high-throughput plant phenotyping –a boost for genomics in the 21st century

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Abstract: Due to the development of highly automated genetic analysis, plant genomics has immensely enlarged our understanding of the genetic structure of plants over the last two decades. The fast evolving need to identify interactions between genes and environmental factors (biotic and abiotic) that brings about a certain plant phenome made it necessary to develop quantitative, reproducible and highly automated plant phenotyping systems for large plant numbers.

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High-throughput Plant Phenotyping –

a Boost for Genomics in the 21st Century

Dr. Jörg Vandenhirtz · CMO LemnaTec Head of Research and Development joerg.vandenhirtz@lemnatec.com

LemnaTec

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 2

founded May 1998 in Aachen, Germany

interdisciplinary team (biology, physics, engineering)

12 years of experience with image based biological measurement

in-house development of image processing software

in-house development of hardware

development of comprehensive and integrated solutions

compliance with international standards

The human eye

How many legs does this elephant have?

The human eye

Count the number of black dots…

The human eye

…are exactly the same size

The circles in the centers…

The human eye

Accelerating plant phenomics

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 7

opening new prospects

high-throughput screens

multiple camera units

non-destructive measurement

quantitative analysis

monitor growth dynamics

stress assessment

link to genomics

scanalyzer3D accelerating plant phenomics

Hardware

Hardware scanalyzer3D

Finding the needle in the haystack

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 10

The LemnaTec Scanalyzer System discovers THE ONE which is relevant…

For example, THE plant with 3% increase in value is worth $50m if it obtains 10% market penetration and released 2 years earlier

Automatic screening 10x more efficient than human screening

+3%

Modern breeding or GMO methods are able to produce THOUSANDS of new varieties per week

scanalyzer3D multiple imaging

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 11

1. top View

2. side View

3. side View 90°

4. more views optional

3D imaging of full plants

1. 2.

3.

scanalyzer3D scanning modes

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 12

1.

2. 3.

4. 5.

1. RGB visible light

2. NIR near infrared light

3. FLUOR fluorescent light

4. IR infrared light

5. ROOT NIR / RGB

Scanning in different wavelength and modes

scanalyzer3D VIS imaging – shape

Scanning – visible light

High-resolution colour images for comprehensive morphological and growth phenotyping

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 13

1.

scanalyzer3D VIS imaging – colour

Scanning – visible light

Plant colour classification – Key to plant health, stress, nutrients and senescence

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 14

infrared light

visible light

near infrared light for roots

near infrared light

fluorescent light

1.

Scanning – visible light

Plant skeleton analysis – key to growth dynamics, morphology and architecture

scanalyzer3D VIS imaging – architecture

separation of stem and leaves

information about nodes, length of leaves

morphology

plant growth phases

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 15

1.

scanalyzer3D VIS imaging – architecture

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 16

2.

Scanning – visible light Phenotyping of growth under field-density growth conditions

Phenotyping based on complex morphological criteria like: structure orientation, momentum of inertia, height, width, roundness, compactness. 0,00

0,50

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axis momentum ratio

be

nd

ing

in

de

x

medium heigt, wide,

kinked leaves

high,straight upright

compact straight,

intermediate height

compact curved

1.

scanalyzer3D NIR imaging – shoot

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 17

2. 0 h

4 h 8 h 16 h

• wheat dried down over 16 h at elevated temperature • LemnaTec NIR imaging and analysis can cover the whole water dynamics of the drying for wheat • LemnaTec NIR cameras are suitable for detection of fine cereals structures of fully grown wheat

0%

10%

20%

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100%

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

% N

IR a

bso

rptio

n c

lass

hours

scanalyzer3D flourescence imaging

Scanning – flourescent light

blue light (< 500 nm) visualises any related fluorescence

chlorophyll (cont., flash)

green fluo. Protein (GFP)

phenolics

autofluorescence

custom solutions

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 18

3.

scanalyzer3D IR imaging

Scanning – infrared light

Quantify temperature differences (e. g. within leaves and between plants)

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 19

4.

scanalyzer3D IR imaging

Scanning – infrared light

Quantify temperature differences (e. g. within leaves and between plants)

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 20

4.

scanalyzer3D RGB imaging – root

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 21

Static root density profiles – growth pattern

Bangui Sweet Corn Prelude Bangui

scanalyzer3D RGB imaging – root

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 22

Dynamic root growth - pattern Day 0 12 25 35 60 100

image and data show the development of roots of one plant in time.

scanalyzer3D NIR imaging – root

Scanning – near infrared light for roots

spatial distribution of water content in soil

0h 2h 4h 6h 8h

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 23

5.

scanalyzer3D NIR imaging – root

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 24

Corn plant previously shown was grown in a transparent 8 cm polyacryl column Results of NIR monitoring allow measurement of spatial distribution water content in soil

0h 2h 4h 6h 8h 0

500

1000

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0 h 2 h 4 h 6 h 8 hare

a c

lassific

atio

n p

ixe

l

total development of drying over time

0 25 50 75 100 125 150 175

L01K01J01I01

H01G01F01E01D01C01B01A01

area classified pixels

drying dynamics based on soil layer

0 h 8 h

0 25 50 75 100 125 150 175

L01

K01

J01

I01

H01

G01

F01

E01

D01

C01

B01

A01

area classified pixels

Hardware Future Developments?

Non invasive imaging of plants and plant roots in substrate with Nuclear Magnetic Resonant Imaging (NMRI) and (Sub) Terahertz Imaging (THz)

Dr. Axel Jung Ricarda van der Heijden Beta Klinik Bonn

Dr. Karsten Specht University of Bergen Prof. Dr. Martin Koch IHF TU Braunschweig

Dr. Jörg Vandenhirtz Matthias Eberius Dirk Vandenhirtz LemnaTec

Goal of this joint study:

Can Nuclear Magnetic Resonant Imaging (NMRI) or (Sub) Terahertz Imaging (Thz) be used for obtaining

non invasive and valuable information about plant roots in substrate?

• 3D root morphologie • 3D root dynamics • 3D root activity (water distribution) • etc...

movingfield linking greenhouse to field

Hardware Future Developments?

Used Plants: Corn Plants in transparent pots:

Hardware Future Developments?

Siemens MAGNETOM Espree 1.5T

NMR settings:

B = 1.5 T

TE = 12 ms

TR = 5.0 s

FoV = 250 x 250

ST = 1 mm

Price: 1.5 Mio Euro

Hardware Future Developments?

First Results of Dr. Axel Jung, Beta-Klinik, Bonn

Hardware Future Developments?

First Results of Dr. Axel Jung, Beta-Klinik, Bonn

Hardware Future Developments?

Problems: • artefacts (debris) in substrate hide roots (H2O + Fe) • low contrast between roots and background • correct setting of NMR has to be researched

-> Contact to Dr. Karsten Specht Department of Biological and Medical Psychology University of Bergen, Norway Research on NMRI

Dr. Specht tried NMRI of his office Plant...

Hardware Future Developments?

First Results of Dr. Karsten Specht, Uni Bergen:

Hardware Future Developments?

First Results of Dr. Karsten Specht, Uni Bergen:

Hardware Future Developments?

Complementary option for non invasive and penetrating plant sensors:

Prof. Dr. Martin Koch Institut für Hochfrequenztechnik TU Braunschweig

THz radation can be considered as high-frequency microwaves or very long waved light.

Infrared Microwaves Ultraviolet X-Ray

Visible

10 9

10 11

10 12

10 13

10 14

10 15

10 16

10 17

10 10

Frequency (Hz)

“THz” Radiowaves

10 8

10 7

Hardware Future Developments?

wat

er c

once

ntra

tion

Fresh leaf 48 hours after cut

B.B. Hu und M.C. Nuss, Opt. Lett. 20, 1716 (1995)

First THz image of a leaf with high resolution

Bell Labs Lucent Technologies

Hardware Future Developments?

First Results of Prof. Dr. Martin Koch, Braunschweig:

THz transmitter

THz detector

moving a coffee plant between a sub THz transmitter and detector…

…and measuring the transmission of the sub THz signal

z f THz

transmitter

detector

Hardware Future Developments?

The lift and rotating devices of the LemnaTec Phenotyping Platform are already integrated for 3D THz scans in cylindrical coordinates:

Combining 3D THz transmission measurements with visible 3D RGB image data of the LemnaTec Phenotyping Platform

Hardware Future Developments?

Conclusions:

NMRI THz

hardware standard of the shelf technology to be developed

software to be developed to be developed

throughput ca. 20 min. per plant ca. 2 min. per plant

resolution ca. 1 mm ca. 1 cm

hazards high B-Field none

price ca. 1.5 Mio. Euro ca. 150,000 Euro

target roots and shoots shoots (roots?)

Software Future Developments?

EuroStar Project PhenoFab January 2009 – January 2011 Project sum ca. 1.000.000 Euro

Development of a software interface between the output database format of DNA sequencers (e.g. La Roche) and the output database format of the image processing of the LemnaTec High Throughput Phenotyping Platform for multidimensional correlation analysis between the 2 datasets

Software Future Developments?

Plant

CORRELATION ???

Genomics

Phenomics

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 40

Contractracted Phenotyping Research

Date: 9/23/2011 Speaker: Dr. Joerg Vandenhirtz – CMO Slide 41

Ph

References Most Global Players in Breeding and GMO are already using the LemnaTec Technology: • BASF • Pioneer Dupont • Bayer CropScience • Monsanto • Syngenta • Dow Agro • Keygene • IPK • CSIRO • ACPFG • etc.

Visit our website www.lemnatec.com

Thank you for your attention!

Visit our website www.lemnatec.com

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