Applications of Biotechnology in

Plant Breeding
Sadanand Dhekney, PhD

University of Florida / IFAS
Mid-Florida Research & Education Center
Apopka, FL 32703

Limitations of Conventional Breeding


Extreme heterozygosity and pronounced
inbreeding depression in plant species

F1 hybrid produced is of intermediate quality

Long juvenile period of sp. makes screening of
new selections tedious and time consuming
Plant biotechnology
What is plant biotechnology?

Products on the market

Benefits of biotechnology

Plant biotechnology
A precise process in which scientific techniques
are adopted to develop useful and beneficial
plants.

Desired gene
Traditional plant breeding
DNA is a strand of genes,
much like a strand of
pearls. Traditional plant
breeding combines many
genes at once.
Traditional donor Commercial variety New variety
Desired Gene
X =
(crosses)
(many genes are transferred)
Plant biotechnology
Using plant biotechnology,
a single gene may be
added to the strand.
Desired gene Commercial variety New variety
(transfers)
=
Desired gene
(only desired gene is transferred)
What is Plant Tissue Culture?

Totipotency

Single cell has the ability to regenerate into
a whole plant
What conditions do plant
cells need to multiply in
vitro?

Freedom from competition
Nutrients and removal of waste
products
A controlled environment
Applications
Seed culture
Increasing efficiency of germination of
seeds that are difficult to germinate in vivo
Precocious germination by application of
plant growth regulators
Production of clean seedlings for explants
or meristem culture
Embryo culture
Overcoming embryo abortion due to
incompatibility barriers
Overcoming seed dormancy and self-
sterility of seeds
Embryo rescue in distant (interspecific or
intergeneric) hybridization where
endosperm development is poor
Shortening of breeding cycle

Ovary or ovule culture
Production of haploid plants
A common explant for the initiation of somatic
embryogenic cultures
Overcoming abortion of embryos of wide hybrids
at very early stages of development due to
incompatibility barriers
In vitro fertilization for the production of distant
hybrids avoiding style and stigmatic
incompatibility that inhibits pollen germination
and pollen tube growth
Anther and microspore culture
Production of haploid plants
Production of homozygous diploid lines
through chromosome doubling, thus
reducing the time required to produce
inbred lines
Uncovering mutations or recessive
phenotypes
Organ culture
Any plant organ can serve as an explant to
initiate cultures
Shoot apical meristem culture
Production of virus free germplasm
Mass production of desirable genotypes
Facilitation of exchange between locations
(production of clean material)
Cryopreservation (cold storage) or in vitro
conservation of germplasm
Somatic embryogenesis
One major path of regeneration
Mass multiplication
Production of artificial seeds
As source material for embryogenic
protoplasts
Amenable to mechanization and for
bioreactors
Embryogenic Culture System for Vitis
Initiation from leaves on NB 2 medium (Gray, 1995)
Leaf explant
Embryogenic callus
Embryo development
Embryogenic Non - embryogenic
Organogenesis
One major path of regeneration
Mass multiplication
Conservation of germplasm at either
normal or sub-zero temperatures
Callus Cultures
In some instances it is necessary to go through
a callus phase prior to regeneration via somatic
embryogenesis or organogenesis
For generation of useful somaclonal variants
(genetic or epigenetic)
As a source of protoplasts and suspension
cultures
For production of metabolites
Used in in vitro selection
Initiation from Stamens and Pistils
Embryogenic callus
Callus formation from
connective tissue
Callus formation from
filament tip
Embryo development Embryo germination
Stamen explant
In vitro mutagenesis
Induction of polyploidy
Introduction of genetic variability
Genetic transformation
Introduction of foreign DNA to generate
novel (and typically desirable) genetic
combinations
Many different explants can be used,
depending on the plant species and its
favored method of regeneration as well as
the method of transformation
Used to study the function of genes
Genetic Transformation System
Regenerate transgenic
plants
Co-cultivate
48 h
+
Wash explants
Agro culture
Callus induction
medium
Select transgenic embryos
using GFP/NPT II system
Embryo development
medium
Embryo explants
Applications
Disease resistance
Insect resistance
Salinity tolerance
Heavy metals tolerance
Improved nutritional characteristics

More than 50 biotech food products
have been approved for commercial
use in the United States
Canola (5%)
Corn (21%)
Cotton (12%)
Papaya
Potato
Soybeans (62%)
Squash
Sugarbeets
Sweet corn
Tomato
Advantages & Limitations
Advantages
Food quantity
Food quality
Eco friendly

Limitations
Recent scientific advances
Tangible benefits may not be long term