Nutritional and end-use quality

Haruki IshikawaPlant physiologist

Promising tool for high-throughput phynotypingApplication of optical measurement

Key factors of phenotyping

-Breeders interest:Yield, tolerance, resistance, architecture, physiology, ecology etc…

-Direct measurement parameters: Yield, image-based projected leaf area, chlorophyll fluorescence, stem diameter, plant height/width, compactness, stress pigment concentration, tip burn, internode length, color, leaf angle, leaf rolling, leaf elongation, seed number, seed size, tiller number, flowering time, germination time etc…

High-throughput

-What is “high-throughput”? e.g. a performance per unit time = it can measure 100 plants per day

-Limitation of labor intensive methodTouch-counting system for efficient counting

Grain size and color measurement system For evaluation of Striga seed

germination in Nematoglogy

ICRISAT HQ, FieldScan (precision screening of tens of thousands of plants under filed conditions): phynospex.com

High throughput phenotyping with Spectroscopy

-Fourier Transform Infra-Red (FT-IR) SpectroscopyFT-IR is a method of measuring an infrared absorption spectrum

-What we can doWe can : Qualitative analysis for find particular components in sample, identification of chemical substances, residual pesticides…etc.

Quantitative analysis for protein contents, sugar or starch contents, water contents, particular components…etc.

FT-IR system (JASCO)“The sugar content automatic measurement system for apple”

Photo by http://syoku-yokote.jugem.jp

-Rapid-Low-cost-Labor-effective-Repeatable

Current study -Nitrogen content in the cowpea seed-

1. Sample preparation Breeders field: varieties, growth condition, number of sample…etc. Lab: sample preparation (depend on method of measurement)2. Establishment of calibration curve/model True value (ref. from chemical analysis) and Spectra data require3. Validation of calibration curve

↓4. Quantitative analysis for interest samples

Chemometrics technique

Current study -Nitrogen content in the cowpea seed-

251 germplasms * 3 different locations * 2 fertilizer conditions * 2 years = 3012 samples

Source: S. Muranaka, M. Shono, M. Kumar, H. Takagi, H. Ishikawa (2015)J. Biological Food Science Research. 4(2), pp16-24.

Spectra data acquisition↓

Chemometrics analysis↓

Calibration curve model↓

Validation

Region: MIRSample condition: powderAcquisition time: 2min/sampleThroughput: 150 samples/day

Current study -Nitrogen content in the cowpea seed-

Source: S. Muranaka, M. Shono, M. Kumar, H. Takagi, H. Ishikawa (2015)J. Biological Food Science Research. 4(2), pp16-24.

With cost and time saving characteristics of

the method, these accurate and robust

calibration models for predicting grain N contents should be useful tools for field

agronomic studies and breeding in cowpea.

Region: NIRSample condition: Whole seedAcquisition time: 17 sec/sampleThroughput: 750 samples/day

Current study -Nitrogen content in the cowpea seed-

Non-destructive whole seed measurement

Establishment of the modelin progress

FTIR -apply for other crops-

e.g. Yam or cassava (starch, water content)Collaboration work with JIRCAS

Handy FTIR (NIR) microPhazirTM AG

AccuracyEstablish robust calibration model

with various samples&

Cost performance Does it balance with the data needed?

NIR gunAnalysis for

sugar content

Focus on different region of IR

Middle Infra-red Region (MIR)

Near Infra-red Region (NIR)

25μm~2.5μm (400~4000cm1)

2.5μm~1.0μm (4000~10000cm1)

Quantitative/Qualitative analysisHydroscopic compound → inadequacy

Hydroscopic compound → adequacyQuantitative/non-destructive analysis