Using Crop Databases to Explore Phenotypes: From QTL to Candidate Genes

Seeds, especially those of certain grasses and legumes, provide the majority of the protein and carbohydrates for much of the world’s population. Therefore, improvements in seed quality and yield are important drivers for the development of new crop varieties to feed a growing population. Quantitative Trait Loci (QTL) have been identified for many biologically interesting and agronomically important traits, including many seed quality traits. QTL can help explain the genetic architecture of the traits and can also be used to incorporate traits into new crop cultivars during breeding. Despite the important contributions that QTL have made to basic studies and plant breeding, knowing the exact gene(s) conditioning each QTL would greatly improve our ability to study the underlying genetics, biochemistry and regulatory networks. The data sets needed for identifying these genes are increasingly available and often housed in species- or clade-specific genetics and genomics databases. In this demonstration, we present a generalized walkthrough of how such databases can be used in these studies using SoyBase, the USDA soybean Genetics and Genomics Database, as an example.

A new decade and new data at SoyBase, the USDA-ARS soybean genetics and genomics database

SoyBase, a USDA genetic and genomics database, holds professionally curated soybean genetic and genomic data, which is integrated and made accessible to researchers and breeders. The site holds several reference genome assemblies, as well as genetic maps, thousands of mapped traits, expression and epigenetic data, pedigree information, and extensive variant and genotyping data sets. SoyBase displays include genetic, genomic, and epigenetic maps of the soybean genome. Gene expression data is presented in the genome viewer as heat maps and pictorial and tabular displays in gene report pages. Millions of sequence variants have been added, representing variations across various collections of cultivars. This variant data is explorable using new interactive tools to visualize the distribution of those variants across the genome, between selected accessions. SoyBase holds several reference-quality soybean genome assemblies, accessible via various query tools and browsers, including a new visualization system for exploring the soybean pan-genome. SoyBase also serves as a nexus of announcements pertinent to the greater soybean research community. The database also includes a soybean-specific anatomic and biochemical trait ontology. The database can be accessed at https://soybase.org.

Systematic analysis of 1,298 RNA-Seq samples and construction of a comprehensive soybean (Glycine max) expression atlas

Soybean (Glycine max [L.] Merr.) is a major crop in animal feed and human nutrition, mainly for its rich protein and oil contents. The remarkable rise in soybean transcriptome studies over the past five years generated an enormous amount of RNA-seq data, encompassing various tissues, developmental conditions, and genotypes. In this study, we have collected data from 1,298 publicly available soybean transcriptome samples, processed the raw sequencing reads, and mapped them to the soybean reference genome in a systematic fashion. We found that 94% of the annotated genes (52,737/56,044) had detectable expression in at least one sample. Unsupervised clustering revealed three major groups, comprising samples from aerial, underground, and seed/seed-related parts. We found 452 genes with uniform and constant expression levels, supporting their roles as housekeeping genes. On the other hand, 1,349 genes showed heavily biased expression patterns towards particular tissues. A transcript-level analysis revealed that 95% (70,963/74,490) of the known transcripts overlap with those reported here, whereas 3,256 assembled transcripts represent potentially novel splicing isoforms. The dataset compiled here constitute a new resource for the community, which can be downloaded or accessed through a user-friendly web interface at http://venanciogroup.uenf.br/resources/. This comprehensive transcriptome atlas will likely accelerate research on soybean genetics and genomics.

Herbicide Program Approaches for Managing Glyphosate-Resistant Palmer Amaranth (Amaranthus palmeri) and Waterhemp (Amaranthus tuberculatusandAmaranthus rudis) in Future Soybean-Trait Technologies

Herbicide-resistantAmaranthusspp. continue to cause management difficulties in soybean. New soybean technologies under development, including resistance to various combinations of glyphosate, glufosinate, dicamba, 2,4-D, isoxaflutole, and mesotrione, will make possible the use of additional herbicide sites of action in soybean than is currently available. When this research was conducted, these soybean traits were still regulated and testing herbicide programs with the appropriate soybean genetics in a single experiment was not feasible. Therefore, the effectiveness of various herbicide programs (PRE herbicides followed by POST herbicides) was evaluated in bare-ground experiments on glyphosate-resistant Palmer amaranth and glyphosate-resistant waterhemp (both tall and common) at locations in Arkansas, Illinois, Indiana, Missouri, Nebraska, and Tennessee. Twenty-five herbicide programs were evaluated; 5 of which were PRE herbicides only, 10 were PRE herbicides followed by POST herbicides 3 to 4 wks after (WA) the PRE application (EPOST), and 10 were PRE herbicides followed by POST herbicides 6 to 7 WA the PRE application (LPOST). Programs with EPOST herbicides provided 94% or greater control of Palmer amaranth and waterhemp at 3 to 4 WA the EPOST. Overall, programs with LPOST herbicides resulted in a period of weed emergence in which weeds would typically compete with a crop. Weeds were not completely controlled with the LPOST herbicides because weed sizes were larger (≥ 15 cm) compared with their sizes at the EPOST application (≤ 7 cm). Most programs with LPOST herbicides provided 80 to 95% control at 3 to 4 WA applied LPOST. Based on an orthogonal contrast, using a synthetic-auxin herbicide LPOST improves control of Palmer amaranth and waterhemp over programs not containing a synthetic-auxin LPOST. These results show herbicides that can be used in soybean and that contain auxinic- or HPPD-resistant traits will provide growers with an opportunity for better control of glyphosate-resistant Palmer amaranth and waterhemp over a wide range of geographies and environments.

Studies on efficiency of RAPD primers in developing molecular profiles for genetic purity studies in soybean (Glycine max L.) cultivars

Major advances have recently been made in our understanding of soybean genetics and of the application of new technologies to soybean improvement. Estimates of genetic relationships on the basis of the enzymes and molecular markers have been shown to be consistent with expectations based on origin and pedigree information. To identify efficient markers, that are to be used for genetic purity studies, polymorphism is the basic criterion. RAPD has been found to be an effective and efficient tool to evaluate and reveal genetic polymorphism in several crop species. In present study a total of 80 RAPD primers were screened, out of which 37 gave amplification and only 30 primers showed unambiguous DNA profile. Out of these 30 primers, 22 gave polymorphic banding patterns. It is evident from the result that, 30/80 primers tried (38%) provided unambiguous amplification and out of 30 primers, 22 primers (73%) were found to be polymorphic. Scorable 30 RAPD primers led to amplification of 120 fragments out of which 81 (67.5%) bands were found to be polymorphic. On an average, we got 4 bands per primer and 15 primers (50%) have been found to produce more number of bands than the average value which is encouraging. 8 primers were found to give 100% polymorphisms. Our results are indicative of the efficiency of RAPD primers towards development of molecular profiles.