Menu
June 1, 2021  |  

De novo assembly of a complex panicoid grass genome using ultra-long PacBio reads with P6C4 chemistry

Drought is responsible for much of the global losses in crop yields and understanding how plants naturally cope with drought stress is essential for breeding and engineering crops for the changing climate. Resurrection plants desiccate to complete dryness during times of drought, then “come back to life” once water is available making them an excellent model for studying drought tolerance. Understanding the molecular networks governing how resurrection plants handle desiccation will provide targets for crop engineering. Oropetium thomaeum (Oro) is a resurrection plant that also has the smallest known grass genome at 250 Mb compared to Brachypodium distachyon (300 Mb) and rice (350 Mb). Plant genomes, especially grasses, have complex repeat structures such as telomeres, centromeres, and ribosomal gene cassettes, and high heterozygosity, which makes them difficult to assembly using short read next generation sequencing technologies. Ultra-long PacBio reads using the new P6C4 chemistry and the latest 15kb Blue Pippin size-selection protocol to generate 20kb insert libraries that yielded an average read length of 12kb providing ~72X coverage, and 10X coverage with reads over 20kb. The HGAP assembly covers 98% of the genome with a contig N50 of 2.4 Mb, which makes it one of the highest quality and most complete plant genomes assembled to date. Oro has a compact genome structure compared to other grasses with only 16% repeat sequences but has very good collinearity with other grasses. Understanding the genomic mechanisms of extreme desiccation tolerance in resurrection plants like Oro will provide insights for engineering and intelligent breeding of improved food, fuel, and fiber crops.


June 1, 2021  |  

The Wild Vigna

PacBio 2015 User Group Meeting Presentation Slides: Ken Naito of the NIAS Genetic Resource Center presented on whole genome sequencing of the azuki bean (Vigna angularis). Using single molecule real-time (SMRT) sequencing technology, they achieved the best contiguity and coverage among currently assembled legume crops.


June 1, 2021  |  

Characterizing the pan-genome of maize with PacBio SMRT Sequencing

Maize is an amazingly diverse crop. A study in 20051 demonstrated that half of the genome sequence and one-third of the gene content between two inbred lines of maize were not shared. This diversity, which is more than two orders of magnitude larger than the diversity found between humans and chimpanzees, highlights the inability of a single reference genome to represent the full pan-genome of maize and all its variants. Here we present and review several efforts to characterize the complete diversity within maize using the highly accurate long reads of PacBio Single Molecule, Real-Time (SMRT) Sequencing. These methods provide a framework for a pan-genomic approach that can be applied to studies of a wide variety of important crop species.


June 1, 2021  |  

Structural variant detection in crops using low-fold coverage long-read sequencing

Genomics studies have shown that the insertions, deletions, duplications, translocations, inversions, and tandem repeat expansions in the structural variant (SV) size range (>50 bp) contribute to the evolution of traits and often have significant associations with agronomically important phenotypes. However, most SVs are too small to detect with array comparative genomic hybridization and too large to reliably discover with short-read DNA sequencing. While de novo assembly is the most comprehensive way to identify variants in a genome, recent studies in human genomes show that PacBio SMRT Sequencing sensitively detects structural variants at low coverage. Here we present SV characterization in the major crop species Oryza sativa subsp. indica (rice) with low-fold coverage of long reads. In addition, we provide recommendations for sequencing and analysis for the application of this workflow to other important agricultural species.


February 5, 2021  |  

PAG Conference: Dawn of the crop pangenome era

To make improvements to crops like corn, soybeans, and canola, scientists at Corteva are building a compendium of crop genomics resources to provide actionable sequence info for genetic discovery, gene-editing,…


April 21, 2020  |  

The bracteatus pineapple genome and domestication of clonally propagated crops.

Domestication of clonally propagated crops such as pineapple from South America was hypothesized to be a ‘one-step operation’. We sequenced the genome of Ananas comosus var. bracteatus CB5 and assembled 513?Mb into 25 chromosomes with 29,412 genes. Comparison of the genomes of CB5, F153 and MD2 elucidated the genomic basis of fiber production, color formation, sugar accumulation and fruit maturation. We also resequenced 89 Ananas genomes. Cultivars ‘Smooth Cayenne’ and ‘Queen’ exhibited ancient and recent admixture, while ‘Singapore Spanish’ supported a one-step operation of domestication. We identified 25 selective sweeps, including a strong sweep containing a pair of tandemly duplicated bromelain inhibitors. Four candidate genes for self-incompatibility were linked in F153, but were not functional in self-compatible CB5. Our findings support the coexistence of sexual recombination and a one-step operation in the domestication of clonally propagated crops. This work guides the exploration of sexual and asexual domestication trajectories in other clonally propagated crops.


April 21, 2020  |  

Chromosome-level reference genome of X12, a highly virulent race of the soybean cyst nematode Heterodera glycines.

Soybean cyst nematode (SCN, Heterodera glycines) is a major pest of soybean that is spreading across major soybean production regions worldwide. Increased SCN virulence has recently been observed in both the United States and China. However, no study has reported a genome assembly for H. glycines at the chromosome scale. Herein, the first chromosome-level reference genome of X12, an unusual SCN race with high infection ability, is presented. Using whole-genome shotgun (WGS) sequencing, PacBio sequencing, Illumina paired-end sequencing, 10X Genomics linked reads and high-throughput chromatin conformation capture (Hi-C) genome scaffolding techniques, a 141.01-Mb assembled genome was obtained with scaffold and contig N50 sizes of 16.27 Mb and 330.54 kb, respectively. The assembly showed high integrity and quality, with over 90% of Illumina reads mapped to the genome. The assembly quality was evaluated using Core Eukaryotic Genes Mapping Approach (CEGMA) and Benchmarking Universal Single-Copy Orthologs (BUSCO). A total of 11,882 genes were predicted using De novo, Homolog and RNAseq data generated from eggs, second-stage juveniles (J2), third-stage juveniles (J3) and fourth-stage juveniles (J4) of X12, and 79.0% of homologous sequences were annotated in the genome. These high-quality X12 genome data will provide valuable resources for research in a broad range of areas, including fundamental nematode biology, SCN-plant interactions and coevolution, and also contribute to the development of technology for overall SCN management. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.


Talk with an expert

If you have a question, need to check the status of an order, or are interested in purchasing an instrument, we're here to help.