“CSI” Plant Style: From the Laboratory to the Wheat Field

1. “CSI” Plant Style: From the Laboratory to the Wheat Field

2. CAP Grant • Discover new, valuable, genes and QTLs • Deploy them into adapted varieties around the country • Use marker-assisted selection strategies to enhance gene deployment

3. CAP Grant • 16 institutions around the country coordinate breeding efforts • 4 year grant started in 2006 • Over 20 cultivars/germplasm already released (http://maswheat.ucdavis.edu)

4. Impact of Wheat in Washington In 2006 • Excess of 2.2 million acres planted • 81% winter wheat, 19% spring wheat • Over 140 million bushels harvested • Net return of $615 million

5. From Problem to Lab to Wheat Field

6. What Traditional Problems Do We Face

7. New Characteristics Needed • Drought tolerance • Disease and Insect resistance/tolerance • Herbicide resistance/tolerance • High yield • Excellent end-use quality (High GPC)

8. DNA—Deoxyribo nucleic acid that carries the genetic information in a cell Gene—a sequence of DNA that occupies a specific location and determines a particular characteristic Definitions

9. Definitions • Allele-one member of a pair of genes that occupies a specific location on DNA • A= a= • = homozygous dominant • = homozygous recessive • = heterozygous

10. Punett Square

11. Crossing • Plants are identified which have the trait and which need the trait • Cross is made to introgress trait Survivor, no game X Rock-star yielder, dies 10 Seeds (all ) 25 ( ) 50 ( ) 25 ( ) Self pollination

33. Crossing • Cross is made to introgress trait Survivor, no game X Rock-star yielder, dies 10 Seeds (all ) 25 ( ) 50 ( ) 25 ( ) Self pollination Survivor, Rock-star yielder?

34. How Do We Determine Which Seeds Carry The Trait We Are Interested In?

36. So, which of you plants are resistant to fungal diseases?

37. I am! I am! I am! I am! I am! I am!

38. Definitions • Molecular Markers—DNA segments that can be used as flags to track genes • Linkage—occurs when particular genes are inherited jointly • Linked Markers—markers which are closely associated with a gene

39. DNA Gene Gene Marker We know the DNA sequence of the marker Sometimes we do not know the sequence of the gene we are looking for We can use the marker to indirectly select for the gene of interest

40. Tightly Linked Marker Gene Marker Gene 95% confident Non-linked Marker Marker Gene Gene 50% confident

41. Marker-Assisted Selection • Identify marker which is linked to gene • We can extract DNA from the plants and look for the presence of the marker • This will tell us which have the trait and which do not

43. Picture of harvesting tissue

44. 1 2 3 4

45. + 95% Ethanol

47. PCR Explanation • Polymerase Chain Reaction • Used to amplify DNA via enzymatic replication • Allows a small amount of DNA to be used for analysis • PCR reaction http://www.danquinnart.com/wheatcap/index.htm

49. Gel Electrophoresis • Allows separation of PCR product • DNA inserted into agarose gel • DNA fragments travel with current • Smaller fragments will travel faster than large fragments