genetics, measurement, and standardization in the potato

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Max FeldmanWA/OR Potato Conference

Kennewick, WAJanuary 23rd, 2020

Genetics, Measurement, and Standardization in the Potato

Genetics Lab at USDA-ARS Prosser

Research Background

Research TechnicianUniversity of Arizona(2004-2006)

Graduate Research AssistantWashington State University(2006-2012)

Postdoctoral Research AssociateDonald Danforth Plant Science Center(2013-2018)

0

5

10

15

20

height_27_BP14

Chromosome

lod

1 2 3 4 5 6 7 8 9

1@75.9

2@98.5

3@27.4

5@100.29@34.9

9@111.71@78.3

2@98.5

5@100.4

9@34.9

2@44.6

2@96.9

3@47.4

3@71.8

4@52.9

5@100.4

6@44.2

7@2.2

7@99.9

9@34.9

9@107.0

2@96.3

3@47.2

5@100.2

7@94.1

9@34.9

9@107.0

Sabelli PA, .. Feldman M.. (2013) DOI: 10.1073/pnas.1304903110

Feldman MJ et al., 2015DOI: 10.1007/s00425-015-2322-4

Feldman MJ et al., 2017DOI: 10.1371/journal.pgen.1006841

Mission: Breed a More Profitable and Sustainable Potato

Mission: Breed a More Profitable and Sustainable Potato

Identify and introgress disease resistance alleles into production clones

(Guenthner 2009)

Fumigation is a major expense

Mission: Breed a More Profitable and Sustainable Potato

Identify and introgress disease resistance alleles into production clones

(Guenthner 2009)

Fumigation is a major expense

Mission: Breed a More Profitable and Sustainable Potato

Identify and introgress disease resistance alleles into production clones

Pathogens

Columbia Root Knot Nematode (CRKN)

Stubby Root Nematode (SRN)

Tobacco rattle virus (TRV)

Potato virus Y (PVY)

(Guenthner 2009)

Mission: Breed a More Profitable and Sustainable Potato

Develop specialty potato clones for fresh market and chipping industry

Yield, size, disease resistance, chemical composition, storage and skin quality are important traits

Purple Pelisse AmaRosa Yukon Nugget

Genetics

Tri-State Breeding Program

20-hills, 2 repsKlamath (SI)

Single Hills Klamath Falls (~60,000 in Field)

20-hills, 2 repsOntario

20-hills, 2 repsHermiston

4-hills (~700)Hermiston

CrossesAberdeen, ID, Prosser, WA and Oregon

Seedling tuber productionCorvallis (~60,000 in GH)

15-hills (~700)Klamath Falls (SI)

Oregon Statewide trials3 Locations

Tri-State TrialsID, OR, WA

W. Regional TrialsCA, CO, ID, OR, TX, WA

NameTri-State release, PVP

Klamath Falls (SI)

Yr. 1

Yr. 2

Yr. 3

Yr. 4-6

Yr. 6-8

Yr. 8-10

Yr. 10-12

Field Generation

Potato is a Clonal Crop

Slide credit: Sagar Sathuvalli

Tri-State Breeding Program

20-hills, 2 repsKlamath (SI)

Single Hills Klamath Falls (~60,000 in Field)

20-hills, 2 repsOntario

20-hills, 2 repsHermiston

4-hills (~700)Hermiston

CrossesAberdeen, ID, Prosser, WA and Oregon

Seedling tuber productionCorvallis (~60,000 in GH)

15-hills (~700)Klamath Falls (SI)

Oregon Statewide trials3 Locations

Tri-State TrialsID, OR, WA

W. Regional TrialsCA, CO, ID, OR, TX, WA

NameTri-State release, PVP

Klamath Falls (SI)

Yr. 1

Yr. 2

Yr. 3

Yr. 4-6

Yr. 6-8

Yr. 8-10

Yr. 10-12

Field Generation

Potato is a Clonal Crop

Slide credit: Sagar Sathuvalli

Single Hill Selection

Single Hill Selection

Tri-State Breeding Program

20-hills, 2 repsKlamath (SI)

Single Hills Klamath Falls (~60,000 in Field)

20-hills, 2 repsOntario

20-hills, 2 repsHermiston

4-hills (~700)Hermiston

CrossesAberdeen, ID, Prosser, WA and Oregon

Seedling tuber productionCorvallis (~60,000 in GH)

15-hills (~700)Klamath Falls (SI)

Oregon Statewide trials3 Locations

Tri-State TrialsID, OR, WA

W. Regional TrialsCA, CO, ID, OR, TX, WA

NameTri-State release, PVP

Klamath Falls (SI)

Yr. 1

Yr. 2

Yr. 3

Yr. 4-6

Yr. 6-8

Yr. 8-10

Yr. 10-12

Field Generation

Potato is a Clonal Crop

Slide credit: Sagar Sathuvalli

Tri-State Breeding Program

20-hills, 2 repsKlamath (SI)

Single Hills Klamath Falls (~60,000 in Field)

20-hills, 2 repsOntario

20-hills, 2 repsHermiston

4-hills (~700)Hermiston

CrossesAberdeen, ID, Prosser, WA and Oregon

Seedling tuber productionCorvallis (~60,000 in GH)

15-hills (~700)Klamath Falls (SI)

Oregon Statewide trials3 Locations

Tri-State TrialsID, OR, WA

W. Regional TrialsCA, CO, ID, OR, TX, WA

NameTri-State release, PVP

Klamath Falls (SI)

Yr. 1

Yr. 2

Yr. 3

Yr. 4-6

Yr. 6-8

Yr. 8-10

Yr. 10-12

Field Generation

Potato is a Clonal Crop

Slide credit: Sagar Sathuvalli

Can Quantitative Genetics Improve Efficiency?

Can Quantitative Genetics Improve Efficiency?

Will estimation of combining ability for important clones help us narrow our search space for cross evaluation?

Example 1: Columbia Root Knot Nematode Resistance

Columbia Root Knot Nematode (CRKN) is an Important Pathogen

Meloidogyne chitwoodi

External

InternalSlide credit: Russ Ingham (OSU)

PA99N82-4 is a CRKN Resistant Clone Developed by Our Program

Dr. Charles Brown

PA99N82-4 Still Has Not Made it to Market

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5

10

15

Yield Tuber_SizeTrait

Value

Data from Rosyara et al., 2016

How can we pick the best crosses to make and families to focus on?

PAN99N82-4

Female Parent

X

Male Parent

Innovator

Mountain Gem

Katahdin

Clearwater

Western

AO6084-1TE

Markies

AO2060-1TE

POR08BD1-3

Currently growing out ten F1 families (~100 clones each) for minitubers

How can we pick the best crosses to make and families to focus on?

Harvest F1s as bulk samples at 10 hill intervals

Emergence/Maturity (UAV)

Yield, Tuber Size (Potato sorter)

Specific gravity Hollow

HeartGrowth Cracks

How can we pick the best crosses to make and families to focus on?

PAN99N82-4

Female Parent

X

Male Parent

Innovator

Mountain Gem

Katahdin

Clearwater

Western

AO6084-1TE

Markies

AO2060-1TE

POR08BD1-3

Select Families Based Upon Mean

and Variance

How can we pick the best crosses to make and families to focus on?

PAN99N82-4

Female Parent

X

Male Parent

Innovator

Mountain Gem

Katahdin

Clearwater

Western

AO6084-1TE

Markies

AO2060-1TE

POR08BD1-3

Select Families Based Upon Mean

and Variance

Can Quantitative Genetics Improve Efficiency?

Can we identify genetic markers linked to important traits? What are the costs and benefits of utilizing this technology?

Example 2: Identify and deploy molecular markers to select TRV and/or PVY resistant clones

Cause of Corky Ringspot DiseaseTobacco rattle

virus (TRV)

Stubby Root Nematode

(Paratrichodorusallius)

Weeds

Crops

Why Is Corky Ringspot Disease a Problem?

Incidence of 6% renders tubers unmarketable

(Hafez and Sundararaj 2009)

25

30

35

40

45

50

−120 −100 −80Longitude

Latitude

Castle Russet: A Success of the Tri-State Breeding Program

Fusarium dry rot resistantErwinia soft rot resistantCold sweetening resistant

TRV resistantPVY resistantPMTV resistant

Long, medium-to-late russet, suitable for fresh or processing market

50%25%

12.5%

6%

X

[ ]

Can we identify QTL associated with CRS resistance?

Next step is identifying genetic markers associated with traits

Castle Russet Susceptible

Gen

oty

pes

Phen

oty

pes

Castle Russet CRS Disease Scores 2018

810.7 12.8

25.1

32.1

9.11.1

1.1

0

5

10

15

20

25

30

35

40

45

0-1 1.1-5 5.1-10 10+

Perc

ent

Average DSI

Phenotypic Evaluation of 185 Progenyusing RT-PCR to Verify Absence or Presence

of TRV

RT-PCRNegRT-PCR Pos

Extreme Resistance

Moderate Resistance

Moderate Susceptible

Susceptible

Castle Russet x Susceptible

Castle Russet

Susceptible

Richard Quick Launa Cimrhakl

Genotyped Population by Genotyping-By-Sequencing (GBS)

Heather Tuttle Laura Shannon

Library Prep and Sequencing at University of

Minnesota

Loga

rithm

of o

dds

(LO

D) s

core

Genome location

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−14

−13

AA BBS9_6724364

wet.d13C_

BP14

S9_6724364●

AA

BB

How Do We Find Genetic Markers?

Castle Russet

Susceptible

Jeff Endelman

Used gwaspoly to do association, will use TetraploidSNPMapand polymapR once implemented

01

23

45

ave_dsi (additive)

Chromosome

−log

10(p)

1 2 3 4 5 6 7 8 9 10 11 12

Castle Russet

Susceptible

We Detect/Verify CRS QTL on Chromosome 9!

Chr09:52647396_T/C

Sagar Sathuvalli

David Douches

NeilGudmestad

Loga

rithm

of o

dds

(LO

D) s

core

Genome location

We can apply this to more traits simultaneously[

[…

Castle Russet

Susceptible

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−14

−13

AA BBS9_6724364

wet.d13C_

BP14

S9_6724364●

AA

BB

Loga

rithm

of o

dds

(LO

D) s

core

Genome location

We can apply this to more traits simultaneously[

[…

Disease Resistance

Castle Russet

Susceptible

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−14

−13

AA BBS9_6724364

wet.d13C_

BP14

S9_6724364●

AA

BB

Loga

rithm

of o

dds

(LO

D) s

core

Genome location

We can apply this to more traits simultaneously[

[…

Disease Resistance Yield

Castle Russet

Susceptible

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−14

−13

AA BBS9_6724364

wet.d13C_

BP14

S9_6724364●

AA

BB

Loga

rithm

of o

dds

(LO

D) s

core

Genome location

We can apply this to more traits simultaneously[

[…

Disease Resistance Yield Tuber

composition

Castle Russet

Susceptible

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−14

−13

AA BBS9_6724364

wet.d13C_

BP14

S9_6724364●

AA

BB

Loga

rithm

of o

dds

(LO

D) s

core

Genome location

Will Help Select Best Progeny

[

[…

Disease Resistance Yield Tuber

composition

Castle Russet

Susceptible

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−14

−13

AA BBS9_6724364

wet.d13C_

BP14

S9_6724364●

AA

BB

Measurement

Need to take more measurements with less personal time

Yield Characteristics

Tuber Properties

Plant/Foliar Growth (UAV)

Digger + Bagger

Sorter

Need to take more measurements with less personal time

Yield Characteristics

Tuber Properties

Digger + Bagger

Sorter

Plant/Foliar Growth (UAV)

What size pot to use in controlled environment assays?

Four pot sizes

Two genotypes- Russet Burbank- Castle Russet

Replicated n=3

1 2 3 4

Katelyn Greeen

How much tuber mass per pot?

Weighted entire tuber lot from each pot

Strong relationship… R2 = 0.65

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100

200

300

400

1 2 3 4Potsize

Wei

ght Genotype

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Castle Russet

Russet Burbank

How many tubers per pot?

Harvested tubers and counted them

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2

4

6

8

1 2 3 4Potsize

Tube

r_C

ount Genotype

●●●●

Castle Russet

Russet Burbank

No significant relationship between pot size and tuber number!

What can we learn using computer vision?

Image acquisition set upPhoto booth: Havox HPB-80D (32” x 32” x 32”)Camera: Nikon D7100Tripod: KF-TM2534T

Image processing pipeline in python

Steps in image processing:

1) Crop image2) Break into sub-images3) Threshold tuber from

background4) Perform

measurements5) Write measurements

to file/object

What % of the trait mean does standard error contribute between sides

Side 1

Side 2

Relationship between Tuber Area, Tuber Number, and Weight

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1e+05

2e+05

3e+05

4e+05

100 200 300 400Weight

Tube

r_Ar

ea Genotype●

Castle Russet

Russet Burbank

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2

4

6

8

100 200 300 400Weight

Tube

r_C

ount Genotype

Castle Russet

Russet Burbank

Total tuber area is more reflective of weight than number of tubers

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Red Green Blue

Length Width L/W Ratio

Area Perimeter Eccentricity

1 2 3 4 1 2 3 4 1 2 3 4

1 2 3 4 1 2 3 4 1 2 3 4

1 2 3 4 1 2 3 4 1 2 3 4

0.2

0.4

0.6

0.8

1.0

1.5

2.0

2.5

40

50

60

70

80

90

500

1000

1500

2000

100

200

300

60

80

100

0

50000

100000

150000

200

400

600

80

100

120

140

160

Potsize

Valu

e

Genotype●●

●●

Castle Russet

Russet Burbank

Characteristics of 118 individual

tubers

Size of tubers increases with pot size

Can see genotypic size differences at pot size 3 but not at pot size 2

Why are RGB values so different for pot size 4?

Color spectrum: Burbank in pot 4 is rotten

Burbank Pot 3

Burbank Pot 4

Color spectrum: Burbank vs. Castle (Pot 3)

Castle Pot 3

Burbank Pot 3

Can discriminate between genotypes

Current efforts

Rich Novy and Jaebum Park

Current efforts

Quantification of disease symptoms

Current efforts

Columbia Root Knot Nematode

Quantification of disease symptoms

Current efforts

Columbia Root Knot Nematode

Quantification of disease symptoms

Powdery scab galls

Other measurements types:Hyperspectral imaging to classify genotypes and disease symptoms (Daniel Gleason & Sushma Thapa)

Other measurements types:Hyperspectral imaging to classify genotypes and disease symptoms (Daniel Gleason & Sushma Thapa)

UAV and integrated field sensors for data collection

Standardization

Electronic record keeping

Electronic record keeping

Collecting data in a consistent manner

across programs and making it publicly

accessible will make our efforts more valuable

and create larger impact

Electronic record keeping

Thank you for your attention. Questions?

Richard Quick Katelyn GreeneLauna Cimrhakl Esteban Caldron

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