Detection of Scab‐Damaged Hard Red Spring Wheat Kernels by Near‐Infrared Reflectance

Scab (Fusarium head blight) is a fungal disease that has become increasingly prevalent in North American wheat during the past 15 years. It is of concern to growers, processors, and the consumers because of depressed yields, poor flour quality, and the potential for elevated concentrations of the mycotoxin, deoxynivalenol (DON). Both wheat breeder and wheat inspector must currently deal with the assessment of scab in harvested wheat by manual human inspection. The study described herein examined the accuracy of a semi‐automated wheat scab inspection system that is based on near‐infrared (NIR) reflectance (1,000–1,700 nm) of individual kernels. Using statistical classification techniques such as linear discriminant analysis and nonparametric (k‐nearest‐neighbor) classification, upper limits of accuracy for NIR‐based classification schemes of ≈88% (cross‐validation) and 97% (test) were determined. An exhaustive search of the most suitable wavelength pairs for the spectral difference, [log(1/R)_λ1 ‐ log(1/R)_λ2], revealed that the slope of the low‐wavelength side of a broad carbohydrate absorption band (centered at ≈1,200 nm) was very effective at discriminating between healthy and scab‐damaged kernels with test set accuracies of 95%. The achieved accuracy levels demonstrate the potential for the use of NIR spectroscopy in commercial sorting and inspection operations for wheat scab.     

Hardness Measurement of Bulk Wheat by Single‐Kernel Visible and Near‐Infrared Reflectance Spectroscopy

Reflectance spectra (400 to 1700 nm) of single wheat kernels collected using the Single Kernel Characterization System (SKCS) 4170 were analyzed for wheat grain hardness using partial least squares (PLS) regression. The wavelengths (650 to 700, 1100, 1200, 1380, 1450, and 1670 nm) that contributed most to the ability of the model to predict hardness were related to protein, starch, and color differences. Slightly better prediction results were observed when the 550–1690 nm region was used compared with 950–1690 nm region across all sample sizes. For the 30‐kernel mass‐averaged model, the hardness prediction for 550–1690 nm spectra resulted in a coefficient of determination (R²) = 0.91, standard error of cross validation (SECV) = 7.70, and relative predictive determinant (RPD) = 3.3, while the 950–1690 nm had R² = 0.88, SECV = 8.67, and RPD = 2.9. Average hardness of hard and soft wheat validation samples based on mass‐averaged spectra of 30 kernels was predicted and compared with the SKCS 4100 reference method (R² = 0.88). Compared with the reference SKCS hardness classification, the 30‐kernel (550–1690 nm) prediction model correctly differentiated (97%) between hard and soft wheat. Monte Carlo simulation technique coupled with the SKCS 4100 hardness classification logic was used for classifying mixed wheat samples. Compared with the reference, the prediction model correctly classified mixed samples with 72–100% accuracy. Results confirmed the potential of using visible and near‐infrared reflectance spectroscopy of whole single kernels of wheat as a rapid and nondestructive measurement of bulk wheat grain hardness.     

High‐Throughput Near‐Infrared Reflectance Spectroscopy for Predicting Quantitative and Qualitative Composition Phenotypes of Individual Maize Kernels

Near‐infrared reflectance (NIR) spectroscopy can be used for fast and reliable prediction of organic compounds in complex biological samples. We used a recently developed NIR spectroscopy instrument to predict starch, protein, oil, and weight of individual maize (Zea mays) seeds. The starch, protein, and oil calibrations have reliability equal or better to bulk grain NIR analyzers. We also show that the instrument can differentiate quantitative and qualitative seed composition mutants from normal siblings without a specific calibration for the constituent affected. The analyzer does not require a specific kernel orientation to predict composition or to differentiate mutants. The instrument collects a seed weight and a spectrum in 4–6 sec and can collect NIR data alone at a 20‐fold faster rate. The spectra are acquired while the kernel falls through a glass tube illuminated with broad spectrum light. These results show significant improvements over prior single‐kernel NIR systems, making this instrument a practical tool to collect quantitative seed phenotypes at high throughput. This technology has multiple applications for studying the genetic and physiological influences on seed traits.     

Selecting and Sorting Waxy Wheat Kernels Using Near‐Infrared Spectroscopy

An automated single kernel near‐infrared (NIR) sorting system was used to separate single wheat (Triticum aestivum L.) kernels with amylose‐free (waxy) starch from reduced‐amylose (partial waxy) or wild‐type wheat kernels. Waxy kernels of hexaploid wheat are null for the granule‐bound starch synthase alleles at all three Wx gene loci; partial waxy kernels have at least one null and one functional allele. Wild‐type kernels have three functional alleles. Our results demonstrate that automated single kernel NIR technology can be used to select waxy kernels from segregating breeding lines or to purify advanced breeding lines for the low‐amylose kernel trait. Calibrations based on either amylose content or the waxy trait performed similarly. Also, a calibration developed using the amylose content of waxy, partial waxy, and wild‐type durum (T. turgidum L. var durum) wheat enabled adequate sorting for hard red winter and hard red spring wheat with no modifications. Regression coefficients indicated that absorption by starch in the NIR region contributed to the classification models. Single kernel NIR technology offers significant benefits to breeding programs that are developing wheat with amylose‐free starches.     

Differentiating Vitreous and Nonvitreous Durum Wheat Kernels by Using Near‐Infrared Spectroscopy

The vitreousnss of durum wheat is used by the wheat industry as an indicator of milling and cooking quality. The current visual method of determining vitreousness is subjective, and classification results between inspectors and countries vary widely. Thus, the use of near‐infrared (NIR) spectroscopy to objectively classify vitreous and nonvitreous single kernels was investigated. Results showed that classification of obviously vitreous or nonvitreous kernels by the NIR procedure agreed almost perfectly with inspector classifications. However, when difficult‐to‐classify vitreous and nonvitreous kernels were included in the analysis, the NIR procedure agreed with inspectors on only 75% of kernels. While the classification of difficult kernels by NIR spectroscopy did not match well with inspector classifications, this NIR procedure quantifies vitreousness and thus may provide an objective classification means that could reduce inspector‐to‐inspector variability. Classifications appear to be due, at least in part, to scattering effects and to starch and protein differences between vitreous and nonvitreous kernels.     

Detection of Sprouted and Midge‐Damaged Wheat Kernels Using Near‐Infrared Hyperspectral Imaging

Sprout damage which results in poor breadmaking quality due to enzymatic activity of α‐amylase is one of the important grading factors of wheat in Canada. Potential of near‐infrared (NIR) hyperspectral imaging was investigated to detect sprouting of wheat kernels. Artificially sprouted, midge‐damaged, and healthy wheat kernels were scanned using NIR hyperspectral imaging system in the range of 1000–1600 nm at 60 evenly distributed wavelengths. Multivariate image analysis (MVI) technique based on principal components analysis (PCA) was applied to reduce the dimensionality of the hyperspectral data. Three wavelengths 1101.7, 1132.2, and 1305.1 nm were identified as significant and used in analysis. Statistical discriminant classifiers (linear, quadratic, and Mahalanobis) were used to classify sprouted, midge‐damaged, and healthy wheat kernels. The discriminant classifiers gave maximum accuracy of 98.3 and 100% for classifying healthy and damaged kernels, respectively.     

An Automated Near‐Infrared System for Selecting Individual Kernels Based on Specific Quality Characteristics

An automated sorting system was developed that nondestructively measured quality characteristics of individual kernels using near‐infrared (NIR) spectra. This single‐kernel NIR system was applied to sorting wheat (Triticum aestivum L.) kernels by protein content and hardness, and proso millet (Panicum miliaceum L.) into amylose‐bearing and amylose‐free fractions. Single wheat kernels with high protein content could be sorted from pure lines so that the high‐protein content portion was 3.1 percentage points higher than the portion with the low‐protein kernels. Likewise, single wheat kernels with specific hardness indices could be removed from pure lines such that the hardness index in the sorted samples was 29.4 hardness units higher than the soft kernels. The system was able to increase the waxy, or amylose‐free, millet kernels in segregating samples from 94% in the unsorted samples to 98% in the sorted samples. The portion of waxy millet kernels in segregating samples was increased from 32% in the unsorted samples to 55% after sorting. Thus, this technology can be used to enrich the desirable class within segregating populations in breeding programs, to increase the purity of heterogeneous advanced or released lines, or to measure the distribution of quality within samples during the marketing process.     

Detecting and Segregating Black Tip‐Damaged Wheat Kernels Using Visible and Near‐Infrared Spectroscopy

Detection of individual wheat kernels with black tip symptom (BTS) and black tip damage (BTD) was demonstrated with near‐infrared reflectance spectroscopy (NIRS) and silicon light‐emitting‐diode (LED) based instruments. The two instruments tested, a single‐kernel NIRS instrument (SKNIRS) and a silicon LED‐based single‐kernel high‐speed sorter (SiLED‐SKS) were both developed by the Stored Product Insect and Engineering Research Unit, Center for Grain and Animal Health Research, USDA Agricultural Research Service. BTD was classified into four levels for the study ranging from sound, symptomatic (BTS) at two levels, and damaged (BTD). Discriminant analysis models for the SKNIRS instrument could distinguish sound undamaged kernels well, correctly classifying kernels 80% of the time. Damaged kernels were classified with 67% accuracy and symptomatic kernels at about 44%. Higher classification accuracy (81–87%) was obtained by creating only two groupings: 1) combined sound and lightly symptomatic kernels and 2) combined heavily symptomatic and damaged kernels. A linear regression model was developed from the SiLED‐SKS sorted fractions to predict the percentage of combined BTS and BTD kernels in a sample. The model had an R² of 0.64 and a standard error of prediction of 7.4%, showing it had some measurement ability for BTS and BTD. The SiLED‐SKS correctly classified and sorted out 90% of BTD and 66% of BTS for all 28 samples after three passes through the sorter. These instruments can serve as important tools for plant breeders and grading facilities of the wheat industry that require timely and objective determination and sorting of different levels of black tip present in wheat samples.     

Authentication of Rice Using Near‐Infrared Reflectance Spectroscopy

The authentication of rice (Korean domestic rice vs. foreign rice) has been attempted using near‐infrared spectroscopy (NIRS). Two sample sets (n₁ = 280 and n₂ = 200) were used to obtain calibration equations and the spectral regions used for this study were 500–600 nm, 700–900nm, and 980–2,498 nm. Modified partial least square (MPLS) regression was used to develop the prediction model. The standard error of cross validation (SECV) and the r² were 0.165 and 0.91 respectively for 1st calibration set and 0.165 and 0.93 for 2nd calibration set respectively. The results of the independent validation (n₃ = 80) showed that all of 80 samples were identified correctly. Even though authentication of rice was performed successfully using NIRS, the calibration statistics in this study showed that further effort is needed for implementation of NIRS for authentication of rice for industry purposes.     

Environmental Effects on Developing Wheat as Sensed by Near‐Infrared Reflectance of Mature Grains

For 30 years, near‐infrared (NIR) spectroscopy has routinely been applied to the cereal grains for the purpose of rapidly measuring concentrations of constituents such as protein and moisture. The research described herein examined the ability of NIR reflectance spectroscopy on harvested wheat to determine weather‐related, quality‐determining properties that occurred during plant development. Twenty commercial cultivars or advanced breeding lines of hard red winter and hard white wheat (Triticum aestivum L.) were grown in 10 geographical locations under prevailing natural conditions of the U.S. Great Plains. Diffuse reflectance spectra (1,100–2,498 nm) of ground wheat from these samples were modeled by partial least squares one (PLS1) and multiple linear regression algorithms for the following properties: SDS sedimentation volume, amount of time during grain fill in which the temperature or relative humidity exceeded or was less than a threshold level (i.e., >30, >32, >35, <24°C; >80%, <40% rh). Rainfall values associated with four pre‐ and post‐planting stages also were examined heuristically by PLS2 analysis. Partial correlation analysis was used to statistically remove the contribution of protein content from the quantitative NIR models. PLS1 models of 9–11 factors on scatter‐corrected and (second order) derivatized spectra produced models whose dimensionless error (RPD, ratio of standard deviation of the property in a test set to the model standard error for that property) ranged from 2.0 to 3.3. Multiple linear regression models, involving the sum of four second‐derivative terms with coefficients, produced models of slightly higher error compared with PLS models. For both modeling approaches, partial correlation analysis demonstrated that model success extends beyond an intercorrelation between property and protein content, a constituent that is well‐modeled by NIR spectroscopy. With refinement, these types of NIR models may have the potential to provide grain handlers, millers, and bakers a tool for identifying the cultural environment under which the purchased grain was produced.     

Development and Evaluation of a Near‐Infrared Instrument for Single‐Seed Compositional Measurement of Wheat Kernels

A single‐kernel, near‐infrared reflectance instrument was designed, built, and tested for its ability to measure composition and traits in wheat kernels. The major objective of the work was targeted at improving an existing design concept of an instrument used for larger seeds such as soybeans and corn but in this case designed for small seeds. Increases in throughput were sought by using a vacuum to convey seeds without compromising measurement accuracy. Instrument performance was evaluated by examining measurement accuracy of wheat kernel moisture, protein content, and kernel mass. Spectral measurements were obtained on individual wheat kernels as they were conveyed by air through an illuminated tube. Partial least squares (PLS) prediction models for these constituents were then developed and evaluated. PLS single‐kernel moisture predictions had a root mean square error of prediction (RMSEP) around 0.5% MC wet basis; protein prediction models had an RMSEP near 0.70%. Prediction of mass was not as good but still provided a reasonable estimate of single‐kernel mass, with RMSEP values of 2.8–4 mg. Data showed that kernel mass and protein content were not correlated, in contrast to some previous research. Overall, results showed the instrument performed comparably to other single‐seed instruments or methods based on accuracy but with an increased throughput at a rate of at least 4 seeds/s.     

Discriminating Oat and Groat Kernels from Other Grains Using Near‐Infrared Spectroscopy

Oats and groats can be discriminated from other grains such as barley, wheat, rye, and triticale (nonoats) with near‐infrared spectroscopy. The two instruments tested herein were the manual version of the United States Department of Agriculture–Agricultural Research Service single‐kernel near‐infrared (SKNIR) instrument and the automated QualySense QSorter Explorer high‐speed sorter, both used in similar near‐infrared spectral ranges. Three linear discriminate self‐prediction models were developed: 1) oats versus groats + nonoats, 2) oats + groats versus nonoats, and 3) groats versus nonoats. For all three models, the SKNIR instrument showed high correct classification of oats or groats (94.5–100%), which was similar to results of the QSorter Explorer at 95.0–99.4%. The amount of nonoats that were misclassified as oats or groats was low for both instruments at 0–0.2% for the SKNIR instrument and 0.8–3.7% for the QSorter Explorer. Linear discriminate models from independent prediction and validation sets yielded classification accuracies of 91.6–99.3% (SKNIR) and 90.5–97.8% (QSorter Explorer). Small differences in classification accuracy were attributed to processing speeds between the two instruments: 3 kernels/s for the SKNIR instrument and 35 kernels/s for the QSorter Explorer. This indicated that both instruments are useful for quantifying grain sample compositions of oat and groat samples and that both could be useful tools for meeting consumer demand for gluten‐free or low‐gluten products. Discrimination between grains will help producers and manufacturers meet various regulatory requirements. Examples include requirements such as those from the U.S. Food and Drug Administration and the Commission of European Communities, in which gluten‐free oats or other products can only be labeled as nongluten if they contain gluten at less than 20 ppm, the established safe consumption limit for people with celiac disease. The QSorter Explorer is currently being used to meet these requirements.     

Bulk Functionality Diversification by Unsupervised Single‐Kernel Near‐Infrared (SKNIR) Sorting of Wheat

We explored the effects of fractioning heterogeneous bulk wheat by fast unsupervised single‐kernel near‐infrared (SKNIR) sorting according to an internal complex NIR functionality trait using a fast prototype kernel sorter designed for postharvest bulk sorting. Sorting into three functionality fractions was performed on low quality lots from an organic field experiment from two growth years and two locations. Sorted lots were mixtures originally diversified by three different preceding catch crops. The resulting 12 fractions, as well as the 12 original wheat lots were characterized by 20 standard quality variables of grains and flours. The data was analyzed by principal component analysis (PCA) and analysis of variance (ANOVA). Within each year and location/cultivar, the SKNIR fractionation had significant positive effect on bulk grain density, protein, wet gluten content, Zeleny sedimentation volume, farinograph water absorption, farinograph softening, falling number, gelatinization temperature, and hardness index. Using the NIR fingerprint directly for sorting without calibration to a univariate reference showed that the resulting fractions were based on the major variance in the entire physicochemical quality trait within each lot as expressed by NIR. This novel unsupervised approach may become a powerful tool for sorting according to complex functionality traits, thus increasing overall quality, applicability, and value of the sorted crop.     

Determining Weight and Moisture Properties of Sound and Fusarium‐Damaged Single Wheat Kernels by Near‐Infrared Spectroscopy

Single kernel moisture content (MC) is important in the measurement of other quality traits in single kernels because many traits are expressed on a dry weight basis. MC also affects viability, storage quality, and price. Also, if near‐infrared (NIR) spectroscopy is used to measure grain traits, the influence of water must be accounted for because water is a strong absorber throughout the NIR region. The feasibility of measurement of MC, fresh weight, dry weight, and water mass of single wheat kernels with or without Fusarium damage was investigated using two wheat cultivars with three visually selected classes of kernels with Fusarium damage and a range of MC. Calibration models were developed either from all kernel classes or from only undamaged kernels of one cultivar that were then validated using all spectra of the other cultivar. A calibration model developed for MC when using all kernels from the wheat cultivar Jagalene had a coefficient of determination (R²) of 0.77 and standard error of cross validation (SECV) of 1.03%. This model predicted the MC of the wheat cultivar 2137 with R² of 0.81 and a standard error of prediction (SEP) of 1.02% and RPD of 2.2. Calibration models developed using all kernels from both cultivars predicted MC, fresh weight, dry weight, or water mass in kernels better than models that used only undamaged kernels from both cultivars. Single kernel water mass was more accurately estimated using the actual fresh weight of kernels and MC predicted by calibrations that used all kernels or undamaged kernels. The necessity for evaluating and expressing constituent levels in single kernels on a mass/kernel basis rather than a percentage basis was elaborated. The need to overcome the effects of kernel size and water mass on single kernel spectra before using in calibration model development was also highlighted.     

Comparison of Near‐Infrared Reflectance Spectroscopy and Texture Analyzer for Measuring Wheat Bread Changes in Storage

Bread staling affects bread texture properties and is one of the most common problems in bread storage. Bread firmness, as measured in compression mode by a texture analyzer (TA) has been commonly used to measure bread staling. This study investigated the potential of visible and near‐infrared reflectance spectroscopy (NIRS) to detect bread changes during storage by comparing NIRS results with those obtained by TA. Twenty‐five loaves of commercial wheat white pan bread from one batch were studied over five days. NIRS and TA measurements were made on the same slice at approximately the same time. The experiment was repeated five times using the same kind of commercial samples from five different batches. NIRS measurements of slices, loaf averages, and daily averages were compared with TA measurements. NIRS spectra had a high correlation to TA firmness. NIRS measurements correlated better with the actual storage time and had smaller standard deviations than the TA measurements. The batch differences had less effect on NIRS measurements than on the TA measurements. The results indicate that NIRS could follow bread changes during storage more accurately than the TA. NIRS is probably based on both physical and chemical changes during bread staling, unlike the TA method that only measures bread firmness, which is only one aspect of the staling phenomenon.     

Evaluation of a Near‐Infrared Reflectance Spectrometer as a Granulation Sensor for First‐Break Ground Wheat: Studies with Six Wheat Classes

In flour milling, a granulation sensor for ground wheat is needed for automatic control of a roller mill's roll gap. A near‐infrared (NIR) reflectance spectrometer was evaluated as a potential granulation sensor of first‐break ground wheat using offline methods. Sixty wheat samples, ground independently, representing six classes and five roller mill gaps, were each used for calibration and validation sets. Partial least squares regression was used to develop the models with cumulative mass of size fraction as the reference value. Combinations of four data pretreatments (log (1/R), baseline correction, unit area normalization, and derivatives) and three wavelength regions (700–1,500, 800–1,600, and 600–1,700 nm) were evaluated. Unit area normalization combined with baseline correction or second derivative yielded models that predicted well each size fraction of first‐break ground wheat. Standard errors of performance of 4.07, 1.75, 1.03, and 1.40 and r² of 0.93, 0.90, 0.88, and 0.38 for the >1,041‐, >375‐, >240‐, and >136‐μm size ranges, respectively, were obtained for the best model. Results indicate that the granulation sensing technique based on NIR reflectance is ready for online evaluation.     

Near‐Infrared Spectroscopic Method for Identification of Fusarium Head Blight Damage and Prediction of Deoxynivalenol in Single Wheat Kernels

Fusarium Head Blight (FHB), or scab, can result in significant crop yield losses and contaminated grain in wheat (Triticum aestivum L.). Growing less susceptible cultivars is one of the most effective methods for managing FHB and for reducing deoxynivalenol (DON) levels in grain, but breeding programs lack a rapid and objective method for identifying the fungi and toxins. It is important to estimate proportions of sound kernels and Fusarium‐damaged kernels (FDK) in grain and to estimate DON levels of FDK to objectively assess the resistance of a cultivar. An automated single kernel near‐infrared (SKNIR) spectroscopic method for identification of FDK and for estimating DON levels was evaluated. The SKNIR system classified visually sound and FDK with an accuracy of 98.8 and 99.9%, respectively. The sound fraction had no or very little accumulation of DON. The FDK fraction was sorted into fractions with high or low DON content. The kernels identified as FDK by the SKNIR system had better correlation with other FHB assessment indices such as FHB severity, FHB incidence and kernels/g than visual FDK%. This technique can be successfully employed to nondestructively sort kernels with Fusarium damage and to estimate DON levels of those kernels. Single kernels could be predicted as having low (<60 ppm) or high (>60 ppm) DON with ≈96% accuracy. Single kernel DON levels of the high DON kernels could be estimated with R² = 0.87 and standard error of prediction (SEP) of 60.8 ppm. Because the method is nondestructive, seeds may be saved for generation advancement. The automated method is rapid (1 kernel/sec) and sorting grains into several fractions depending on DON levels will provide breeders with more information than techniques that deliver average DON levels from bulk seed samples.     

Predicting Wheat Quality Characteristics and Functionality Using Near‐Infrared Spectroscopy

The accuracy of using near‐infrared spectroscopy (NIRS) for predicting 186 grain, milling, flour, dough, and breadmaking quality parameters of 100 hard red winter (HRW) and 98 hard red spring (HRS) wheat and flour samples was evaluated. NIRS shows the potential for predicting protein content, moisture content, and flour color b* values with accuracies suitable for process control (R² > 0.97). Many other parameters were predicted with accuracies suitable for rough screening including test weight, average single kernel diameter and moisture content, SDS sedimentation volume, color a* values, total gluten content, mixograph, farinograph, and alveograph parameters, loaf volume, specific loaf volume, baking water absorption and mix time, gliadin and glutenin content, flour particle size, and the percentage of dark hard and vitreous kernels. Similar results were seen when analyzing data from either HRW or HRS wheat, and when predicting quality using spectra from either grain or flour. However, many attributes were correlated to protein content and this relationship influenced classification accuracies. When the influence of protein content was removed from the analyses, the only factors that could be predicted by NIRS with R² > 0.70 were moisture content, test weight, flour color, free lipids, flour particle size, and the percentage of dark hard and vitreous kernels. Thus, NIRS can be used to predict many grain quality and functionality traits, but mainly because of the high correlations of these traits to protein content.     

Properties of Starches from Near‐Isogenic Wheat Lines with Different Wx Protein Deficiencies

To determine the effect of amylose content on the starch properties, the amylose content, pasting properties, swelling power, enzymatic digestibility, and thermal properties of partial and perfect waxy types along with their wild‐type parent were analyzed. As expected, amylose content decreases differently in response to the loss of each Wx gene, showing the least response to Wx‐A1a. Most of the characteristics, except the thermal properties of the amylose‐lipid complex in differential scanning calorimetry (DSC), differed significantly among the tested types. Furthermore, the breakdown, setback, and pasting temperatures from the Rapid Visco Analyser (RVA) and the enzymatic digestibility, swelling power, peak temperature, and enthalpy of starch gelatinization from DSC showed a correlation with the amylose content. The relationships between the peak viscosity from the RVA and the onset temperature of starch gelatinization determined by DSC with amylose content of the tested materials were not clear. Waxy starch, which has no amylose, showed a contrasting behavior in starch gelatinization compared with nonwaxy starches. Among the nonwaxy starches, lower setback, lower pasting temperature, higher enzyme digestibility, higher peak temperature, higher enthalpy of starch gelatinization, and higher swelling were generally associated with low amylose starches.