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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

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applications of biotechnology in plant breeding

Applications of Biotechnology inPlant Breeding

Sadanand Dhekney, PhD

University of Florida / IFAS

Mid-Florida Research & Education Center

Apopka, FL 32703

limitations of conventional breeding
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
Plant biotechnology

What is plant biotechnology?

Products on the market

Benefits of biotechnology

plant biotechnology4
Plant biotechnology

A precise process in which scientific techniques

are adopted to develop useful and beneficial plants.

slide5

Commercial variety

New variety

Desired gene

Using plant biotechnology, a single gene may be added to the strand.

(only desired gene is transferred)

=

(transfers)

Desired gene

Traditional plant breeding

Commercial variety

New variety

Traditional donor

DNA is a strand of genes, much like a strand of pearls. Traditional plant breeding combines many genes at once.

(many genes are transferred)

=

X

(crosses)

Desired Gene

Desired gene

Plant biotechnology

what is plant tissue culture
What is Plant Tissue Culture?

Totipotency

Single cell has the ability to regenerate into a whole plant

slide7

What conditions do plant cells need to multiply in vitro?

  • Freedom from competition
  • Nutrients and removal of waste products
  • A controlled environment
slide9

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
slide10

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
slide11

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
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
Organ culture

Any plant organ can serve as an explant to initiate cultures

shoot apical meristem culture
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
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

Embryogenic

Non - embryogenic

Leaf explant

Embryogenic callus

Embryo development

Embryogenic Culture System for Vitis

Initiation from leaves on NB 2 medium (Gray, 1995)

organogenesis
Organogenesis
  • One major path of regeneration
  • Mass multiplication
  • Conservation of germplasm at either normal or sub-zero temperatures
callus cultures
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

Stamen explant

Callus formation from

connective tissue

Callus formation from

filament tip

Embryogenic callus

Embryo development

Embryo germination

Initiation from Stamens and Pistils
in vitro mutagenesis
In vitro mutagenesis
  • Induction of polyploidy
  • Introduction of genetic variability
genetic transformation
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
Genetic Transformation System

+

Co-cultivate

48 h

Callus induction

medium

Embryo explants

Wash explants

Agro culture

Select transgenic embryos

using GFP/NPT II system

Embryo development

medium

Regenerate transgenic

plants

applications
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
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 & Limitations

Advantages

  • Food quantity
  • Food quality
  • Eco friendly

Limitations

  • Recent scientific advances
  • Tangible benefits may not be long term