ID NO:65), a Ysc/FliF family protein, is comprise a plastid targeting sequence; wherein the transgenic expressed under control of the Super promoter, transgenic plant demonstrates increased yield as compared to a wildtype pinats are larger than control plants. The Ysc J/FliF family plant of the same variety which does not comprise the expres proteins stabilize membrane proteins and complexes such as sion cassette. transporters, which provide cells with important compounds 0117. As shown in Tables 21 and 22 below, when the for cell growth. Ysc J/FliF family proteins are characterized, Synechocystis gene SLL 1282 (SEQ ID NO:71), a lumazine in part, by the presence of a YSc.J. FliF signature sequence. synthase, is expressed under control of the PcUbi promoter, 0112 The transgenic plant of this embodiment may com transgenic plants demonstrate increased yield as compared to prise any polynucleotide encoding an Ysc J/FliF family pro a wild type plant of the same variety which does not comprise tein. Preferably, the transgenic plant of this embodiment com the SLL 1282 gene. Lumazine synthase enzymes are charac prises a polynucleotide encoding a full-length polypeptide terized, in part, by the presence of a DMRL synthase signa having Ysc J/FliFactivity, wherein the polypeptide comprises ture sequence. Such signature sequences are exemplified in a first YscJ/FliF signature sequence comprising amino acids the lumazine synthase proteins set forth in FIG. 13. 17 to 225 of SEQID NO:66 and a secondYsc/FliF signature 0118. The expression cassette employed in the transgenic sequence comprising amino acids 250 to 429 of SEQ ID plant of this embodiment may comprise any polynucleotide NO:66. Most preferably, the transgenic plant of this embodi encoding a lumazine synthase polypeptide which does not ment comprises a polynucleotide encoding aYsc J/FliF family comprise a Subcellular targeting peptide. Preferably, the protein having a sequence comprising amino acids 1 to 552 of transgenic plant of this embodiment comprises a polynucle SEQID NO:66. otide encoding a full-length polypeptide having lumazine synthase activity, wherein the polypeptide comprises a P. Riboflavin Biosynthetic Genes DMRL synthase signature sequence selected from the group 0113. In another embodiment, the invention provides a consisting of amino acids 12 to 158 of SEQID NO:72; amino transgenic plant transformed with an expression cassette acids 83 to 226 of SEQID NO:74; amino acids 70 to 213 of comprising, in operative association, a promoter and an iso SEQID NO:76, amino acids 70 to 215 of SEQID NO:78. In lated polynucleotide encoding a full length GTP cyclohydro accordance with the invention, when any of the plant lase II polypeptide which does not comprise a Subcellular lumazine synthases set forth in SEQID NOs: 100,76, or 78 is targeting sequence; wherein the transgenic plant demon employed in the expression cassette, the polynucleotide strates increased yield as compared to a wild type plant of the sequences encoding the Subcellular targeting peptide (amino same variety that does not comprise the expression cassette. acids 1 to 53 of SEQID NO:74; amino acids 1 to 56 of SEQ 0114. As shown in Table 20 below, when the Synechocys ID NO:76, and amino acids 1 to 80 of SEQ ID NO:78) are tis gene SLL 1894 (SEQID NO:67), a GTP cyclohydrolase II, deleted. More preferably, the transgenic plant of this embodi was expressed under control of the PcUbi promoter with no ment comprises a polynucleotide encoding a lumazine Syn Subcellular targeting, transgenic plants were larger than con thase comprising amino acids 1 to 164 of SEQ ID NO:72, trol plants under water limited conditions. GTP cyclohydrase amino acids 54 to 228 of SEQID NO:74, amino acids 57 to II enzymes are characterized, in part, by the presence of a 217 of SEQID NO:76, or amino acids 81 to 217 of SEQID DHBP synthase signature sequence in the N-terminus and a NO:78. GTP cyclohydrolase II signature sequence in the C-terminus. Such signature sequences are exemplified in the GTP cyclo Q. Vitamin B6 Biosynthetic Genes hydrolase II proteins set forth in FIG. 12. 0119. In another embodiment, the invention provides a 0115 The expression cassette employed in the transgenic transgenic plant transformed with an expression cassette plant of this embodiment may comprise any polynucleotide comprising, in operative association, an isolated polynucle encoding a full-length polypeptide having GTP cyclohydro otide encoding a promoter capable of enhancing gene expres lase II activity which does not comprise a Subcellular target Sionin roots and shoots and an isolated polynucleotide encod ing peptide. Preferably, the GTP cyclohydrolase II polypep ing a full length polypeptide which is capable of enhancing tide comprises a DHBP synthase domain selected from the PLP synthesis. In this embodiment, the expression cassette groups consisting of amino acids 5 to 202 of SEQID NO:68 may optionally further comprise an isolated polynucleotide and amino acids 120 to 317 of SEQ ID NO:70 and a GTP encoding a mitochondrial or plastid transit peptide. Poly cyclohydrolase II domain selected from the group consisting nucleotides comprising any gene of the vitamin B6 synthetic of amino acids 207 to 377 of SEQID NO:68 and amino acids pathway are suitable for use in this embodiment of the inven 322 to 491 of SEQID NO:70. In accordance with the inven tion. For example, the vitamin B6 synthetic gene may be tion, when the G. max GTP cyclohydrolase II set forth in SEQ pdxY (e.g., SEQ ID NO:79), pdxH (e.g., SEQ ID NO:81), ID NO:70 is employed in the expression cassette, polynucle pdxK (e.g., SEQID NOs:83, 85, and 87), yfei (e.g., SEQID otide sequence encoding the Subcellular targeting peptide NOs: 89 and 91), Yn8fp (SEQID NO:93): pdxJ (e.g., SEQID (amino acids 1 to 49 of SEQ ID NO:70) is deleted. More NOs: 95 and 97), pdx1/sor1 (e.g., SEQIS NOs: 99, 101, 103, preferably, the transgenic plant of this embodiment com 105, 107, 109, 111, 113, 115, 117, or 119). prises a polynucleotide encoding a GTP cyclohydrolase II I0120 When the transgenic plant comprises the vitamin B6 polypeptide having a sequence comprising amino acids 1 to biosynthetic gene PdxY, the promoter is preferably a root- or 556 of SEQID NO:68 or amino acids 50 to 544 of SEQID shoot-specific promoter and the expression cassette further NO: 70. comprises a polynucleotide encoding a plastid transit peptide. US 2011/O 145948 A1 Jun. 16, 2011
As shown in Tables 23 and 24 below, when the E. coli gene the isolated polynucleotide of the invention is an isolated b1636 (SEQID NO:79), designated pdxY, is targeted to the polynucleotide encoding a polypeptide having an amino acid plastid under control of the Super promoter, transgenic plants sequence selected from the group consisting of SEQ ID were larger than control plants under well watered and water NO:4: SEQID NO:6; SEQID NO:10, SEQID NO:12; SEQ limited conditions. ID NO:14: SEQID NO:18: SEQID NO:20: SEQID NO:24: 0121 The PdxY gene encodes a PL kinase which com SEQ ID NO:26: SEQ ID NO:30; SEQ ID NO:32: SEQ ID prises a phosphomethyl-pyrimidine kinase domain. The NO:34: SEQ ID NO:38: SEQ ID NO:54: SEQ ID NO:60; transgenic plant of this embodiment may comprise any poly SEQ ID NO:62; SEQ ID NO:64; SEQ ID NO:70, SEQ ID nucleotide encoding a PL kinase. Preferably, the transgenic NO:74; SEQ ID NO:76; and SEQ ID NO:78. A polynucle plant of this embodiment comprises a polynucleotide encod otide of the invention can be isolated using standard molecu ing a full-length polypeptide having PL kinase activity, lar biology techniques and the sequence information pro wherein the polypeptide comprises a phosphomethyl-pyrimi vided herein, for example, using an automated DNA dine kinase signature sequence Such as amino acids 61 to 259 synthesizer. of SEQ ID NO:80. More preferably, the transgenic plant of 0.125. The isolated polynucleotides of the invention this embodiment comprises a polynucleotide encoding a PL include homologs of the polynucleotides of Table 1. kinase having a sequence comprising amino acids 1 to 287 of “Homologs are defined herein as two nucleic acids or SEQID NO:80. polypeptides that have similar, or Substantially identical, 0122. As shown in Table 25 below, when the Synechocys nucleotide or amino acid sequences, respectively. Homologs tis gene SLL1779 (SEQ ID NO:95), designated pdx.J., is include allelic variants, analogs, and orthologs, as defined expressed under control of the PcUbi promoter, with or with below. As used herein, the term “analogs’ refers to two out Subcellular targeting, transgenic plants are larger than nucleic acids that have the same or similar function, but that control plants under well-watered conditions. Table 26 shows have evolved separately in unrelated organisms. As used that when SLL1779 (SEQID NO:95) is targeted to mitochon herein, the term “orthologs’ refers to two nucleic acids from dria under control of the PCUbi promoter, transgenic plants different species, but that have evolved from a common are larger than control plants when tested under water-limited ancestral gene by speciation. The term homolog further conditions. Accordingly, when the transgenic plant comprises encompasses nucleic acid molecules that differ from one of the Pdx.J. gene, the expression cassette may optionally further the nucleotide sequences shown in Table 1 due to degeneracy comprise a polynucleotide encoding a mitochondrial or plas of the genetic code and thus encode the same polypeptide. tid transit peptide. As set forth above, the Pdx.J enzyme acts in 0.126 To determine the percent sequence identity of two a concerted manner with the PdxA enzyme to form PNP. amino acid sequences (e.g., one of the polypeptide sequences Amino acids 2 to 218 of SEQID NO:96 represent a signature of Table 1 and a homolog thereof), the sequences are aligned sequence of the Pdx.J. gene from Synechocystis sp. PCC6803. for optimal comparison purposes (e.g., gaps can be intro Preferably, the Pdx.J. gene employed in the expression cassette duced in the sequence of one polypeptide for optimal align of this embodiment encodes a polypeptide amino acids 1 to ment with the other polypeptide or nucleic acid). The amino 221 of SEQ ID NO:96 or amino acids 1 to 243 of SEQ ID acid residues at corresponding amino acid positions are then NO:98. compared. When a position in one sequence is occupied by 0123. The invention further provides a seed which is true the same amino acid residue as the corresponding position in breeding for the expression cassettes (also referred to herein the other sequence then the molecules are identical at that as “transgenes') described herein, wherein transgenic plants position. The same type of comparison can be made between grown from said seed demonstrate increased yield as com two nucleic acid sequences. pared to a wild type variety of the plant. The invention also I0127 Preferably, the isolated amino acid homologs, ana provides a product produced by or from the transgenic plants logs, and orthologs of the polypeptides of the present inven expressing the polynucleotide, their plant parts, or their seeds. tion are at least about 50-60%, preferably at least about The product can be obtained using various methods well 60-70%, and more preferably at least about 70-75%, 75-80%, known in the art. As used herein, the word “product' includes, 80-85%, 85-90%, or 90-95%, and most preferably at least but not limited to, a foodstuff, feedstuff, a food supplement, about 96%, 97%, 98%, 99%, or more identical to an entire feed Supplement, fiber, cosmetic or pharmaceutical. Food amino acid sequence identified in Table 1. In another pre stuffs are regarded as compositions used for nutrition or for ferred embodiment, an isolated nucleic acid homolog of the Supplementing nutrition. Animal feedstuffs and animal feed invention comprises a nucleotide sequence which is at least Supplements, in particular, are regarded as foodstuffs. The about 40-60%, preferably at least about 60-70%, more pref invention further provides an agricultural product produced erably at least about 70-75%, 75-80%, 80-85%, 85-90%, or by any of the transgenic plants, plant parts, and plant seeds. 90-95%, and even more preferably at least about 95%, 96%, Agricultural products include, but are not limited to, plant 97%, 98%, 99%, or more identical to a nucleotide sequence extracts, proteins, amino acids, carbohydrates, fats, oils, shown in Table 1. polymers, vitamins, and the like. I0128. For the purposes of the invention, the percent 0.124. The invention also provides an isolated polynucle sequence identity between two nucleic acid or polypeptide otide which has a sequence selected from the group consist sequences is determined using Align 2.0 (Myers and Miller, ing of SEQID NO:3: SEQID NO:5; SEQID NO:9; SEQID CABIOS (1989) 4:11-17) with all parameters set to the NO:11; SEQ ID NO:13; SEQ ID NO:17; SEQ ID NO:19; default settings or the Vector NTI Advance 10.3.0 (PC) soft SEQ ID NO:23: SEQ ID NO:25; SEQ ID NO:29; SEQ ID ware package (Invitrogen, 1600 Faraday Ave., Carlsbad, NO:31: SEQ ID NO:33; SEQ ID NO:37: SEQ ID NO:45; Calif. 92008). For percent identity calculated with Vector SEQ ID NO:53: SEQ ID NO:59; SEQ ID NO:61; SEQ ID NTI, a gap opening penalty of 15 and a gap extension penalty NO:63; SEQ ID NO:65; SEQ ID NO:69; SEQ ID NO:73; of 6.66 are used for determining the percent identity of two SEQ ID NO:75; and SEQ ID NO:77. Also encompassed by nucleic acids. A gap opening penalty of 10 and a gap exten US 2011/O 145948 A1 Jun. 16, 2011 sion penalty of 0.1 are used for determining the percent iden acyl-carrier protein; and c) an expression cassette compris tity of two polypeptides. All other parameters are set at the ing, in operative association, an isolated polynucleotide default settings. For purposes of a multiple alignment (Clustal encoding a promoter, an isolated polynucleotide encoding a W algorithm), the gap opening penalty is 10, and the gap Subcellular targeting peptide; and an isolated polynucleotide extension penalty is 0.05 with blosum62 matrix. It is to be encoding an acyltransferase polypeptide. understood that for the purposes of determining sequence 0.132. In another embodiment, the recombinant expression identity when comparing a DNA sequence to an RNA vector of the invention comprises an isolated polynucleotide sequence, a thymidine nucleotide is equivalent to a uracil having a sequence selected from the group consisting of SEQ nucleotide. ID NO:4: SEQ ID NO:6; SEQ ID NO:10, SEQ ID NO:12; 0129. Nucleic acid molecules corresponding to homologs, SEQ ID NO:14: SEQ ID NO:18: SEQ ID NO:20: SEQ ID analogs, and orthologs of the polypeptides listed in Table 1 NO:24: SEQ ID NO:26: SEQ ID NO:30; SEQ ID NO:32: can be isolated based on their identity to said polypeptides, SEQ ID NO:34: SEQ ID NO:38: SEQ ID NO:54: SEQ ID using the polynucleotides encoding the respective polypep NO:60; SEQ ID NO:62; SEQ ID NO:64; SEQ ID NO:70; tides or primers based thereon, as hybridization probes SEQ ID NO:74; SEQ ID NO:76; and SEQ ID NO:78. In according to standard hybridization techniques under Strin addition, the recombinant expression vector of the invention gent hybridization conditions. As used herein with regard to comprises an isolated polynucleotide encoding a polypeptide hybridization for DNA to a DNA blot, the term “stringent having an amino acid sequence selected from the group con conditions’ refers to hybridization overnight at 60° C. in sisting SEQID NO:4: SEQID NO:6; SEQ ID NO:10: SEQ 10xDenhart's solution, 6xSSC, 0.5% SDS, and 100 g/ml ID NO:12; SEQID NO:14: SEQID NO:18: SEQID NO:20; denatured salmon sperm DNA. Blots are washed sequentially SEQ ID NO:24: SEQ ID NO:26: SEQ ID NO:30; SEQ ID at 62° C. for 30 minutes each time in 3xSSC/0.1% SDS, NO:32: SEQ ID NO:34: SEQ ID NO:38: SEQ ID NO:54; followed by 1XSSC/0.1% SDS, and finally 0.1xSSC/0.1% SEQ ID NO:60; SEQ ID NO:62; SEQ ID NO:64; SEQ ID SDS. As also used herein, in a preferred embodiment, the NO:70; SEQID NO:74; SEQID NO:76; and SEQID NO:78. phrase “stringent conditions” refers to hybridization in a I0133. The recombinant expression vector of the invention 6xSSC solution at 65° C. In another embodiment, “highly may also include one or more regulatory sequences, selected stringent conditions' refers to hybridization overnight at 65° on the basis of the host cells to be used for expression, which C. in 10xDenhart's solution, 6xSSC, 0.5% SDS and 100 is in operative association with the isolated polynucleotide to ug/ml denatured salmon sperm DNA. Blots are washed be expressed. As used herein with respect to a recombinant sequentially at 65° C. for 30 minutes each time in 3xSSC/0. expression vector, “in operative association” or “operatively 1% SDS, followed by 1XSSC/0.1% SDS, and finally 0.1x linked' means that the polynucleotide of interest is linked to SSC/0.1% SDS. Methods for performing nucleic acid hybrid the regulatory sequence(s) in a manner which allows for izations are well known in the art. expression of the polynucleotide when the vector is intro 0130. The isolated polynucleotides employed in the inven duced into the host cell (e.g., in a bacterial or plant host cell). tion may be optimized, that is, genetically engineered to The term “regulatory sequence' is intended to include pro increase its expression in a given plant or animal. To provide moters, enhancers, and other expression control elements plant optimized nucleic acids, the DNA sequence of the gene (e.g., polyadenylation signals). can be modified to: 1) comprise codons preferred by highly 0.134. As set forth above, certain embodiments of the expressed plant genes; 2) comprise an A+T content in nucle invention employ promoters that are capable of enhancing otide base composition to that Substantially found in plants; gene expression in leaves. In some embodiments, the pro 3) form a plant initiation sequence; 4) to eliminate sequences moter is a leaf-specific promoter. Any leaf-specific promoter that cause destabilization, inappropriate polyadenylation, may be employed in these embodiments of the invention. degradation and termination of RNA, or that form secondary Many such promoters are known, for example, the USP pro structure hairpins or RNA splice sites; or 5) elimination of moter from Vicia faba (SEQID NO:123 or SEQID NO:124, antisense open reading frames. Increased expression of Baeumlein et al. (1991) Mol. Gen. Genet. 225, 459-67), pro nucleic acids in plants can be achieved by utilizing the distri moters of light-inducible genes Such as ribulose-1.5-bispho bution frequency of codon usage in plants in general or in a sphate carboxylase (rbcS promoters), promoters of genes particular plant. Methods for optimizing nucleic acid expres encoding chlorophyll a?b-binding proteins (Cab), Rubisco sion in plants can be found in EPA 0359472: EPA 0385962: activase, B-subunit of chloroplast glyceraldehyde 3-phos PCT Application No. WO 91/16432; U.S. Pat. No. 5,380,831; phate dehydrogenase from A. thaliana, (Kwon et al. (1994) U.S. Pat. No. 5,436,391; Perlack et al., 1991, Proc. Natl. Plant Physiol. 105,357-67) and other leaf specific promoters Acad. Sci. USA 88:3324-3328; and Murray et al., 1989, such as those identified in Aleman, I. (2001) Isolation and Nucleic Acids Res. 17:477-498. characterization of leaf-specific promoters from alfalfa 0131 The invention further provides a recombinant (Medicago sativa), Masters thesis, New Mexico State Uni expression vector which comprises an expression cassette versity, Los Cruces, N. Mex., and the like a constitutive pro selected from the group consisting of a) an expression cas moter. Constitutive promoters are active under most condi sette comprising, in operative association, an isolated poly tions. Examples of constitutive promoters Suitable for use in nucleotide encoding a promoter, an isolated polynucleotide these embodiments include the parsley ubiquitin promoter encoding a Subcellular targeting peptide, and an isolated from Petroselinum crispum described in WO 2003/102198 polynucleotide encoding a full-length phosphatidate cytidy (SEQID NO:121); the CaMV 19S and 35S promoters, the SX lyltransferase polypeptide; b) an expression cassette compris CaMV 35S promoter, the Sep 1 promoter, the rice actin pro ing, in operative association, an isolated polynucleotide moter, the Arabidopsis actin promoter, the maize ubiquitin encoding a promoter capable of enhancing expression in promoter, pEmu, the figwort mosaic virus 35S promoter, the leaves, an isolated polynucleotide encoding a mitochondrial Smas promoter, the super promoter (U.S. Pat. No. 5,955,646), transit peptide, and an isolated polynucleotide encoding an the GRP1-8 promoter, the cinnamyl alcohol dehydrogenase US 2011/O 145948 A1 Jun. 16, 2011 promoter (U.S. Pat. No. 5,683.439), promoters from the transgenic corn seed of the invention may be made using T-DNA of Agrobacterium, Such as mannopine synthase, Agrobacterium transformation, as described in U.S. Pat. Nos. nopaline synthase, and octopine synthase, the Small Subunit 5,591,616; 5,731,179; 5,981,840; 5,990,387; 6,162,965; of ribulose biphosphate carboxylase (ssuRUBISCO) pro 6.420,630, U.S. patent application publication number 2002/ moter, and the like. 01.04132, and the like. Transformation of soybean can be 0135. In other embodiments of the invention, a root or performed using for example any of the techniques described shoot specific promoter is employed. For example, the Super in European Patent No. EP0424047, U.S. Pat. No. 5,322,783, promoter (SEQID NO:122) provides high level expression in European Patent No. EP 0397 687, U.S. Pat. No. 5,376,543, both root and shoots (Ni et al. (1995) Plant J. 7: 661-676). or U.S. Pat. No. 5,169,770. A specific example of wheat Other root specific promoters include, without limitation, the transformation can be found in PCT Application No. WO TobRB7 promoter (Yamamoto et al. (1991) Plant Cell 3, 93/07256. Cotton may be transformed using methods dis 371-382), the roll) promoter (Leach et al. (1991) Plant Sci closed in U.S. Pat. Nos. 5,004,863; 5,159,135; 5,846,797, and ence 79, 69-76); CaMV 35S Domain A (Benfey et al. (1989) the like. Rice may be transformed using methods disclosed in Science 244, 174-181), and the like. U.S. Pat. Nos. 4,666,844; 5,350,688: 6,153,813; 6,333,449; 0136. In accordance with the invention, a chloroplast tran sit sequence refers to a nucleotide sequence that encodes a 6.288,312; 6,365,807; 6,329,571, and the like. Canola may be chloroplast transit peptide. Chloroplast targeting sequences transformed, for example, using methods such as those dis are known in the art and include the chloroplast small subunit closed in U.S. Pat. Nos. 5,188,958; 5,463,174; 5,750,871; ofribulose-1,5-bisphosphate carboxylase (Rubisco) (de Cas EP1566443; WO02/00900; and the like. Other plant transfor tro Silva Filho et al. (1996) Plant Mol. Biol. 30:769-780; mation methods are disclosed, for example, in U.S. Pat. Nos. Schnell et al. (1991) J. Biol. Chem. 266(5):3335-3342): 5,932,782; 6,153,811: 6,140,553; 5,969,213; 6,020,539, and 5-(enolpyruvyl)shikimate-3-phosphate synthase (EPSPS) the like. Any plant transformation method suitable for insert (Archer et al. (1990).J. Bioenerg. Biomemb. 22(6):789-810); ing a transgene into a particular plant may be used in accor tryptophan synthase (Zhao et al. (1995) J. Biol. Chem. 270 dance with the invention. (11):6081-6087); plastocyanin (Lawrence et al. (1997) J. 0.139. According to the present invention, the introduced Biol. Chem. 272(33):20357-20363); chorismate synthase polynucleotide may be maintained in the plant cell stably if it (Schmidt et al. (1993).J. Biol. Chem. 268(36):27447-27457); is incorporated into a non-chromosomal autonomous repli ferredoxin NADP+ oxidoreductase (Jansen et al. (1988) Curr. con or integrated into the plant chromosomes. Alternatively, Genetics 13:517-522) (SEQ ID NO:13): nitrite reductase the introduced polynucleotide may be present on an extra (Back et al (1988) MGG 212:20-26) and the light harvesting chromosomal non-replicating vector and may be transiently chlorophyll a?b binding protein (LHBP) (Lamppa et al. expressed or transiently active. (1988) J. Biol. Chem. 263:14996-14999). See also Von Heijneet al. (1991) Plant Mol. Biol. Rep. 9:104-126: Clarket 0140. The invention is also embodied in a method of pro al. (1989) J. Biol. Chem. 264:17544-17550; Della-Cioppa et ducing a transgenic plant comprising at least one polynucle al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) otide listed in Table 1, wherein expression of the polynucle Biochem. Biophys. Res. Commun. 196:1414-1421; and Shah otide in the plant results in the plants increased growth and/or et al. (1986) Science 233:478-481. yield under normal or water-limited conditions and/or 0.137 As defined herein, a mitochondrial transit sequence increased tolerance to an environmental stress as compared to refers to a nucleotide sequence that encodes a mitochondrial a wild type variety of the plant comprising the steps of: (a) presequence and directs the protein to mitochondria. introducing into a plant cell an expression cassette described Examples of mitochondrial presequences include groups above, (b) regenerating a transgenic plant from the trans consisting of ATPase subunits, ATP synthase subunits, formed plant cell; and selecting higher-yielding plants from Rieske-FeS protein, Hsp60, malate dehydrogenase, citrate the regenerated plant sells. The plant cell may be, but is not synthase, aconitase, isocitrate dehydrogenase, pyruvate limited to, a protoplast, gamete producing cell, and a cell that dehydrogenase, malic enzyme, glycine decarboxylase, serine regenerates into a whole plant. As used herein, the term hydroxymethyl transferase, isovaleryl-CoA dehydrogenase “transgenic’ refers to any plant, plant cell, callus, plant tissue, and Superoxide dismutase. Such transit peptides are known in or plant part, that contains the expression cassette described the art. See, for example, Von Heijne et al. (1991) Plant Mol. above. In accordance with the invention, the expression Biol. Rep. 9:104-126: Clark et al. (1989) J. Biol. Chem. casette is stably integrated into a chromosome or stable extra 264:17544-17550; Romer et al. (1993) Biochem. Biophys. chromosomal element, so that it is passed on to Successive Res. Commun. 196:1414-1421: Faivre-Nitschke etal (2001) generations. Eur J Biochem 268 1332-1339 and Shah et al. (1986) Science 233: 478-481. 0.141. The effect of the genetic modification on plant 0.138. In a preferred embodiment of the present invention, growth and/or yield and/or stress tolerance can be assessed by the polynucleotides listed in Table 1 are expressed in plant growing the modified plant under normal and/or less than cells from higher plants (e.g., the spermatophytes, such as Suitable conditions and then analyzing the growth character crop plants). A polynucleotide may be “introduced into a istics and/or metabolism of the plant. Such analytical tech plant cell by any means, including transfection, transforma niques are well known to one skilled in the art, and include tion or transduction, electroporation, particle bombardment, measurements of dry weight, wet weight, seed weight, seed agroinfection, and the like. Suitable methods for transform number, polypeptide synthesis, carbohydrate synthesis, Syn ing or transfecting plant cells are disclosed, for example, thesis, evapotranspiration rates, general plant and/or crop using particle bombardment as set forth in U.S. Pat. Nos. yield, flowering, reproduction, seed setting, root growth, res 4,945,050; 5,036,006; 5,100,792; 5,302,523: 5,464,765; piration rates, photosynthesis rates, metabolite composition, 5,120,657; 6,084,154; and the like. More preferably, the and the like. US 2011/O 145948 A1 Jun. 16, 2011
0142. The invention is further illustrated by the following 1997, Nucleic Acids Res. 25:3389-3402). One homolog from examples, which are not to be construed in any way as impos sunflower were identified. The amino acid relatedness of ing limitations upon the scope thereof. these sequences is shown in FIG. 6. 0150. The b3803 gene from E. coli encodes a uroporphy Example 1 rin-III C-methyltransferase. The full-length amino acid sequence of the functional homologs of b3803 from Table 11 Characterization of Genes were blasted against proprietary databases of cDNAS at an e 0143 YBR029C (SEQ ID NO:1), YKL192C (SEQ ID value of e-10 (Altschulet al., 1997, Nucleic Acids Res. 25: NO:7), YDRO18C (SEQ ID NO:15), B2341 (SEQ ID 3389-3402). One homolog from Brassica napus, one NO:21), B0452 (SEQ ID NO:27), YNL202W (SEQ ID homolog from Glycine max, and one homolog from Zea mays NO:35), YKL140W (SEQ ID NO:39), SLL1023 (SEQ ID were identified. Homolog cDNAs were isolated from propri NO:41), SLR0252 (SEQIDNO:43), b3803 (SEQID NO:45), etary libraries of the respective species using known methods. b3209 (SEQ ID NO:53), b2578 (SEQ ID NO:59), b2682 Sequences were processed and annotated using bioinformat (SEQ ID NO:61), b3285 (SEQ ID NO:63), b1938 (SEQ ID ics analyses. The amino acid relatedness of these sequences is NO:65), SLL1894 (SEQ ID NO:67), SLL 1282 (SEQ ID shown in FIG. 7. NO:5), b1636 (SEQ ID NO:79) and SLR1779 (SEQ ID 0151. The b3209 gene from E. coli encodes an isoprenoid NO: 95) were cloned using standard recombinant techniques. biosynthesis protein. The full-length amino acid sequence of The functionality of each lead gene was predicted by com functional homologs of b3209 from Table 15 were blasted paring the amino acid sequence of the gene with other genes against proprietary databases of cDNAs at an evalue of e-10 of known functionality. Homolog cDNAs were isolated from (Altschul et al., 1997, Nucleic Acids Res. 25: 3389-3402). proprietary libraries of the respective species using known One homolog from Glycine max and one homolog from methods. Sequences were processed and annotated using bio Helianthus annuus were identified. The amino acid related informatics analyses. ness of these sequences is shown in FIG. 8. 0144. The YBR029C (SEQ ID NO:2) from S. cerevisiae 0152 The SLL 1894 (SEQ ID NO:67) gene from Syn encodes a phosphatidate cytidylyltransferase. The DNA echocystis sp. PCC 6803 encodes a GTP cyclohydrase II. The sequence of this gene was blasted against proprietary data full-length DNA sequence of SLL 1894 was blasted against a bases of canola and maize cDNAs at an e value of e-10 proprietary databases of soybean cloNAs at an evalue of e-10 (Altschul et al., 1997, Nucleic Acids Res. 25: 3389-3402). (Altschulet al., 1997, Nucleic Acids Res. 25:3389-3402) and One homolog from canola and one homolog from maize were one homolog from soybean was identified. The amino acid identified. The amino acid relatedness of these sequences is relatedness of these sequences is shown in FIG. 12. indicated in the alignments shown in FIG. 1. 0153. The SLL 1282 gene (SEQ ID NO:71) from Syn (0145 The YKL192O (SEQ ID NO:8) from S. cerevisiae echocystis sp. PCC 6803 encodes a lumazine synthase. The encodes an acyl-carrier protein. The DNA sequence of this full-length DNA sequence of this gene was blasted against gene was blasted against proprietary databases of plant proprietary databases of Soybean and maize cDNAS at an e cDNAs at an evalue of e-10 (Altschul et al., 1997, Nucleic value of e-10 (Altschulet al., 1997, Nucleic Acids Res. 25: Acids Res. 25: 3389-3402). One homolog each from canola, 3389-3402). One homolog from soybean and two homologs soybean, and sunflower were identified. The amino acid relat from maize were identified. The amino acid relatedness of edness of these sequences is indicated in the alignments these sequences is shown in FIG. 13. shown in FIG. 2. 0146 The ofYDR018C (SEQID NO:16) from S. cerevi Example 2 siae encodes an acyltransferase protein. The DNA sequence of this gene was blasted against proprietary databases of Overexpression of Lead Genes in Plants soybean and maize cDNAs at an evalue of e-10 (Altschulet 0154 Each of the genes described in Example 1 was al., 1997, Nucleic Acids Res. 25: 3389-3402). One homolog ligated into an expression cassette using known methods. from soybean and one from maize were identified. The amino Four different promoters were used to control expression of acid relatedness of these sequences is shown in FIG. 3. the transgenes in Arabidopsis: the parsley ubiquitin promoter 0147 The B2341 (SEQ ID NO:21) gene from E. coli (SEQID NO:121) designated “PCUbi” in Tables 2 to 26: the encodes a bifunctional anaerobic fatty acid oxidation com super promoter (SEQ ID NO: 122) designated “Super in plex polypeptide. The DNA sequence of this gene was blasted Tables 2 to 26: the USP promoter from Vicia faba (SEQ ID against a proprietary maize cDNA database at an evalue of NO:124), designated “USP” in Tables 2 to 26 was used for e-10 (Altschul et al., 1997, Nucleic Acids Res. 25: 3389 expression of genes from prokaryotes or SEQID NO:123 was 3402). Two homologs from maize were identified. The amino used for expression of genes from S. cerevisiae). For selective acid relatedness of these sequences is shown in FIG. 4. targeting of the polypeptides, the mitochondrial transit pep 0148. The B0452 (SEQ ID NO:28) from E. coli encodes tide from an A. thaliana gene encoding mitochondrial isov an acyl-CoA thioesterase protein. The DNA sequence of this aleryl-CoA dehydrogenase designated “Mit” in Tables 2 to gene was blasted against proprietary databases of plant 26, SEQID NO:126 was used for expression of genes from cDNAs at an evalue of e-10 (Altschul et al., 1997, Nucleic prokaryotes or SEQID NO:128 was used for expression of Acids Res. 25: 3389-3402). Two homologs from canola and genes from S. cerevisiae. In addition, for selective targeting of one from soybean were identified. The amino acid relatedness polypeptides to the chloroplast, the transit peptide of an of these sequences is shown in FIG. 5. Spinacia oleracea gene encoding ferredoxin nitrite reductase 0149 YNL202W (SEQ ID NO:36) from S. cerevisiae designated “Chlor” in Tables 2 to 26: SEQ ID NO:130 was encodes a 2,4-dienoyl-CoA reductase protein. The DNA used. sequence of this gene was blasted against proprietary data 0155 The Arabidopsis ecotype C24 was transformed with bases of plant cDNAs at an evalue of e-10 (Altschul et al., constructs containing the lead genes described in Example 1 US 2011/O 145948 A1 Jun. 16, 2011
using known methods. Seeds from T2 transformed plants measurement of the transgenic relative to the control plants as were pooled on the basis of the promoter driving the expres a percentage of the control non-transgenic plants; p value is Sion, gene source species and type of targeting (chloroplastic, the statistical significance of the difference between trans mitochondrial and cytoplasmic). The seed pools were used in genic and control plants based on a T-test comparison of all the primary screens for biomass underwell watered and water independent events where NS indicates not significant at the limited growth conditions. Hits from pools in the primary 5% level of probability: “No. of events' indicates the total screen were selected, molecular analysis performed and seed number of independent transgenic events tested in the experi ment: “No. of positive events’ indicates the total number of collected. The collected seeds were then used for analysis in independent transgenic events that were larger than the con secondary Screens where a larger number of individuals for trol in the experiment: “No. of negative events’ indicates the each transgenic event were analyzed. If plants from a con total number of independent transgenic events that were struct were identified in the secondary Screen as having smaller than the control in the experiment. NS indicates not increased biomass compared to the controls, it passed to the significant at the 5% level of probability. tertiary screen. In this screen, over 100 plants from all trans genic events for that construct were measured under well A. Phosphatidate Cytidylyltransferase watered and drought growth conditions. The data from the 0159. The phosphatidate cytidylyltransferase designated transgenic plants were compared to wild type Arabidopsis as YBR029C (SEQ ID NO:2) was expressed in Arabidopsis plants or to plants grown from a pool of randomly selected using three constructs: in one constructYBR029C expression transgenic Arabidopsis seeds using standard Statistical pro was controlled by the PCUbi promoter (SEQID NO:121) and cedures. targeted to chloroplasts; in another construct, YBR029C 0156 Plants that were grown under well watered condi expression was controlled by the Super promoter (SEQ ID tions were watered to soil saturation twice a week. Images of NO:122) and targeted to chloroplasts; and in the third con the transgenic plants were taken at 17 and 21 days using a struct YBR029C expression was controlled by the USP pro commercial imaging system. Alternatively, plants were moter (SEQ ID NO:123) and targeted to the mitochondria. grown under water limited growth conditions by watering to Table 2 sets forth biomass and health index data obtained soil saturation infrequently which allowed the soil to dry from the Arabidopsis plants transformed with these con between watering treatments. In these experiments, water structs and tested under water-limiting conditions.
TABLE 2
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events YBRO29C PCUbi Chlor Biomass at day -16.7 OOOOO 6 1 5 2O PCUbi Chlor Biomass at day -17.2 OOOOO 6 O 6 27 YBRO29C PCUbi Chlor Health index -9.S. O.OOS6 6 1 5 YBRO29C Super Chlor Biomass at day 8.2 O.O340 6 4 2 2O YBRO29C Super Chlor Biomass at day 19.4 OOOOO 6 5 1 27 YBRO29C Super Chlor Health index O6 NS 6 4 2 YBRO29C USP Mit Biomass at day -10.1 O.OO40 8 2 6 2O YBRO29C USP Mit Biomass at day -9.5 O.OO34 8 2 6 27 YBRO29C USP Mit Health index -1.8 NS 8 2 6 was given on days 0, 8, and 19 after sowing. Images of the 0160 Table 2 shows that, under water limiting conditions, transgenic plants were taken at 20 and 27 days using a com Arabidopsis plants expressing the YBR029C (SEQID NO:1) mercial imaging System. gene under control of the Super promoter with targeting of the 0157 Image analysis software was used to compare the protein product to the chloroplast and grown were signifi images of the transgenic and control plants grown in the same cantly larger than the control plants. Table 2 also shows that experiment. The images were used to determine the relative the majority of independent Super promoter/chloroplast-tar geted transgenic events were larger than the controls. Table 2 size or biomass of the plants as pixels and the color of the also shows that Arabidopsis plants grown under water limit plants as the ratio of dark green to total area. The latter ratio, ing conditions and expressing the YBR029C gene under con termed the health index, was a measure of the relative amount trol of either the USP promoter with targeting of the protein of chlorophyll in the leaves and therefore the relative amount product to mitochondria or the PCUbi promoter with target of leaf senescence or yellowing and was recorded at day 27 ing of the protein product to the chloroplast were significantly only. Variation exists among transgenic plants that contain the smaller than the control plants that did not express YBR029C. various lead genes, due to different sites of DNA insertion and Table 2 also shows that the majority of independent trans other factors that impact the level or pattern of gene expres genic events were Smaller than the controls. Sion. 0.161 Table 3 sets forth biomass and health index data 0158 Tables 2 to 26 show the comparison of measure obtained from the Arabidopsis plants transformed with these ments of the Arabidopsis plants. Percent change indicates the constructs and tested under well watered conditions. US 2011/O 145948 A1 Jun. 16, 2011 18
TABLE 3
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events
YBRO29C PCUbi Chlor Biomass at day 9.S. O.OOO1 6 5 1 7 YBRO29C PCUbi Chlor Biomass at day S.S. O.OO66 6 5 1 21 YBRO29C PCUbi Chlor Health index -9.6 0.0052 6 O 6 YBRO29C Super Chlor Biomass at day S.2 O.OSO 6 4 2 7 YBRO29C Super Chlor Biomass at day 6.7 O.OO3S 6 4 2 21 YBRO29C Super Chlor Health index -10.7 O.OO12 6 O 6 YBRO29C USP Chlor Biomass at day 6.2. O.O378 6 5 1 7 YBRO29C USP Chlor Biomass at day 6.2 O.O174 6 5 1 21 YBRO29C USP Chlor Health index -2.7 NS 6 2 4 YBRO29C USP Mi Biomass at day 14.7 O.OOOO 8 7 1 7 YBRO29C USP M Biomass at day 9.6 O.OOOO 8 8 O 21 YBRO29C USP Mi Health index 1.2 NS 8 4 4
0162 Table 3 shows that, when grown under well-watered B. Acyl-Carrier Protein conditions, Arabidopsis plants expressing YBR029C were 0163 The acyl-carrier protein designated as YKL 192O (SEQ ID NO:8) was expressed in Arabidopsis using a con larger than the control plants that did not express YBR029C. struct wherein the acyl-carrier protein expression was con The increase in biomass occurred with the PCUbi, Super and trolled by the USP promoter (SEQID NO:123) and targeted Ubi promoters and the effect of the transgene was observed in to the mitochondria. Table 4 sets forth biomass and health index data obtained from the Arabidopsis plants transformed constructs where the YBR029C protein product (SEQ ID with these constructs and tested under water-limiting condi NO:2) was targeted to the mitochondria or the chloroplast. tions.
TABLE 4
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events
YKL192C USP Mit Biomass at day 29.8 OOOOO 5 5 O 2O YKL192C USP Mit Biomass at day 16.6 OOOOO 5 4 1 27 YKL192C USP Mit Health index 14.2 OOOOO 5 4 1 US 2011/O 145948 A1 Jun. 16, 2011 19
0164. Table 4 shows that Arabidopsis plants expressing constructs: in one construct, YDR018C (SEQ ID NO:15) the YKL192O (SEQID NO:7) gene under control of the USP gene expression was controlled by the Super promoter (SEQ promoter with targeting to the mitochondria that were grown under water limiting conditions were significantly larger than ID NO:122) and the YDR018C protein product was targeted the control plants that did not express YKL 192O. Table 4 also to the choloroplast, and in the second construct, YDR018C shows that the majority of independent transgenic events expression was controlled by the USP promoter (SEQ ID were larger and healthier than the controls. NO:123) and targeted to the mitochondria. Table 5 sets forth C. Acyltransferase biomass and health index data obtained from the Arabidopsis (0165. The acyltransferase designated as YDR018C (SEQ plants transformed with these constructs and tested under ID NO:16) was expressed in Arabidopsis using two different water-limiting conditions.
TABLE 5
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events
YDRO18C Super Chlor Biomass at day 6.8 O.0486 6 5 1 2O YDRO18C Super Chlor Biomass at day 21.1 OOOOO 6 5 1 27 YDRO18C Super Chlor Health index 3.0 NS 6 4 2 YDRO18C USP Mit Biomass at day 11.5 O.O240 6 4 2 2O YDRO18C USP Mit Biomass at day 7.1 NS 6 4 2 27 YDRO18C USP Mit Health index 1.7 NS 6 4 2
0166 Table 5 shows that Arabidopsis plants expressing YDRO18C tended to be significantly larger than the control plants that did not express YDRO18C when they were grown under water limiting conditions. Table 5 also shows that the majority of independent transgenic events were larger than the controls. 0.167 Table 6 sets forth biomass and health index data obtained from the Arabidopsis plants transformed with these constructs and tested under well watered conditions.
TABLE 6
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events
YDRO18C Super Chlor Biomass at -0.2 NS 6 3 3 day 17 YDRO18C Super Chlor Biomass at 1.7 NS 6 3 3 day 21 YDRO18C Super Chlor Health index -7.2 O.O375 6 O 6 YDRO18C USP Mit Biomass at 20.9 OOOOO 6 6 O day 17 YDRO18C USP Mit Biomass at 20.3 OOOOO 6 5 1 day 21 YDRO18C USP Mit Health index 2.4 NS 6 3 3 US 2011/O 145948 A1 Jun. 16, 2011 20
0168 Table 6 shows that Arabidopsis plants expressing D. Bifunctional Anaerobic Fatty Acid Oxidation Complex YDR018C under control of the USP promoter with targeting Polypeptide to mitochondria were larger than the control plants that did 0169. The bifunctional anaerobic fatty acid oxidation not express YDRO18C when grown under well watered con complex polypeptide designated as B2341 (SEQID NO:22) ditions. However, when the YDR018C gene was expressed was expressed in Arabidopsis using a construct wherein under control of the Super promoter and the protein product B2341 expression was controlled by the USP promoter (SEQ was targeted to the chloroplast, the plants were not signifi ID NO:124) and the protein product was targeted to the mito cantly larger than control plants that did not express chondria. Table 7 sets forth biomass and health index data YDRO18C, when the plants were grown under well watered obtained from the Arabidopsis plants transformed with these conditions. constructs and tested under water-limiting conditions.
TABLE 7
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events b2341 USP Mit Biomass at day 24.2 OOOOO 6 5 1 2O b2341 USP Mit Biomass at day 15.9 O.OOO9 6 5 1 27 b2341 USP Mit Health index 10.6 O.OOO7 6 5 1
0170 Table 7 shows that Arabidopsis plants expressing the B2341 gene (SEQ ID NO:21) were significantly larger than the control plants that did not express B2341, when grown under water limiting conditions. Table 7 also shows that the majority of independent transgenic events were larger than the controls. (0171 Table 8 sets forth biomass and health index data obtained from Arabidopsis plants transformed with these constructs and tested under well watered conditions.
TABLE 8
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events
b2341 USP Mit Biomass at 8.5 O.OO67 6 6 O day 20 b2341 USP Mit Biomass at 1.6 NS 6 5 1 day 27 b2341 USP Mit Health index 14.2 O.O1OO 6 5 1
0172 Table 8 shows that the majority of independent transgenic events expressing B2341 were larger than the con trol plants that did not express B2341 when grown under well-watered conditions. Table 8 also shows that the plants expressing B2341 were significantly darker green than the controls. E. Acyl-CoA Thioesterase (0173 B0452 (SEQ ID NO:27) was expressed in Arabi dopsis under control of the USP promoter (SEQID NO:124) and the protein product (SEQID NO:28) was targeted to the mitochondria. Table 9 sets forth biomass and health index data obtained from the Arabidopsis plants transformed with this construct and tested under cycling drought (CD) or well watered (WW) conditions. US 2011/O 145948 A1 Jun. 16, 2011 21
TABLE 9 Assay % p No. of Positive Negative Type Gene Promoter Targeting Measurement Change Value Events Events Events CD BO452 USP Mit Day 20 36.56 OOOOO 4 4 O CD BO452 USP Mit Day 27 21.03 OOOOO 4 4 O CD BO452 USP Mit Health 15.14 O.OOO2 4 4 O Index WW BO452 USP Mit Day 17 30.57 OOOOO 5 5 O WW BO452 USP Mit Day 21 15.39 O.OOOO 5 5 O WW BO452 USP Mit Health 2.59 NS 5 3 2 Index
0.174 Table 9 shows that transgenic plants expressing the and WW2) under well-watered conditions than the control B0452 gene under the control of the USP promoter with plants that did not express the YNL202W gene. In these targeting to the mitochondria were significantly larger under experiments, all the independent transgenic events with mito either well-watered or drought conditions than the control chondria targeting were larger than the controls in both the plants that did not express the B0452 gene. The difference cycling drought and well-watered environments. The trans was even more striking at earlier stages, indicating a positive genic plants expressing the YNL202W gene under control of effect on seedling vigor. In these experiments, all of the the USP promoter with targeting to the mitochondria were independent transgenic events were larger than the controls in also significantly healthier in one experiment under well both the cycling drought and well-watered environments. The watered conditions than the control plants that did not express growth advantage of the transgenic plants was even more the YNL202W gene. prominent under cycling drought conditions. The transgenic 0177 Table 10 shows that transgenic plants expressing the plants expressing the B0452 gene stayed significantly YNL202W gene under control of the PCUbi promoter with healthier than the wild-type control under the cycling drought targeting to the plastids were Smaller but not significantly conditions. different compared to the control plants that did not express F. 2,4-dienoyl-CoA Reductase the YNL202W gene, under cycling drought conditions. (0175 YNL202W (SEQID NO:35) was expressed in Ara bidopsis using two constructs, one of which transcription was G. Sterol Esterase under control of the USP promoter (SEQID NO:123) and the (0178 YKL140W (SEQID NO:39) was expressed in Ara protein productYNL202W (SEQID NO:36) was targeted to bidopsis using three different constructs: in one construct, the mitochondria, and the other construct in which transcrip expression was controlled by the PCUbi promoter (SEQ ID tional expression was under control of the PCUbi promoter NO:121) and the protein product (SEQ ID NO:40) was tar (SEQID NO:121) and the protein product was targeted to the geted to the mitochondria, expression in the second construct plastids, respectively. Table 10 sets forth biomass and health was also under the control of the PCUbi promoter, but the index data obtained from the Arabidopsis plants transformed protein product was targeted to plastids; and expression in the with these constructs and tested under cycling drought (CD) third construct was controlled by the USP promoter (SEQID and well-watered (WW) conditions. NO:123) with the protein product being targeted to plastids.
TABLE 10 Assay % No. of Positive Negative Type Gene Promoter Targeting Measurement Change Value Events Events Events CD YNL2O2W PCUbi Chlor Day 20 -0.77 0.8773 6 3 3 CD YNL2O2W PCUbi Chlor Day 27 -13.24 OO646 6 O 6 CD YNL2O2W PCUbi Chlor Health -0.78 O.8045 6 3 3 index CD YNL2O2W USP Mit Day 20 58.29 OOOOO 6 6 O CD YNL2O2W USP Mit Day 27 27.33 OOOOO 6 6 O CD YNL2O2W USP Mit Health O.36 0.9046 6 2 4 index WW1 YNL2O2W USP Mit Day 17 33.64 OOOOO 8 8 O WW1 YNL2O2W USP Mit Day 21 25.52 O.OOOO 8 8 O WW1 YNL2O2W USP Mit Health O.16 0.9592 8 5 3 index WW2 YNL2O2W USP Mit Day 17 31.51 O.OOOO 6 6 O WW2 YNL2O2W USP Mit Day 21 21.82 O.OOOO 6 6 O WW2 YNL2O2W USP Mit Health 12.62 O.OOO2 6 5 1 index
0176 Table 10 shows that transgenic plants expressing the Table 11 sets forth biomass and health index data obtained YNL202W gene under control of the USP promoter with from the Arabidopsis plants transformed with these con targeting to the mitochondria were significantly larger under structs and tested under cycling drought or well-watered con cycling drought conditions and in two experiments (WW1 ditions. US 2011/O 145948 A1 Jun. 16, 2011 22
TABLE 11
Assay % No. of Positive Negative Type Gene Promoter Targeting Measurement Change p-Value Events Events Events
CD YKL14OW USP Mit Day 20 13.86 O.OOO3 6 6 O CD YKL14OW USP Mit Day 27 -2.03 O.S931 6 3 CD YKL14OW USP Mit Health S.65 0.0969 6 5 1 Index CD YKL14OW USP Chlor Day 20 -37.76 O.OOOO 5 O 5 CD YKL14OW USP Chlor Day 27 -21.06 OOOOO 5 O 5 CD YKL14OW USP Chlor Health -10.81 O.OOO1 5 O 5 Index WW YKL14OW PCUbi Chlor Day 17 42.97 OOOOO 5 5 WW YKL14OW PCUbi Chlor Day 21 26.82 O.OOOO 5 WW YKL14OW PCUbi Chlor Health -109 0.7119 5 2 Index
0179 Table 11 shows that transgenic plants expressing the 0181 Table 12 shows that transgenic plants expressing the YKL140W gene under control of the PCUbi promoter with SLL1023 gene under control of the PCUbi promoter with targeting to the plastids were significantly larger under well targeting to the plastids were significantly larger under watered conditions than the control plants that did not express the YKL140W gene. In these experiments, all of the indepen cycling drought and well-watered conditions than the control dent transgenic events with plastid targeting were larger than plants that did not express the SLL 1023 gene. In these experi the controls in the cycling drought environment. Transgenic ments, all the independent transgenic events with plastid tar plants expressing the YKL140W gene under control of the geting Were larger than the controls under both cycling USP promoter with targeting to the mitochondria were also drought and well-watered conditions. The transgenic plants significantly larger under cycling drought conditions than the control wild-type plants. Table 11 shows that transgenic expressing the SLL 1023 gene under control of the PCUbi plants expressing the YKL140W gene under control of the promoter with targeting to the plastids were also significantly USP promoter with targeting to the plastid were significantly healthier under well-watered conditions than the control Smaller under drought conditions than the control plants that plants. The presence of the SLL 1023 protein in the plastids did not express the YKL140W gene. Additionally, these promoted plant growth under both well-watered and drought transgenic plants had lower health index scores relative to the conditions. control in water-limited conditions. H. Succinate-CoA Ligase I. Cobalt-precorrin-6A Reductase 0180. The Synechocystis gene SLL 1023 (SEQID NO:41) 0182. The Synechocystis gene SLR0252 (SEQID NO:43) was expressed in Arabidopsis under control of the PCUbi was expressed in Arabidopsis under control of the PCUbi promoter (SEQID NO:121) and targeted to the plastids. Table promoter (SEQID NO:121). Table 13 sets forth biomass and 12 sets forth biomass and health index data obtained from health index data obtained from the Arabidopsis plants trans Arabidopsis plants transformed with this construct and tested formed with this construct and tested under cycling drought under cycling drought and well-watered conditions. conditions.
TABLE 12
Assay % p No. of Positive Negative Type Gene Promoter Targeting Measurement Change Value Events Events Events
CD SLL1023 PCUbi Chlor Day 20 43.O2 OOOOO 6 6 O CD SLL1023 PCUbi Chlor Day 27 37.78 O.OOOO 6 6 O CD SLL1023 PCUbi Chlor Health 0.61 0.8357 6 3 3 Index WW SLL1023 PCUbi Chlor Day 17 24.08 OOOOO 6 6 O WW SLL1023 PCUbi Chlor Day 21 16.79 OOOOO 6 6 O WW SLL1023 PCUbi Chlor Health 9.33 O.OO38 6 5 1 Index US 2011/O 145948 A1 Jun. 16, 2011
TABLE 13
Assay % p- No. of Positive Negative Type Gene Promoter Targeting Measurement Change Value Events Events Events CD SLRO2S2 PCUbi None Day 20 20.20 OOOOO 5 5 O CD SLRO2S2 PCUbi None Day 27 16.30 OOOO1 5 5 O CD SLRO2S2 PCUbi None Health -4.07 O.OO76 5 O 5 Index
0183 Table 13 shows that transgenic plants expressing the J. Uroporphyrin-III C-methyltransferase SLR0252 gene under control of the PCUbi promoter were significantly larger under cycling drought conditions than the (0.184 The E. coli gene designated b3803 (SEQ ID control plants that did not express the SLR0252 gene. In these NO:45), encoding a uroporphyrin-III C-methyltransferase, experiments, all the independent transgenic events were was expressed in Arabidopsis using two different constructs: larger than the controls in the cycling drought environment. As evidenced by the observation that the transgenic plants constructs controlled by the USP promoter (SEQID NO:124) were larger than the control under cycling drought conditions, and constructs controlled by the super promoter (SEQ ID the presence of the SLR0252 protein in the cytoplasm enabled NO:122). Table 14 sets forth biomass data obtained from the transgenic plant to growth better under water-limited con Arabidopsis plants transformed with these constructs and ditions. tested under well-watered conditions.
TABLE 1.4
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b38O3 USP Biomass at Wt 15.31 OOOOO 6 6 O day 20 b3803 Super Biomass at Wt -1.87 O.S346 6 3 3 day 20 b3803 Super Biomass at Wt -1.25 O.6151 6 3 3 day 27 b38O3 USP Biomass at Wt 14.86 OOOOO 6 6 O day 27 b3803 Super Biomass at Superpool 8.89 O.O139 6 5 1 day 20 b3803 Super Biomass at Superpool 2.22 O.4364 6 3 3 day 27
0185. Table 14 shows that Arabidopsis plants expressing b3803 under control of the USP promoter that were grown under well-watered conditions were significantly larger than the control plants that did not express b3803. Table 14 also shows that the majority of independent transgenic events were larger than the controls. K. Isoprenoid Biosynthesis Protein 0186. The gene designated b3209 (SEQ ID NO:53), end coding an isoprenoid biosynthesis protein, was expressed in Arabidopsis using a construct controlled by the Super pro moter (SEQ ID NO:122). Table 15 sets forth biomass data obtained from Arabidopsis plants transformed with these constructs and tested under well-watered conditions.
TABLE 1.5
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b3209 Super Biomass at Wt 17.71 O.OOOO 7 7 O day 20 b3209 Super Biomass at Wt 14.47 OOOOO 7 7 O day 27 US 2011/O 145948 A1 Jun. 16, 2011 24
TABLE 15-continued
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b3209 Super Biomass at Superpool 29.46 OOOOO 7 7 O day 20 b3209 Super Biomass at Superpool 22.10 O.OOOO 7 7 O day 27
0187 Table 15 shows that Arabidopsis plants expressing b3209 under control of the super promoter that were grown under well-watered conditions were significantly larger than the control plants that did not express b3209. Table 15 also shows that the majority of independent transgenic events were larger than the controls. L. LysE Type Translocator 0188 The E. coli gene designated b2578 (SEQ ID NO:59), encoding a LysE type translocator, was expressed in Arabidopsis using a construct controlled by the Super pro moter (SEQ ID NO:122). Table 16 sets forth biomass and health index data obtained from Arabidopsis plants trans formed with these constructs and tested under well-watered conditions.
TABLE 16
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b2578 Super Health index Wt -6.74 O.O221 7 1 6 b2578 Super Biomass at Wt 7.66 O.O352 7 5 2 day 20 b2578 Super Biomass at Wt 8.22 O.OO61 7 6 1 day 27 b2578 Super Health index Superpool 8.17 O.O2OO 7 6 b2578 Super Biomass at Superpool 1841 OOOOO 7 6 day 20 b2578 Super Biomass at Superpool 1544 OOOOO 7 6 day 27 0189 Table 16 shows that Arabidopsis plants expressing b2578 that were grown under well-watered conditions were significantly larger than the control plants that did not express b2578. Table 16 also shows that the majority of independent transgenic events were larger than the controls. M. Branched-Chain Amino Acid Transporter (0190. The gene designated b2682 (SEQ ID NO:61), encoding a branched-chain amino acid transporter, was expressed in Arabidopsis using a construct controlled by the super promoter (SEQID NO:122). Table 17 sets forth biom ass and health index data obtained from the Arabidopsis plants transformed with these constructs and tested under well-watered conditions.
TABLE 17
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b2682 Super Health index Wt -0.95 0.7523 7 3 4 b2682 Super Biomass at Wt 16.28 OOOOO 7 7 O day 20 US 2011/O 145948 A1 Jun. 16, 2011 25
TABLE 17-continued
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b2682 Super Biomass at Wt 7.49 O.OO73 7 7 O day 27 b2682 Super Health index Superpool 14.89 OOOOO 7 6 1 b2682 Super Biomass at Superpool 27.89 O.OOOO 7 7 O day 20 b2682 Super Biomass at Superpool 14.66 OOOOO 7 7 O day 27
0191 Table 17 shows that Arabidopsis plants expressing b2682 under control of the super promoter that were grown under well-watered conditions were significantly larger than the control plants that did not express b2682. Table 17 also shows that the majority of independent transgenic events were larger than the controls. N. DNA-Binding Protein (0192. The gene designated b3285 (SEQ ID NO:63), encoding a DNA-binding protein, was expressed in Arabi dopsis using a construct controlled by the Super promoter (SEQ ID NO:122). Table 18 sets forth biomass and health index data obtained from Arabidopsis plants transformed with these constructs and tested under well-watered condi tions.
TABLE 18
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b3285 Super Health index Wt -6.46 O.O351 7 2 5 b3285 Super Biomass at Wt 16.84 OOOOO 7 6 1 day 20 b3285 Super Biomass at Wt 14.41 OOOOO 7 7 O day 27 b3285 Super Health index Superpool 849 O.O2OO 7 5 2 b3285 Super Biomass at Superpool 28.51 O.OOOO 7 7 O day 20 b3285 Super Biomass at Superpool 21.75 O.OOOO 7 7 O day 27
0193 Table 18 shows that Arabidopsis plants expressing b3285 under control of the super promoter that were grown under well-watered conditions were significantly larger than the control plants that did not express b3285. Table 18 also shows that the majority of independent transgenic events were larger than the controls. O.Y.Sc.J/FliF Protein (0194 The gene designated b1938 (SEQ ID NO:65), encoding a YscJ/FliF protein, was expressed in Arabidopsis using a construct controlled by the super promoter (SEQ ID NO: 122). Table 19 sets forth biomass and health index data obtained from the Arabidopsis plants transformed with these constructs and tested under well-watered conditions. TABLE 19
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b1938 Super Biomass at Wt 5.64 O. 1074 7 5 2 day 20 US 2011/O 145948 A1 Jun. 16, 2011 26
TABLE 19-continued
No of No. of Control % p- No. of Positive Negative Gene Promoter Measurement type Change Value Events Events Events b1938 Super Biomass at Wt 7.41 O.O123 7 5 2 day 27 b1938 Super Biomass at Superpool 16.18 OOOOO 7 6 1 day 20 b1938 Super Biomass at Superpool 14.57 O.OOOO 7 5 2 day 27
Table 19 shows that Arabidopsis plants expressing b1938 under control of the Super promoter that were grown under well-watered conditions were significantly larger than the control plants that did not express b1938. Table 19 also shows that the majority of independent transgenic events were larger than the controls. P. Riboflavin Biosynthetic Genes (0195 The Synechocystis gene designated SLL 1894 (SEQ ID NO:67) encoding GTP cyclohydrolase II was expressed in Arabidopsis using three different constructs controlled by the parsley ubiquitin promoter (SEQ ID NO:121): constructs with no subcellular targeting, constructs targeted to the chlo roplast, and constructs targeted to mitochondria. Table 20 sets forth biomass and health index data obtained from Arabidop sis plants transformed with SLL 1894 under control of the parsley ubiquitin promoter with and without subcellular tar geting, and tested under water limited conditions.
TABLE 20
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events S1894 PCUbi None Biomass at 12.2 O.OO16 7 6 1 20 days S1894 PCUbi None Biomass at 7.8 0.0499 7 5 2 27 days S1894 PCUbi None Health Index 9.4 O.OO92 7 5 2 S1894 PCUbi Mito Biomass at -2.1 NS 6 3 3 20 days S1894 PCUbi Mito Biomass at -21.8 O.OOO2 6 O 6 27 days S1894 PCUbi Mito Health Index 2.5 NS 6 5 1 S1894 PCUbi Chlor Biomass at -1.3 NS 6 3 3 20 days S1894 PCUbi Chlor Biomass at -0.9 NS 6 4 2 27 days S1894 PCUbi Chlor Health Index 7.1 0.0327 6 5 1
0196. Transgenic plants expressing the SLL 1894 gene plants that did not express the SLL 1894 gene. Transgenic with no Subcellular targeting were significantly larger under plants expressing the SLL 1894 gene with subcellular target water limited conditions than the control plants that did not ing to the plastid were similar in size under water limited express the SLL 1894 gene. In addition, the transgenic plants conditions to the control plants that did not express the were darker green in color than the controls under water SLL1894 gene, but had a larger health index. limited conditions as shown by the increased health index. In 0198 The Synechocystis gene designated gene SLL 1282 these experiments, the majority of the independent transgenic (SEQID NO:71) encoding lumazine synthase was expressed events were larger than the controls in the water limited in Arabidopsis using a construct controlled by the parsley environment. ubiquitin promoter (SEQ ID NO:121) with no subcellular 0.197 Transgenic plants expressing the SLL 1894 gene targeting. Table 21 sets forth biomass and health index data with Subcellular targeting to the mitochondria were signifi obtained from Arabidopsis plants transformed with this con cantly smaller under water limited conditions than the control struct and tested under well-watered conditions. US 2011/O 145948 A1 Jun. 16, 2011 27
TABLE 21
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events S1282 PCUbi None Biomass at 2.2 NS 6 3 3 17 days S1282 PCUbi None Biomass at -0.6 NS 6 2 4 21 days S1282 PCUbi None Health Index -1.5 NS 6 3 3
0199 The growth of the Arabidopsis plants expressing the SLL 1282 gene controlled by the PCUbi promoter and with no Subcellular targeting was similar to control plants under well watered conditions. 0200 Table 22 sets forth biomass and health index data obtained from the Arabidopsis plants transformed with SLL 1282 under control of the parsley ubiquitin promoter, with and without Subcellular targeting, and tested underwater limited conditions.
TABLE 22
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events S1282 PCUbi None Biomass at 27.5 O.OOOO 6 6 O 20 days S1282 PCUbi None Biomass at 34.5 O.OOOO 6 6 O 27 days S1282 PCUbi None Health Index 9.S. O.OOO3 6 6 O S1282 PCUbi Mito Biomass at 8.7 O.O166 7 4 3 20 days S1282 PCUbi Mito Biomass at 10.7 O.OO29 7 5 2 27 days S1282 PCUbi Mito Health Index -3.1 NS 7 3 4 S1282 PCUbi Chlor Biomass at -16.9 OOOOO 6 1 5 20 days S1282 PCUbi Chlor Biomass at -4.6 NS 6 1 5 27 days S1282 PCUbi Chlor Health Index -6.8 O.OO44 6 1 5
0201 Arabidopsis plants that were grown under water Q. Vitamin B6 Biosynthetic Genes limited conditions were significantly larger than the control plants that did not express the SLL 1282 gene at two measur ing times, if the protein did not contain a Subcellular targeting (0202) The E. coli gene designated b1636 (SEQID NO:79) sequence. If a mitochondrial targeting sequence was encoding the pyridoxal kinase PdxY was expressed in Ara included, the gene was less effective, but did provide some bidopsis using a construct controlled by the Super promoter improvement relative to the control. Targeting the protein to the plastid resulted in reduced growth. In these experiments, (SEQ ID NO:122) targeted to the chloroplast. Table 23 sets all independent transgenic events with no Subcellular target forth biomass and health index data obtained from Arabidop ing were larger than the controls in the water limited environ sis plants transformed with these constructs and tested under ment. well-watered conditions.
TABLE 23
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events b1636 Super Chlor Biomass at 10.7 O.OOO1 6 5 1 17 days b1636 Super Chlor Biomass at 10.0 OOOOO 6 5 1 21 days b1636 Super Chlor Health Index -10.7 O.OOO9 6 1 5 US 2011/O 145948 A1 Jun. 16, 2011 28
0203 Arabidopsis plants that were grown under well watered conditions were significantly larger than the control plants that did not express the b1636 gene at two measuring times. In these experiments, the majority of the independent transgenic events were larger than the controls in the well watered environment. 0204 Table 24 sets forth biomass and health index data obtained from the Arabidopsis plants transformed with b1636 controlled by the super promoter targeted to the chloroplast and tested under water limited conditions.
TABLE 24
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events b1636 Super Chlor Biomass at 8.9 O.O243 6 4 2 20 days b1636 Super Chlor Biomass at 21.9 O.OOOO 6 3 3 27 days b1636 Super Chlor Health Index 1.6 NS 6 4 2
0205 Arabidopsis plants that were grown under water protein Pdx.J was expressed in Arabidopsis using three differ limited conditions were significantly larger than the control ent constructs controlled by the parsley ubiquitin promoter plants that did not express the b1636 gene at two measuring times. In these experiments, three or four of the six indepen (SEQ ID NO:121): constructs with no subcellular targeting, dent transgenic events were larger than the controls in the constructs targeted to the chloroplast, and constructs targeted water limited environment. to mitochondria. Table 25 sets forth biomass and health index 0206. The Synechocystis gene designated SLR1779 (SEQ data obtained from the Arabidopsis plants transformed with ID NO:95) encoding the pyridoxal phosphate biosynthetic these constructs and tested under well-watered conditions.
TABLE 25
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events Sr1779 PCUbi None Biomass at 23.8 OOOOO 8 8 O 17 days Sr1779 PCUbi None Biomass at 20.6 OOOOO 8 8 O 21 days Sr1779 PCUbi None Health Index -1.1 NS 8 4 4 Sr1779 PCUbi Mito Biomass at 18.8 OOOOO 6 6 O 17 days Sr1779 PCUbi Mito Biomass at 7.6 O.OOO8 6 6 O 21 days Sr1779 PCUbi Mito Health Index 8.7 O.O104 6 6 O Sr1779 PCUbi Chlor Biomass at 27.6 OOOOO 6 6 O 17 days Sr1779 PCUbi Chlor Biomass at 15.1 OOOOO 6 6 O 21 days Sr1779 PCUbi Chlor Health Index -1.0 NS 6 3 3
0207 Table 26 sets forth biomass and health index data of the Arabidopsis plants transformed with the SLR1779 gene controlled by the parsley ubiquitin promoter with subcellular targeting and tested under water limited conditions.
TABLE 26
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events Sr1779 PCUbi Mito Biomass at -6.5 O.OOS8 6 4 2 20 days Sr1779 PCUbi Mito Biomass at -47 O.O3O2 6 3 3 27 days US 2011/O 145948 A1 Jun. 16, 2011 29
TABLE 26-continued
No of No. of % p- No. of Positive Negative Gene Promoter Targeting Measurement Change Value Events Events Events Sr1779 PCUbi Mito Health Index -0.4 NS 6 3 3 Sr1779 PCUbi Chlor Biomass at 16.6 O.OOOO 6 5 1 20 days Sr1779 PCUbi Chlor Biomass at 8.5 O.OOO2 6 4 2 27 days Sr1779 PCUbi Chlor Health Index 13.2 O.OOOO 6 5 1
0208 Transgenic plants expressing the SLR 1779 gene genic plants were darker green in color than the controls were larger under well watered conditions than the control under water limited conditions as shown by the increased plants that did not express the SLR 1779 gene. This effect was health index. In these experiments, the majority of the inde observed with all three constructs that had no subcellular targeting or with Subcellular targeting either to the mitochon pendent transgenic events were larger than the controls in the dria or to the plastid. water limited environment. 0209 Transgenic plants expressing the SLR 1779 gene 0210 Transgenic plants expressing the SLR 1779 gene with Subcellular targeting to the plastid were significantly with Subcellular targeting to the mitochondria were signifi larger under water limited conditions than the control plants cantly smaller under water limited conditions than the control that did not express the SLR 1779 gene. In addition, the trans plants that did not express the SLR 1779 gene.
SEQUENCE LISTING
<16 O NUMBER OF SEQ ID NOS: 130
<21 Os SEQ ID NO 1 LENGTH: 1374 TYPE: DNA ORGANISM: Saccharomyces cerevisiae FEATURE; NAME/KEY: CDS LOCATION: (1) ... (1374) OTHER INFORMATION: phosphatidate cytidylyltransferase (YRO29C)
< 4 OOs SEQUENCE: 1.
atg tot gac aac cott gag atg a.a.a. C Ca Cat ggt acg agc aag gag att 48 Met Ser Asp Asn Pro Glu Met Pro His Gly Thir Ser Lys Glu Ile 1. 5 1O 15
gtg gag tcg gtt act gac gcc a CC toa aag gcg att gat a.a.a. ttg Cala 96 Wall Glu Ser Wall Thir Asp Ala Thir Ser Lys Ala Ile Asp Lys Lell Glin 2O 25 3 O
gaa gaa citc. CaC aag gac gcc agc gaa t cc gt C acg cc.g gtg a CC aag 144 Glu Glu Lell His Lys Asp Ala Ser Glu Ser Wall Thir Pro Wall Thir Lys 35 4 O 45
gaa agc act gct gct aca aag gaa agc agg a.a.a. tac aac titt tto att 192 Glu Ser Thir Ala Ala Thir Lys Glu Ser Arg Lys Tyr Asn Phe Phe Ile SO 55 60
aga a Ca gtt tgg acg titt gtt atg atc. agt ttic ttic atc. a CC tta 24 O Arg Thir Wall Trp Thir Phe Wall Met Ile Ser Phe Phe le Thir Lell 65 70 8O
gca tcg ggit Cat gca tgg tgt ata gtg Ctg att ttg ggc tgc Cala att 288 Ala Ser Gly His Ala Trp Cys Ile Wall Lell Ile Luell Gly Cys Glin Ile 85 90 95
gct act titt a.a.a. gag tgt att gcc gta a Ca agt gca tot ggt gaa 336 Ala Thir Phe Lys Glu Cys Ile Ala Wall Thir Ser Ala Ser Gly Arg Glu 1OO 105 110
aag aat ttg CC a ttg aca aag acg ttg aac tgg tac citt citc. tto a CC 384 Lys Asn Lell Pro Luell Thir Lys Thir Lell Asn Trp Luell Tell Phe Thir 115 12O 125
US 2011/O 145948 A1 Jun. 16, 2011 31
- Continued ata at a to a gog aag aat titt gala aag ttg got gac atc titt aat gtg 1344 Ile Ile Ser Ala Lys Asn. Phe Glu Lys Lieu Ala Asp Ile Phe Asin Val 435 44 O 445 acc aag aaa to a titg acc aat cac tot toga 1374 Thr Lys Llys Ser Lieu. Thir Asn His Ser 450 45.5
<210s, SEQ ID NO 2 &211s LENGTH: 457 212. TYPE: PRT <213> ORGANISM: Saccharomyces cerevisiae
<4 OOs, SEQUENCE: 2 Met Ser Asp Asn Pro Glu Met Llys Pro His Gly Thr Ser Lys Glu Ile 1. 5 1O 15 Val Glu Ser Val Thr Asp Ala Thir Ser Lys Ala Ile Asp Llys Lieu. Glin 2O 25 3O Glu Glu Lieu. His Lys Asp Ala Ser Glu Ser Val Thr Pro Val Thir Lys 35 4 O 45 Glu Ser Thr Ala Ala Thr Lys Glu Ser Arg Llys Tyr Asn Phe Phe Ile SO 55 6 O Arg Thr Val Trp Thr Phe Val Met Ile Ser Gly Phe Phe Ile Thr Lieu. 65 70 7s 8O Ala Ser Gly His Ala Trp Cys Ile Val Lieu. Ile Lieu. Gly Cys Glin Ile 85 90 95 Ala Thr Phe Lys Glu. Cys Ile Ala Val Thir Ser Ala Ser Gly Arg Glu 1OO 105 11 O Lys Asn Lieu Pro Lieu. Thir Lys Thr Lieu. Asn Trp Tyr Lieu. Lieu. Phe Thr 115 12 O 125 Thir Ile Tyr Tyr Lieu. Asp Gly Lys Ser Leu Phe Llys Phe Phe Glin Ala 13 O 135 14 O Thr Phe Tyr Glu Tyr Pro Val Lieu. Asn Phe Ile Val Thr Asn His Lys 145 150 155 160 Phe Ile Cys Tyr Cys Lieu. Tyr Lieu Met Gly Phe Val Lieu Phe Val Cys 1.65 17O 17s Ser Lieu. Arg Lys Gly Phe Lieu Lys Phe Glin Phe Gly Ser Lieu. Cys Val 18O 185 19 O
Thir His Met Wall Leu Lleu Lleu. Wal Wall Phe Glin Ala His Lieu. Ile Ile 195 2OO 2O5 Lys Asn Val Lieu. Asn Gly Lieu. Phe Trp Phe Lieu Lleu Pro Cys Gly Lieu. 21 O 215 22O Val Ile Val Asn Asp Ile Phe Ala Tyr Lieu. Cys Gly Ile Thr Phe Gly 225 23 O 235 24 O Llys Thir Lys Lieu. Ile Glu Ile Ser Pro Llys Llys Thr Lieu. Glu Gly Phe 245 250 255 Lieu. Gly Ala Trp Phe Phe Thr Ala Lieu Ala Ser Ile Ile Lieu. Thir Arg 26 O 265 27 O Ile Leu Ser Pro Tyr Thr Tyr Lieu. Thr Cys Pro Val Glu Asp Lieu. His 27s 28O 285 Thr Asn Phe Phe Ser Asn Lieu. Thr Cys Glu Lieu. Asn Pro Val Phe Leu 29 O 295 3 OO Pro Glin Val Tyr Arg Lieu Pro Pro Ile Phe Phe Asp Llys Val Glin Ile 3. OS 310 315 32O US 2011/O 145948 A1 Jun. 16, 2011 32
- Continued
Asn Ser Ile Thr Val Llys Pro Ile Tyr Phe His Ala Lieu. Asn Lieu Ala 3.25 330 335 Thr Phe Ala Ser Lieu. Phe Ala Pro Phe Gly Gly Phe Phe Ala Ser Gly 34 O 345 35. O Lieu Lys Arg Thr Phe Llys Wall Lys Asp Phe Gly His Ser Ile Pro Gly 355 360 365 His Gly Gly Ile Thr Asp Arg Val Asp Cys Glin Phe Ile Met Gly Ser 37 O 375 38O Phe Ala Asn Lieu. Tyr Tyr Glu Thr Phe Ile Ser Glu. His Arg Ile Thr 385 390 395 4 OO Val Asp Thr Val Lieu. Ser Thir Ile Lieu Met Asn Lieu. Asn Asp Llys Glin 4 OS 41O 415 Ile Ile Glu Lieu. Ile Asp Ile Lieu. Ile Arg Phe Lieu. Ser Lys Lys Gly 42O 425 43 O Ile Ile Ser Ala Lys Asn. Phe Glu Lys Lieu Ala Asp Ile Phe Asin Val 435 44 O 445 Thr Lys Llys Ser Lieu. Thir Asn His Ser 450 45.5
<210s, SEQ ID NO 3 &211s LENGTH: 1104 &212s. TYPE: DNA <213> ORGANISM: Brassica napus 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (1104) <223> OTHER INFORMATION: phosphatidate cytidylyltransferase (BNO4MC3O805)
<4 OOs, SEQUENCE: 3 atg gag gag gag agc aac gitg acc agc acc agc ccg agc acc ccg gtg 48 Met Glu Glu Glu Ser Asn. Wall. Thir Ser Thir Ser Pro Ser Thr Pro Wall 1. 5 1O 15
Cag agg Ctg agg cac agg aag agg agc agc acc gag gtg Ctg gaC ggc 96 Glin Arg Lieu. Arg His Arg Lys Arg Ser Ser Thr Glu Val Lieu. Asp Gly 2O 25 3O gac aag gtgaac goc agc ccg ctg. Ctg gtgaac gac agg aac aag tac 144 Asp Llys Val Asn Ala Ser Pro Lieu. Lieu Val Asn Asp Arg Asn Llys Tyr 35 4 O 45 aag agc titc at g g to agg acc tac agc acc ctg tdg atg at C goc ggc 192 Lys Ser Phe Met Val Arg Thr Tyr Ser Thr Lieu. Trp Met Ile Ala Gly SO 55 6 O ttic gtg atg gtg gtg tac atg ggc cac ctg tac atc acc gcc atg gtg 24 O Phe Val Met Val Val Tyr Met Gly His Leu Tyr Ile Thr Ala Met Val 65 70 7s 8O
Ctg gtg at C cag atc titc atg gcc aag gag ctg titc aac Ctg ctg agg 288 Lieu Val Ile Glin Ile Phe Met Ala Lys Glu Lieu. Phe Asn Lieu. Lieu. Arg 85 90 95 aag gcc ccg gag gac aag tec Ctg ccg ggc atc aag cag ctgaac tig 336 Lys Ala Pro Glu Asp Llys Cys Lieu Pro Gly Ile Lys Glin Lieu. Asn Trp 1OO 105 11 O cac ttic titc ttic acc gcc atg citg titc gtg tac ggc agg at c ctd agc 384 His Phe Phe Phe Thr Ala Met Leu Phe Val Tyr Gly Arg Ile Leu Ser 115 12 O 125
Cag agg ctg gCC aac acc gtg acc gcc gaC cag titc Ctg tac agg Ctg 432 Glin Arg Lieu Ala Asn Thr Val Thir Ala Asp Glin Phe Lieu. Tyr Arg Lieu. 13 O 135 14 O
US 2011/O 145948 A1 Jun. 16, 2011 34
- Continued
Phe Val Met Val Val Tyr Met Gly His Leu Tyr Ile Thr Ala Met Val 65 70 7s 8O Lieu Val Ile Glin Ile Phe Met Ala Lys Glu Lieu. Phe Asn Lieu. Lieu. Arg 85 90 95 Lys Ala Pro Glu Asp Llys Cys Lieu Pro Gly Ile Lys Glin Lieu. Asn Trp 1OO 105 11 O His Phe Phe Phe Thr Ala Met Leu Phe Val Tyr Gly Arg Ile Leu Ser 115 12 O 125 Glin Arg Lieu Ala Asn Thr Val Thir Ala Asp Glin Phe Lieu. Tyr Arg Lieu. 13 O 135 14 O Val Ser Gly Lieu. Ile Llys Tyr His Met Ala Ile Cys Tyr Phe Lieu. Asn 145 150 155 160 Ile Ile Gly Phe Met Trp Phe Ile Lieu. Thir Lieu Lys Lys Llys Met Tyr 1.65 17O 17s Lys Tyr Glin Phe Gly Glin Tyr Ala Trp Thr His Met Ile Lieu. Ile Val 18O 185 19 O Val Phe Thr Glin Ser Ser Phe Thr Val Ala Asn Ile Phe Glu Gly Ile 195 2OO 2O5 Phe Trp Phe Leu Lleu Pro Ala Ser Lieu. Ile Ile Ile Asn Asp Ile Phe 21 O 215 22O Ala Tyr Ile Phe Gly Phe Phe Phe Gly Arg Thr Pro Leu. Ile Llys Lieu. 225 23 O 235 24 O Ser Pro Llys Llys Thr Trp Glu Gly Phe Ile Gly Ala Ser Val Thir Thr 245 250 255 Ile Ile Ser Ala Phe Val Lieu Ala Asn Val Lieu. Gly Arg Phe Pro Trp 26 O 265 27 O Lieu. Thir Cys Pro Arg Glin Asp Lieu. Ser Thr Gly Trp Lieu. Glin Cys Asp 27s 28O 285 Ala Asp Pro Leu Phe Llys Pro Glu Pro Phe Thr Lieu Pro Ala Trp Ile 29 O 295 3 OO Pro Gly Trp Phe Pro Trp Lys Glu Met Glu Val Lieu Pro Val Glin Trp 3. OS 310 315 32O His Ala Lieu. Cys Lieu. Gly Lieu. Phe Ala Ser Ile Ile Ala Pro Phe Gly 3.25 330 335 Gly Phe Phe Ala Ser Gly Phe Lys Arg Ala Phe Lys Ile Lys Asp Phe 34 O 345 35. O Gly Asp Ser Ile Pro Gly. His Gly Gly Ile Thr Asp Arg Met Asp 355 360 365
<210s, SEQ ID NO 5 &211s LENGTH: 1278 &212s. TYPE: DNA <213> ORGANISM: Zea mays 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (1278 <223> OTHER INFORMATION: phosphatidate cytidylyltransferase (ZMO6MC3 O283)
<4 OOs, SEQUENCE: 5 atg cag agg gaC acc agc agc agc gac gitg agc gcc agc cac gtg ggc 48 Met Glin Arg Asp Thr Ser Ser Ser Asp Val Ser Ala Ser His Val Gly 1. 5 1O 15 agg gtg agg agg agg aag cac ccg agc gag gCC agc acc gaC ggc aac 96
US 2011/O 145948 A1 Jun. 16, 2011 36
- Continued Ala Lieu Ala Lieu. Gly Lieu. Phe Ala Ser Ile Ile Ala Pro Phe Gly Gly 3.25 330 335 ttic titc gcc agc ggc titc aag agg gcc titc aag atc aag gaC titC ggc O56 Phe Phe Ala Ser Gly Phe Lys Arg Ala Phe Lys Ile Lys Asp Phe Gly 34 O 345 35. O gac agc at C ccg ggC Cac ggc ggc at C acc gac agg atg gaC tec cag 104 Asp Ser Ile Pro Gly. His Gly Gly Ile Thr Asp Arg Met Asp Cys Glin 355 360 365 atg gtg atg goc gtg titc gcc tac atc tac cac cag agc titc atc gcc 152 Met Val Met Ala Val Phe Ala Tyr Ile Tyr His Glin Ser Phe Ile Ala 37 O 375 38O ccg cag aac tt C agc gtg gag at C at C ctg gaC cag atc Ctg agg aac 2OO Pro Glin Asn. Phe Ser Val Glu Ile Ile Lieu. Asp Glin Ile Lieu. Arg Asn 385 390 395 4 OO
Ctg acc tac gag gag cag aag tac Ctg tac gag cag Ctg ggc gag atg 248 Lieu. Thir Tyr Glu Glu Gln Lys Tyr Lieu. Tyr Glu Gln Leu Gly Glu Met 4 OS 41O 415 ttic cac gag agg cag Ctg ggc cag agc tiga 278 Phe His Glu Arg Glin Lieu. Gly Glin Ser 42O 425
<210s, SEQ ID NO 6 &211s LENGTH: 425 212. TYPE: PRT <213> ORGANISM: Zea mays <4 OOs, SEQUENCE: 6 Met Glin Arg Asp Thr Ser Ser Ser Asp Val Ser Ala Ser His Val Gly 1. 5 1O 15 Arg Val Arg Arg Arg Llys His Pro Ser Glu Ala Ser Thr Asp Gly Asn 2O 25 3O Arg Ala Asn Gly Glin Pro Lieu. Lieu Val Asn Asp Glin Asn Llys Tyr Lys 35 4 O 45 Ser Met Lieu. Ile Arg Thr Tyr Ser Thr Val Trp Met Ile Gly Gly Phe SO 55 6 O Ala Phe Ile Ile Tyr Val Gly His Leu Tyr Ile Trp Ala Met Val Val 65 70 7s 8O Val Ile Glin Ile Tyr Met Ala Arg Glu Lieu. Phe Asn Lieu. Lieu. Arg Llys 85 90 95 Ser Ser Glu Glu Lys Glin Lieu Pro Gly Phe Arg Lieu. Lieu. Asn Trp His 1OO 105 11 O Phe Phe Phe Thr Ala Met Leu Tyr Thr Tyr Gly Arg Phe Leu Ser Arg 115 12 O 125 Glin Lieu Val Asn Thr Val Thir Ser Asp His Lieu. Lieu. Tyr Llys Val Val 13 O 135 14 O Ser Gly Lieu. Ile Llys Tyr Gln Met Phe Ile Cys Tyr Phe Leu Tyr Ile 145 150 155 160 Ala Gly Phe Val Trp Phe Ile Lieu. Thir Lieu Lys Llys Llys Thr Tyr Lys 1.65 17O 17s Tyr Glin Phe Lys Glin Tyr Ala Trp Thr His Met Ile Leu Lieu. Thr Val 18O 185 19 O Phe Ala Glin Ser Ala Phe Thr Val Ala Asn Ile Phe Glu Gly Ile Phe 195 2OO 2O5 Trp Phe Lieu. Lieu Pro Ala Ser Lieu. Ile Val Ile Asn Asp Ile Phe Ala 21 O 215 22O US 2011/O 145948 A1 Jun. 16, 2011 37
- Continued
Tyr Lieu Phe Gly Phe Phe Leu Gly Arg Thr Pro Leu. Ile Llys Lieu Ser 225 23 O 235 24 O Pro Llys Llys Thr Trp Glu Gly Phe Ile Gly Ala Ser Val Thir Thir Ile 245 250 255 Ile Ser Ala Phe Lieu. Lieu Ala Asn. Wal Met Gly Arg Phe Glin Trp Lieu. 26 O 265 27 O Thir Cys Pro Arg Lys Asp Lieu. Ser Thr Gly Trp Lieu. Tyr Cys Asp Pro 27s 28O 285 Gly Pro Met Phe Llys Pro Glu. His Tyr Ser Leu Gly Glu Ser Val Pro 29 O 295 3 OO His Trp Phe Pro Trp Lys Asp Leu Ala Ile Met Pro Val Glin Trp His 3. OS 310 315 32O Ala Lieu Ala Lieu. Gly Lieu. Phe Ala Ser Ile Ile Ala Pro Phe Gly Gly 3.25 330 335 Phe Phe Ala Ser Gly Phe Lys Arg Ala Phe Lys Ile Lys Asp Phe Gly 34 O 345 35. O Asp Ser Ile Pro Gly. His Gly Gly Ile Thr Asp Arg Met Asp Cys Glin 355 360 365 Met Val Met Ala Val Phe Ala Tyr Ile Tyr His Glin Ser Phe Ile Ala 37 O 375 38O Pro Glin Asn. Phe Ser Val Glu Ile Ile Lieu. Asp Glin Ile Lieu. Arg Asn 385 390 395 4 OO Lieu. Thir Tyr Glu Glu Gln Lys Tyr Lieu. Tyr Glu Gln Leu Gly Glu Met 4 OS 41O 415 Phe His Glu Arg Glin Lieu. Gly Glin Ser 42O 425
<210s, SEQ ID NO 7 &211s LENGTH: 378 &212s. TYPE: DNA <213> ORGANISM: Saccharomyces cerevisiae 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (378) <223> OTHER INFORMATION: acyl-carrier protein (YKL192C)
<4 OO > SEQUENCE: 7 atgttt aga toc gtt togc cqc att to t t c c cqc gtg gca cct tct gcg 48 Met Phe Arg Ser Val Cys Arg Ile Ser Ser Arg Val Ala Pro Ser Ala 1. 5 1O 15 tac cqc act ata atg ggc cqt to c gtt atgtcc aac acc at a ctic goa 96 Tyr Arg Thr Ile Met Gly Arg Ser Val Met Ser Asn. Thir Ile Leu Ala 2O 25 3O caa aga titt tat t cit gca aac titg agc aaa gat cag gtt tot caa agg 144 Glin Arg Phe Tyr Ser Ala Asn Lieu. Ser Lys Asp Glin Val Ser Glin Arg 35 4 O 45 gtc att gat gtt atc aag gog titt gat aag aac tot coc aac att go c 192 Val Ile Asp Val Ile Lys Ala Phe Asp Lys Asn. Ser Pro Asn. Ile Ala SO 55 6 O aac aag caa at c toc agc gat acc caa titt cac aag gat ttgggg ttg 24 O Asn Lys Glin Ile Ser Ser Asp Thr Glin Phe His Lys Asp Lieu. Gly Lieu 65 70 7s 8O gac to C titg gac act gtc. gag ctg. Ct c gta gct att gaa gala gala titt 288 Asp Ser Lieu. Asp Thr Val Glu Lieu. Lieu Val Ala Ile Glu Glu Glu Phe 85 90 95 US 2011/O 145948 A1 Jun. 16, 2011 38
- Continued gat att gala at C cct gac aaa gtg gct gat gag titg aga agt gtt ggit 336 Asp Ile Glu Ile Pro Asp Llys Val Ala Asp Glu Lieu. Arg Ser Val Gly 1OO 105 11 O gaa acg gt c gat tat atc gct tcc aat coc gac gca aac taa 378 Glu Thr Val Asp Tyr Ile Ala Ser Asn Pro Asp Ala Asn 115 12 O 125
<210s, SEQ ID NO 8 &211s LENGTH: 125 212. TYPE: PRT <213> ORGANISM: Saccharomyces cerevisiae <4 OOs, SEQUENCE: 8 Met Phe Arg Ser Val Cys Arg Ile Ser Ser Arg Val Ala Pro Ser Ala 1. 5 1O 15 Tyr Arg Thr Ile Met Gly Arg Ser Val Met Ser Asn. Thir Ile Leu Ala 2O 25 3O Glin Arg Phe Tyr Ser Ala Asn Lieu. Ser Lys Asp Glin Val Ser Glin Arg 35 4 O 45 Val Ile Asp Val Ile Lys Ala Phe Asp Lys Asn. Ser Pro Asn. Ile Ala SO 55 6 O Asn Lys Glin Ile Ser Ser Asp Thr Glin Phe His Lys Asp Lieu. Gly Lieu 65 70 7s 8O Asp Ser Lieu. Asp Thr Val Glu Lieu. Lieu Val Ala Ile Glu Glu Glu Phe 85 90 95 Asp Ile Glu Ile Pro Asp Llys Val Ala Asp Glu Lieu. Arg Ser Val Gly 1OO 105 11 O Glu Thr Val Asp Tyr Ile Ala Ser Asn Pro Asp Ala Asn 115 12 O 125
<210s, SEQ ID NO 9 &211s LENGTH: 354 &212s. TYPE: DNA <213> ORGANISM: Brassica napus 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (354) <223> OTHER INFORMATION: acyl-carrier protein (BN1004 MS.43616414)
<4 OOs, SEQUENCE: 9 atg cac to at C agg agc agc at C ctg cag cac Ctg agg ctg agg gtg 48 Met His Cys Ile Arg Ser Ser Ile Lieu Gln His Lieu. Arg Lieu. Arg Val 1. 5 1O 15 ccg gtg agg agc gtg Ctg ctg. Ctg gag aag gag aac gtg Ctg at C agc 96 Pro Val Arg Ser Val Lieu. Lieu. Lieu. Glu Lys Glu Asn Val Lieu. Ile Ser 2O 25 3O aag atgaac ttic acc agc ggc ggc ggc cag gaC cag gtg Ctg agc aag 144 Llys Met Asn. Phe Thir Ser Gly Gly Gly Glin Asp Glin Val Lieu. Ser Lys 35 4 O 45 gtg at C gag ctg gtg aag aag tac gaC acc acc agc gcc agc aag gtg 192 Val Ile Glu Lieu Val Llys Llys Tyr Asp Thir Thr Ser Ala Ser Llys Val SO 55 6 O acc gag acc gcc gac titc aag aag gac ctg agc Ctg gac agc ctg. gac 24 O Thr Glu Thir Ala Asp Phe Llys Lys Asp Lieu. Ser Lieu. Asp Ser Lieu. Asp 65 70 7s 8O agg gtg gag at C gtg atg gcc at C gag gag gag titc agc gtg gag at C 288 Arg Val Glu Ile Val Met Ala Ile Glu Glu Glu Phe Ser Val Glu Ile 85 90 95 US 2011/O 145948 A1 Jun. 16, 2011 39
- Continued ccg gac gag aag gcc gac aag Ctg acc tec tec goc gac at C goc agc 336 Pro Asp Glu Lys Ala Asp Llys Lieu. Thir Cys Cys Ala Asp Ile Ala Ser 1OO 105 11 O titc at C gtg agc gag ta 3.54 Phe Ile Wal Ser Glu 115
<210s, SEQ ID NO 10 &211s LENGTH: 117 212. TYPE: PRT <213> ORGANISM: Brassica napus <4 OOs, SEQUENCE: 10 Met His Cys Ile Arg Ser Ser Ile Lieu Gln His Lieu. Arg Lieu. Arg Val 1. 5 1O 15 Pro Val Arg Ser Val Lieu. Lieu. Lieu. Glu Lys Glu Asn Val Lieu. Ile Ser 2O 25 3O Llys Met Asn. Phe Thir Ser Gly Gly Gly Glin Asp Glin Val Lieu. Ser Lys 35 4 O 45 Val Ile Glu Lieu Val Llys Llys Tyr Asp Thir Thr Ser Ala Ser Llys Val SO 55 6 O Thr Glu Thir Ala Asp Phe Llys Lys Asp Lieu. Ser Lieu. Asp Ser Lieu. Asp 65 70 7s 8O Arg Val Glu Ile Val Met Ala Ile Glu Glu Glu Phe Ser Val Glu Ile 85 90 95 Pro Asp Glu Lys Ala Asp Llys Lieu. Thir Cys Cys Ala Asp Ile Ala Ser 1OO 105 11 O
Phe Ile Wal Ser Glu 115
<210s, SEQ ID NO 1 &211s LENGTH: 384 &212s. TYPE: DNA <213> ORGANISM: Glycine max 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (384) <223> OTHER INFORMATION: acyl-carrier protein (GMO6MCO7589)
<4 OOs, SEQUENCE: 1 atg caa agc at a agg aaa tict at C titg act cqt gtgaat ttg agg aga 48 Met Glin Ser Ile Arg Llys Ser Ile Lieu. Thir Arg Val Asn Lieu. Arg Arg 1. 5 1O 15 tca acc gala aga tigg ttt ttg acc agg gat gag gtt gtg cat atg caa 96 Ser Thr Glu Arg Trp Phe Lieu. Thr Arg Asp Glu Val Val His Met Glin 2O 25 3O ttg aga tigt togg togc tict tca aca gct gtc agc tict gat caa at a citg 144 Lieu. Arg Cys Trp Cys Ser Ser Thr Ala Val Ser Ser Asp Glin Ile Lieu. 35 4 O 45 gac ca gtg att gca Citg gcc aag aaa tat gat aaa att gat ggc tica 192 Asp Arg Val Ile Ala Lieu Ala Lys Llys Tyr Asp Llys Ile Asp Gly Ser SO 55 6 O aag gt C act gala aca gct gat titt caa aaa gac ttgaac Ctg gaC agt 24 O Llys Val Thr Glu Thir Ala Asp Phe Glin Lys Asp Lieu. Asn Lieu. Asp Ser 65 70 7s 8O ttg gac cqa gtg gaa citc att atg gcc citt gaa gaa gaa ttt tot att 288 Lieu. Asp Arg Val Glu Lieu. Ile Met Ala Lieu. Glu Glu Glu Phe Ser Ile 85 90 95 US 2011/O 145948 A1 Jun. 16, 2011 40
- Continued gaa at C cct gat gag aag gct gat aag Ctt gct tcc tit gct gat att 336 Glu Ile Pro Asp Glu Lys Ala Asp Llys Lieu Ala Cys Cys Ala Asp Ile 1OO 105 11 O gca aaa tac at a gca gag gtt gat cag aaa aac Ctg gala aag ccc tra 384 Ala Lys Tyr Ile Ala Glu Val Asp Gln Lys Asn Lieu. Glu, Llys Pro 115 12 O 125
<210s, SEQ ID NO 12 &211s LENGTH: 127 212. TYPE: PRT <213> ORGANISM: Glycine max <4 OOs, SEQUENCE: 12 Met Glin Ser Ile Arg Llys Ser Ile Lieu. Thir Arg Val Asn Lieu. Arg Arg 1. 5 1O 15 Ser Thr Glu Arg Trp Phe Lieu. Thr Arg Asp Glu Val Val His Met Glin 2O 25 3O Lieu. Arg Cys Trp Cys Ser Ser Thr Ala Val Ser Ser Asp Glin Ile Lieu. 35 4 O 45 Asp Arg Val Ile Ala Lieu Ala Lys Llys Tyr Asp Llys Ile Asp Gly Ser SO 55 6 O Llys Val Thr Glu Thir Ala Asp Phe Glin Lys Asp Lieu. Asn Lieu. Asp Ser 65 70 7s 8O Lieu. Asp Arg Val Glu Lieu. Ile Met Ala Lieu. Glu Glu Glu Phe Ser Ile 85 90 95 Glu Ile Pro Asp Glu Lys Ala Asp Llys Lieu Ala Cys Cys Ala Asp Ile 1OO 105 11 O Ala Lys Tyr Ile Ala Glu Val Asp Gln Lys Asn Lieu. Glu, Llys Pro 115 12 O 125
<210s, SEQ ID NO 13 &211s LENGTH: 360 &212s. TYPE: DNA <213> ORGANISM; Helianthus annuus 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (360) <223> OTHER INFORMATION: acyl-carrier protein (HA1004 MS66693619
<4 OOs, SEQUENCE: 13 atg gcc gcc agg aac gcc ctg. Ctg aag tac Ctg agg gtgaac gitg acc 48 Met Ala Ala Arg Asn Ala Lieu. Lieu Lys Tyr Lieu. Arg Val Asin Val Thr 1. 5 1O 15 ccg gcc ctg cag agc agc agc gcc cag aac ccg agc gcc at C ggc ggc 96 Pro Ala Lieu. Glin Ser Ser Ser Ala Glin Asn Pro Ser Ala Ile Gly Gly 2O 25 3O ggc ctg at C cag Ctg titc agg agg cac tt C agc gag gag gtg agg ggc 144 Gly Lieu. Ile Glin Lieu. Phe Arg Arg His Phe Ser Glu Glu Val Arg Gly 35 4 O 45 agc titc ctg gac aag agc gag gtg acc gac agg gtg gtg acc tic gtg 192 Ser Phe Lieu. Asp Llys Ser Glu Val Thr Asp Arg Val Val Thir Cys Val SO 55 6 O aag aac tt C cag aag gtg gaC ccg agc aag gtg acc ccg acc gcc cac 24 O Lys Asn Phe Gln Lys Val Asp Pro Ser Llys Val Thr Pro Thr Ala His 65 70 7s 8O ttic cag aac gac Ctg ggc Ctg gac agc ctg gac acc gtg gag gtg gtg 288 Phe Glin Asn Asp Lieu. Gly Lieu. Asp Ser Lieu. Asp Thr Val Glu Val Val 85 90 95 US 2011/O 145948 A1 Jun. 16, 2011 41
- Continued atg gcc ctg gag gag gag titc ggc titc gag atc cc.g. gac aac gag gCC 336 Met Ala Lieu. Glu Glu Glu Phe Gly Phe Glu Ile Pro Asp Asn. Glu Ala 1OO 105 11 O gac aag at C agc agc atc gag tea 360 Asp Llys Ile Ser Ser Ile Glu 115
SEQ ID NO 14 LENGTH: 119 TYPE PRT ORGANISM: Helianthus annuus
< 4 OOs SEQUENCE: 14 Met Ala Ala Arg Asn Ala Lieu. Lieu Lys Tyr Lieu. Arg Val Asin Val Thr 1. 5 1O 15 Pro Ala Lieu. Glin Ser Ser Ser Ala Glin Asn Pro Ser Ala Ile Gly Gly
Gly Lieu. Ile Glin Lieu. Phe Arg Arg His Phe Ser Glu Glu Val Arg Gly 35 4 O 45 Ser Phe Lieu. Asp Llys Ser Glu Val Thr Asp Arg Val Val Thir Cys Val SO 55 6 O Lys Asn Phe Gln Lys Val Asp Pro Ser Llys Val Thr Pro Thr Ala His 65 Phe Glin Asn Asp Lieu. Gly Lieu. Asp Ser Lieu. Asp Thr Val Glu Val Val 85 90 95 Met Ala Lieu. Glu Glu Glu Phe Gly Phe Glu Ile Pro Asp Asn. Glu Ala 1OO 105 11 O Asp Llys Ile Ser Ser Ile Glu 115
SEO ID NO 15 LENGTH: 1191. TYPE: DNA ORGANISM: Saccharomyces cerevisiae FEATURE: NAME/KEY: CDS LOCATION: (1) ... (1191) OTHER INFORMATION: acyltransferase (YDRO18C)
SEQUENCE: 15 atg aag cat tcc caa aaa tac cqt agg tat gga att tat gala aag act 48 Met Lys His Ser Gln Lys Tyr Arg Arg Tyr Gly Ile Tyr Glu Lys Thr 1. ggit aat coc titt ata aaa ggg ttg caa agg ctg. Ctt atc gct tcc ttg 96 Gly Asn Pro Phe Ile Llys Gly Lieu. Glin Arg Lieu. Lieu. Ile Ala Cys Lieu. titc att to a ggc ticg citg agt att gtc gtt titt cag atc tigt cta cag 144 Phe Ile Ser Gly Ser Leu Ser Ile Val Val Phe Glin Ile Cys Lieu. Glin 35 4 O 45 gtg citt ct c cot togg agc aag att aga titt caa aat gigt ata aat caa 192 Val Lieu. Lieu Pro Trp Ser Lys Ile Arg Phe Glin Asn Gly Ile Asin Glin SO 55 6 O agt aag aag got titt atc gtt tta tta tigc atg atc ttgaac at g g td 24 O Ser Lys Lys Ala Phe Ile Val Lieu. Lieu. Cys Met Ile Lieu. Asn Met Val 65 gct coc tot tot ttgaat gtc act titt gala aca tog cqg cca ttgaag 288 Ala Pro Ser Ser Lieu. Asn Val Thr Phe Glu Thir Ser Arg Pro Leu Lys 85 90 95
US 2011/O 145948 A1 Jun. 16, 2011 43
- Continued
<210s, SEQ ID NO 16 &211s LENGTH: 396 212. TYPE: PRT <213> ORGANISM: Saccharomyces cerevisiae <4 OOs, SEQUENCE: 16 Met Lys His Ser Gln Lys Tyr Arg Arg Tyr Gly Ile Tyr Glu Lys Thr 1. 5 1O 15 Gly Asn Pro Phe Ile Llys Gly Lieu. Glin Arg Lieu. Lieu. Ile Ala Cys Lieu. 2O 25 3O Phe Ile Ser Gly Ser Leu Ser Ile Val Val Phe Glin Ile Cys Lieu. Glin 35 4 O 45 Val Lieu. Lieu Pro Trp Ser Lys Ile Arg Phe Glin Asn Gly Ile Asin Glin SO 55 6 O Ser Lys Lys Ala Phe Ile Val Lieu. Lieu. Cys Met Ile Lieu. Asn Met Val 65 70 7s 8O Ala Pro Ser Ser Lieu. Asn Val Thr Phe Glu Thir Ser Arg Pro Leu Lys 85 90 95 Asn Ser Ser Asn Ala Lys Pro Cys Phe Arg Phe Lys Asp Arg Ala Ile 1OO 105 11 O Ile Ile Ala Asn His Glin Met Tyr Ala Asp Trp Ile Tyr Lieu. Trp Trp 115 12 O 125 Lieu. Ser Phe Val Ser Asn Lieu. Gly Gly ASn Val Tyr Ile Ile Lieu Lys 13 O 135 14 O Lys Ala Lieu. Glin Tyr Ile Pro Lieu. Lieu. Gly Phe Gly Met Arg Asn. Phe 145 150 155 160 Llys Phe Ile Phe Lieu. Ser Arg Asn Trp Gln Lys Asp Glu Lys Ala Lieu. 1.65 17O 17s Thir Asn. Ser Lieu Val Ser Met Asp Lieu. Asn Ala Arg Cys Lys Gly Pro 18O 185 19 O Lieu. Thir Asn Tyr Lys Ser Cys Tyr Ser Lys Thr Asn Glu Ser Ile Ala 195 2OO 2O5 Ala Tyr Asn Lieu. Ile Met Phe Pro Glu Gly Thr Asn Lieu. Ser Lieu Lys 21 O 215 22O Thir Arg Glu Lys Ser Glu Ala Phe Cys Glin Arg Ala His Lieu. Asp His 225 23 O 235 24 O Val Glin Lieu. Arg His Lieu Lleu Lieu Pro His Ser Lys Gly Lieu Lys Phe 245 250 255 Ala Val Glu Lys Lieu Ala Pro Ser Lieu. Asp Ala Ile Tyr Asp Val Thr 26 O 265 27 O Ile Gly Tyr Ser Pro Ala Lieu. Arg Thr Glu Tyr Val Gly Thr Llys Phe 27s 28O 285 Thir Lieu Lys Lys Ile Phe Lieu Met Gly Val Tyr Pro Glu Lys Val Asp 29 O 295 3 OO Phe Tyr Ile Arg Glu Phe Arg Val Asn. Glu Ile Pro Lieu. Glin Asp Asp 3. OS 310 315 32O Glu Val Phe Phe Asn Trp Lieu. Lieu. Gly Val Trp Llys Glu Lys Asp Glin 3.25 330 335 Lieu. Lieu. Glu Asp Tyr Tyr Asn Thr Gly Glin Phe Llys Ser Asn Ala Lys 34 O 345 35. O Asn Asp Asn Glin Ser Ile Val Val Thir Thr Glin Thr Thr Gly Phe Glin 355 360 365
US 2011/O 145948 A1 Jun. 16, 2011 45
- Continued Ile Pro Llys Ser Ser Pro Ala Pro Thr Met Leu Arg Lieu. Phe Arg Gly 225 23 O 235 24 O aag agc agc ctg gtg cac gtg cac at C aag agg cac gcc atg aag gac 768 Llys Ser Ser Lieu Val His Val His Ile Lys Arg His Ala Met Lys Asp 245 250 255
Ctg cc.g gag gag gac gag gcc gtg gCC Cag tig tic agg gaC gttgttc 816 Lieu Pro Glu Glu Asp Glu Ala Val Ala Glin Trp Cys Arg Asp Val Phe 26 O 265 27 O gtg gCC aag gaC gcc Ctg Ctg gaC aag cac atc gcc gag gaC acc titc 864 Val Ala Lys Asp Ala Lieu. Lieu. Asp Llys His Ile Ala Glu Asp Thir Phe 27s 28O 285 agc gaC cag gag Ctg cag gac acc ggc agg ccg atc aag agc ctg. Ctg 912 Ser Asp Glin Glu Lieu. Glin Asp Thr Gly Arg Pro Ile Llys Ser Lieu. Lieu. 29 O 295 3 OO gtg gtg at C agc tigg gtg to ctg gtg gtg gcc ggc agc gtg aag titc 96.O Val Val Ile Ser Trp Val Cys Lieu Val Val Ala Gly Ser Val Llys Phe 3. OS 310 315 32O
Ctg cag agg agc agc ctg. Ctg agc agc tigg aag ggc gtg gCC titc agc OO8 Lieu. Glin Arg Ser Ser Lieu Lleu Ser Ser Trp Llys Gly Val Ala Phe Ser 3.25 330 335 gcc ttic ggc ctg gcc gtg gtg acc gcc ctg atg cag atc ctg at C cag O56 Ala Phe Gly Lieu Ala Val Val Thir Ala Lieu Met Glin Ile Lieu. Ile Glin 34 O 345 35. O ttic agc cag agc gag agg agc aac ccg gcc aag atc gtg ccg gCC aag 104 Phe Ser Glin Ser Glu Arg Ser ASn Pro Ala Lys Ile Val Pro Ala Lys 355 360 365 agc aag aac aag ggc ggc cag Ctg gag gCC agg aac gac aag cag cag 152 Ser Lys Asn Lys Gly Gly Glin Lieu. Glu Ala Arg Asn Asp Llys Glin Glin 37 O 375 38O tga 155
<210s, SEQ ID NO 18 &211s LENGTH: 384 212. TYPE: PRT <213> ORGANISM: Glycine max
<4 OOs, SEQUENCE: 18 Met Thr Ala Val Val Val Val Pro Leu Gly Lieu Lleu Phe Phe Ala Ser 1. 5 1O 15 Gly Lieu. Ile Val Asn Lieu. Ile Glin Ala Ile Cys Tyr Val Val Val Arg 2O 25 3O Pro Val Ser Lys Asn Lieu. Tyr Arg Arg Met Asn Arg Val Val Ala Glu 35 4 O 45 Lieu. Lieu. Trp Lieu. Glu Lieu Val Trp Ile Ile Asp Trp Trp Ala Gly Val SO 55 6 O Llys Val Glin Val Phe Thr Asp Pro Glu Thr Phe His Ser Met Gly Lys 65 70 7s 8O Glu. His Ala Lieu Val Ile Ser Asn His Arg Ser Asp Ile Asp Trp Lieu. 85 90 95 Val Gly Trp Val Lieu Ala Glin Arg Ser Gly Cys Lieu. Gly Ser Thr Lieu 1OO 105 11 O Ala Val Met Lys Lys Ser Ser Llys Phe Leu Pro Val Ile Gly Trp Ser 115 12 O 125 Met Trp Phe Ser Glu Tyr Lieu. Phe Lieu. Glu Arg Ser Trp Ala Lys Asp 13 O 135 14 O US 2011/O 145948 A1 Jun. 16, 2011 46
- Continued Glu Arg Thr Lieu Lys Ser Gly Lieu. Glin Glin Lieu. Arg Asp Phe Pro Lieu. 145 150 155 160 Pro Phe Trp Leu Ala Leu Phe Val Glu Gly Thr Arg Phe Thr Glin Ala 1.65 17O 17s Llys Lieu. Lieu Ala Ala Glin Glu Tyr Ala Ala Ser Ala Gly Lieu Pro Val 18O 185 19 O Pro Arg Asn Val Lieu. Ile Pro Arg Thr Lys Gly Phe Val Ser Ala Val 195 2OO 2O5 Asn His Met Arg Ser Phe Val Pro Ala Ile Tyr Asp Val Thr Val Ala 21 O 215 22O Ile Pro Llys Ser Ser Pro Ala Pro Thr Met Leu Arg Lieu. Phe Arg Gly 225 23 O 235 24 O Llys Ser Ser Lieu Val His Val His Ile Lys Arg His Ala Met Lys Asp 245 250 255 Lieu Pro Glu Glu Asp Glu Ala Val Ala Glin Trp Cys Arg Asp Val Phe 26 O 265 27 O Val Ala Lys Asp Ala Lieu. Lieu. Asp Llys His Ile Ala Glu Asp Thir Phe 27s 28O 285 Ser Asp Glin Glu Lieu. Glin Asp Thr Gly Arg Pro Ile Llys Ser Lieu. Lieu. 29 O 295 3 OO Val Val Ile Ser Trp Val Cys Lieu Val Val Ala Gly Ser Val Llys Phe 3. OS 310 315 32O Lieu. Glin Arg Ser Ser Lieu Lleu Ser Ser Trp Llys Gly Val Ala Phe Ser 3.25 330 335 Ala Phe Gly Lieu Ala Val Val Thir Ala Lieu Met Glin Ile Lieu. Ile Glin 34 O 345 35. O Phe Ser Glin Ser Glu Arg Ser Asn Pro Ala Lys Ile Val Pro Ala Lys 355 360 365 Ser Lys Asn Lys Gly Gly Glin Lieu. Glu Ala Arg Asn Asp Llys Glin Glin 37 O 375 38O
<210s, SEQ ID NO 19 &211s LENGTH: 999 &212s. TYPE: DNA <213> ORGANISM: Zea mays 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (999) <223> OTHER INFORMATION: acyltransferase (ZMO6MCO4863)
<4 OOs, SEQUENCE: 19 atg gag tec agc aac agc acc agc agc cag ggc cac cac gtg ggc aag 48 Met Glu. Cys Ser Asn Ser Thr Ser Ser Glin Gly His His Val Gly Lys 1. 5 1O 15 tgg aac cag gtg cag atc atc gaC ccg agc atc gtg gag atg ggc cc.g 96 Trp Asn Glin Val Glin Ile Ile Asp Pro Ser Ile Val Glu Met Gly Pro 2O 25 3O agc cac ctg. cc.g. citg agc titc atg agg agg tec agg ggc gtg ctg. tc 144 Ser His Lieu Pro Lieu. Ser Phe Met Arg Arg Cys Arg Gly Val Lieu. Cys 35 4 O 45
Ctg gtg at C atg atc Ctg acc gcc tt C atg atg atg gtg tac Ctg agc 192 Lieu Val Ile Met Ile Lieu. Thir Ala Phe Met Met Met Val Tyr Lieu Ser SO 55 6 O ccg gtg acc acc titc Ctg gtg agg ctg. itt C agc gtg cac tac agc agg 24 O Pro Val Thir Thr Phe Leu Val Arg Lieu Phe Ser Val His Tyr Ser Arg 65 70 7s 8O
US 2011/O 145948 A1 Jun. 16, 2011 48
- Continued Trp Asn Glin Val Glin Ile Ile Asp Pro Ser Ile Val Glu Met Gly Pro 2O 25 3O Ser His Lieu Pro Lieu. Ser Phe Met Arg Arg Cys Arg Gly Val Lieu. Cys 35 4 O 45 Lieu Val Ile Met Ile Lieu. Thir Ala Phe Met Met Met Val Tyr Lieu Ser SO 55 6 O Pro Val Thir Thr Phe Leu Val Arg Lieu Phe Ser Val His Tyr Ser Arg 65 70 7s 8O Lys Ser Thr Cys Phe Leu Phe Gly Met Trp Leu Ala Met Trp Pro Phe 85 90 95 Lieu. Phe Glu Lys Ile Asn Lys Thr Arg Phe Val Phe Ser Gly Glu Ser 1OO 105 11 O Val Pro Ala Lys Glu Arg Val Lieu. Lieu. Phe Ala Asn His Arg Thr Glu 15 12 O 125 Val Asp Trp Met Tyr Lieu. Trp Asp Phe Ala Lieu. Arg Lys Gly Arg Lieu 13 O 135 14 O Glin Cys Ile Llys Tyr Ile Lieu Lys Llys Ser Lieu Met Lys Lieu Pro Val 145 150 155 160 Phe Asn Trp Ala Phe His Ile Ile Glu Phe Ile Pro Val Glu Arg Lys 1.65 17O 17s Trp Glu Ile Asp Glu Ala Ile Ile Arg Ser Arg Lieu. Ser Glu Phe Lys 18O 185 19 O Asn Pro Lys Asp Pro Lieu. Trp Lieu Ala Val Phe Pro Glu Gly. Thir Asp 95 2OO 2O5 Tyr Thr Glu Lys Lys Cys Ile Llys Ser Glin Glu Tyr Ala Ala Glu. His 21 O 215 22O Gly Lieu Pro Val Lieu Lys Asn Val Lieu. Lieu Pro Llys Thr Lys Gly Phe 225 23 O 235 24 O Asn. Cys Cys Lieu. Glin Val Lieu. Arg Ser Thir Ile Asp Ala Val Tyr Asp 245 250 255 Ile Thir Ile Ala Tyr Lys His Arg Pro Pro Thr Phe Lieu. Asp Asn Val 26 O 265 27 O Tyr Gly Val Gly Lieu. Arg Llys Ser Ser Ser Thir Ser Ala Ala Ser Arg 27s 28O 285 Ser Pro Thr Tyr Arg Arg Pro Llys Thr Gly Trp Leu Ala Gly Trp Trp 29 O 295 3 OO Ser Gly Ser Gly Ser Arg Thr Ser Cys Cys Pro Ala Ser Pro Arg Trp 3. OS 310 315 32O Ala Thr Ser Pro Thr Lys Gly Pro Pro Arg Gly Thr 3.25 330
<210s, SEQ ID NO 21 &211s LENGTH: 2142 &212s. TYPE: DNA <213> ORGANISM: Escherichia coli 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (2142) <223> OTHER INFORMATION: bifunctional anaerobic fatty acid oxidation complex protein (B2341)
<4 OOs, SEQUENCE: 21 atg gaa atg aca. tca gcg titt acc citt aat gtt cqt ctd gac aac att 48 Met Glu Met Thir Ser Ala Phe Thr Lieu. Asn Val Arg Lieu. Asp Asn. Ile 1. 5 1O 15
US 2011/O 145948 A1 Jun. 16, 2011 51
- Continued atg ctgaat gala gca gta C9t togt gtt gat gag cag gtt at C cqt agc 1920 Met Lieu. Asn. Glu Ala Val Arg Cys Val Asp Glu Glin Val Ile Arg Ser 625 630 635 64 O gtg cqt gaC ggg gat att ggc gcg gta titt ggc att ggt titt CC g cca 1968 Val Arg Asp Gly Asp Ile Gly Ala Val Phe Gly Ile Gly Phe Pro Pro 645 650 655 titt Ct c ggt gga ccg titc. c9c tat at c gat tct Ctc ggc gcg ggc gala 2O16 Phe Lieu. Gly Gly Pro Phe Arg Tyr Ile Asp Ser Lieu. Gly Ala Gly Glu 660 665 67 O gtg gtt gca at a atg caa cla Ctt gcc acg cag tat ggit to C cqt titt 2O64 Val Val Ala Ile Met Glin Arg Lieu Ala Thr Glin Tyr Gly Ser Arg Phe 675 68O 685 acc cct tc gag cqt ttg gtc gag atg ggc gcg cqt ggg gala agt titt 2112 Thr Pro Cys Glu Arg Lieu Val Glu Met Gly Ala Arg Gly Glu Ser Phe 69 O. 695 7 OO tgg aaa aca act gca act gac ct g caa taa 2142 Trp Llys Thr Thr Ala Thr Asp Leu Gln 7 Os 71O
<210s, SEQ ID NO 22 &211s LENGTH: 713 212. TYPE: PRT <213> ORGANISM: Escherichia coli
<4 OOs, SEQUENCE: 22 Met Glu Met Thir Ser Ala Phe Thr Lieu. Asn Val Arg Lieu. Asp Asn. Ile 1. 5 1O 15 Ala Val Ile Thir Ile Asp Val Pro Gly Glu Lys Met Asn. Thir Lieu Lys 2O 25 3O Ala Glu Phe Ala Ser Glin Val Arg Ala Ile Ile Llys Glin Lieu. Arg Glu 35 4 O 45 Asn Lys Glu Lieu. Arg Gly Val Val Phe Val Ser Ala Lys Pro Asp Asn SO 55 6 O Phe Ile Ala Gly Ala Asp Ile Asn Met Ile Gly Asn. Cys Llys Thr Ala 65 70 7s 8O Glin Glu Ala Glu Ala Lieu Ala Arg Glin Gly Glin Glin Lieu Met Ala Glu 85 90 95 Ile His Ala Lieu Pro Ile Glin Val Ile Ala Ala Ile His Gly Ala Cys 1OO 105 11 O Lieu. Gly Gly Gly Lieu. Glu Lieu Ala Lieu Ala Cys His Gly Arg Val Cys 115 12 O 125 Thir Asp Asp Pro Llys Thr Val Lieu. Gly Lieu Pro Glu Val Glin Lieu. Gly 13 O 135 14 O Lieu. Lieu Pro Gly Ser Gly Gly Thr Glin Arg Lieu Pro Arg Lieu. Ile Gly 145 150 155 160 Val Ser Thir Ala Lieu. Glu Met Ile Lieu. Thr Gly Lys Glin Lieu. Arg Ala 1.65 17O 17s Lys Glin Ala Lieu Lys Lieu. Gly Lieu Val Asp Asp Val Val Pro His Ser 18O 185 19 O Ile Lieu. Lieu. Glu Ala Ala Val Glu Lieu Ala Lys Lys Glu Arg Pro Ser 195 2OO 2O5 Ser Arg Pro Lieu Pro Val Arg Glu Arg Ile Lieu Ala Gly Pro Lieu. Gly 21 O 215 22O Arg Ala Lieu. Lieu. Phe Llys Met Val Gly Lys Llys Thr Glu. His Llys Thr US 2011/O 145948 A1 Jun. 16, 2011 52
- Continued
225 23 O 235 24 O Gln Gly Asn Tyr Pro Ala Thr Glu Arg Ile Leu Glu Val Val Glu Thr 245 250 255 Gly Lieu Ala Glin Gly. Thir Ser Ser Gly Tyr Asp Ala Glu Ala Arg Ala 26 O 265 27 O Phe Gly Glu Lieu Ala Met Thr Pro Glin Ser Glin Ala Lieu. Arg Ser Ile 27s 28O 285 Phe Phe Ala Ser Thr Asp Wall Lys Lys Asp Pro Gly Ser Asp Ala Pro 29 O 295 3 OO Pro Ala Pro Lieu. Asn. Ser Val Gly Ile Lieu. Gly Gly Gly Lieu Met Gly 3. OS 310 315 32O Gly Gly Ile Ala Tyr Val Thr Ala Cys Lys Ala Gly Ile Pro Val Arg 3.25 330 335 Ile Lys Asp Ile Asn. Pro Glin Gly Ile Asn His Ala Lieu Lys Tyr Ser 34 O 345 35. O Trp Asp Gln Lieu. Glu Gly Llys Val Arg Arg Arg His Lieu Lys Ala Ser 355 360 365 Glu Arg Asp Llys Glin Lieu Ala Lieu. Ile Ser Gly. Thir Thr Asp Tyr Arg 37 O 375 38O Gly Phe Ala His Arg Asp Lieu. Ile Ile Glu Ala Val Phe Glu Asn Lieu. 385 390 395 4 OO Lieu Lys Glin Gln Met Val Ala Glu Val Glu Gln Asn. CyS Ala Ala His 4 OS 41O 415 Thir Ile Phe Ala Ser Asn Thr Ser Ser Leu Pro Ile Gly Asp Ile Ala 42O 425 43 O Ala His Ala Thr Arg Pro Glu Glin Val Ile Gly Lieu. His Phe Phe Ser 435 44 O 445 Pro Val Glu Lys Met Pro Leu Val Glu Ile Ile Pro His Ala Gly Thr 450 45.5 460 Ser Ala Glin Thir Ile Ala Thir Thr Val Lys Lieu Ala Lys Lys Glin Gly 465 470 47s 48O Llys Thr Pro Ile Val Val Arg Asp Lys Ala Gly Phe Tyr Val Asn Arg 485 490 495 Ile Lieu Ala Pro Tyr Ile Asn. Glu Ala Ile Arg Met Lieu. Thr Glin Gly SOO 505 51O Glu Arg Val Glu. His Ile Asp Ala Ala Lieu Val Llys Phe Gly Phe Pro 515 52O 525 Val Gly Pro Ile Glin Lieu. Lieu. Asp Glu Val Gly Ile Asp Thr Gly Thr 53 O 535 54 O Lys Ile Ile Pro Val Lieu. Glu Ala Ala Tyr Gly Glu Arg Phe Ser Ala 5.45 550 555 560 Pro Ala Asn Val Val Ser Ser Ile Lieu. Asn Asp Asp Arg Lys Gly Arg 565 st O sts Lys Asn Gly Arg Gly Phe Tyr Lieu. Tyr Gly Glin Lys Gly Arg Llys Ser 58O 585 59 O Llys Lys Glin Val Asp Pro Ala Ile Tyr Pro Lieu. Ile Gly. Thr Glin Gly 595 6OO 605 Glin Gly Arg Ile Ser Ala Pro Glin Val Ala Glu Arg Cys Wal Met Lieu. 610 615 62O Met Lieu. Asn. Glu Ala Val Arg Cys Val Asp Glu Glin Val Ile Arg Ser 625 630 635 64 O
US 2011/O 145948 A1 Jun. 16, 2011 56
- Continued
Asp Wall Lys Ala Ile Val Val Thr Gly Lys Gly Gly Llys Phe Ser Gly SO 55 6 O Gly Phe Asp Ile Ser Ser Phe Gly Gly Val Glin Gly Gly Glin Thr Met 65 70 7s 8O Glin Pro Llys Val Gly Tyr Ile Ala Ile Asp Ile Lieu. Thir Asp Thr Val 85 90 95 Glu Ala Ala Thr Llys Pro Ser Val Ala Ala Ile Asp Gly Lieu Ala Lieu. 1OO 105 11 O Gly Gly Gly Lieu. Glu Val Ala Met Ala Cys His Ala Arg Ile Ala Thr 115 12 O 125 Pro Thr Ala Glin Lieu. Gly Lieu Pro Glu Lieu Gln Leu Gly Ile Ile Pro 13 O 135 14 O Gly Phe Gly Gly Thr Glin Arg Lieu Pro Arg Lieu Val Gly Lieu. Thir Lys 145 150 155 160 Ser Lieu. Glu Met Met Lieu Lleu Ser Llys Pro Ile Lys Gly Gly Glu Ala 1.65 17O 17s His Glin Lieu. Gly Lieu Val Asp Ala Lieu Val Ser Pro Asn Asp Lieu Val 18O 185 19 O Asn. Thir Ala Arg Gln Trp Ala Lieu. Asp Ile Tyr Glu. Cys Arg Arg Pro 195 2OO 2O5 Trp Ile Llys Ser Lieu. Tyr Llys Thr Asp Llys Lieu. Glu Pro Lieu. Gly Glu 21 O 215 22O Ala Arg Glu Ile Lieu Lys Phe Ala Arg Ala Glin Ala Gln Lys Glin Ala 225 23 O 235 24 O Ala Asn Lieu. His His Pro Lieu Val Cys Ile Asp Val Ile Glu Glu Gly 245 250 255 Ile Val Ala Gly Pro Arg Ala Gly Lieu. Trp Llys Glu Ala Thir Ser Phe 26 O 265 27 O Gln Glu Lieu. Leu Phe Ser Asp Thr Cys Llys Ser Lieu Val His Val Phe 27s 28O 285 Phe Ser Glin Arg Ala Thir Ser Lys Ile Pro Gly Ala Thr Asp Lieu. Gly 29 O 295 3 OO Lieu Met Pro Arg Lys Ile Thr Llys Val Ala Ile Lieu. Gly Gly Gly Lieu. 3. OS 310 315 32O Met Gly Ser Gly Ile Ala Thr Ala Met Val Leu Ser Asn Tyr Pro Val 3.25 330 335 Lieu. Lieu Lys Glu Val Asn. Glu Lys Phe Lieu. Thir Ala Gly Ile Asn Arg 34 O 345 35. O Ile Glin Ala Asn Lieu. Glin Ser Arg Val Llys Lys Gly Llys Met Thr Glu 355 360 365 Glu Arg Tyr Glu Lys Ala Met Ser Lieu Val Thr Gly Val Lieu. Asp Tyr 37 O 375 38O Glu Arg Phe Lys Asp Wall Asp Lieu Val Ile Glu Ala Val Ile Glu Asn 385 390 395 4 OO Val Llys Lieu Lys Glin Glin Ile Phe Ser Asp Lieu. Glu Lys Tyr Cys Pro 4 OS 41O 415 Ser His Cys Ile Lieu Ala Thr Asn. Thir Ser Thir Ile Asp Lieu. Asn Lieu. 42O 425 43 O Ile Gly Glu Lys Thr Lys Ala Glin Asp Arg Ile Ala Gly Ala His Phe 435 44 O 445 US 2011/O 145948 A1 Jun. 16, 2011 57
- Continued Phe Ser Pro Ala His Val Met Pro Leu Lieu. Glu Ile Val Arg Thr Glin 450 45.5 460 His Thir Ser Pro Glin Val Val Val Asp Lieu. Lieu. Asp Val Gly Llys Llys 465 470 47s 48O Ile Llys Llys Thr Pro Ile Val Val Gly Asn Cys Thr Gly Phe Ala Val 485 490 495 Asn Arg Met Phe Phe Pro Tyr Thr Glin Ser Ala Leu Phe Tyr Val Asp SOO 505 51O Lieu. Gly Met Asp Val Tyr Lys Ile Asp Arg Ala Cys Thr Llys Phe Gly 515 52O 525 Met Pro Met Gly Pro Phe Arg Lieu Ala Asp Leu Val Gly Phe Gly Val 53 O 535 54 O Ala Val Ala Thr Gly Met Glin Tyr Lieu. Glu Asn Phe Pro Glu Arg Val 5.45 550 555 560 Tyr Lys Ser Met Lieu Lleu Pro Lieu Met Met Glu Gly Asn Arg Ala Gly 565 st O sts Glu Ala Thr Glin Lys Gly Phe Tyr Lys Tyr Glu Gly Lys Arg Lys Ala 58O 585 59 O Thr Pro Asp Pro Glu Ile Met Lys Tyr Ile Glu Lys Ser Arg Ser Met 595 6OO 605 Ala Gly Val Thr Pro Asp Pro Glu Lieu Met Lys Lieu. Ser Glu Lys Asp 610 615 62O Ile Val Glu Met Val Phe Phe Pro Val Ile Asn Glu Ala Cys Arg Val 625 630 635 64 O Lieu. Asp Glu Gly Ile Ala Wall Lys Ala Ser Asp Lieu. Asp Ile Ala Ser 645 650 655 Ile Phe Gly Met Gly Phe Pro Pro Tyr Arg Gly Gly Val Met His Trp 660 665 67 O Ala Asp Ser Ile Gly Ala Lys Tyr Ile His Gly Llys Lieu. Glu Glu Trip 675 68O 685 Thr Lys Arg Tyr Gly Gly Phe Phe Llys Pro Cys Ser Tyr Lieu. Ala Glu 69 O. 695 7 OO Arg Ala Ala Lys Gly Ile Pro Lieu. Ser Ala Pro Thr Llys Llys Val Glin 7 Os 71O 71s 72O Ala Arg Lieu.
<210s, SEQ ID NO 25 &211s LENGTH: 218.4 &212s. TYPE: DNA <213> ORGANISM: Zea mays 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (2184) <223> OTHER INFORMATION: bifunctional anaerobic fatty acid oxidation complex protein (ZMO6MC15742)
<4 OOs, SEQUENCE: 25 atg gcc gcc ggc agc atc agg gtg acc atg gag gtg ggc gcc gaC ggc 48 Met Ala Ala Gly Ser Ile Arg Val Thr Met Glu Val Gly Ala Asp Gly 1. 5 1O 15 gtg gCC Ctg at C acc atc gcc aac ccg ccg gtgaac gcc ctg. cac cc.g 96 Wall Ala Lieu. Ile Thir Ile Ala ASn Pro Pro Wall Asn Ala Lieu. His Pro 2O 25 3O atc at C at C goc ggc ctgaag gaC aag tac goc gag gcc ctg agg agg 144 Ile Ile Ile Ala Gly Lieu Lys Asp Llys Tyr Ala Glu Ala Lieu. Arg Arg
US 2011/O 145948 A1 Jun. 16, 2011 60
- Continued
645 650 655 gtg Ctg ggc atgggc titc ccg aag tac agg ggc ggc Ctg gtg tt C tig 2O16 Val Lieu. Gly Met Gly Phe Pro Llys Tyr Arg Gly Gly Lieu Val Phe Trp 660 665 67 O gcc gaC acc gtgggc gcc ccg tac at C cac agc aag Ctg agc aag tig 2O64 Ala Asp Thr Val Gly Ala Pro Tyr Ile His Ser Llys Lieu. Ser Lys Trip 675 68O 685 gcc gag at C tac ggC ccg titc tt C aag ccg agc agc tac ctg gag cag 2112 Ala Glu Ile Tyr Gly Pro Phe Phe Llys Pro Ser Ser Tyr Lieu. Glu Gln 69 O. 695 7 OO agg gcc aag agc ggc gtg ccg Ctg agc gcc ccg ggc gcc agc cag cag 216 O Arg Ala Lys Ser Gly Val Pro Lieu. Ser Ala Pro Gly Ala Ser Glin Glin 7 Os 71O 71s 72O ggc agc gcc agg agc agg atg tda 21.84 Gly Ser Ala Arg Ser Arg Met 72
<210s, SEQ ID NO 26 &211s LENGTH: 727 212. TYPE: PRT <213> ORGANISM: Zea mays <4 OOs, SEQUENCE: 26 Met Ala Ala Gly Ser Ile Arg Val Thr Met Glu Val Gly Ala Asp Gly 1. 5 1O 15
Wall Ala Lieu. Ile Thir Ile Ala ASn Pro Pro Wall Asn Ala Lieu. His Pro 2O 25 3O Ile Ile Ile Ala Gly Lieu Lys Asp Llys Tyr Ala Glu Ala Lieu. Arg Arg 35 4 O 45 Asp Asp Wall Lys Ala Ile Val Lieu. Thr Gly Ala Gly Gly Llys Phe Cys SO 55 6 O Gly Gly Phe Asp Ile Asn Val Phe Thr Llys Val His Glin Thr Gly Asp 65 70 7s 8O Val Ser Leu Met Pro Asp Val Ser Val Glu Lieu Val Ser Asn Met Met 85 90 95 Glu Glu Gly Lys Llys Pro Ser Val Ala Ala Ile Glin Gly Lieu Ala Lieu. 1OO 105 11 O Gly Gly Gly Lieu. Glu Lieu. Thir Met Gly Cys His Ala Arg Ile Ser Thr 115 12 O 125 Pro Glu Ala Glin Lieu. Gly Lieu Pro Glu Lieu. Thir Lieu. Gly Ile Ile Pro 13 O 135 14 O Gly Phe Gly Gly Thr Glin Arg Lieu Pro Arg Lieu Val Gly Lieu Pro Llys 145 150 155 160 Ala Ile Glu Met Met Lieu. Glin Ser Llys Phe Ile Thr Ala Lys Glu Gly 1.65 17O 17s Lys Glu Arg Gly Lieu. Ile Asp Ala Lieu. Cys Ser Pro Asp Glu Lieu. Ile 18O 185 19 O Llys Thir Ser Arg Lieu. Trp Ala Lieu. Glu Ile Ala Asn. Cys Arg Llys Pro 195 2OO 2O5 Trp Met Arg Ser Lieu. Gly Arg Thr Asp Arg Lieu. Gly Pro Lieu. Ser Glu 21 O 215 22O Ala Arg Ala Val Lieu. Asn Ala Ala Arg Glin Glin Ala Met Lys Ile Ala 225 23 O 235 24 O Pro Asn Met Pro Glin Asn Glin Ala Cys Lieu. Asp Wal Met Glu Glu Gly US 2011/O 145948 A1 Jun. 16, 2011 61
- Continued
245 250 255
Ile Luell Gly Gly Glin Ala Gly Wall Luell Lys Glu Ala Met Wall Phe 26 O 265 27 O
Glu Luell Wall Ile Ala Pro Thir Ser Ala Lell Wall His Wall Phe 27s 28O 285
Phe Ala Glin Arg Ser Thir Thir Wall Pro Gly Wall Thir Asp Wall Glin 29 O 295 3 OO
Lell Pro Arg Pro Ile Arg Wall Ala Wall Ile Gly Gly Gly Luell 3. OS 310 315
Met Gly Ser Gly Ile Ala Thir Ser Luell Luell Wall Ser Asn Ile Ser Wall 3.25 330 335
Wall Luell Glu Wall Asn Pro Glin Phe Luell Glin Arg Gly Glu Thir 34 O 345 35. O
Ile Ala Gly Asn Lell Glu Gly Luell Wall Arg Ser Pro Thir 355 360 365
Asp Arg Met His Ala Met Ala Luell Luell Lys Ala Luell Asp 37 O 375
Ser Asp Phe Asp Wall Asp Met Wall Ile Glu Wall Ile Glu Lys 385 390 395 4 OO
Ile Pro Luell Glin Ser Ile Phe Ala Asp Ile Ile Cys Pro 4 OS 415
His Ile Lell Ala Thir Asn Thir Ser Thir Asp Luell Asn Wall 425 43 O
Wall Gly Lys Thir Asn Ser Glin Asp Arg Ile Gly Ala His Phe 435 44 O 445
Phe Ser Pro Ala His Ile Met Pro Luell Luell Glu Wall Arg Thir Glu 450 45.5
Lys Thir Ser Pro Glin Ala Ile Luell Asp Luell Ile Ile Gly Ile 465 470
Ile Wall Pro Ile Wall Wall Gly Asn Cys Thir Gly Phe Ala Wall 485 490 495
Asn Luell Thir Phe Phe Pro Thir Glin Gly Ser His Lell Luell Wall Ser SOO 505
Lell Gly Ile Asp Wall Phe Arg Ile Asp Arg Wall Ile Ser Thir Phe Gly 515 525
Met Pro Met Gly Pro Phe Glin Luell Glin Asp Wall Ala Gly Tyr Gly Wall 53 O 535 54 O
Ala Luell Ala Wall Lys Asp Ile Ala Asp Ala Phe Gly Glu Arg Asn 5.45 550 555 560
Lell Asp Ser Asp Lell Wall Asp Luell Met Wall Asp Gly Arg Glin Gly 565 st O sts
Wall Asn Gly Lys Gly Tyr Ile Glu Gly Gly Lys Pro 585 59 O
Pro Asp Pro Ser Wall His Wall Ile Glu Glu Tyr Arg His 595 6OO 605
Ala Asn Thir Met Pro Gly Gly Lys Pro Wall Thir Lell Thir Asp Glin Asp 610 615 62O
Ile Luell Glu Met Ile Phe Phe Pro Wall Wall ASn Glu Ala Arg Wall 625 630 635 64 O
Met Asp Glu Asn Wall Wall Ile Arg Ala Ser Asp Lell Asp Ile Ala Ser 645 650 655
US 2011/O 145948 A1 Jun. 16, 2011 63
- Continued cgc gtt cat cag tat citg ctic ggit tac got tot gat citt aac titc ct g 624 Arg Val His Glin Tyr Lieu. Lieu. Gly Tyr Ala Ser Asp Lieu. Asn. Phe Lieu 195 2OO 2O5 ccg gta gct ct a cag ccg cac ggc at C ggit ttt Ctc gaa ccg ggg att 672 Pro Val Ala Leu Gln Pro His Gly Ile Gly Phe Lieu. Glu Pro Gly Ile 21 O 215 22O cag att gcc acc att gac cat tcc atg togg titc cat cqc cog titt aat 72 O Glin Ile Ala Thr Ile Asp His Ser Met Trp Phe His Arg Pro Phe Asn 225 23 O 235 24 O ttgaat gala tig citg Ctg tat agc gtg gag agc acc ticg gCd to C agc 768 Lieu. Asn Glu Trp Lieu. Leu Tyr Ser Val Glu Ser Thr Ser Ala Ser Ser 245 250 255 gca cqt ggc titt gtg cqc ggt gag titt tat acc Caa gac ggc gta ctg 816 Ala Arg Gly Phe Val Arg Gly Glu Phe Tyr Thr Glin Asp Gly Val Lieu 26 O 265 27 O gtt gcc tog acc gtt Cag gaa ggg gtg atg cgt aat cac aat taa 861 Val Ala Ser Thr Val Glin Glu Gly Val Met Arg Asn His Asn 27s 28O 285
<210s, SEQ ID NO 28 &211s LENGTH: 286 212. TYPE: PRT <213> ORGANISM: Escherichia coli
<4 OOs, SEQUENCE: 28 Met Ser Glin Ala Lieu Lys Asn Lieu. Lieu. Thr Lieu. Lieu. Asn Lieu. Glu Lys 1. 5 1O 15 Ile Glu Glu Gly Lieu. Phe Arg Gly Glin Ser Glu Asp Lieu. Gly Lieu. Arg 2O 25 3O Glin Val Phe Gly Gly Glin Val Val Gly Glin Ala Lieu. Tyr Ala Ala Lys 35 4 O 45 Glu Thr Val Pro Glu Glu Arg Lieu Val His Ser Phe His Ser Tyr Phe SO 55 6 O Lieu. Arg Pro Gly Asp Ser Llys Llys Pro Ile Ile Tyr Asp Val Glu Thir 65 70 7s 8O Lieu. Arg Asp Gly Asn. Ser Phe Ser Ala Arg Arg Val Ala Ala Ile Glin 85 90 95 Asn Gly Llys Pro Ile Phe Tyr Met Thr Ala Ser Phe Glin Ala Pro Glu 1OO 105 11 O Ala Gly Phe Glu. His Gln Lys Thr Met Pro Ser Ala Pro Ala Pro Asp 115 12 O 125 Gly Lieu Pro Ser Glu Thr Glin Ile Ala Glin Ser Lieu Ala His Lieu. Lieu. 13 O 135 14 O Pro Pro Val Lieu Lys Asp Llys Phe Ile Cys Asp Arg Pro Lieu. Glu Val 145 150 155 160 Arg Pro Val Glu Phe His Asn Pro Leu Lys Gly His Val Ala Glu Pro 1.65 17O 17s His Arg Glin Val Trp Ile Arg Ala Asn Gly Ser Val Pro Asp Asp Lieu. 18O 185 19 O Arg Val His Glin Tyr Lieu. Lieu. Gly Tyr Ala Ser Asp Lieu. Asn. Phe Lieu 195 2OO 2O5 Pro Val Ala Leu Gln Pro His Gly Ile Gly Phe Lieu. Glu Pro Gly Ile 21 O 215 22O Glin Ile Ala Thr Ile Asp His Ser Met Trp Phe His Arg Pro Phe Asn 225 23 O 235 24 O US 2011/O 145948 A1 Jun. 16, 2011 64
- Continued
Lieu. Asn Glu Trp Lieu. Leu Tyr Ser Val Glu Ser Thr Ser Ala Ser Ser 245 250 255 Ala Arg Gly Phe Val Arg Gly Glu Phe Tyr Thr Glin Asp Gly Val Lieu 26 O 265 27 O Val Ala Ser Thr Val Glin Glu Gly Val Met Arg Asn His Asn 27s 28O 285
<210s, SEQ ID NO 29 &211s LENGTH: 747 &212s. TYPE: DNA <213> ORGANISM: Brassica napus 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (747) <223> OTHER INFORMATION: Acyl-CoA thioesterase (BN42634969) <4 OOs, SEQUENCE: 29 atg aag tita gt c cat agt tta cac goc tat titc citt citt tot gga gat 48 Met Lys Lieu Val His Ser Lieu. His Ala Tyr Phe Lieu Lleu Ser Gly Asp 1. 5 1O 15 act aat att coc atc at a tat gala gtt agc cqc tta cqc gat ggc aac 96 Thir Asn. Ile Pro Ile Ile Tyr Glu Val Ser Arg Lieu. Arg Asp Gly Asn 2O 25 3O aat titt gcc acc ca aga gta gat gca aga cag aaa gga aaa acc at a 144 Asn Phe Ala Thr Arg Arg Val Asp Ala Arg Glin Lys Gly Lys Thir Ile 35 4 O 45 titc acc titg titc gca toa titt cag aga gag caa caa got titt gat cac 192 Phe Thr Lieu Phe Ala Ser Phe Glin Arg Glu Gln Glin Gly Phe Asp His SO 55 6 O cag gag tog aac atg cct cat atg ct a cct cott gaa acg citt gta to a 24 O Glin Glu Ser Asn Met Pro His Met Leu Pro Pro Glu. Thir Lieu. Wal Ser 65 70 7s 8O agg gat gala atg Ctt Caa C9g Cdt atg act gaC cat Ctg at a cct agg 288 Arg Asp Glu Met Lieu. Glin Arg Arg Met Thr Asp His Lieu. Ile Pro Arg 85 90 95 tat tac cqa aat aaa gtt gca acc caa tat act gcc cca citg cct at a 336 Tyr Tyr Arg Asn Llys Val Ala Thr Glin Tyr Thr Ala Pro Leu Pro Ile 1OO 105 11 O gat att cqa ttt togt gag cca aat tac tot aca gaa gag aga aag tot 384 Asp Ile Arg Phe Cys Glu Pro Asn Tyr Ser Thr Glu Glu Arg Llys Ser 115 12 O 125
Cct tca aga ttgaac tat td ttt aag gCa agg gga aaa citt tot gat 432 Pro Ser Arg Lieu. Asn Tyr Trp Phe Lys Ala Arg Gly Llys Lieu. Ser Asp 13 O 135 14 O gac caa got tta cac cqa tdt gtg gtt goa titt gct tca gat citg at a 48O Asp Glin Ala Lieu. His Arg Cys Val Val Ala Phe Ala Ser Asp Lieu. Ile 145 150 155 160 titt gca tot at C ggt tta aac cct cac cqt aga aag ggc atg agt gca 528 Phe Ala Cys Ile Gly Lieu. Asn Pro His Arg Arg Lys Gly Met Ser Ala 1.65 17O 17s gct gct citt agc cta gac cac tog titg togg titc. cac cqa colt cita aga 576 Ala Ala Lieu. Ser Lieu. Asp His Ser Lieu. Trp Phe His Arg Pro Lieu. Arg 18O 185 19 O gct gat gac togg citt citc titt gtg at g g tog aat coc aca to a titc cag 624 Ala Asp Asp Trp Leu Lleu Phe Val Met Val Asn Pro Thr Ser Phe Glin 195 2OO 2O5 agt cqt ggit ctg acc act gga gala atgttcaac aga aaa gga gag ct a 672 Ser Arg Gly Lieu. Thir Thr Gly Glu Met Phe Asn Arg Lys Gly Glu Lieu. US 2011/O 145948 A1 Jun. 16, 2011 65
- Continued
21 O 215 22O gtg gta to a ttg acg Cala gaa gca ttg cta aaa gala gcg gtg acg att 72 O Wall Wall Ser Luell Thir Glin Glu Ala Lieu. Lieu Lys Glu Ala Val Thir Ile 225 23 O 235 24 O aag cc c at C ttic ggc gcc aag citc. taa 747 Lys Pro Ile Phe Gly Ala Lys Luell 245
SEQ ID NO 3 O LENGTH: 248 TYPE : PRT ORGANISM: Brassica napus
< 4 OOs SEQUENCE: 3 O
Met Lys Lieu Val His Ser Lell His Ala Tyr Phe Lell Lell Ser Gly Asp 1. 5 15
Thir Asn Ile Pro Ile Ile Glu Wall Ser Arg Lell Arg Asp Gly Asn 25
Asn Phe Ala Thir Arg Wall Asp Ala Arg Glin Lys Gly Thir Ile 35 4 O 45
Phe Thir Luell Phe Ala Ser Phe Glin Arg Glu Glin Glin Gly Phe Asp His SO 55 6 O
Glin Glu Ser Asn Met Pro His Met Luell Pro Pro Glu Thir Luell Wall Ser 65 70 8O
Arg Asp Glu Met Lell Glin Arg Arg Met Thir Asp His Lell Ile Pro Arg 85 90 95
Arg Asn Lys Wall Ala Thir Glin Tyr Thir Ala Pro Luell Pro Ile 105 11 O
Asp Ile Arg Phe Cys Glu Pro Asn Ser Thir Glu Glu Arg Ser 115 12 O 125
Pro Ser Arg Luell Asn Trp Phe Ala Arg Gly Luell Ser Asp 13 O 135 14 O
Asp Glin Ala Luell His Arg Wall Wall Ala Phe Ala Ser Asp Luell Ile 145 150 155 160
Phe Ala Ile Gly Lell Asn Pro His Arg Arg Lys Gly Met Ser Ala 1.65 17O 17s
Ala Ala Luell Ser Lell Asp His Ser Luell Trp Phe His Arg Pro Luell Arg 18O 185 19 O
Ala Asp Asp Trp Lell Lell Phe Wall Met Wall ASn Pro Thir Ser Phe Glin 195
Ser Arg Gly Luell Thir Thir Gly Glu Met Phe ASn Arg Gly Glu Luell 21 O 215
Wall Wall Ser Luell Thir Glin Glu Ala Luell Luell Lys Glu Ala Wall Thir Ile 225 23 O 235 24 O
Pro Ile Phe Gly Ala Luell 245
SEQ ID NO 31 LENGTH: 639 TYPE: DNA ORGANISM: Brassica napus FEATURE: NAME/KEY: CDS LOCATION: (1) . . (639) OTHER INFORMATION: Acyl-CoA thioesterase (BNP53 O23 O) US 2011/O 145948 A1 Jun. 16, 2011 66
- Continued <4 OOs, SEQUENCE: 31 atgtct cat cat tdt tot ttg citt gac acc aac cct gtc. tcc gat coa 48 Met Ser His His Cys Ser Lieu. Lieu. Asp Thr Asn Pro Val Ser Asp Pro 1. 5 1O 15 gct aaa gat citt gag acc ccc tdt citt gtg gaa cqc att ttgtct ct c 96 Ala Lys Asp Lieu. Glu Thr Pro Cys Lieu Val Glu Arg Ile Lieu. Ser Lieu 2O 25 3O cat coa tta tat titg act citt titt cqa ggit titc act at a ccc aat gct 144 His Pro Leu Tyr Lieu. Thir Lieu Phe Arg Gly Phe Thr Ile Pro Asn Ala 35 4 O 45 ata acc titt ggc aag gtt ttt gga ca caa tta gtt gga cag gCa Ctt 192 Ile Thr Phe Gly Llys Val Phe Gly Arg Glin Lieu Val Gly Glin Ala Lieu. SO 55 6 O gcc gca ccg aca aac act gat gala tot acc aag att gtt cat agt tta 24 O Ala Ala Pro Thr Asn Thr Asp Glu Ser Thr Lys Ile Val His Ser Leu 65 70 7s 8O cac toc tat titc cta citc gtt gga gat at a act att coc at c ct a tac 288 His Ser Tyr Phe Lieu. Leu Val Gly Asp Ile Thir Ile Pro Ile Leu Tyr 85 90 95 gaa gtt aac cac tta cqt gat ggc aac aac titt gcc acc cda aga gtt 336 Glu Val Asn His Lieu. Arg Asp Gly Asn. Asn. Phe Ala Thr Arg Arg Val 1OO 105 11 O gat gct aga cag aaa gcc aaa acc att titc atc ttg titt got to a titt 384 Asp Ala Arg Glin Lys Gly Llys Thir Ile Phe Ile Lieu. Phe Ala Ser Phe 115 12 O 125
Cag aga gat cala caa ggt titc gat cac caa gag ticg aac atg cct Ctt 432 Glin Arg Asp Glin Glin Gly Phe Asp His Glin Glu Ser Asn Met Pro Lieu. 13 O 135 14 O atg tca cct cot gala acg Ctt gta aca aga gag gag atg att gala C9g 48O Met Ser Pro Pro Glu Thir Lieu Val Thr Arg Glu Glu Met Ile Glu Arg 145 150 155 160 cgt atg act gac cct ctd cta cct agg gat tac cqa aac aaa att goa 528 Arg Met Thir Asp Pro Lieu Lleu Pro Arg Asp Tyr Arg Asn Lys Ile Ala 1.65 17O 17s gct gala aaa atc ctic aca togg cct at a gac att cqa titt tdt gag cca 576 Ala Glu Lys Ile Lieu. Thir Trp Pro Ile Asp Ile Arg Phe Cys Glu Pro 18O 185 19 O agt tat tat aca gaa cat aca aag tot cot coa aga ttgaac tat tigg 624 Ser Tyr Tyr Thr Glu. His Thr Lys Ser Pro Pro Arg Lieu. Asn Tyr Trp 195 2OO 2O5 titt aag gCa agg ta 639 Phe Lys Ala Arg 21 O
<210s, SEQ ID NO 32 &211s LENGTH: 212 212. TYPE: PRT <213> ORGANISM: Brassica napus <4 OOs, SEQUENCE: 32 Met Ser His His Cys Ser Lieu. Lieu. Asp Thr Asn Pro Val Ser Asp Pro 1. 5 1O 15 Ala Lys Asp Lieu. Glu Thr Pro Cys Lieu Val Glu Arg Ile Lieu. Ser Lieu 2O 25 3O His Pro Leu Tyr Lieu. Thir Lieu Phe Arg Gly Phe Thr Ile Pro Asn Ala 35 4 O 45 Ile Thr Phe Gly Llys Val Phe Gly Arg Glin Lieu Val Gly Glin Ala Lieu. US 2011/O 145948 A1 Jun. 16, 2011 67
- Continued
SO 55 6 O Ala Ala Pro Thr Asn Thr Asp Glu Ser Thr Lys Ile Val His Ser Leu 65 70 7s 8O His Ser Tyr Phe Lieu. Leu Val Gly Asp Ile Thir Ile Pro Ile Leu Tyr 85 90 95 Glu Val Asn His Lieu. Arg Asp Gly Asn. Asn. Phe Ala Thr Arg Arg Val 1OO 105 11 O Asp Ala Arg Glin Lys Gly Llys Thir Ile Phe Ile Lieu. Phe Ala Ser Phe 115 12 O 125 Glin Arg Asp Glin Glin Gly Phe Asp His Glin Glu Ser Asn Met Pro Lieu. 13 O 135 14 O Met Ser Pro Pro Glu Thir Lieu Val Thr Arg Glu Glu Met Ile Glu Arg 145 150 155 160 Arg Met Thir Asp Pro Lieu Lleu Pro Arg Asp Tyr Arg Asn Lys Ile Ala 1.65 17O 17s Ala Glu Lys Ile Lieu. Thir Trp Pro Ile Asp Ile Arg Phe Cys Glu Pro 18O 185 19 O Ser Tyr Tyr Thr Glu. His Thr Lys Ser Pro Pro Arg Lieu. Asn Tyr Trp 195 2OO 2O5 Phe Lys Ala Arg 21 O
<210s, SEQ ID NO 33 &211s LENGTH: 594 &212s. TYPE: DNA <213> ORGANISM: Glycine max 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (594) <223> OTHER INFORMATION: Acyl-CoA thioesterase (GMsaee Of 11) <4 OOs, SEQUENCE: 33 atg gct tca ttt caa aaa gaa gala to a ggg atg gtc. cac cala gag gta 48 Met Ala Ser Phe Gln Lys Glu Glu Ser Gly Met Val His Glin Glu Val 1. 5 1O 15 gct at a cca tot gtc. cct gct coa gat aag Ctt Ctg ccg atg gala gag 96 Ala Ile Pro Ser Val Pro Ala Pro Asp Llys Lieu Lleu Pro Met Glu Glu 2O 25 3O cta cqg gag aga cqt citt act gac cct cqt tta cca ata acc tat cqg 144 Lieu. Arg Glu Arg Arg Lieu. Thir Asp Pro Arg Lieu Pro Ile Thir Tyr Arg 35 4 O 45 aac aaa gta gct aca tot caa titc atc cca togg ccc at a gag at a cqa 192 Asn Llys Val Ala Thr Ser Glin Phe Ile Pro Trp Pro Ile Glu Ile Arg SO 55 6 O tta tdt gala tat gaa act gca aca aat atg aca aaa tot cot coc agt 24 O Lieu. Cys Glu Tyr Glu Thir Ala Thr Asn Met Thr Lys Ser Pro Pro Ser 65 70 7s 8O ttg aga tac tig titt aga gcc aag gga aaa citt to a gat gat cala gCC 288 Lieu. Arg Tyr Trp Phe Arg Ala Lys Gly Lys Lieu. Ser Asp Asp Glin Ala 85 90 95 ttg cat agg tdt gtg gta gca tat aca to a gat cita atc ttic citt caa 336 Lieu. His Arg Cys Val Val Ala Tyr Thir Ser Asp Lieu. Ile Phe Lieu. Glin 1OO 105 11 O gtg agt ttgaac C cc aac ct agg aag gga agg aag gct cqt gct gtg 384 Val Ser Lieu. Asn Pro Asn Arg Arg Lys Gly Arg Lys Ala Arg Ala Val 115 12 O 125 US 2011/O 145948 A1 Jun. 16, 2011 68
- Continued agt ctd gac cac toc atg togg titt cac aga cct tta aga got gat gat 432 Ser Lieu. Asp His Ser Met Trp Phe His Arg Pro Lieu. Arg Ala Asp Asp 13 O 135 14 O tgg at a ct a titt gtg atc titt agt cct act gcc aat aat gcc cqc ggc 48O Trp Ile Leu Phe Val Ile Phe Ser Pro Thr Ala Asn Asn Ala Arg Gly 145 150 155 160 tat gt C act ggc caa atgttcaat cag aag gga gag cat Ctt gtg tot 528 Tyr Val Thr Gly Gln Met Phe Asin Gln Lys Gly Glu. His Leu Val Ser 1.65 17O 17s gtg gtt Caa gala ggit gta atg agg gala gtt att tot gct aag to a gCC 576 Val Val Glin Glu Gly Val Met Arg Glu Val Ile Ser Ala Lys Ser Ala 18O 185 19 O atc aaa tot aat cita tda 594 Ile Llys Ser Asn Lieu. 195
<210s, SEQ ID NO 34 &211s LENGTH: 197 212. TYPE: PRT <213> ORGANISM: Glycine max <4 OOs, SEQUENCE: 34 Met Ala Ser Phe Gln Lys Glu Glu Ser Gly Met Val His Glin Glu Val 1. 5 1O 15 Ala Ile Pro Ser Val Pro Ala Pro Asp Llys Lieu Lleu Pro Met Glu Glu 2O 25 3O Lieu. Arg Glu Arg Arg Lieu. Thir Asp Pro Arg Lieu Pro Ile Thir Tyr Arg 35 4 O 45 Asn Llys Val Ala Thr Ser Glin Phe Ile Pro Trp Pro Ile Glu Ile Arg SO 55 6 O Lieu. Cys Glu Tyr Glu Thir Ala Thr Asn Met Thr Lys Ser Pro Pro Ser 65 70 7s 8O Lieu. Arg Tyr Trp Phe Arg Ala Lys Gly Lys Lieu. Ser Asp Asp Glin Ala 85 90 95 Lieu. His Arg Cys Val Val Ala Tyr Thir Ser Asp Lieu. Ile Phe Lieu. Glin 1OO 105 11 O Val Ser Lieu. Asn Pro Asn Arg Arg Lys Gly Arg Lys Ala Arg Ala Val 115 12 O 125 Ser Lieu. Asp His Ser Met Trp Phe His Arg Pro Lieu. Arg Ala Asp Asp 13 O 135 14 O Trp Ile Leu Phe Val Ile Phe Ser Pro Thr Ala Asn Asn Ala Arg Gly 145 150 155 160 Tyr Val Thr Gly Gln Met Phe Asin Gln Lys Gly Glu. His Leu Val Ser 1.65 17O 17s Val Val Glin Glu Gly Val Met Arg Glu Val Ile Ser Ala Lys Ser Ala 18O 185 19 O Ile Llys Ser Asn Lieu. 195
<210s, SEQ ID NO 35 &211s LENGTH: 888 &212s. TYPE: DNA <213> ORGANISM: Saccharomyces cerevisiae 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) ... (888) <223> OTHER INFORMATION: 2, 4-dienoyl-CoA reductase (YNL2O2W)
US 2011/O 145948 A1 Jun. 16, 2011 70
- Continued Lys Ser Met Thr Ser Lys Lieu. 29 O 295
<210s, SEQ ID NO 36 &211s LENGTH: 295 212. TYPE: PRT <213> ORGANISM: Saccharomyces cerevisiae
<4 OOs, SEQUENCE: 36 Met Asp Thr Met Asn Thr Ala Asn Thr Lieu. Asp Gly Llys Phe Val Thr 1. 5 1O 15 Glu Gly Ser Trp Arg Pro Asp Lieu. Phe Lys Gly Llys Val Ala Phe Val 2O 25 3O Thr Gly Gly Ala Gly. Thir Ile Cys Arg Val Glin Thr Glu Ala Leu Val 35 4 O 45 Lieu. Lieu. Gly Cys Lys Ala Ala Ile Val Gly Arg Asp Glin Glu Arg Thr SO 55 6 O Glu Glin Ala Ala Lys Gly Ile Ser Glin Lieu Ala Lys Asp Lys Asp Ala 65 70 7s 8O Val Lieu Ala Ile Ala Asn Val Asp Val Arg Asn. Phe Glu Glin Val Glu 85 90 95 Asn Ala Wall Lys Llys Thr Val Glu Lys Phe Gly Lys Ile Asp Phe Val 1OO 105 11 O Ile Ala Gly Ala Ala Gly ASn Phe Val Cys Asp Phe Ala Asn Lieu. Ser 115 12 O 125 Pro Asn Ala Phe Llys Ser Val Val Asp Ile Asp Lieu. Lieu. Gly Ser Phe 13 O 135 14 O Asn. Thir Ala Lys Ala Cys Lieu Lys Glu Lieu Lys Llys Ser Lys Gly Ser 145 150 155 160 Ile Leu Phe Val Ser Ala Thr Phe His Tyr Tyr Gly Val Pro Phe Glin 1.65 17O 17s Gly His Val Gly Ala Ala Lys Ala Gly Ile Asp Ala Lieu Ala Lys Asn 18O 185 19 O Lieu Ala Val Glu Lieu. Gly Pro Lieu. Gly Ile Arg Ser Asn. Cys Ile Ala 195 2OO 2O5 Pro Gly Ala Ile Asp Asn Thr Glu Gly Lieu Lys Arg Lieu Ala Gly Lys 21 O 215 22O Llys Tyr Lys Glu Lys Ala Lieu Ala Lys Ile Pro Lieu. Glin Arg Lieu. Gly 225 23 O 235 24 O Ser Thr Arg Asp Ile Ala Glu Ser Thr Val Tyr Ile Phe Ser Pro Ala 245 250 255 Ala Ser Tyr Val Thr Gly Ala Val Lieu Val Val Asp Gly Gly Met Trp 26 O 265 27 O His Leu Gly Thr Tyr Phe Gly His Glu Lieu. Tyr Pro Glu Ala Lieu. Ile 27s 28O 285 Lys Ser Met Thr Ser Lys Lieu. 29 O 295
<210s, SEQ ID NO 37 &211s LENGTH: 795 &212s. TYPE: DNA <213> ORGANISM; Helianthus annuus 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (795)
US 2011/O 145948 A1 Jun. 16, 2011 72
- Continued
<213> ORGANISM; Helianthus annuus
<4 OOs, SEQUENCE: 38 Met Ala Lieu Pro Ser Lieu. Ser Trp Asp Wall Ala Lys Pro Ser Lieu. Thir 1. 5 1O 15 Lieu Pro Thr Arg Pro Ser Ser His Leu Val Phe Arg Lieu Ser Ala Phe 2O 25 3O Ala Gly Asp Val Arg Lys Glin Glu Asp Ala Lys Arg Val Ile Glu Ser 35 4 O 45 Thr Val Llys His Phe Gly Llys Lieu. Asp Ile Lieu Val Asn. Ser Ala Ala SO 55 6 O Gly Asn. Phe Lieu Val Ser Pro Glu Asp Lieu. Ser Pro Asn Gly Phe Lys 65 70 7s 8O Thr Val Met Asp Ile Asp Ser Val Gly Thr Phe Thr Met Cys His Glu 85 90 95 Ala Ile Asn Tyr Lieu Lys Lys Gly Gly Pro Gly Arg Ser Ser Asp Asp 1OO 105 11 O Ser Gly Gly Ile Ile Lieu. Asn Ile Ser Ala Thr Lieu. His Tyr Thr Ala 115 12 O 125 Thir Trp Tyr Glin Ile His Val Ala Ala Ala Lys Ala Ala Val Asp Ala 13 O 135 14 O Ile Thr Arg Asn Lieu Ala Lieu. Glu Trp Gly. Thir Asp Tyr Asp Ile Arg 145 150 155 160 Val Asn Gly Ile Ala Pro Gly Pro Ile Gly Asp Thr Ala Gly Val Arg 1.65 17O 17s Llys Lieu. Gly Pro Ala Glu Ile Llys Ser Gly Thr Arg Glu Ile Met Pro 18O 185 19 O Lieu. Phe Llys Lieu. Gly Glu Lys Trp Asp Ile Ala Val Ala Ala Val Tyr 195 2OO 2O5 Lieu Ala Ser Asp Ala Gly Llys Phe Val Asn Gly. Thir Thr Lieu Val Val 21 O 215 22O Asp Gly Gly Glin Trp Lieu. Ser Gly Pro Arg His Met Pro Lys Glu Glu 225 23 O 235 24 O Val Lys Met Lieu. Ser Arg Val Val Glu Lys Arg Ala Arg Val Ala Pro 245 250 255 Thr Gly Val Pro Ser Ser Lys Lieu. 26 O
<210s, SEQ ID NO 39 &211s LENGTH: 1647 &212s. TYPE: DNA <213> ORGANISM: Saccharomyces cerevisiae 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (1647) <223> OTHER INFORMATION: Sterol esterase (YKL14. OW)
<4 OOs, SEQUENCE: 39 atg tac titc ccc titt tta ggc aga tta tog ata aca gat tac att at a 48 Met Tyr Phe Pro Phe Leu Gly Arg Lieu Ser Ile Thr Asp Tyr Ile Ile 1. 5 1O 15 gtc git c ttg gta tac at a gaa agc att att to a tica gtc. citt aaa citt 96 Val Val Lieu Val Tyr Ile Glu Ser Ile Ile Ser Ser Val Lieu Lys Lieu ata cca caa cca atg att aac citt tt c gaa togg cta ata aat titc. tca 144 US 2011/O 145948 A1 Jun. 16, 2011 73
- Continued
Ile Pro Glin Pro Met Ile Asn Luell Phe Glu Trp Lell Ile Asn Phe Ser 35 4 O 45 acg to c to c gat gac aat a CC at C gala gala aag tta agg to a gct CC a 192 Thir Ser Ser Asp Asp Asn Thir Ile Glu Glu Lys Lell Arg Ser Ala Pro SO 55 6 O act at C Cat gala atg tgt gca att titt gat ata tot gtc gag gat Cat 24 O Thir Ile His Glu Met Cys Ala Ile Phe Asp Ile Ser Wall Glu Asp His 65 70 7s 8O ttg gta aga att gaa gac aat tat at C ttg aca Ctg Cat aga at C CC a 288 Lell Wall Arg Ile Glu Asp Asn Tyr Ile Luell Thir Lell His Arg Ile Pro 85 90 95 cc.g att tot a.a.a. aac aga titt aac aat a.a.a. gtg gta tac tta Cat CaC 336 Pro Ile Ser Lys Asn Arg Phe Asn Asn Wall Wall Luell His His 1OO 105 11 O ggit citt ttg atg tgt tot gat gtt tgg tgt tgc aat att gala aga Cat 384 Gly Luell Luell Met Cys Ser Asp Wall Trp Cys Cys Asn Ile Glu Arg His 115 12 O 125 a.a.a. aac tta cc.g titt gta ttg Cat gat tta ggt tac gac gtC tgg atg 432 Asn Luell Pro Phe Wall Lell His Asp Luell Gly Tyr Asp Wall Trp Met 13 O 135 14 O gga aat aat aga ggit aat a.a.a. tat to a act gcc CaC ttg aac a.a.a. CC a Gly Asn Asn Arg Gly Asn Ser Thir Ala His Lell Asn Pro 145 150 155 160
C Ca a.a.a. tog aac aag titt tgg gat titt tot atc. gac gaa titt gcg ttic 528 Pro Ser Asn Lys Phe Trp Asp Phe Ser Ile Asp Glu Phe Ala Phe 1.65 17O 17s titt gac att CC a aac toa att gala ttic at C tta gat ata aca a.a.a. gtg 576 Phe Asp Ile Pro Asn Ser Ile Glu Phe Ile Luell Asp Ile Thir Wall 18O 185 19 O gac aag gt C at C tgc atc. gga ttic tot Cala ggc tot gct Cala atg titt 624 Asp Lys Wall Ile Cys Ile Gly Phe Ser Glin Gly Ser Ala Glin Met Phe 195 2OO 2O5 gct gca titt tog ttg agt gaa a.a.a. ttg aat cga a.a.a. gtc to c Cat titt 672 Ala Ala Phe Ser Lell Ser Glu Lys Luell Asn Arg Lys Wall Ser His Phe 21 O 215 22O ata gcc at a gca c cc gct atg act CC a aag 999 ttg CaC aac aga att 72 O Ile Ala Ile Ala Pro Ala Met Thir Pro Lys Gly Lell His Asn Arg Ile 225 23 O 235 24 O gtc gat acc ttg gcc a.a.a. toa tog cc c ggc titt atg tat citt ttic titt 768 Wall Asp Thir Luell Ala Ser Ser Pro Gly Phe Met Luell Phe Phe 245 250 255 ggit agg a.a.a. att gtg tta cott to a gct gt C att tgg Cala aga act tta 816 Gly Arg Ile Wall Lell Pro Ser Ala Wall Ile Trp Glin Arg Thir Luell 26 O 265 27 O
Cat CC a aca citt tto aat ttg at C gat atc. gca aac aag at a Ctg 864 His Pro Thir Luell Phe Asn Lell Ile Asp Ile Ala Asn Lys Ile Luell 27s 285 tto aat tgg aag tcg titt aac att Cta cc.g aga Cala aag att gct tot 912 Phe Asn Trp Lys Ser Phe Asn Ile Luell Pro Arg Glin Lys Ile Ala Ser 29 O 295 3 OO tac gca a.a.a. citt tat toa acg acc agt gta a.a.a. t cc att gtt CaC tgg 96.O Tyr Ala Luell Tyr Ser Thir Thir Ser Wall Lys Ser Ile Wall His Trp 3. OS 310 315 tto Cala at a tta aga tot Cag a.a.a. titt Cala atg titt gaa gag tot gat 1008 Phe Glin Ile Luell Arg Ser Glin Lys Phe Glin Met Phe Glu Glu Ser Asp 3.25 330 335 aac atg Cta aat t cc tta act agg cott tac Cala att gct aat titt cott 1056 US 2011/O 145948 A1 Jun. 16, 2011 74
- Continued Asn Met Lieu. Asn Ser Lieu. Thir Arg Pro Tyr Glin Ile Ala Asn Phe Pro 34 O 345 35. O act aga aca aat atc aag att coc att citt tta att tat ggit ggc at a 104 Thr Arg Thr Asn Ile Lys Ile Pro Ile Leu Lieu. Ile Tyr Gly Gly Ile 355 360 365 gat tot tta gtt gat att gat gtg atgaaa aaa aat cita ccc titc aac 152 Asp Ser Lieu Val Asp Ile Asp Wal Met Lys Lys Asn Lieu Pro Phe Asn 37 O 375 38O tcc gtc titt gat gtt aaa gtt gac aat tat gaa cac ctd gat titg att 2OO Ser Val Phe Asp Wall Lys Val Asp Asn Tyr Glu. His Lieu. Asp Lieu. Ile 385 390 395 4 OO tgg ggc aaa gat gcc gat acc ttg gtc att gct aaa gtc titg agg titt 248 Trp Gly Lys Asp Ala Asp Thr Lieu Val Ile Ala Lys Val Lieu. Arg Phe 4 OS 41O 415 att gala titt titt aac cct ggit aat gtt to a gtg aag act aac cag tta 296 Ile Glu Phe Phe Asn Pro Gly Asn Val Ser Val Lys Thr Asn Gln Leu 42O 425 43 O
Cta cca to a gca agt ctg gtt gag gala tta cca agc acg aca tog aag 344 Lieu Pro Ser Ala Ser Lieu Val Glu Glu Lieu Pro Ser Thr Thr Trp Llys 435 44 O 445 aca act cat coa acg cat gigt ct c agt tat aga act cac tog goa gac 392 Thir Thr His Pro Thr His Gly Lieu Ser Tyr Arg Thr His Ser Ala Asp 450 45.5 460 cgt tot cog titg tct gta Caa gct gat gala gC9 gat gag gtc. cac aat 44 O Arg Ser Pro Lieu. Ser Val Glin Ala Asp Glu Ala Asp Glu Val His Asn 465 470 47s 48O gct gac aat tca agg titc tta aga cqa gtg ttt tot act agt gcc at a 488 Ala Asp Asn. Ser Arg Phe Lieu. Arg Arg Val Phe Ser Thir Ser Ala Ile 485 490 495 gac gag gac aac gala aat gag cac cag gat gat aca gaa gat caa at C 536 Asp Glu Asp Asn. Glu Asn. Glu. His Glin Asp Asp Thr Glu Asp Glin Ile SOO 505 51O
Cat aag gag cag caa aga C9g tta agt gtC tat Ctg gaa to a to C aaa 584 His Lys Glu Glin Glin Arg Arg Lieu. Ser Val Tyr Lieu. Glu Ser Ser Lys 515 52O 525 gat tta cd a caa cita gat gcc aac tot tog aca act gcg citg gat gct 632 Asp Lieu. Arg Glin Lieu. Asp Ala Asn. Ser Ser Thir Thr Ala Lieu. Asp Ala 53 O 535 54 O cta aat aaa gaa toga 647 Lieu. Asn Lys Glu 5.45
<210s, SEQ ID NO 4 O &211s LENGTH: 548 212. TYPE: PRT <213> ORGANISM: Saccharomyces cerevisiae <4 OOs, SEQUENCE: 4 O Met Tyr Phe Pro Phe Leu Gly Arg Lieu Ser Ile Thr Asp Tyr Ile Ile 1. 5 1O 15 Val Val Lieu Val Tyr Ile Glu Ser Ile Ile Ser Ser Val Lieu Lys Lieu 2O 25 3O Ile Pro Gln Pro Met Ile Asn Lieu Phe Glu Trp Lieu. Ile Asin Phe Ser 35 4 O 45 Thir Ser Ser Asp Asp Asn. Thir Ile Glu Glu Lys Lieu. Arg Ser Ala Pro SO 55 6 O Thir Ile His Glu Met Cys Ala Ile Phe Asp Ile Ser Val Glu Asp His US 2011/O 145948 A1 Jun. 16, 2011 75
- Continued
Lieu Val Arg Ile Glu Asp Asn Tyr Ile Lieu. Thir Lieu. His Arg Ile Pro 85 90 95 Pro Ile Ser Lys Asn Arg Phe Asin Asn Llys Val Val Tyr Lieu. His His 1OO 105 11 O Gly Lieu. Lieu Met Cys Ser Asp Val Trp Cys Cys Asn. Ile Glu Arg His 115 12 O 125 Lys Asn Lieu Pro Phe Val Lieu. His Asp Lieu. Gly Tyr Asp Val Trp Met 13 O 135 14 O Gly Asn. Asn Arg Gly Asn Llys Tyr Ser Thir Ala His Lieu. Asn Llys Pro 145 150 155 160 Pro Llys Ser Asn Llys Phe Trp Asp Phe Ser Ile Asp Glu Phe Ala Phe 1.65 17O 17s Phe Asp Ile Pro Asn Ser Ile Glu Phe Ile Lieu. Asp Ile Thr Llys Val 18O 185 19 O Asp Llys Val Ile Cys Ile Gly Phe Ser Glin Gly Ser Ala Gln Met Phe 195 2OO 2O5 Ala Ala Phe Ser Lieu. Ser Glu Lys Lieu. Asn Arg Llys Val Ser His Phe 21 O 215 22O Ile Ala Ile Ala Pro Ala Met Thr Pro Lys Gly Lieu. His Asn Arg Ile 225 23 O 235 24 O Val Asp Thr Lieu Ala Lys Ser Ser Pro Gly Phe Met Tyr Lieu. Phe Phe 245 250 255 Gly Arg Lys Ile Val Lieu Pro Ser Ala Val Ile Trp Glin Arg Thr Lieu. 26 O 265 27 O His Pro Thir Lieu. Phe Asn Lieu. Cys Ile Asp Ile Ala Asn Lys Ile Lieu. 27s 28O 285 Phe Asn Trp Llys Ser Phe Asn. Ile Lieu Pro Arg Glin Lys Ile Ala Ser 29 O 295 3 OO Tyr Ala Lys Lieu. Tyr Ser Thr Thr Ser Val Lys Ser Ile Val His Trp 3. OS 310 315 32O Phe Glin Ile Leu Arg Ser Gln Llys Phe Glin Met Phe Glu Glu Ser Asp 3.25 330 335 Asn Met Lieu. Asn Ser Lieu. Thir Arg Pro Tyr Glin Ile Ala Asn Phe Pro 34 O 345 35. O Thr Arg Thr Asn Ile Lys Ile Pro Ile Leu Lieu. Ile Tyr Gly Gly Ile 355 360 365 Asp Ser Lieu Val Asp Ile Asp Wal Met Lys Lys Asn Lieu Pro Phe Asn 37 O 375 38O Ser Val Phe Asp Wall Lys Val Asp Asn Tyr Glu. His Lieu. Asp Lieu. Ile 385 390 395 4 OO Trp Gly Lys Asp Ala Asp Thr Lieu Val Ile Ala Lys Val Lieu. Arg Phe 4 OS 41O 415 Ile Glu Phe Phe Asn Pro Gly Asn Val Ser Val Lys Thr Asn Gln Leu 42O 425 43 O Lieu Pro Ser Ala Ser Lieu Val Glu Glu Lieu Pro Ser Thr Thr Trp Llys 435 44 O 445 Thir Thr His Pro Thr His Gly Lieu Ser Tyr Arg Thr His Ser Ala Asp 450 45.5 460 Arg Ser Pro Lieu. Ser Val Glin Ala Asp Glu Ala Asp Glu Val His Asn 465 470 47s 48O
US 2011/O 145948 A1 Jun. 16, 2011 78
- Continued
Glu Tyr Pro Glu Val Ile Lieu Ala Glu Ala Arg Tyr Asp Ala Glin Ser 85 90 95 Glu Lieu. Phe Lieu Ala Ile Val Lieu. Asp Tyr Glin Arg Glin Cys Pro Val 1OO 105 11 O Lieu Met Gly Ser Ser Glu Gly Gly Ile Asp Val Glu Thir Lieu. Lieu. Glu 115 12 O 125 Gln Met Glin Ser Val Ser Lieu. Arg Thr Asn Phe Ser Pro Tyr Lieu Ala 13 O 135 14 O Arg Arg Lieu Ala Val Llys Met Gly Lieu. Thr Gly Pro Lieu Val Thir Ala 145 150 155 160 Val Ser Gly Ile Ile Gly Lys Met Tyr Glu Lieu Phe Val Thr Tyr Asp 1.65 17O 17s Lieu. Asp Val Ile Glu Ile Asn Pro Lieu. Gly Ile Ser Ala Asp Gly Glu 18O 185 19 O Val Met Ala Lieu. Asp Gly Lys Ile Thr Val Asn Asp Thr Ala Ile Asn 195 2OO 2O5 Arg His Pro Asp Lieu. Ile Asn Trp Arg Ser Glu Gln Trp Thr Gly His 21 O 215 22O Ser Trp Lieu Pro Gly Asn Lieu Ala Glin Gly Glin Ile Gly Lieu. Ile Cys 225 23 O 235 24 O ASn Ser Glu Gly Lieu. Ala Lieu. Ser Thr Trp Asp Lieu. Lieu. ASn Ser Phe 245 250 255 Gly Ile Thr Gly Ala Tyr Lieu. Lieu. Asp Glu Gly Arg Ser Asp Ile Thr 26 O 265 27 O Lieu. Gly Glu Gln Lieu. Glu Lieu Ala Phe Asn His Lieu. Ser Glin Ala Pro 27s 28O 285 Asn Lieu Lys Gly Ile Phe Val Asn Lieu Ala Thr Arg Ala Thr Asp Thr 29 O 295 3 OO Ser Ala Lieu Ala Glu Asp Lieu. Arg Ser Phe Lieu Pro Lieu Pro Pro Asn 3. OS 310 315 32O Lieu. Ser Ser Glu Asp Arg Ser Lieu. Arg Gly Thr Gly Pro Ser Lieu Pro 3.25 330 335 Glin Arg Glin Arg Val Pro Glin Arg Glin Thr Tyr Thr Gly Glu Val Lieu. 34 O 345 35. O Pro Val Val Ile Arg Phe Ser Glin Gly Asn Lieu. Asp His Lieu. Glin Glin 355 360 365 Met Tyr Asp Asn. Ser Lieu Val His Trp His Asn Asp Lieu. Glu Thir Ala 37 O 375 38O Ile Ala Lys Met Lieu. Ser Lieu Met Pro Glin Glu Thir Ala Lieu Ala Glin 385 390 395 4 OO Gly
<210s, SEQ ID NO 43 &211s LENGTH: 786 &212s. TYPE: DNA <213> ORGANISM: Synechocystis sp. 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (786) <223> OTHER INFORMATION: cobalt-precorrin-6A reductase (SLRO252)
<4 OOs, SEQUENCE: 43
US 2011/O 145948 A1 Jun. 16, 2011 80
- Continued Met Val Asp Ser Lieu Ala Thr Val Trp Lieu. Ile Gly Gly Thr Val Asp 1. 5 1O 15 Ser Arg Ala Val Ala Glu Gly Lieu. Ile Ala Glin Gly Ile Asn. Cys Lieu. 2O 25 3O Val Thr Val Thir Thr Ser Glu Ala Lys His Leu Tyr Pro Ile His Glin 35 4 O 45 Cys Lieu. Thr Val His Val Gly Ala Lieu. Thr Pro Glin Glu Ile Pro Llys SO 55 6 O Phe Lieu Lys Arg His Ser Ile Ala Val Ile Val Asp Ala Ser His Pro 65 70 7s 8O Phe Ala Ala Glin Ile Thir Thr Thr Val Thr Ala Ile Ala Lys Glu Gln 85 90 95 Glin Ile Pro Tyr Ile Arg Phe Glu Arg Pro Pro Lieu Ala Lieu. Gly Lys 1OO 105 11 O Asn. Thir Lieu. Glu Val Pro Asp Ile Glin Ser Lieu. Thir Arg Gly Lys Tyr 115 12 O 125 Glin Pro Tyr Lieu. Arg Gly Lys Arg Val Lieu. Lieu. Thr Val Gly Ala Arg 13 O 135 14 O Trp Lieu. Ser His Phe Ser Lieu. Lieu. Glin Asp Glu Ala Val Lieu. Phe Ala 145 150 155 160 Arg Ile Lieu Pro Tyr Pro Glin Ala Lieu Ala Glin Ala Ile Ala Ala Gly 1.65 17O 17s Phe Thir Ser Asp Arg Ile Ile Ala Lieu. Arg Pro Pro Val Ala Glu Pro 18O 185 19 O Lieu. Glu Lys Ala Lieu. Trp Glin Glin Trp Glin Ile Glin Gly Val Val Thr 195 2OO 2O5 Lys Ala Ser Gly Ala Glin Gly Gly Glu Lieu Val Lys Glin Llys Val Ala 21 O 215 22O Glu Ala Lieu. Gly Val Asn Lieu. Ile Arg Ile Ala Arg Pro Glin Thir Ile 225 23 O 235 24 O Pro Gly Glin Ile Thr Asp Asp Leu Ser Glin Ile Asn Glin Phe Cys Glin 245 250 255 Arg His Lieu Pro Ser 26 O
<210s, SEQ ID NO 45 &211s LENGTH: 1182 &212s. TYPE: DNA <213> ORGANISM: Escherichia coli 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (1182) <223> OTHER INFORMATION: uroporphyrin III C-methyltransferase (b3803)
<4 OOs, SEQUENCE: 45 atg acg gaa caa gaa aaa acc to C gcc gtg gtt gaa gag acc agg gag 48 Met Thr Glu Gln Glu Lys Thir Ser Ala Val Val Glu Glu Thr Arg Glu 1. 5 1O 15 gcc gtg gaC acc acg tca caa cct gtC goa aca gala aaa aag agt aag 96 Ala Val Asp Thir Thr Ser Glin Pro Val Ala Thr Glu Lys Llys Ser Lys 2O 25 3O aac aat acc gca ttg att citc agc gcg gtg got atc gct att gct ctd 144 Asn Asn. Thir Ala Lieu. Ile Lieu. Ser Ala Wall Ala Ile Ala Ile Ala Lieu. 35 4 O 45 gcg gCd ggc at C ggit ttg tat ggc tigg ggit aaa Caa cag gCC gtC aat 192
US 2011/O 145948 A1 Jun. 16, 2011 82
- Continued Lieu. Glu Lys Lieu Met Glin Thr Arg Val Arg Asn Lieu. Lieu Ala Glin Pro 355 360 365 gca gCd ggg aca acg gaa got aaa cct gca cct gca ccg caa gCt gat 1152 Ala Ala Gly. Thir Thr Glu Ala Lys Pro Ala Pro Ala Pro Glin Ala Asp 37 O 375 38O act cog gca gcc gcg ccg caa gga gala taa 1182 Thr Pro Ala Ala Ala Pro Glin Gly Glu 385 390
<210s, SEQ ID NO 46 &211s LENGTH: 393 212. TYPE: PRT <213> ORGANISM: Escherichia coli
<4 OOs, SEQUENCE: 46 Met Thr Glu Gln Glu Lys Thir Ser Ala Val Val Glu Glu Thr Arg Glu 1. 5 1O 15 Ala Val Asp Thir Thr Ser Glin Pro Val Ala Thr Glu Lys Llys Ser Lys 2O 25 3O
Asn Asn. Thir Ala Lieu. Ile Lieu. Ser Ala Wall Ala Ile Ala Ile Ala Lieu. 35 4 O 45 Ala Ala Gly Ile Gly Lieu. Tyr Gly Trp Gly Lys Glin Glin Ala Val Asn SO 55 6 O Glin Thr Ala Thir Ser Asp Ala Lieu Ala Asn. Glin Lieu. Thir Ala Lieu. Glin 65 70 7s 8O Lys Ala Glin Glu Ser Glin Lys Ala Glu Lieu. Glu Gly Ile Ile Lys Glin 85 90 95 Glin Ala Ala Glin Lieu Lys Glin Ala Asn Arg Glin Glin Glu Thir Lieu Ala 1OO 105 11 O Lys Glin Lieu. Asp Glu Val Glin Glin Llys Val Ala Thir Ile Ser Gly Ser 115 12 O 125 Asp Ala Lys Thir Trp Lieu. Lieu Ala Glin Ala Asp Phe Lieu Val Lys Lieu 13 O 135 14 O Ala Gly Arg Llys Lieu. Trp Ser Asp Glin Asp Val Thir Thr Ala Ala Ala 145 150 155 160 Lieu. Lieu Lys Ser Ala Asp Ala Ser Lieu Ala Asp Met Asn Asp Pro Ser 1.65 17O 17s Lieu. Ile Thr Val Arg Arg Ala Ile Thr Asp Asp Ile Ala Ser Lieu. Ser 18O 185 19 O Ala Val Ser Glin Val Asp Tyr Asp Gly Ile Ile Lieu Lys Lieu. Asn Glin 195 2OO 2O5 Lieu. Ser Asn Glin Val Asp Asn Lieu. Arg Lieu Ala Asp Asn Asp Ser Asp 21 O 215 22O Gly Ser Pro Met Asp Ser Asp Gly Glu Glu Lieu Ser Ser Ser Ile Ser 225 23 O 235 24 O Glu Trp Arg Ile Asn Lieu Gln Lys Ser Trp Glin Asn. Phe Met Asp Asn 245 250 255 Phe Ile Thir Ile Arg Arg Arg Asp Asp Thir Ala Val Pro Lieu. Lieu Ala 26 O 265 27 O Pro Asn Glin Asp Ile Tyr Lieu. Arg Glu Asn. Ile Arg Ser Arg Lieu. Lieu. 27s 28O 285 Val Ala Ala Glin Ala Val Pro Arg His Glin Glu Glu Thir Tyr Arg Glin 29 O 295 3 OO US 2011/O 145948 A1 Jun. 16, 2011 83
- Continued Ala Lieu. Glu Asn Val Ser Thir Trp Val Arg Ala Tyr Tyr Asp Thir Asp 3. OS 310 315 32O Asp Ala Thir Thr Lys Ala Phe Lieu. Asp Glu Val Asp Gln Lieu. Ser Glin 3.25 330 335 Gln Asn Ile Ser Met Asp Leu Pro Glu Thir Lieu. Glin Ser Glin Ala Met 34 O 345 35. O Lieu. Glu Lys Lieu Met Glin Thr Arg Val Arg Asn Lieu. Lieu Ala Glin Pro 355 360 365 Ala Ala Gly. Thir Thr Glu Ala Lys Pro Ala Pro Ala Pro Glin Ala Asp 37 O 375 38O Thr Pro Ala Ala Ala Pro Glin Gly Glu 385 390
<210s, SEQ ID NO 47 &211s LENGTH: 1107 &212s. TYPE: DNA <213> ORGANISM: Brassica napus 22 Os. FEATURE: <221s NAME/KEY: CDS <222s. LOCATION: (1) . . (1107 <223> OTHER INFORMATION: uroporphyrin III C-methyltransferase (BN51286476)
<4 OOs, SEQUENCE: 47 atg got citt gtc. cag cqg att coc agt titc. tca toc aat citt cqa aac 48 Met Ala Lieu Val Glin Arg Ile Pro Ser Phe Ser Ser Asn Lieu. Arg ASn 1. 5 1O 15 tgg aag agc acc aat tcc aca aac cat aag cct gtt tot tot ct c cat 96 Trp Llys Ser Thr Asn Ser Thr Asn His Llys Pro Val Ser Ser Lieu. His 2O 25 3O tac aaa act cag atc act gct tct tct tcc tog ccc titc acg gag aag 144 Tyr Lys Thr Glin Ile Thr Ala Ser Ser Ser Ser Pro Phe Thr Glu Lys 35 4 O 45 cac tot gtt gag aga tac caa aga gat caa tig citg tac aaa gCatcC 192 His Ser Val Glu Arg Tyr Glin Arg Asp Gln Trp Lieu. Tyr Lys Ala Ser SO 55 6 O act coa tot coat ct coa tog ccg cc.g. tcg aat cag caa gat gala gtc 24 O Thr Pro Ser Pro Ser Pro Ser Pro Pro Ser Asn Glin Glin Asp Glu Val 65 70 7s 8O titt gtt agg gala aac gac atc gca tog cag ctg. cct gag ctgaag aag 288 Phe Val Arg Glu Asn Asp Ile Ala Ser Glin Lieu Pro Glu Lieu Lys Llys 85 90 95
Ct c ttg gag gtt Ctg aga gag aag aga gag agt gga tigC aga ggc ggit 336 Lieu. Lieu. Glu Val Lieu. Arg Glu Lys Arg Glu Ser Gly Cys Arg Gly Gly 1OO 105 11 O gat tic gga cca gga gat gtg titt Ctg gt C ggg aca gga ccg gga gat 384 Asp Cys Gly Pro Gly Asp Val Phe Lieu Val Gly Thr Gly Pro Gly Asp 115 12 O 125 cct gag citt ttg act ttgaaa got gtc aga gtc att caa agc gct gat 432 Pro Glu Lieu. Lieu. Thir Lieu Lys Ala Val Arg Val Ile Glin Ser Ala Asp 13 O 135 14 O citt ctd citt tac gac agg citt gtc. tct aat gat gtt ctd gag titg gtt 48O Lieu. Lieu. Lieu. Tyr Asp Arg Lieu Val Ser Asn Asp Val Lieu. Glu Lieu Val 145 150 155 160 gct cot gat got agg citt citc tat gtc ggc aaa act gct ggc tat cat 528 Ala Pro Asp Ala Arg Lieu. Lieu. Tyr Val Gly Lys Thr Ala Gly Tyr His 1.65 17O 17s agc aga act cag gag gag at a cat gaa ct a ctic Ctt agt titt gct gala 576 US 2011/O 145948 A1 Jun. 16, 2011 84
- Continued Ser Arg Thr Glin Glu Glu Ile His Glu Lieu. Lieu Lleu Ser Phe Ala Glu 18O 185 19 O gct ggit gct act gtt gtg agg ctit aaa ggt gga gat cct ttg gtt titt 624 Ala Gly Ala Thr Val Val Arg Lieu Lys Gly Gly Asp Pro Lieu Val Phe 195 2OO 2O5 gga C9a ggt ggg gala gag atg gaC titt Ctg caa Caa caa ggg att Ca 672 Gly Arg Gly Gly Glu Glu Met Asp Phe Lieu. Glin Glin Glin Gly Ile Arg 21 O 215 22O gtt aaa gtt at C cca ggg att aca gC9 gog tot ggg at a gca gca gag 72 O Val Llys Val Ile Pro Gly Ile Thr Ala Ala Ser Gly Ile Ala Ala Glu 225 23 O 235 24 O
Ctt ggg att coc tta acg cat ca ggt gtt gca act agt gtg agg tt C 768 Lieu. Gly Ile Pro Lieu. Thir His Arg Gly Val Ala Thr Ser Val Arg Phe 245 250 255
Ct c act ggit cat tcc agg aaa gga ggg acc gat cct Ctc ttic gt C gca 816 Lieu. Thr Gly. His Ser Arg Lys Gly Gly. Thir Asp Pro Lieu. Phe Val Ala 26 O 265 27 O gag aat gca gct gat cct gac acg aca. Ctt gtt gtt tat atg ggit ttg 864 Glu Asn Ala Ala Asp Pro Asp Thir Thr Lieu Val Val Tyr Met Gly Lieu. 27s 28O 285 gga act tta cct tct citt gcg cag aaa citg atg gac cac ggit ct a cct 912 Gly. Thir Lieu Pro Ser Lieu Ala Glin Llys Lieu Met Asp His Gly Lieu Pro 29 O 295 3 OO tgt gaC aca cca gct gtt gcg gtt gaa cqt gga acc act cot Ctt cag 96.O Cys Asp Thr Pro Ala Val Ala Val Glu Arg Gly Thr Thr Pro Leu Gln 3. OS 310 315 32O cgt aat gtt ttt gct gag Ctt aaa gac titt gcg act gag att cag gca 1008 Arg Asn Val Phe Ala Glu Lieu Lys Asp Phe Ala Thr Glu Ile Glin Ala 3.25 330 335 gct gga ttg gtg tca cca acg ct C at C at C at a ggg aaa gtC gtC gag 1056 Ala Gly Lieu Val Ser Pro Thr Lieu. Ile Ile Ile Gly Llys Val Val Glu 34 O 345 35. O cta tot cot tta togg cct cat tdc acg aag gaa tac titt ttg citt gtt 1104 Lieu. Ser Pro Leu Trp Pro His Cys Thr Lys Glu Tyr Phe Lieu. Leu Val 355 360 365 taa 1107
<210s, SEQ ID NO 48 &211s LENGTH: 368 212. TYPE: PRT <213> ORGANISM: Brassica napus <4 OOs, SEQUENCE: 48 Met Ala Lieu Val Glin Arg Ile Pro Ser Phe Ser Ser Asn Lieu. Arg Asn 1. 5 1O 15 Trp Llys Ser Thr Asn Ser Thr Asn His Llys Pro Val Ser Ser Lieu. His 2O 25 3O Tyr Lys Thr Glin Ile Thr Ala Ser Ser Ser Ser Pro Phe Thr Glu Lys 35 4 O 45 His Ser Val Glu Arg Tyr Glin Arg Asp Gln Trp Lieu. Tyr Lys Ala Ser SO 55 6 O Thr Pro Ser Pro Ser Pro Ser Pro Pro Ser Asn Glin Glin Asp Glu Val 65 70 7s 8O Phe Val Arg Glu Asn Asp Ile Ala Ser Glin Lieu Pro Glu Lieu Lys Llys 85 90 95 Lieu. Lieu. Glu Val Lieu. Arg Glu Lys Arg Glu Ser Gly Cys Arg Gly Gly