Lecture 3 - Plant Transformation - Casson

Question 1

Why is breeding a genetics problem?

Answer 1

You must understand linkage. Unwanted traits may be inherited too.
It is also a numbers game - the more you can screen, the higher chance you will ultimately generate the plant you want.

Question 2

What are the three main techniques of introducing foreign DNA into plants?

Answer 2

1. Vector (Agrobacterium) - mediated
2. Direct methods (gene gun)
3. Pollen magnofection

Question 3

What are the three critical steps to make a GM plant?

Answer 3

1. Introduction of the construct (using any of the techniques)
2. Selection of transformed cells
3. Regeneration of a whole plant

Question 4

What is the principle behind plant regeneration?

Answer 4

Regeneration of a whole adult plant from tissue taken from a donor

Question 5

What tissues can be used?

Answer 5

• Embryo culture
• Apical meristem (stem cells of plants)
• Leaf tissue (contains meristematic tissue)
• Cell culture eg suspension cultures or protoplasts

Question 6

What is the problem?

Answer 6

Some plants are difficult to regenerate, eg cotton.

Question 7

Describe the steps in plant regeneration (and conditions)

Answer 7

1. Induce proliferation - callus formation (auxin and cytokinin)
2. Induce shoot formation (auxin and CYTOKININ)
3. Induce root formation (AUXIN and cytokinin)
4. Transplant to soil

Question 8

What are common methods of selection?

Answer 8

Antibiotic resistance

Herbicide tolerance