Field Phenomics: Will It Enable Crop Improvement?

Field phenomics has been identified as a promising enabling technology to assist plant breeders with the development of improved cultivars for farmers. Yet, despite much investment, there are few examples demonstrating the application of phenomics within a plant breeding program. We review recent progress in field phenomics and highlight the importance of targeting breeders' needs, rather than perceived technology needs, through developing and enhancing partnerships between phenomics researchers and plant breeders.

Impact of Varying Light and Dew on Ground Cover Estimates from Active NDVI, RGB, and LiDAR.

Canopy ground cover (GC) is an important agronomic measure for evaluating crop establishment and early growth. This study evaluates the reliability of GC estimates, in the presence of varying light and dew on leaves, from three different ground-based sensors: (1) normalized difference vegetation index (NDVI) from the commercially available GreenSeeker®; (2) RGB images from a digital camera, where GC was determined as the portion of pixels from each image meeting a greenness criterion (i.e., (Green - Red)/(Green + Red) > 0); and (3) LiDAR using two separate approaches: (a) GC from LiDAR red reflectance (whereby red reflectance less than five was classified as vegetation) and (b) GC from LiDAR height (whereby height greater than 10 cm was classified as vegetation). Hourly measurements were made early in the season at two different growth stages (tillering and stem elongation), among wheat genotypes highly diverse for canopy characteristics. The active NDVI showed the least variation through time and was particularly stable, regardless of the available light or the presence of dew. In addition, between-sample-time Pearson correlations for NDVI were consistently high and significant (P < 0.0001), ranging from 0.89 to 0.98. In comparison, GC from LiDAR and RGB showed greater variation across sampling times, and LiDAR red reflectance was strongly influenced by the presence of dew. Excluding times when the light was exceedingly low, correlations between GC from RGB and NDVI were consistently high (ranging from 0.79 to 0.92). The high reliability of the active NDVI sensor potentially affords a high degree of flexibility for users by enabling sampling across a broad range of acceptable light conditions.

Ground-Based LiDAR Improves Phenotypic Repeatability of Above-Ground Biomass and Crop Growth Rate in Wheat.

Highly repeatable, nondestructive, and high-throughput measures of above-ground biomass (AGB) and crop growth rate (CGR) are important for wheat improvement programs. This study evaluates the repeatability of destructive AGB and CGR measurements in comparison to two previously described methods for the estimation of AGB from LiDAR: 3D voxel index (3DVI) and 3D profile index (3DPI). Across three field experiments, contrasting in available water supply and comprising up to 98 wheat genotypes varying for canopy architecture, several concurrent measurements of LiDAR and AGB were made from jointing to anthesis. Phenotypic correlations at discrete events between AGB and the LiDAR-derived biomass indices were significant, ranging from 0.31 (P < 0.05) to 0.86 (P < 0.0001), providing confidence in the LiDAR indices as effective surrogates for AGB. The repeatability of the LiDAR biomass indices at discrete events was at least similar to and often higher than AGB, particularly under water limitation. The correlations between calculated CGR for AGB and the LiDAR indices were moderate to high and varied between experiments. However, across all experiments, the repeatabilities of the CGR derived from the LiDAR indices were appreciably greater than those for AGB, except for the 3DPI in the water-limited environment. In our experiments, the repeatability of either LiDAR index was consistently higher than that of AGB, both at discrete time points and when CGR was calculated. These findings provide promising support for the reliable use of ground-based LiDAR, as a surrogate measure of AGB and CGR, for screening germplasm in research and wheat breeding.

Genetic variation for photosynthetic capacity and efficiency in spring wheat.

One way to increase yield potential in wheat is screening for natural variation in photosynthesis. This study uses measured and modelled physiological parameters to explore genotypic diversity in photosynthetic capacity (Pc, Rubisco carboxylation capacity per unit leaf area at 25 °C) and efficiency (Peff, Pc per unit of leaf nitrogen) in wheat in relation to fertilizer, plant stage, and environment. Four experiments (Aus1, Aus2, Aus3, and Mex1) were carried out with diverse wheat collections to investigate genetic variation for Rubisco capacity (Vcmax25), electron transport rate (J), CO2 assimilation rate, stomatal conductance, and complementary plant functional traits: leaf nitrogen, leaf dry mass per unit area, and SPAD. Genotypes for Aus1 and Aus2 were grown in the glasshouse with two fertilizer levels. Genotypes for Aus3 and Mex1 experiments were grown in the field in Australia and Mexico, respectively. Results showed that Vcmax25 derived from gas exchange measurements is a robust parameter that does not depend on stomatal conductance and was positively correlated with Rubisco content measured in vitro. There was significant genotypic variation in most of the experiments for Pc and Peff. Heritability of Pc reached 0.7 and 0.9 for SPAD. Genotypic variation and heritability of traits show that there is scope for these traits to be used in pre-breeding programmes to improve photosynthesis with the ultimate objective of raising yield potential.

Evaluation of the Phenotypic Repeatability of Canopy Temperature in Wheat Using Continuous-Terrestrial and Airborne Measurements.

Infrared canopy temperature (CT) is a well-established surrogate measure of stomatal conductance. There is ample evidence showing that genotypic variation in stomatal conductance is associated with grain yield in wheat. Our goal was to determine when CT repeatability is greatest (throughout the season and within the day) to guide CT deployment for research and wheat breeding. CT was measured continuously with ArduCrop wireless infrared thermometers from post-tillering to physiological maturity, and with airborne thermography on cloudless days from manned helicopter at multiple times before and after flowering. Our experiments in wheat, across two years contrasting for water availability, showed that repeatability for CT was greatest later in the season, during grain-filling, and usually in the afternoon. This was supported by the observation that repeatability for ArduCrop, and more so for airborne CT, was significantly associated (P < 0.0001) with calculated clear-sky solar radiation and to a lesser degree, vapor pressure deficit. Adding vapor pressure deficit to a model comprising either clear-sky solar radiation or its determinants, day-of-year and hour-of-day, made little to no improvement to the coefficient of determination. Phenotypic correlations for airborne CT afternoon sampling events were consistently high between events in the same year, more so for the year when soil water was plentiful (r = 0.7 to 0.9) than the year where soil water was limiting (r = 0.4 to 0.9). Phenotypic correlations for afternoon airborne CT were moderate between years contrasting in soil water availability (r = 0.1 to 0.5) and notably greater on two separate days following irrigation or rain in the drier year, ranging from r = 0.39 to 0.53 (P < 0.0001) for the midday events. For ArduCrop CT the pattern of phenotypic correlations, within a given year, was similar for both years: phenotypic correlations were higher during the grain-filling months of October and November and for hours-of-day from 11 onwards. The lowest correlations comprised events from hours-of-day 8 and 9 across all months. The capacity for the airborne method to instantaneously sample CT on hundreds of plots is more suited to large field experiments than the static ArduCrop sensors which measure CT continuously on a single experimental plot at any given time. Our findings provide promising support for the reliable deployment of CT phenotyping for research and wheat breeding, whereby the high repeatability and high phenotypic correlations between afternoon sampling events during grain-filling could enable reliable screening of germplasm from only one or two sampling events.

Field crop phenomics: enabling breeding for radiation use efficiency and biomass in cereal crops.

Plant phenotyping forms the core of crop breeding, allowing breeders to build on physiological traits and mechanistic science to inform their selection of material for crossing and genetic gain. Recent rapid progress in high-throughput techniques based on machine vision, robotics, and computing (plant phenomics) enables crop physiologists and breeders to quantitatively measure complex and previously intractable traits. By combining these techniques with affordable genomic sequencing and genotyping, machine learning, and genome selection approaches, breeders have an opportunity to make rapid genetic progress. This review focuses on how field-based plant phenomics can enable next-generation physiological breeding in cereal crops for traits related to radiation use efficiency, photosynthesis, and crop biomass. These traits have previously been regarded as difficult and laborious to measure but have recently become a focus as cereal breeders find genetic progress from 'Green Revolution' traits such as harvest index become exhausted. Application of LiDAR, thermal imaging, leaf and canopy spectral reflectance, Chl fluorescence, and machine learning are discussed using wheat and sorghum phenotyping as case studies. A vision of how crop genomics and high-throughput phenotyping could enable the next generation of crop research and breeding is presented.

High Throughput Determination of Plant Height, Ground Cover, and Above-Ground Biomass in Wheat with LiDAR.

Crop improvement efforts are targeting increased above-ground biomass and radiation-use efficiency as drivers for greater yield. Early ground cover and canopy height contribute to biomass production, but manual measurements of these traits, and in particular above-ground biomass, are slow and labor-intensive, more so when made at multiple developmental stages. These constraints limit the ability to capture these data in a temporal fashion, hampering insights that could be gained from multi-dimensional data. Here we demonstrate the capacity of Light Detection and Ranging (LiDAR), mounted on a lightweight, mobile, ground-based platform, for rapid multi-temporal and non-destructive estimation of canopy height, ground cover and above-ground biomass. Field validation of LiDAR measurements is presented. For canopy height, strong relationships with LiDAR (r2 of 0.99 and root mean square error of 0.017 m) were obtained. Ground cover was estimated from LiDAR using two methodologies: red reflectance image and canopy height. In contrast to NDVI, LiDAR was not affected by saturation at high ground cover, and the comparison of both LiDAR methodologies showed strong association (r2 = 0.92 and slope = 1.02) at ground cover above 0.8. For above-ground biomass, a dedicated field experiment was performed with destructive biomass sampled eight times across different developmental stages. Two methodologies are presented for the estimation of biomass from LiDAR: 3D voxel index (3DVI) and 3D profile index (3DPI). The parameters involved in the calculation of 3DVI and 3DPI were optimized for each sample event from tillering to maturity, as well as generalized for any developmental stage. Individual sample point predictions were strong while predictions across all eight sample events, provided the strongest association with biomass (r2 = 0.93 and r2 = 0.92) for 3DPI and 3DVI, respectively. Given these results, we believe that application of this system will provide new opportunities to deliver improved genotypes and agronomic interventions via more efficient and reliable phenotyping of these important traits in large experiments.

Methodology for High-Throughput Field Phenotyping of Canopy Temperature Using Airborne Thermography.

Lower canopy temperature (CT), resulting from increased stomatal conductance, has been associated with increased yield in wheat. Historically, CT has been measured with hand-held infrared thermometers. Using the hand-held CT method on large field trials is problematic, mostly because measurements are confounded by temporal weather changes during the time required to measure all plots. The hand-held CT method is laborious and yet the resulting heritability low, thereby reducing confidence in selection in large scale breeding endeavors. We have developed a reliable and scalable crop phenotyping method for assessing CT in large field experiments. The method involves airborne thermography from a manned helicopter using a radiometrically-calibrated thermal camera. Thermal image data is acquired from large experiments in the order of seconds, thereby enabling simultaneous measurement of CT on potentially 1000s of plots. Effects of temporal weather variation when phenotyping large experiments using hand-held infrared thermometers are therefore reduced. The method is designed for cost-effective and large-scale use by the non-technical user and includes custom-developed software for data processing to obtain CT data on a single-plot basis for analysis. Broad-sense heritability was routinely >0.50, and as high as 0.79, for airborne thermography CT measured near anthesis on a wheat experiment comprising 768 plots of size 2 × 6 m. Image analysis based on the frequency distribution of temperature pixels to remove the possible influence of background soil did not improve broad-sense heritability. Total image acquisition and processing time was ca. 25 min and required only one person (excluding the helicopter pilot). The results indicate the potential to phenotype CT on large populations in genetics studies or for selection within a plant breeding program.

SensorDB: a virtual laboratory for the integration, visualization and analysis of varied biological sensor data.

BACKGROUND: To our knowledge, there is no software or database solution that supports large volumes of biological time series sensor data efficiently and enables data visualization and analysis in real time. Existing solutions for managing data typically use unstructured file systems or relational databases. These systems are not designed to provide instantaneous response to user queries. Furthermore, they do not support rapid data analysis and visualization to enable interactive experiments. In large scale experiments, this behaviour slows research discovery, discourages the widespread sharing and reuse of data that could otherwise inform critical decisions in a timely manner and encourage effective collaboration between groups. RESULTS: In this paper we present SensorDB, a web based virtual laboratory that can manage large volumes of biological time series sensor data while supporting rapid data queries and real-time user interaction. SensorDB is sensor agnostic and uses web-based, state-of-the-art cloud and storage technologies to efficiently gather, analyse and visualize data. CONCLUSIONS: Collaboration and data sharing between different agencies and groups is thereby facilitated. SensorDB is available online at http://sensordb.csiro.au.