An ASAE/CSAE Meeting Presentation

Paper Number: 043065

Estimating Water Stress in Plants Using Hyperspectral Sensing

Carol L. Jones, Research Engineer
Biosystems and Agricultural Engineering, Oklahoma State University, Stillwater, OK

Paul R. Weckler, Assistant Professor

Biosystems and Agricultural Engineering, Oklahoma State University

Niels O. Maness, Professor

Horticulture and Landscape Architecture, Oklahoma State University

Marvin L. Stone, Regents Professor

Biosystems and Agricultural Engineering, Oklahoma State University

Roshani Jayasekara, Research Engineer

Biosystems and Agricultural Engineering, Oklahoma State University

Written for presentation at the 2004 ASAE/CSAE Annual International Meeting Sponsored by ASAE/CSAE Fairmont Chateau Laurier, The Westin, Government Centre Ottawa, Ontario, Canada 1 - 4 August 2004
Abstract. The ability to estimate plant water content may provide valuable information to environmental and irrigation system managers to relieve dehydration symptoms and prevent permanent growth and production damage. A portable spectroradiometer was used to gather hyperspectral reflectance data from three plant species (corn, spinach and snap beans) grown in a greenhouse and subjected to different watering conditions to instigate different levels of moisture deficiency. Spectral bands at 950-970, 1150-1260, 1450, 1950, and 2250 nm and five indices (WI, NDVI, SIPI, fWBI, and WI/NDVI) were analyzed to determine the best method of nondestructively estimating plant water 2 content. In corn and snap beans, the 1450 nm band data provided the best estimate (r = 0.67 and 0.50). In spinach, reflectance data from the 950-970 nm bands were the most useful (r2 = 0.94). Keywords. Reflectance, hyperspectral, moisture content, water stress, spectroradiometer, spectral indices, plants, crops

Introduction
An accurate estimate of plant water content is significant in making management decisions for irrigation, wildfire risk, and drought assessment (Chandler, et al., 1983; Penuelas, et al., 1993; Penuelas, et al., 1996). A method to nondestructively assess in real time plant water content (PWC) in plants may eliminate the tedium and delay associated with oven-drying (Allen, 1989). Variations in reflectance related to OH bonding of water may provide a method of nondestructively estimating plant water content in certain plant species (Palmer and Williams, 1974). Absorption of infrared radiation has been tested as a method of estimating plant water concentration (Carter, 1991; Danson, et al., 1992; Jackson, 1984; Hatfield and Pinter Jr., 1993) Researchers have investigated different spectral bands and for water sensitivity. The spectral bands at 950-970, 1150-1260, 1450, 1950, and 2250 nm have shown promise in estimating water content in certain species (Sims and Gamon, 2003). Water content sensitive spectral indices are typically combinations between reflectance or intensity at wavelengths where water absorbs energy at different magnitudes. Spectral indices that have been developed using these water bands (water band indices) are: relative water content (Penuelas, et al., 1993; Penuelas, et al., 1996), leaf water potential (Penuelas, et al., 1993; Penuelas, et al., 1996; Pierce, et al., 1990; Riggs and Running, 1991), water index (WI) (Penuelas, et al., 1997), normalized difference vegetation index (NDVI) (Penuelas, et al., 1997; Rouse, et al., 1974), water index divided by normalized difference vegetation index (WI/NDVI)(Penuelas, et al., 1997), structure-independent pigment index (SIPI) (Penuelas, et al., 1995), and floating-position water band index (fWBI) (Strachan, et al., 2002). Each of these indices has been successful with different plant species and is sensitive to the structure of the plants and the method in which the data were collected (Sims and Gamon, 2003). The wavelength regions considered in this research were 950-970, 1150-1260, 1450, 1950, and 2250 nm and the indices investigated included WI, NDVI, SIPI, fWBI, (Table 1) and WI normalized with NDVI (WI/NDVI). Table 1. Summary of investigated indices. Rxxx = reflectance. Subscript wavelengths represent a narrow range of 10 nm.
Index Water Index (WI) Equation
WI = R 900 R 970

Normalized difference vegetation index (NDVI)

NDVI =

Structure-independent pigment index (SIPI)

SIPI =

(R 800 (R 800

(R 800 (R 800

R 445 ) R 680 )

+ R 640 )

R 640 )

Floating-position water band index (fWBI)

fWBI =

R 900 min(R 930 R 980 )

Reference and Notes (Penuelas, et al., 1997; Sims and Gamon, 2003) -significantly correlated with plant water content when a wide range of water content is considered. (Rouse, et al., 1974; Sims and Gamon, 2003) -found less sensitive to water content than other water band indices (Penuelas, et al., 1995) -uses blue and red wavelengths to assess proportion of total photosynthetic pigments to chlorophyll. (Strachan, et al., 2002) -position of spectral water absorption trough shown to be dynamic during stress conditions, reflected in the range in the denominator

The objectives of the research reported here are to: Assess the accuracy of hyperspectral reflectance information in estimating plant water content in corn, spinach and snap beans. These crops were chosen because of their inherent differences in foliage presentation. Identify the reflectance index that provides the best precision in providing an estimate of plant water content in each species.

Methods and materials

Plant Sampling
Corn, spinach and snap beans were grown in a greenhouse in standard flats. One flat of each species was watered according to best practices and used as a control. Remaining flats were subjected to desiccation culminating in a permanent wilt stage. Beginning at the post cotyledon stage in spinach and snap beans and the V2 stage in corn, plants were chosen randomly every other day for sampling. At least ten hyperspectral measurements from different locations throughout the foliage of each chosen plant were recorded using a spectroradiometer with a self-contained illumination system. The readings were taken directly at the leaf surface with care taken to exclude any external light introduction. The vegetative portions of the plants were subsequently harvested and moisture content was determined using oven methods. Weights were recorded before the plants were placed in the oven (70C) and at scheduled intervals until no change in weight was observed. Weight change was assumed to be due to loss of water content. PWC was determined using the weights obtained from the oven testing method and computed using the following equation: PWC (%) = ((WF WD)/WD)100 where: WF = wet weight, g WD = dry weight, g (1)

Spectroradiometer
Hyperspectral data were collected using a Field Spec Pro JR portable spectroradiometer (Analytical Devices, Boulder, Colorado) connected serially to a laptop computer. This spectrometer was equipped with a high intensity contact probe that housed a halogen bulb and provided a 10 mm spot. The spectroradiometer was calibrated before and after readings for each plant using a barium sulfate coated reflectance plate (white plate). Both dark and white readings were used for calibration according to the manufacturers directions. The Field Spec Pro JR gathers irradiance, computes reflectance by subtracting dark current and dividing by the white plate reading and records the reflectance in the 300 to 2400 nm wave bands. Using ViewSpec Pro, version 3.06 software (Analytical Devices, Boulder, Colorado) to assemble and interpret data, files were converted and imported into statistical software for further analysis.

Experiment Structure
This study was conducted as a split plot design with the plant species as the main plots and the collection days as the subplots. Samples were collected on seven different days for each plant species. Within the subplots, two plants were randomly removed each day from growing flats that were watered according to best practices for maximum production and two plants were randomly removed from flats that had received no water subsequent to the beginning of the

study at post-cotyledon stage for spinach and snap beans and V2 for corn. Hyperspectral data were collected from ten random locations in the vegetative portion of each plant.

Data Analysis
Spreadsheet software was used to organize data. Reflectance data at 435 - 455, 630 - 650, 670 690, 790 810, 890 910, 930 980, 950-970, 1150-1260, 1450, 1950, and 2250 nm were separated from the 300 to 2400 nm data collected by the spectroradiometer. The indices listed in Table 1 were computed. The indices and reflectance data were imported into SAS (S.A.S. Institute Inc., Cary, North Carolina) for linear regression and correlation to plant water content.

Results
Corn and spinach had similar reflectance response when placed under water deficiency stress, while snap beans exhibited less reflectance change (Table 2). Figures 1, 2 and 3 show examples of the hyperspectral reflectance profile of each species at two different plant water content levels. Table 2. Example of differences in reflectance for corn, spinach, and snap beans placed under water deficiency stress. Sample responses are typical of study population. Difference in reflectance (percentage points) Wave band Corn Spinach Snap Bean 630-650 2.50 4.0 4.0 950-970 3.0 11.0 1.0 1150-1260 2.5 14.0 1.0 1450 8.0 11.0 2.0 1950 6.5 6.0 2.0 2250 10.0 20.0 2.0

Corn 40.0 Reflectance, % 25.0 30.0 25.0 20.0 15.0 10.0 5.0 0.0 350 600 850 1100 1350 1600 1850 2100 2350 Wavelength, nm PWC = 84% PWC = 93%

Figure 1. Typical hyperspectral reflectance profile for corn at two plant water content levels.

Spinach 60.0 50.0 Reflectance, % 40.0 30.0 20.0 10.0 0.0 350 600 850 1100 1350 1600 1850 2100 2350 Wavelength, nm PWC = 92% PWC = 66%

Figure 2. Typical hyperspectral reflectance profile for spinach at two plant water content levels.

Snap Beans 60.0 PWC =89% 50.0 Reflectance, % 40.0 30.0 20.0 10.0 0.0 350 600 850 1100 1350 1600 1850 2100 2350 Wavelength, nm PWC =78%

Figure 3. Typical hyperspectral reflectance profile for snap beans at two plant water content levels. Significant correlations (p < 0.01) occurred between PWC and 1450 nm, 2250 nm, WI, and fWBI in corn, 950 - 970 nm, 1150 1250 nm, 1450 nm, 1950 nm, 2250 nm, and WI in spinach, and 1450 nm and WI in snap beans. Figures 4, 5 and 6 indicate the relationship between each of these significant water bands or indices and plant water content. The statistical linear regression significance and coefficients of determination of each water band and index correlated to the plant water content are presented in Table 3.

Plant Water Content vs. Water Band Reflectance, Corn

96.0 Plant Water Content, % 94.0 92.0 90.0 88.0 86.0 84.0 82.0 0.00 5.00 10.00 15.00 20.00 25.00 Reflectance, % 1450 2250

Plant Water Content vs. Index Value, Corn

Plant Water Content, %

96.0 94.0 92.0 90.0 88.0 86.0 84.0 82.0 1.03 1.04 1.05 1.06 1.07 Index Value

WI fWBI

1.08

Figure 4. Relationship between plant water content, water band reflectance, and indices value for Corn

Plant Water Content vs. Water Bands, Spinach 950-970 1150-1260 90.0 Plant Water Content, % 85.0 80.0 75.0 70.0 65.0 60.0 0.0 10.0 20.0 30.0 Reflectance, % 40.0 50.0 60.0 1450 1950 2250

95.0

Plant Water Content vs. WI, Spinach

y = 9.47WI - 9.019 r 2 = 0.70 90.0

Plant Water Content, %

85.0 80.0 75.0 70.0 65.0 60.0 1.02 1.025 1.03 1.035 1.04 1.045 1.05 1.055 1.06
Index Value, WI

Figure 5. Relationship between plant water content, water band reflectance, and indices value for Spinach

Plant Water Content vs. Average R1450, Snap Beans

Plant Water Content, %

95.0 90.0 85.0 80.0 75.0 70.0 65.0 60.0 5.0 10.0

y = -0.871R1450 + 0.962 r 2 = 0.50

15.0

20.0

25.0

Reflectance at 1450 nm, %

Plant Water Content vs. WI, Snap Beans y = 3.442WI - 2.713 r 2 = 0.35

Plant Water Content, %

95.0 90.0 85.0 80.0 75.0 70.0 65.0 60.0 1.01 1.02 1.03

1.04

1.05

Index Value, WI

Figure 6. Relationship between plant water content, water band reflectance, and indices value for Snap Beans

Table 3. Coefficients of determination (r2) of plant water content with average spectral reflectance and indices Coefficient of Variation (r2) between PWC and average spectral reflectance 960 10 nm 1150 1260 1450 10 1950 10 nm 2250 10 nm nm nm 0.29 (3) Corn 0.18 (4) 0.32 (3) 0.67 (1) 0.61 (1) (1) (2) (2) (2) Spinach 0.93 0.85 0.80 0.79 (2) 0.94 (3) (4) (3) (1) 0.22 0.07 0.02 (4) Snap Beans 0.14 0.50 Coefficient of Variation (r2) between PWC and indices WI NDVI SIPI (4) (2) 0.0002 Corn 0.09 (4) 0.43 (4) 0.11 Spinach 0.51 (3) 0.70 (2) Snap Beans 0.06 (4) 0.18 (3) 0.35 (1)
r2 statistical significance p: 1p< 0.001, 2p < 0.01, 3p<0.05, 4p>0.05.

fWBI 0.42 (2) 0.03 (4) 0.0006 (4)

WI/NDVI 0.002 (4) 0.002 (4) 0.02 (4)

Visual inspection of the desiccating snap bean flats revealed indications of wilt after five days (second sampling day) with an 85% PWC and permanent wilt occurred after seven days (third sampling day) with a 79% PWC. Permanent wilt in corn was observed on the second sampling day with an 85% PWC. Permanent wilt in spinach was observed on the third sampling day with an 88% PWC.

Discussion and Conclusions

Corn, spinach and snap beans follow the expected reflectance profile by exhibiting a higher reflectance level in the water absorption bands at lower plant water content (Figures 1 3). Corn and spinach exhibit a more distinct difference in reflectance compared to snap beans. The percent PWC loss in corn and snap beans were similar (9% and 11%, respectively). For corn and snap beans, the water absorption band at 1450 nm appears to provide the most accurate estimate of plant water content (r2 = 0.67 and 0.50, respectively). The weak correlations at other bands indicate that this spectral response may be a weak indicator of plant water content. Reflectance measurements in spinach are quite sensitive to plant water content, particularly in the 950 970 nm and the 1150 1260 nm bands (r2 = 0.94 and 0.93). No improvement could be found for these three species for estimating plant water content by using indices or ratios over average reflectance data. While NDVI may be a good indicator of nitrogen content and biomass at the canopy level, at the plant level it does not provide an estimate of plant water content in these three species (r2 < 0.12). WI and fWBI provided essentially the same estimate of plant water content in corn (r2 = 0.43 and 0.42). Little shift occurred in spectral absorption during plant desiccation. However, in spinach and snap beans, WI and fWBI were significantly different with WI having a stronger correlation with plant water content (r2 = 0.70 and 0.35). SIPI and WI normalized with NDVI were not significant indicators of plant water content in any of these three species in this study. The use of reflectance data appears to have promise for use at the plant level in estimating plant water content in corn and spinach but less promising in snap beans. This technique combined with a physiological study of the plant water content at the permanent wilt stage may allow growers to improve decisions concerning the application of water and other nutrients in these commercial crops.

Acknowledgements
The authors recognize and appreciate support through funding from the United States Department of Agriculture in the form of a Special Research Grant number 2003-06134. The use of trade names is only meant to provide specific information to the reader, and does not constitute endorsement by Oklahoma State University.

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