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Genetic
Engineering
PLANT DISEASE RESISTANCE
AND GENETIC ENGINEERING
What is a plant disease?
 A plant disease is any abnormal condition
that alters the abnormal growth or function
of a plant. Disease may also reduce yield and
quality of harvested product.
 Plant diseases are classified in 2 categories:
a) Abiotic
b) Biotic
Abiotic Diseases
 Are caused by (non-living) environmental
conditions such as frost, hail, and
chemical burn.
 Damage caused by chronic exposure to
air pollutants such as nitrogen dioxide,
sulfur dioxide etc.
Biotic Diseases
 Are caused by living organisms such as
fungi, bacteria, viruses, nematodes, etc.
 Pathogens may infect all types of plant
tissues to include leaves, shoots, roots,
fruit, seeds etc.
The Disease Triangle
 For a biotic disease to occur there must be
a susceptible host plant, the pathogen,
optimum environmental conditions.
The Disease Cycle
 The development of visual disease
symptoms on a plant requires that the
pathogen must
(a) come into contact with a susceptible
host
(b) gain entrance or penetrate the host
through either a wound, a natural opening
or via direct penetration of the host
(c) establish itself within the host
(d) grow and reproduce within or on the
host
Biotic Components
Fungi:-
 They damage plants by killing cells or causing
plant stress.
 Sources are infected seed, soil, crop debris,
nearby crops and weed, which spread by
wind and water splash, and through the
movement of contaminated soil etc.
 They enter plants through natural openings
such as stomata and through wounds caused
by pruning, harvesting, hail, insects, other
diseases, and mechanical damage.
Common fungal diseases
 White blister/White rust
 Clubroot
 Botrytis rots
 Anthracnose
 Tuber diseases
Viral Infections
 Viruses cause many plant diseases.
The spread of most viruses is very
difficult to control.
 Viruses are often transmitted from
plant to plant by insects.
 Normally, when a RNA virus attacks a
cell, it will produce enormous number
of copies of itself. The copies, in turn,
produce viral protein, which can help
to disable the cells defenses to the
virus.
 One way of preventing viral infections is
by giving a plant a viral gene encoding
the virus' 'coat protein'. The plant then
produces this viral protein before
the virus infects the plant. If the
virus arrives, it is not able to reproduce.
 This is called co-suppression. When a
foreign viral DNA enters the plant cell,
viral coat protein is produced, and it
eventually shuts down the viral protein's
expression. When the virus tries to infect
the plant, the production of its
essential coat protein is already blocked.
 VCPs encapsulate the viral nucleic
acid and are thought to be important in
nearly every stage of viral infection
including replication, movement
throughout an infected plant, and
transport from plant to plant
 Alternatively, apical or axillary
meristems are generally free from viral
particles, which has helped the
scientists to produce virus free plants,
by culturing small meristems collected
from virus infected plants.
Gene Transfer in plants
 Vector used: Ti plasmid of
Agrobacterium Tumefaciens.
 Ti Plasmid- Tumor Inducing Plasmid
with Transfer DNA.
 Strategy:
Collect leaf discs
 Infect the tissue with Agrobacterium
carrying recombinant Ti plasmid.
 The infected tissue is then raised in Shoot
regeneration medium for 2-3 days, so that
transfer of T-DNA along with gene of
interest takes place.
 Then the transformed tissues are
transferred onto selection cum plant
regeneration medium supplemented with
usually lethal concentration of an antibiotic.
 This medium also contains a bacteriostatic
agent, which suppresses the
Agrobacterium present with the
transformed tissues.
 After 3-5 weeks, the regenerated shoots
are transferred to root inducing medium.
 After another 3-4 weeks, complete plants
are obtained, which are transferred to soil,
following the hardening of regenerated
plants.
Late Blight in Potato
 produce millions of spores from infected
plants under the wet weather conditions
that favor the disease.
 Spores produced on infected potatoes can
travel through the air, land on infected
plants, and if the weather is sufficiently
wet, cause new infections.
 Late blight is caused by the oomycete
Phytophthora infestans.
 Several R genes originating from
introgressions of S. demissum have been
mapped to potato chromosomes using DNA
markers.
 the molecular cloning of R1 gene for
resistance to late blight that is located in the
resistance hot spot on potato chromosome V.
 R1 among plant resistance genes containing
a conserved nucleotide binding domain
(NBS), a leucine-rich repeat domain (LRR)
and a leucine zipper motif.
 300 Restriction fragment length
polymorphism(RFLP markers)
 Race specific and hyper sensitive to
p.infectants
 Those groups were R1, R3, R4, R10 and
groups with a larger amount of
accumulated R alleles and 90 different
clones belonging to the species S.
demissum, S. tuberosum ssp. andigena,
S. phureja, S. bulbocastanum and S.
stoloni
 Remaining clones in the physical map are
BACs with lengths between 70 and 100
kb.
 Grey bars: BACs from the chromosome
carrying r1.
 Solid black bars: BACs from the
chromosome carrying R1.
 Mapped BAC ends are indicated by the
number of recombinants separating the
BAC end from R1.
References
 http://ohioline.osu.edu/hyg-
fact/3000/pdf/PP401_01.pdf
 The%20R1%20gene%20for%20potato%20
resistance%20to%20late%20blight%20(Phy
tophthora%20infestans)%20belongs%20to
%20the%20leucine%20zipper,NBS,LRR%2
0class%20of%20plant%20resistance%20g
enes.%20(4).pdf
 http://www.gmeducation.org/environment/p
190974-thirsty-plants-rely-on-fungus-for-
help.html
Plant Disease Resistant And Genetic Engineering

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Plant Disease Resistant And Genetic Engineering

  • 2. PLANT DISEASE RESISTANCE AND GENETIC ENGINEERING
  • 3. What is a plant disease?  A plant disease is any abnormal condition that alters the abnormal growth or function of a plant. Disease may also reduce yield and quality of harvested product.  Plant diseases are classified in 2 categories: a) Abiotic b) Biotic
  • 4. Abiotic Diseases  Are caused by (non-living) environmental conditions such as frost, hail, and chemical burn.  Damage caused by chronic exposure to air pollutants such as nitrogen dioxide, sulfur dioxide etc.
  • 5. Biotic Diseases  Are caused by living organisms such as fungi, bacteria, viruses, nematodes, etc.  Pathogens may infect all types of plant tissues to include leaves, shoots, roots, fruit, seeds etc.
  • 6. The Disease Triangle  For a biotic disease to occur there must be a susceptible host plant, the pathogen, optimum environmental conditions.
  • 7. The Disease Cycle  The development of visual disease symptoms on a plant requires that the pathogen must (a) come into contact with a susceptible host (b) gain entrance or penetrate the host through either a wound, a natural opening or via direct penetration of the host (c) establish itself within the host (d) grow and reproduce within or on the host
  • 8. Biotic Components Fungi:-  They damage plants by killing cells or causing plant stress.  Sources are infected seed, soil, crop debris, nearby crops and weed, which spread by wind and water splash, and through the movement of contaminated soil etc.  They enter plants through natural openings such as stomata and through wounds caused by pruning, harvesting, hail, insects, other diseases, and mechanical damage.
  • 9. Common fungal diseases  White blister/White rust  Clubroot  Botrytis rots  Anthracnose  Tuber diseases
  • 10. Viral Infections  Viruses cause many plant diseases. The spread of most viruses is very difficult to control.  Viruses are often transmitted from plant to plant by insects.  Normally, when a RNA virus attacks a cell, it will produce enormous number of copies of itself. The copies, in turn, produce viral protein, which can help to disable the cells defenses to the virus.
  • 11.  One way of preventing viral infections is by giving a plant a viral gene encoding the virus' 'coat protein'. The plant then produces this viral protein before the virus infects the plant. If the virus arrives, it is not able to reproduce.  This is called co-suppression. When a foreign viral DNA enters the plant cell, viral coat protein is produced, and it eventually shuts down the viral protein's expression. When the virus tries to infect the plant, the production of its essential coat protein is already blocked.
  • 12.  VCPs encapsulate the viral nucleic acid and are thought to be important in nearly every stage of viral infection including replication, movement throughout an infected plant, and transport from plant to plant
  • 13.  Alternatively, apical or axillary meristems are generally free from viral particles, which has helped the scientists to produce virus free plants, by culturing small meristems collected from virus infected plants.
  • 14. Gene Transfer in plants  Vector used: Ti plasmid of Agrobacterium Tumefaciens.  Ti Plasmid- Tumor Inducing Plasmid with Transfer DNA.  Strategy: Collect leaf discs  Infect the tissue with Agrobacterium carrying recombinant Ti plasmid.
  • 15.  The infected tissue is then raised in Shoot regeneration medium for 2-3 days, so that transfer of T-DNA along with gene of interest takes place.  Then the transformed tissues are transferred onto selection cum plant regeneration medium supplemented with usually lethal concentration of an antibiotic.  This medium also contains a bacteriostatic agent, which suppresses the Agrobacterium present with the transformed tissues.
  • 16.  After 3-5 weeks, the regenerated shoots are transferred to root inducing medium.  After another 3-4 weeks, complete plants are obtained, which are transferred to soil, following the hardening of regenerated plants.
  • 17. Late Blight in Potato  produce millions of spores from infected plants under the wet weather conditions that favor the disease.  Spores produced on infected potatoes can travel through the air, land on infected plants, and if the weather is sufficiently wet, cause new infections.
  • 18.  Late blight is caused by the oomycete Phytophthora infestans.  Several R genes originating from introgressions of S. demissum have been mapped to potato chromosomes using DNA markers.  the molecular cloning of R1 gene for resistance to late blight that is located in the resistance hot spot on potato chromosome V.  R1 among plant resistance genes containing a conserved nucleotide binding domain (NBS), a leucine-rich repeat domain (LRR) and a leucine zipper motif.
  • 19.  300 Restriction fragment length polymorphism(RFLP markers)  Race specific and hyper sensitive to p.infectants  Those groups were R1, R3, R4, R10 and groups with a larger amount of accumulated R alleles and 90 different clones belonging to the species S. demissum, S. tuberosum ssp. andigena, S. phureja, S. bulbocastanum and S. stoloni
  • 20.
  • 21.  Remaining clones in the physical map are BACs with lengths between 70 and 100 kb.  Grey bars: BACs from the chromosome carrying r1.  Solid black bars: BACs from the chromosome carrying R1.  Mapped BAC ends are indicated by the number of recombinants separating the BAC end from R1.

Notes de l'éditeur

  1. No matter the weather conditions, there will be problematic diseases every year. Which disease may change from year to year. This presentation provides background on plant diseases resistance and genetic engineering.
  2. It is a physiological process that affects some or all plant functions. Diseases are the result of some disturbance in the normal life process of the plant.
  3. but in fact hail caused the damage to the apple tree.
  4. A susceptible host, a pathogen, and environmental conditions that favor the pathogen must be present in the right mix to yield disease ; for the pathogen to grow, and time for the disease to develop.. If any one of these three components is missing or minimized, disease will not occur.
  5. NATURAL OPENING (stomates, lenticels, hydathodes)
  6. Sources of fungal infections are …Pruning is an essential gardening skill.
  7. Insecticides are sometimes used to control viral infections, but success is very limited.
  8. Because in meristems, cells are actively dividing and cell differeciation isnt there, so the viral particles are unable to cross through plasmodesmata.
  9. Phytophthora infestans causing late blight in potato is the most destructive disease in potato cultivation worldwide. . It encodes a protein of 1293 amino acids with a molecular mass of 149.4 kDa S. demissum, a wild potato species indigenous to Mexico…The gene was identified by a combined positional cloning and candidate gene approach. The molecular structure of the gene allows classification of … A leucine-rich repeat (LRR) is a protein structural motif that forms an α/β horseshoe fold.[1][2] It is composed of repeating 20–30amino acid stretches that are unusually rich in the hydrophobic amino acid leucine. These repeats commonly fold together to form asolenoid protein domain, termed leucine-rich repeat domain. A leucine zipper, aka leucine scissors,[1] is a common three-dimensional structural motif in proteins. The primary function of these motifs appears to be to provide a versatile structural framework for the formation of protein-protein interactions.
  10. --The first allele is dominant and the second allele is recessive.
  11. --two RFLP markers, GP21 and GP179 is a group of loci for the resistance gene controlling among other things the potato resistance against most viruses .. --SPUD237 and AFLP1 are converted AFLP markers .. Genetic distances are given in cM. CosS is a cosmid clone selected with SPUD237. Grey bars: BACs from the chromosome carrying r1. .. Solid black bars: BACs from the chromosome carrying R1. --This marker was converted in CAPS (Cleaved Amplified Polymorphic Sequence) marker more usable for a wider research public. --Cosmid and BAC ends used for chromosome walking are indicated by the vertical arrows. RGL, resistance-gene-like fragment.