Plant Biotechnology

Chapter 6

Motivation for genetically engineered crops

Agriculture is the biggest industrial sector in the world
$1.3 trillion of products/year

Over past 40 years, world population has doubled while agricultural land area has increased by only 10%

Plant transgenics
Transfer of genes to plants directly accelerates selective breeding practices used in the past.
Cotton fiber strength
increased 1.5% per year through conventional breeding Increased 60% by inserting a single gene into the plant

Corn and soybean have been targets of much genetic engineering

Genetic engineering techniques applied to plants

Cloning
Many types of plants can regenerate from a single cell, similar to a bacterium. The resulting plant is a clone or replica of plant from which original cell was taken.

Protoplast fusion
Introducing a gene into a denuded plant cell and generating a new plant

Protoplast fusion

Agrobacterium tumefaciens as a vector for transferring foreign genes into plant chromosome
This bacterium naturally infects plant cells causing cancerous growths - crown gall disease Infection (vir) genes carried on Ti plasmid

Infection Process
Vir genes copy T-DNA Open channel in bacterial cell membrane for T-DNA to pass through T-DNA enters plant through wound, integrates itself into plant chromosome

http://www.bio.davidson.edu/people/kabernd/seminar/2002/method/dsmeth/ds.htm

Leaf fragment technique used to introduce foreign genes into plant

Small discs are cut from plant leaf Discs are cultured to start a new plant Early in the regeneration process, the bacterium Agrobacterium tumefaciens carrying a Ti plasmid is introduced into the culture The plasmid DNA combines with the plant chromosome Discs are treated with hormones to encourage shoot and root development and then the new plant is planted in the soil

Make leaf discs

Leaf fragment technique

Transfer to shoot stimulating medium Agrobacterium with Ti plasmid with foreign gene Briefly culture discs with genetically modified Agrobacterium Transfer to filter paper over nurse cells Culture 2-3 days

Gene gun

Gene guns

Antisense technology
Used to produce the Flavr-Savr tomato in 1994. Enzyme polygalacturonase breaks down structural polysaccharide pectin in wall of a plant. This is part of the natural decay process in a plant Monsanto identified the gene than encodes the enzyme and made another gene that blocked the production of the enzyme.

Antisense molecules

Plant vaccines
Plants are susceptible to diseases caused by viruses (tobacco mosaic virus) Virus surface protein induces an immune response in the plant against the protein/virus Researchers inserted the virus protein into the plant genome using the Ti plasmid/ Agrobacter vector for insertion and expression in the plant

Concerns about genetically modified foods

Human health
Unsuspected allergens What other issues are there?

Environment Messing up the gene pool of non-target species in the environment

Lateral gene transfer Still poorly understood in nature

Summary
Variety of techniques are available to introduce genes into plants and have the plants express the gene Such genetic engineering is used to
Improve disease resistance Flavor of product Nutrition of product Shelf life of product Any other property of plant that improves its value