Advances in imaging technologies for soybean seed analysis

Among grain-producing species, soybean is one of the most important commodities, with increasing demand for production in coming years. Evaluation of soybean seed quality is fundamental for ensuring maximum germination and yield potential. Therefore, effective methods are necessary for examining different properties associated with physical-chemical, physiological, and seed-health changes that affect seed quality. This review focuses on the fundamental principles and on the application of techniques of radiographic imaging, magnetic resonance imaging, multispectral imagining, chlorophyll fluorescence imaging, and infrared thermography to evaluate changes related to loss of soybean seed quality, such as mechanical injury, injury caused by insects, embryonic malformation, and incomplete maturation. Computerized seedling image analysis is also presented for evaluation of seed lot vigor. The examples presented here show the potential of these image analysis techniques for identifying different types of injuries and increasing the efficiency of in-house quality control programs in soybean seed production companies.

Index terms:
Glycine max (L.) Merrill; image spectroscopy; optical sensing; seed technology

Authorship

Fabiano França-Silva

Department of Crop Science, ‘Luiz de Queiroz’ College of Agriculture, University of São Paulo, 13418-900, Piracicaba, SP, Brasil.University of São PauloBrazilPiracicaba, SP, BrazilDepartment of Crop Science, ‘Luiz de Queiroz’ College of Agriculture, University of São Paulo, 13418-900, Piracicaba, SP, Brasil.

https://orcid.org/0000-0002-3805-0084

Francisco Guilhien Gomes-Junior ^* *Corresponding author: francisco1@usp.br

https://orcid.org/0000-0001-9620-6270

Carlos Henrique Queiroz Rego

https://orcid.org/0000-0001-5483-9264

Agide Gimenez Marassi

Physics Institute of São Carlos, University of São Paulo, 13566-590, São Carlos, SP, Brasil., University of São PauloBrazilSão Carlos, SP, BrazilPhysics Institute of São Carlos, University of São Paulo, 13566-590, São Carlos, SP, Brasil.

https://orcid.org/0000-0001-5429-6554

Alberto Tannús

https://orcid.org/0000-0002-1675-1971

*Corresponding author:
francisco1@usp.br

SCIMAGO INSTITUTIONS RANKINGS

Figures

Figures (7)

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Figure 1
Radiographic images of soybean seeds with (a) mechanical damage in the hypocotyl-radicle axis region, (b) microcrack in the cotyledonary region, (c) wrinkling in the seed coat, cotyledons, and axis of the embryo resulting from weathering damage, (d) spots on the cotyledon(s) resulting from insect attacks, (e) malformation of the embryonic axis, and (f) retraction of the cotyledons, with formation of free space between the cotyledons and the seed coat.

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Figure 2
Two-dimensional (2D) radiographic images of soybean seeds with a microcrack (a) and damaged by insects (b) and X-ray micro-computed tomography images of the same seeds radiographed and visualized in 2D sections in the coronal (a1 and b1), transaxial (a2 and b2), and sagittal (a3 and b3) planes; the areas with injuries are indicated with red arrows. c1 and c2 - cotyledons 1 and 2, respectively; e.a - embryonic axis.

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Figure 3
Radiographic images of soybean seeds without damage (a) and seeds damaged by insects (c) with 9% water content, and magnetic resonance imaging (in false color) of the same seeds (b; d) after hydration until reaching 18% water content. In the radiographic image, the spots in slightly darker shades of gray delimited by the red square (c) and black shades (d) are tissues with lower metabolism (dead tissues) resulting from damage caused by insects and, consequently, lower intensity of the magnetic resonance signal.

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Figure 4
Red-green-blue (RGB) image (a) and multispectral image (a1) of soybean seeds with purple spot (C. kikuchii), seed with insect damage observed in RBG image (b) and multispectral image (b1), rupture in the seed coat of soybean observed with RGB image (c) and multispectral image (c1), and visible mechanical damage in the seed coat in the RGB image (d) and multispectral image (d1). In the multispectral images, the false colors red, orange, and yellow indicate the damaged regions (arrows), while the false color blue represents the region of the seed without damage.

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Figure 5
Red-green-blue (RGB) images of soybean seeds of the cultivar SYN 1163 RR with different levels of retained chlorophyll (a), chlorophyll fluorescence images of the same seeds obtained by multispectral equipment operating at the spectral wavelength of 730 nm (b) and 660 nm (c), images of the germination test showing the soybean seedlings coming from the yellow seeds without retained chlorophyll (d), and dead seeds that had chlorophyll retention (e). Spectral differences of the yellow seeds with a mean rate of chlorophyll fluorescence points equal to 4970 (Y) and greenish seeds with mean rates of 11,871 (G1 - Green 1), 17,153 (G2 - Green 2), 24,106 (G3 - Green 3), and 28,424 (G4 - Green 4) (f), with the most expressive differences at the wavelengths of 630 nm, 645 nm, 660 nm, and 690 nm. In the images, the color map was attributed based on the gray levels, where the shades of colors from red to white (730 nm) and from light blue to red (660 nm) represent the greenish seeds with high rates of chlorophyll retention and that emitted greater signals of chlorophyll fluorescence.

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Figure 6
Thermograms of soybean seeds without aging (a and a2) and after artificial aging (a1 and a3) obtained after 6 h (a and a1) and 12 h (a2 and a3) of imbibition at 25 °C. Radiographic image of a soybean seed with insect attack (region defined by the ellipse) (b), and its respective magnetic resonance images in the coronal (b1) and transaxial (b2) sections and thermographic image (b3).

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Figure 7
Seed Vigor Automated Analysis System (Sistema Análise Automatizada do Vigor de Sementes - Vigor-S) for soybean: representation of the step of setting up the germination test (a), of the devices for capturing (metallic box with scanner) and storage/processing (computer) of the seedling images (b), and of the analysis window, showing soybean seedlings digitalized and processed by the software (c), highlighting the first seedling of the sample with the hypocotyl marked in blue and the primary root in red.

Figure 1 Radiographic images of soybean seeds with (a) mechanical damage in the hypocotyl-radicle axis region, (b) microcrack in the cotyledonary region, (c) wrinkling in the seed coat, cotyledons, and axis of the embryo resulting from weathering damage, (d) spots on the cotyledon(s) resulting from insect attacks, (e) malformation of the embryonic axis, and (f) retraction of the cotyledons, with formation of free space between the cotyledons and the seed coat.

Figure 2 Two-dimensional (2D) radiographic images of soybean seeds with a microcrack (a) and damaged by insects (b) and X-ray micro-computed tomography images of the same seeds radiographed and visualized in 2D sections in the coronal (a1 and b1), transaxial (a2 and b2), and sagittal (a3 and b3) planes; the areas with injuries are indicated with red arrows. c1 and c2 - cotyledons 1 and 2, respectively; e.a - embryonic axis.

Figure 3 Radiographic images of soybean seeds without damage (a) and seeds damaged by insects (c) with 9% water content, and magnetic resonance imaging (in false color) of the same seeds (b; d) after hydration until reaching 18% water content. In the radiographic image, the spots in slightly darker shades of gray delimited by the red square (c) and black shades (d) are tissues with lower metabolism (dead tissues) resulting from damage caused by insects and, consequently, lower intensity of the magnetic resonance signal.

Figure 4 Red-green-blue (RGB) image (a) and multispectral image (a1) of soybean seeds with purple spot (C. kikuchii), seed with insect damage observed in RBG image (b) and multispectral image (b1), rupture in the seed coat of soybean observed with RGB image (c) and multispectral image (c1), and visible mechanical damage in the seed coat in the RGB image (d) and multispectral image (d1). In the multispectral images, the false colors red, orange, and yellow indicate the damaged regions (arrows), while the false color blue represents the region of the seed without damage.

Figure 5 Red-green-blue (RGB) images of soybean seeds of the cultivar SYN 1163 RR with different levels of retained chlorophyll (a), chlorophyll fluorescence images of the same seeds obtained by multispectral equipment operating at the spectral wavelength of 730 nm (b) and 660 nm (c), images of the germination test showing the soybean seedlings coming from the yellow seeds without retained chlorophyll (d), and dead seeds that had chlorophyll retention (e). Spectral differences of the yellow seeds with a mean rate of chlorophyll fluorescence points equal to 4970 (Y) and greenish seeds with mean rates of 11,871 (G1 - Green 1), 17,153 (G2 - Green 2), 24,106 (G3 - Green 3), and 28,424 (G4 - Green 4) (f), with the most expressive differences at the wavelengths of 630 nm, 645 nm, 660 nm, and 690 nm. In the images, the color map was attributed based on the gray levels, where the shades of colors from red to white (730 nm) and from light blue to red (660 nm) represent the greenish seeds with high rates of chlorophyll retention and that emitted greater signals of chlorophyll fluorescence.

Figure 6 Thermograms of soybean seeds without aging (a and a2) and after artificial aging (a1 and a3) obtained after 6 h (a and a1) and 12 h (a2 and a3) of imbibition at 25 °C. Radiographic image of a soybean seed with insect attack (region defined by the ellipse) (b), and its respective magnetic resonance images in the coronal (b1) and transaxial (b2) sections and thermographic image (b3).

Figure 7 Seed Vigor Automated Analysis System (Sistema Análise Automatizada do Vigor de Sementes - Vigor-S) for soybean: representation of the step of setting up the germination test (a), of the devices for capturing (metallic box with scanner) and storage/processing (computer) of the seedling images (b), and of the analysis window, showing soybean seedlings digitalized and processed by the software (c), highlighting the first seedling of the sample with the hypocotyl marked in blue and the primary root in red.

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[1] *Corresponding author:
francisco1@usp.br