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Approaches to subspecies diagnostics in big sagebrush (Artemisia tridentata) using an Electronic Nose
Hector Ortiz, Bryce Richardson
USFS-Rocky Mountain Station, Chicago Botanic Garden
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The Chemical Species Profile Discrimination (A Brief Description)
• Smell receptor
• Chemosensors
• Odor sensing,
e-nose applications
• Smell profiles
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Camphene Arthole 1,8-Cineole Methy Santolinate Camphor
Methacrolein or methacroldehyde Molecular formula C4H6O
Big Sagebrush Emission of HIPV’s, GLV’s, VOC’s
- High Sensitivity to Light and Temperature - Herbivore Selection Pressure, caused speciation? - Insects
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Baldwing et, al. 2006
Aerial interaction of the wild tabacco Nicotiana atenuata and sagebrush Artemisia tridentata.
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• Determinate sensitivity to known plant volatiles (methacrolein)
• Develop an standard protocol for volatile
analysis • Assess effectiveness at discriminating
subspecies
Objectives
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• Parameterization with pure methacrolein
• Sensor selection
• Train E-Nose with Artemisia samples
PC-Nose Standardization
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1. Collect and freeze fresh leaves 2. Crush and elute with water 3. Centrifuge 4. Program the settings of the PC Nose 5. Training and sample analysis
Sample Preparation
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Methacroleine 10% 20% 50% 70% 80%
10% 8 0 0 0 0
20% 0 8 0 0 0
50% 0 0 8 0 0
70% 0 0 0 8 0
80% 0 0 0 0 8
Identified As
Trai
ne
d A
s
Correct: 100%
Cross Validation Methacrolein Method
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• First analysis shows, how the volatile profiles within and between samples of the same subspecies are related
- These samples were collected from a common garden
• Second analysis shows differences among subspecies (tridentata, vaseyana and wyomingensis)
• Third analysis was a comparison between subspecies within a small geographic range
Results
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WAT-1
ORT-3 Putative hybrid
NVT-2
NVT-1
MTT-1 IDT-3
Canonical Projection Plot first analysis—within subspecies and individual
variation, A. tridentata
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0.0
5.0
10.0
15.0
20.0
M-
Dis
tan
ce
WAT-1 ORT-3 NVT-2 NVT-1 MTT-1 IDT-3
Potential hybrid
Variation within samples A. tridentata
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MTV-1
IDV-4
IDV-3 Putative hybrid
IDV-2
CAW-3
UTV-1
Canonical Projection Plot first analysis—within subspecies and individual
variation, A. vaseyana
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M-D
ista
nce
0.0
5.0
10.0
15.0
20.0
25.0
MTV-1 IDV-4 IDV-3 IDV-2 CAW-3 UTV-1
Potential hybrid
Variation within samples A. vaseyana
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MTW-3
UTV-2 COW-2
MTW-1
IDW-2
IDW-3
Canonical Projection Plot A. wyominensis
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M-D
ista
nce
0.0
5.0
10.0
20.0
25.0
30.0
35.0
MTW-1 UTV-2 COW-2 IDW-2 IDW-3 MTW-1
Variation within samples A. wyomingensis
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A. wyomingensis
A. vaseyana
A. tridentata
-10.0 -5.0 0.0 5.0 10.0 15.0 20.0 25.0 30.0
-5.0
0.0
5.0
10.0
Fact
or
2
Factor 1
Second analysis comparison among A. tridentata, A. vaseyana, A. wyomingensis
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vaseyana UTV-3
wyomingensis UTW-1
tridentata UTT-1
Factor 1
Fact
or
2
-25.0 -20.0 -15.0 -10.0 -5.0 0.0 5.0 10.0 15.0
-10.0
-5.0
0.0
5.0
10.0
Comparison between subspecies within a small geographic range (<15km)
Location: La Sal, Utah
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• Other factors under consideration: • Environment
• Time of year
• Stems, seeds and flower parts?
On Going Work
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• The e-nose can detect presence or absence of VOC’s, and different concentrations of methacrolein
• The protocol developed appears to capture the important volatiles involved in distinguishing subspecies
• The e-nose was capable of differentiating subspecies from a common environment
Summary