Genetic Enhancement of Biodiesel Feedstocks

Richard F. WilsonOilseeds & Bioscience Consulting, LLC

Raleigh, NC

Energy Costs

Petroleum Biofuel

Vegetable Oil Prices (¢/lb) Oil 2002 Current Peanut 32.565.6 Sunflower 23.358.1 Cotton 18.046.6 Corn 19.243.6 Canola 20.547.7 Soybean 16.543.5 Palm 14.936.2

USDA FAS October 2007

World Vegetable Oil Situation

2006/2007 MMT MMT/yr Supply 132 + 5.5

Demand 122 + 5.3

End Stocks 1010 MMT Oil represents about 28 days supply

World Use of Diesel Fuel = 321 billion gallons in 2007Need 300 MMT of vegetable oil to make B20 Biodiesel

USDA FAS Sept 2007

Growth Rate

How to Ensure Adequate Supply of Vegetable Oils?

• Increase Crush Capacity• Increase Area of Production

• Increase Yield per Area•Develop New Oil Crops•Genetically Enhance Oil Concentration & Quality in Major Oilseed Commodities

Variation in Oil Concentration

•Soybean 7.0 to 26%•Sunflower 9.0 to 53%•Canola 12 to 42%•Cotton 20 to 30%•Peanut 30 to 65%

An increase from 20 to 30% oil in soybean seed equals a 50% gain in oil content per

bushel

Mapping & Marking Oil GenesA1 A2 B1 B2 C1 C2 E D2 H M

Satt 276 Satt 067 Satt 509 Satt 304 Satt 194 Satt 291 Satt 598 Satt 458 Satt 353 Satt 201Satt 593 Satt 177 Satt 251 Satt 474 Satt 399 Satt 457 Satt 573 Satt 486 Satt 192 Satt 150Satt 454 Satt 508 Satt 519 Satt 070 Satt 361 Satt 305 Satt 107 Satt 372 Satt 442 567203Satt 526 Satt 429 Satt 597 Satt 272 Satt 190 Satt 277 Satt 602 Satt 154 Satt 009 Satt 435Satt 364 Satt 421 Satt 359 Satt 122 Satt 294 Satt 365 Satt 045 Satt 397 Satt 052 Satt 540Satt 042 Satt 453 Satt 020 Satt 338 Satt 557 Satt 268 Satt 447 Satt 302 Satt 463Satt 300 G Satt 180 Satt 289 Satt 204 Satt 389 Satt 293 Satt 245Satt 073 Satt 163 D1a D1b Satt 164 Satt 134 Satt 263 Satt 461 Satt 317 Satt 323Satt 591 Satt 038 Satt 184 Satt 216 Satt 100 Satt 231 Satt 311 Satt 181 Satt 220Satt 050 Satt 309 Satt 532 Satt 542 L Satt 460 Satt 553 Satt 514 Satt 434 Satt 536Satt 385 Satt 130 Satt 502 Satt 266 Satt 232 Satt 079 Satt 528 Satt 175Satt 545 Satt 324 Satt 548 Satt 428 Satt 182 Satt 307 F Satt 464 K Satt 551Satt 200 Satt 394 Satt 547 Satt 290 Satt 143 Satt 433 Satt 146 Satt 543 Satt 539 Satt 250Satt 511 Satt 594 Satt 603 Satt 537 Satt 523 Satt 202 Satt 343 Satt 082 Satt 242 Satt 210Satt 236 Satt 115 Satt 179 Satt 005 Satt 278 Satt 371 Satt 569 Satt 574 Satt 102 Satt 336Satt 211 Satt 427 Satt 254 Satt 600 Satt 418 Satt 586 Satt 001 Satt 137 Satt 216

Satt 564 Satt 383 Satt 604 Satt 398 O Satt 030 Satt 301 Satt 178I Satt 138 Satt 267 Satt 189 Satt 284 Satt 500 Satt 269 Satt 022 Satt 555 J

Satt 451 Satt 533 Satt 515 Satt 506 Satt 481 Satt 259 Satt 252 Satt 031 Satt 349 Satt 287Satt 419 Satt 303 Satt 203 Satt 141 Satt 156 Satt 345 Satt 423 Satt 256 Satt 381 Satt 249Satt 587 Satt 352 Satt 580 Satt 350 Satt 010 Satt 094 Satt 348 Satt 337 Satt 285Satt 239 Satt 566 Satt 370 Satt 546 Satt 076 Satt 550 Satt 149 N Satt 552 Satt 456Satt 496 Satt 131 Satt 507 Satt 172 Satt 561 Satt 479 Satt 160 Satt 159 Satt 441 Satt 065Satt 270 Satt 340 Satt 077 Satt 274 Satt 527 Satt 188 Satt 425 Satt 152 Satt 417 Satt 596Satt 292 Satt 501 Satt 436 Satt 069 Satt 166 Satt 262 Satt 374 Satt 584 Satt 046 Satt 414Satt 162 Satt 199 Satt 071 Satt 172 Satt 229 Satt 345 Satt 595 Satt 485 Satt 124 Satt 280Satt 148 Satt 012 Satt 408 Satt 459 Satt 373 Satt 173 Satt 114 Satt 387 Satt 264Satt 440 Satt 503 Satt 147 Satt 477 Satt 335 Satt 521 Satt 167Satt 189 Satt 517 Satt 129 Satt 123 Satt 490 Satt 549 Satt 518

Satt 472 Satt 592 Satt 522 Satt 255 Satt 337Satt 191 Satt 581 Satt 144 Satt 234 Satt 326Satt 117 Satt 153 Satt 257 Satt 273

Satt 260

Developing New Genomic Breeding Tools

New DNA markers in Physical Map

Candidate Genes

Genetic markers Genetic mapQTL

BAC contigs, ESTs

Minimal Tiling Path & BAC-end sequences

Physical Map

Bioinformatic Analyses

Genetic Regulation of Oil Concentration

High Yield x High Oil ↓

F1 Plants ↓

F2 Population ↓ SSD

F3 Population ↓ SSD

F4 Population ↓ Single Plants

F4:5 Families ↓ Select & Bulk Test for Yield & Oil

↓ Retest selections

Breeding for Increased Oil

Concentration & Yielding Ability

Genetic Enhancement of Oil %

World Oil Supply, USDA FAS, Sept 2007

Which one is Crude Peanut Oil?

Oleic acid improves oxidative stability

and ignition properties (cetane index) of biodiesel

KAS IIFAT B

FAD 2FAD 3

D9 DS

Linolenic Acid Pathway

FAD2-1 FAD 3

18:1 18:2 18:3Oleic Linoleic Linolenic

Fad Fan

w-3 desaturasew-6 desaturase

Genetic Regulation of Oleic Acid Concentration in Soybean

MAS with Perfect Gene Markers

1% 18:3

GMFAD3A has 2 recessive alleles (aa29, aacx)GMFAD3B has 1 recessive allele (bb29)GMFAD3C has 2 recessive alleles (cccx, cc29) aa29 bb29 cc29 X aacx BB cccx

F1

F2

aa29 bb29 cccx aacx bb29 cc29

~1% of the F3 generation

3% 18:3

Kristen Bilyeu, Columbia MO

New Sources of Oleic Acid