Don Lee, Agronomy and Horticulture Crop and Pest Genetics ... · RNA . 1. Extract all DNA from...
72
#1: Pest Genetics Survey Says! Selection Pressure Mode of Action #2: RNAi Targets and Stories Don Lee, Agronomy and Horticulture Crop and Pest Genetics applied to Crop Management
Transcript of Don Lee, Agronomy and Horticulture Crop and Pest Genetics ... · RNA . 1. Extract all DNA from...
#1: Pest Genetics Survey Says! Selection Pressure Mode of Action #2: RNAi Targets and Stories
Don Lee, Agronomy and Horticulture
Crop and Pest Genetics applied to
Crop Management
A. 1 B. 2 C. 3 if it is perennial D.Many times
How many times can you kill a weed ?
A. 1 B. 2 C. 3 if it is perennial D.Many ways
How many ways can you kill a weed ?
A.95% B.89% C.57% D.25%
700 farmers from Indiana, Illinois and Iowa completed a survey.
What % of these farmers agreed with the statement that
“farmers have much of the responsibility for glyphosate
resistant weed populations”?
A.95% B.89% C.57% D.25%
What percentage of 700 farmers agreed with the statement that
“industry has much of the responsibility for glyphosate resistant weed populations”?
A. Industry will come out with new herbicides and crops resistant to those herbicides before I need to change my weed management. B. I need to practice more complex weed management now because new options will not be developed fast enough by industry.
What choice below do these 700 farmers agree with THE MOST?
Sensitive Resistant Highly resistant
SS SR SR
more selection pressure? selection pressure
diploid weeds
Frequency in the
populations
Read the title, answer questions #1 and 2.
Table 1. Genotypes and Allele Frequencies in an Experimental European corn borer populations
RR RS SS
Table 1. Genotypes and Allele Frequencies in an Experimental European corn borer populations
RR RS SS
Which Treatment applies the MOST selection pressure for the R allele in the ECB population?
A. ALL the same B. Treatment 1
C. Treatment 2 and 3 D. Treatment 4
Table 1. Genotypes and Allele Frequencies in an Experimental European corn borer populations
RR RS SS
Which Treatment would the farmer like best in the short term?
A. ALL the same B. Treatment 1
C. Treatment 2 and 3 D. Treatment 4
Table 1. Genotypes and Allele Frequencies in an Experimental European corn borer populations
RR RS SS
Which Treatment places more selection pressure for Bt resistant?
A. Treatment 2 B. Treatment 3
C. Both the same
#1: Pest Genetics Survey Says! Selection Pressure Mode of Action #2: RNAi Targets and Stories
Don Lee, Agronomy and Horticulture 1
Crop and Pest Genetics applied to
Crop Management
App = Scalable Online Learning Environment
X
Transgenic Soybeans
Soybean cells
Protein
1. Extract all DNA from
organism
2. Isolate (clone)
the one gene of interest 3. Modify
the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the transgenic
plant with other plants
Gene
Pseudomonas bacteria
Dicamba Resistance
A Design a gene that controls resistance to both B. Transform BOTH resistance genes in a new event C. Breed both into the same variety
Q: What step was used to create a soybean variety with glyphosate +
dicamba resistance?
Field Evaluation
Insect Resistance Disease
Resistance Herbicide
Treatments
A Insects B. Diseases C. Weeds.
Q2: Which of the BIOTIC stresses have been targeted by the fewest
commercially released transgenes?
Bt corn
Cry 3Bb Protein
1. Extract all DNA from
organism
2. Isolate (clone)
the one gene of interest
3. Modify the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the transgenic
plant with other plants
Gene
Bacillus thuringiensis
Dr. Sahay
D. Lee Agronomy and
Horticulture UNL
X
Transgenic Soybeans
Soybean cells
Protein
1. Extract all DNA from
organism
2. Isolate (clone)
the one gene of interest 3. Modify
the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the transgenic
plant with other plants
Gene
Pseudomonas bacteria
Dicamba resistance
Transgenic Virus Res. Squash
Squash cells
Protein
1. Extract all DNA from virus
2. Isolate (clone)
the one gene of interest 3. Modify
the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the
transgenic plant with
other plants
Gene
Mosaic virus
Virus Coat
A. cell exploder B. mystery C. chemical bond cleaver
Q3: Match the transgene encoded protein with its function?
__ virus coat protein __ Cry3Bb __ Dicamba resistance
RNA interference
Transgenic Virus Res. Squash
Squash cells
1. Extract all DNA from virus
2. Isolate (clone)
the one gene of interest 3. Modify
the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the
transgenic plant with
other plants
Gene
Virus Coat
Mirror-image RNA
RNAi
Mosaic virus
A Insects B. Diseases C. Weeds.
Q4: Which of the BIOTIC stresses have populations that have
evolved resistance?
#1: RNAi vs. ‘Standard’ Transgenics #2: RNAi Targets and Stories
Don Lee, Agronomy and Horticulture 1
RNAi and
New Gene Technologies applied to
Crop Management
Field Evaluation
Insect Resistance
RNAi
Crop Genetic Engineering Process
RNA
1. Extract all DNA from
organism
2. Isolate (clone)
the one gene of interest
3. Modify the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the transgenic
plant with other plants
Gene
RNAi
17
8
Root worm Gene
Rootworm mRNA
Discover a critical rootworm gene that encodes a critical rootworm protein
13
Controlled Feeding Studies of CRW
CRW food
CRW food
CRW food
Control +RNAi #1 …..+RNAi 270
Rootworm Gene Modify the rootworm gene So it will be expressed in corn plant root Cells and make “interference RNAs”
Corn Root Preferred Interference RNA
14
Promoter Designed By Nature
A. expressed in the rootworm B. expressed in plant root C. expressed in seed
Q5: Which of the promoter designs should be used for this RNAi transgene?
Crop Genetic Engineering Process
RNA
1. Extract all DNA from
organism
2. Isolate (clone)
the one gene of interest
3. Modify the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the transgenic
plant with other plants
Gene
RNAi
17
Rootworm Gene Modify the rootworm gene So it will be expressed in corn plant root Cells and make “interference RNAs”
Corn Root Preferred Interference RNA
14
Promoter Designed By Nature
RNAi Insert Root worm RNAi
Trangene Into corn
15
Root worm Gene
Rootworm mRNA
RNAi
RNAi
16
Field Evaluation
Disease Resistance
App = Scalable Online Learning Environment
19
Pathogen Gene Discovery by sequence comparison
Plant Root
Microbe
interactive
biology
From S. Kale New Phytologist 2012 193:874
R gene
R protein
Avr protein
effector
MK Bhattacharyya, Iowa State University
Hypersensitive
Disease Progression
From S. Kale New Phytologist 2012 193:874
R gene
R protein
Avr protein
effector
Mutation In the Pathogen’s Avr Gene
• University of Georgia – Oomycete project
From S. Kale New Phytologist 2012 193:874
R gene
R protein
Avr protein
effector
Oomycete Gene
Modify the pathogen gene so it will be expressed in soybean plant root cells and make “interference RNAs”
Root Promoter Interference RNA
14
form Double stranded RNA
From S. Kale New Phytologist 2012 193:874
R gene
R protein
Avr protein
effector
Transgenic For GFP
From S. Kale New Phytologist 2012 193:874
RNAi transgene
Avr protein
effector
Q7 The BEST RNAi HIGS gene discovery would encode a protein used in..?
a. a wide variety of plant microbe interactions. b. one strain of Phytophthora or Fusarium pathogen
MK Bhattacharyya, Iowa State University
Hypersensitive
Disease Progression
Field Evaluation
Disease Resistance
X
Transgenic Soybeans
Soybean cells
1. Extract all DNA from
organism
2. Isolate (clone)
the one gene of interest 3. Modify
the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. Breed the transgenic
plant with other plants
Gene
Oomycete
Gene Knockout
By TASI
RNAi
Field Evaluation
Insect Resistance Disease
Resistance Herbicide
Treatments
LOTS OF Mare’s Tail Survivors
18 RNAi sprays?
RNAi RNAi
RNAi
RNAi
What would be a RNAi
target in this field?
Q8. The the proper
RNAi target would be RNA
expressed from..?
18 RNAi sprays?
A. .. a gene critical for all plants B. ..a gene critical for just Mare’s tail C. ..either would work
Non-GMO vs. GMO
Change a gene
Alter Protein
Control Traits
Insert a new gene
Make New
Protein
Non-GMO vs. GMO
Change a gene
Alter Protein
Change Plant or Seed traits
Insert a new gene
Make New
Protein RNA i transgene
No protein
Q10. Order these RNAi-type technologies from first to be used in the field to last to become a technology. A. RNAi based disease resistance crop B. RNAi based insect resistance crop C. RNAi weed control sprays D. RNAi insect control sprays E. RNAi type of crop
first was …..then ............then...........next........... and last.
gene A gene F
Microbe gene encodes a DNA
editing protein
edited gene Q new function… enhanced function...no function
“CRISPR”
Allergy Free Peanuts
peanut cells
1. Use DNA
sequence
2. Isolate (clone)
the one gene of interest 3. Modify
the gene for plant
expression
4. Transform plant cells
5. Propagate (clone)
plant from cell
6. TEST then Breed the transgenic
plant with other plants
Gene
Transgene Encode Talens
that Target Peanut
Allergen
gene A gene F
Microbe gene encodes a DNA
editing protein
edited Allergen gene ..no function
“Talen”
A A
T
a
BonusQ: Would offspring from this plant That are aa with NO T be GMOs?
