1. Shirley’s Lab Works
the new laboratory facility at UNL

2. Shirley’s Lab Works
Overall process Shirley’s lab goes through to insert certain genes into soybeans using Agrobacterium.

3. Shirley’s Lab Works
Mature soybeans are sterilized, put on medium for germination for 5 days; after 5 days the base of the cotyledon is cut to allow modified Agrobacterium to infect the tissues of the soybean.
Mature soybeans are sterilized, put on medium for germination for 5 days; after 5 days the base of the cotyledon is cut to allow modified Agrobacterium to infect the tissues of the soybean.

4. Shirley’s Lab Works
Callus of soybean

5. Shirley’s Lab Works
Soybean cotyledons, explant source for soybean transformation. (callus cells form at tips & agrobacterium infect & transform.

6. Shirley’s Lab Works
Various stages of tissue culture for soybean transformation with agrobacterium.

7. Shirley’s Lab Works
Illustration of how a gene gun works.

8. Shirley’s Lab Works
Illustration of the process of transformation with Agrobacterium tumefaciens.

9. Shirley’s Lab Works
Photographic illustration of the genetic engineering of wheat.
Photographic illustration of the genetic engineering of wheat. This process may take up to 6 months to go from the starting materials on the left to the final product on the right.

10. Shirley’s Lab Works
This picture illustrates the callus cells with the possible genes growing in a medium that will select those callus cells that have the correct genes. The next petri dish has the callus cells growing in hormonal medium to cause the tissues to develop. The final covered dish is a single plant growing in special medium to enhance growth.

11. Shirley’s Lab Works
The plant in the pot is the result of 6 months or more of work in the lab. The next step is growing this plant in the greenhouse to determine what characteristics this transgenic plant will have.

12. Shirley’s Lab Works
This picture illustrates the starting material in developing transgenic wheat. The beaker contains the starting plants; the next petri dish contains the isolated wheat seed; the next petri dish contains the wheat seed growing in medium to cause callus cells to form. These cells will be bombarded with DNA with the goal of having some of these cells ‘take up’ the new genes.

13. Shirley’s Lab Works
Teachers working with solutions of Agrobacterium and the arabidopsis plant.

14. Shirley’s Lab Works
Preparing mixtures of the Agrobacterium

15. Shirley’s Lab Works
Removing open flowers on the arabidopsis plant

16. Shirley’s Lab Works
The arabidopsis plant targeted for infection from Agrobacterium.

17. Shirley’s Lab Works
Each arabidopsis plant is submerged into a solution of Agrobacterium for about 5 minutes (upsidedown).

18. Shirley’s Lab Works
Prepare solutions of Agrobacterium for dunking the arabidopsis plant.

19. Shirley’s Lab Works
Preparing solutions of Agrobacterium.

20. Shirley’s Lab Works
Dunking the arabidopsis plant into solutions of Agrobacterium.

21. Shirley’s Lab Works
Another target plant for genetic modification: tobacco.

22. Shirley’s Lab Works
Preparing a disinfecting solution for the tobacco leaves.

23. Shirley’s Lab Works
Working carefully with tobacco plants.

24. Shirley’s Lab Works
Careful measurement is extremely important, as Kristine is demonstrating!

25. Shirley’s Lab Works
Practical experience is very important to the professional teacher – Working under the hood with tobacco leaves.

26. Shirley’s Lab Works
Transferring sections of tobacco leaves from one medium to the next. Good technique is extremely important if you hope to get good results.

27. Shirley’s Lab Works
Teamwork, concentration, and dedication to the task are also very important. Delicate work involved in cutting sections from the sterilized tobacco leaves.

28. Shirley’s Lab Works
Illustrating the final product – petri dishes full of specimens we hope will develop into viable transgenic cells.

29. Shirley’s Lab Works
Adding a 10% chlorine solution to the tobacco leaves to disinfect them.

30. Shirley’s Lab Works
Two plants that are model systems for transformation are shown here (these are the white mice of the plant world). With arabidopsis we bypass tissue culture by infecting flowers during seed development resulting in transgenic seed. Tobacco uses traditional tissue culture which is infected with the agrobacterium to transform the cells.

31. Shirley’s Lab Works
Shirley did a great job directing us teachers to perform these lab procedures!

32. Shirley’s Lab Works
Heat disinfecting device used in labs to sterilize metals tools used in culturing new colonies of transgenic cells.

33. Shirley’s Lab Works
Tools used in the lab for developing colonies of tobacco leaves infected with Agrobacterium.

34. Shirley’s Lab Works
Lab technicians working on developing colonies of transgenic cells.

35. Shirley’s Lab Works
Graduate student working on preparing pure solutions of Agrobacteria.

36. Shirley’s Lab Works
Specialized refrigerators that keep specimens in the lab for extended periods of time by reducing the temperature.