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Creators/Authors contains: "Singh, Asheesh"

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  1. Using a reliable and accurate method to phenotype disease incidence and severity is essential to unravel the complex genetic architecture of disease resistance in plants, and to develop disease resistant cultivars. Genome-wide association studies (GWAS) involve phenotyping large numbers of accessions, and have been used for a myriad of traits. In field studies, genetic accessions are phenotyped across multiple environments and replications, which takes a significant amount of labor and resources. Deep Learning (DL) techniques can be effective for analyzing image-based tasks; thus DL methods are becoming more routine for phenotyping traits to save time and effort. This research aims to conduct GWAS on sudden death syndrome (SDS) of soybean [ Glycine max L. (Merr.)] using disease severity from both visual field ratings and DL-based (using images) severity ratings collected from 473 accessions. Images were processed through a DL framework that identified soybean leaflets with SDS symptoms, and then quantified the disease severity on those leaflets into a few classes with mean Average Precision of 0.34 on unseen test data. Both visual field ratings and image-based ratings identified significant single nucleotide polymorphism (SNP) markers associated with disease resistance. These significant SNP markers are either in the proximity of previously reported candidate genes for SDS or near potentially novel candidate genes. Four previously reported SDS QTL were identified that contained a significant SNPs, from this study, from both a visual field rating and an image-based rating. The results of this study provide an exciting avenue of using DL to capture complex phenotypic traits from images to get comparable or more insightful results compared to subjective visual field phenotyping of traits for disease symptoms. 
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  2. Abstract

    The symbiotic relationship between soybean [Glycine maxL. (Merr.)] roots and bacteria (Bradyrhizobium japonicum) lead to the development of nodules, important legume root structures where atmospheric nitrogen (N2) is fixed into bio‐available ammonia (NH3) for plant growth and development. With the recent development of the Soybean Nodule Acquisition Pipeline (SNAP), nodules can more easily be quantified and evaluated for genetic diversity and growth patterns across unique soybean root system architectures. We explored six diverse soybean genotypes across three field year combinations in three early vegetative stages of development and report the unique relationships between soybean nodules in the taproot and non‐taproot growth zones of diverse root system architectures of these genotypes. We found unique growth patterns in the nodules of taproots showing genotypic differences in how nodules grew in count, size, and total nodule area per genotype compared to non‐taproot nodules. We propose that nodulation should be defined as a function of both nodule count and individual nodule area resulting in a total nodule area per root or growth regions of the root. We also report on the relationships between the nodules and total nitrogen in the seed at maturity, finding a strong correlation between the taproot nodules and final seed nitrogen at maturity. The applications of these findings could lead to an enhanced understanding of the plant‐Bradyrhizobiumrelationship and exploring these relationships could lead to leveraging greater nitrogen use efficiency and nodulation carbon to nitrogen production efficiency across the soybean germplasm.

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  3. Abstract

    Insect pests cause significant damage to food production, so early detection and efficient mitigation strategies are crucial. There is a continual shift toward machine learning (ML)‐based approaches for automating agricultural pest detection. Although supervised learning has achieved remarkable progress in this regard, it is impeded by the need for significant expert involvement in labeling the data used for model training. This makes real‐world applications tedious and oftentimes infeasible. Recently, self‐supervised learning (SSL) approaches have provided a viable alternative to training ML models with minimal annotations. Here, we present an SSL approach to classify 22 insect pests. The framework was assessed on raw and segmented field‐captured images using three different SSL methods, Nearest Neighbor Contrastive Learning of Visual Representations (NNCLR), Bootstrap Your Own Latent, and Barlow Twins. SSL pre‐training was done on ResNet‐18 and ResNet‐50 models using all three SSL methods on the original RGB images and foreground segmented images. The performance of SSL pre‐training methods was evaluated using linear probing of SSL representations and end‐to‐end fine‐tuning approaches. The SSL‐pre‐trained convolutional neural network models were able to perform annotation‐efficient classification. NNCLR was the best performing SSL method for both linear and full model fine‐tuning. With just 5% annotated images, transfer learning with ImageNet initialization obtained 74% accuracy, whereas NNCLR achieved an improved classification accuracy of 79% for end‐to‐end fine‐tuning. Models created using SSL pre‐training consistently performed better, especially under very low annotation, and were robust to object class imbalances. These approaches help overcome annotation bottlenecks and are resource efficient.

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  4. Reliable seed yield estimation is an indispensable step in plant breeding programs geared towards cultivar development in major row crops. The objective of this study is to develop a machine learning (ML) approach adept at soybean ( Glycine max L. (Merr.)) pod counting to enable genotype seed yield rank prediction from in-field video data collected by a ground robot. To meet this goal, we developed a multiview image-based yield estimation framework utilizing deep learning architectures. Plant images captured from different angles were fused to estimate the yield and subsequently to rank soybean genotypes for application in breeding decisions. We used data from controlled imaging environment in field, as well as from plant breeding test plots in field to demonstrate the efficacy of our framework via comparing performance with manual pod counting and yield estimation. Our results demonstrate the promise of ML models in making breeding decisions with significant reduction of time and human effort and opening new breeding method avenues to develop cultivars. 
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  5. Abstract

    High‐throughput phenotyping (HTP) with unoccupied aerial systems (UAS), consisting of unoccupied aerial vehicles (UAV; or drones) and sensor(s), is an increasingly promising tool for plant breeders and researchers. Enthusiasm and opportunities from this technology for plant breeding are similar to the emergence of genomic tools ∼30 years ago, and genomic selection more recently. Unlike genomic tools, HTP provides a variety of strategies in implementation and utilization that generate big data on the dynamic nature of plant growth formed by temporal interactions between growth and environment. This review lays out strategies deployed across four major staple crop species: cotton (Gossypium hirsutumL.), maize (Zea maysL.), soybean (Glycine maxL.), and wheat (Triticum aestivumL.). Each crop highlighted in this review demonstrates how UAS‐collected data are employed to automate and improve estimation or prediction of objective phenotypic traits. Each crop section includes four major topics: (a) phenotyping of routine traits, (b) phenotyping of previously infeasible traits, (c) sample cases of UAS application in breeding, and (d) implementation of phenotypic and phenomic prediction and selection. While phenotyping of routine agronomic and productivity traits brings advantages in time and resource optimization, the most potentially beneficial application of UAS data is in collecting traits that were previously difficult or impossible to quantify, improving selection efficiency of important phenotypes. In brief, UAS sensor technology can be used for measuring abiotic stress, biotic stress, crop growth and development, as well as productivity. These applications and the potential implementation of machine learning strategies allow for improved prediction, selection, and efficiency within breeding programs, making UAS HTP a potentially indispensable asset.

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  6. Abstract

    Glycerol-3-phosphate (G3P) is a well-known mobile regulator of systemic acquired resistance (SAR), which provides broad spectrum systemic immunity in response to localized foliar pathogenic infections. We show that G3P-derived foliar immunity is also activated in response to genetically-regulated incompatible interactions with nitrogen-fixing bacteria. Using gene knock-down we show that G3P is essential for strain-specific exclusion of non-desirable root-nodulating bacteria and the associated foliar pathogen immunity in soybean. Grafting studies show that while recognition of rhizobium incompatibility is root driven, bacterial exclusion requires G3P biosynthesis in the shoot. Biochemical analyses support shoot-to-root transport of G3P during incompatible rhizobia interaction. We describe a root-shoot-root signaling mechanism which simultaneously enables the plant to exclude non-desirable nitrogen-fixing rhizobia in the root and pathogenic microbes in the shoot.

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