Fungal Whole Genome de novo Sequencing

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CD Genomics provides fungal whole genome sequencing with the PacBio Sequel system to offer more insights into genetic structure and functions. We are best in the knowledge, practice, and experience.

The Introduction of Fungal Whole Genome de novo Sequencing

Fungi play vital roles in the biosphere and are closely related to human life with important medical and economic value. Fungi are involved in a wide range of activities—some fungi are decomposers, parasites or pathogens of other organisms, and others are beneficial partners in symbiosis with animals, plants or algae. Some fungi affect human health in various ways. Fungi are also the major sources of antibiotics, such as penicillin from the fungus Penicillium.

Obtaining fungal whole genome sequence is the basics of fungi research, and to better understand fungal biodiversity, growth, nutrition, physiology, genetics, metabolism, and ecology. Thus, hundreds of fungal genomes have been sequenced and are publically available today with a significant increase, although these initiatives have typically yielded considerably fragmented genome assemblies, they often lack large contiguous genomic regions. Many important genomic features are contained in intergenic DNA that is often missing in current genome assemblies.

To solve this problem, CD Genomics introduces the PacBio platform. This machine uses single molecule real-time (SMRT) detection technology that achieves real-time sequencing of individual polymerase molecules. SMRT detection is based on the properties of zero-mode waveguides (ZMWs), consisting of DNA polymerases bound to nanophotonic confinement structures. This technology does not require amplification of the genomic DNA, which addresses one of the major problems of second generation sequencing technologies; thus leading to the least degree of bias and longer read lengths (average >15,000 bp, some reads >100,000 bp). With the long reads of SMRT Sequencing on the PacBio System, we can generate complete de novo assemblies for fungal genomes achieve or approximate 0 gaps or N-based errors, contig N50>1 Mb, 99.999% accuracy. Additionally, we provide microbial whole genome sequencing by using next-generation sequencing.

Project Workflow

Our highly experienced expert team executes quality management by following every procedure to ensure comprehensive and accurate results. The general workflow for fungi whole genome sequencing is outlined below.

Sample Requirements:

DNA amount: ≥ 10 μg
DNA Purity: OD260/280 =1.8 ~2.0 without degradation or RNA contamination

Sequencing Strategy:

20 kb Library, ≥ 60X genome coverage depth

Data analysis

  • Genome assembly
  • Gene prediction
  • Gene annotation
  • Comparative species genomes analysis

Analysis pipeline

PacBio, the third-generation sequencers, is highly robust and cost-effective and should be the platform of choice in sequencing fungi genomes, particularly for those that are well-known to be difficult-to-sequence. CD Genomics has extensive experience to offer the fungi whole genome sequencing service. Please contact us for more information and a detailed quote.

1. What are the challenges in fungal sample preparation?

Some samples are prone to degrade, so we suggest a quick library construction and sequencing after DNA extraction. Some fungal strains are difficult to culture, so library construction with a small quantity of DNA or whole-genome amplification is recommended. For samples that are difficult to isolate DNA, you can take methods from literature.

2. Why does the survey of fungal genome is required?

Compared to bacteria, fungal genomes, plasmids, and sample preparation are more complicated. Additionally, only a few fungal genomes have been published. Through genome survey, we can obtain information of genome size, GC content, repetitive sequences, and plasmids, which can pave the way to the substantial sequencing and genome assembly.

Genome Sequence, Assembly and Characterization of Two Metschnikowia fructicola Strains Used as Biocontrol Agents of Postharvest Diseases

Background

The yeast Metschnikowia fructicola was reported as an efficient biological control agent of postharvest diseases of fruits and vegetables, and it is the bases of the commercial formulated product “Shemer.” Researchers assembled the whole genome sequence of two strains of M. fructicola .

Results

Strain 277of M. fructicola was sequenced on PacBio RS II Sequencer (P6-C4 Kit, 20 Kb Library, 24 SMRT cells, target coverage of 20X), yielded a high-quality draft genome consisting of 93 contigs with an N50 of 957,836 bp. The estimated genome size is approximately 26 Mb. Total of 8,629 genes were predicted with MAKER, and 6,262 were successfully annotated with Blast2GO and InterProScan.

M. fructicola AP47 was sequenced used by Illumina MiSeq technology, library insert size 330 bp, paired-end 300 bp. The genome size (_26 Mb) of both M. fructicola strains, as well as the rate of mutation, may suggest that M. fructicola could undergo genomic changes in order to adapt to plant surfaces, tolerate various environmental stresses and survive under restricted nutritional resources.

Reference:

Edoardo P.; et al. Genome Sequence, Assembly and Characterization of Two Metschnikowia fructicola Strains Used as Biocontrol Agents of Postharvest Diseases. Frontiers in Microbiology. 2017, 8:1-15.

For Research Use Only. Not for use in diagnostic procedures.

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