quarTeT is a collection of tools for T2T genome assembly and basic analysis in automatic workflow.
Task include:
1.2.2
1.2.1
1.2.0
1.1.8
1.1.7
1.1.6
1.1.5
1.1.4
1.1.3
1.1.2
1.1.1
1.1.0
1.0.4
1.0.3
1.0.2
1.0.1
1.0.0
quarTeT can be easily accessed on our web server. (Currently its version is lagged.)
quarTeT command-line program is availble for Linux.
All these dependencies can be easily install via conda:
conda create -n quartet --channel conda-forge --channel bioconda Python Minimap2 MUMmer4 trf CD-hit BLAST tidk R R-RIdeogram R-ggplot2 gnuplot
(Recently we discover that using conda to install R will result in blank PNG. However, SVG is correctly generated.)
quarTeT do not require installation.
Just clone this repository, and run python3 {path}/quartet.py
quarTeT: Telomere-to-telomere Toolkit
Usage: python3 quartet.py <module> <parameters>
Modules:
AssemblyMapper | am Assemble draft genome.
GapFiller | gf Fill gaps in draft genome.
TeloExplorer | te Identify telomeres.
CentroMiner | cm Identify centromere candidates.
Use <module> -h for module usage.AssemblyMapper is a reference-guided assemble tool.
A phased contig-level assembly and a close-related reference genome are required as input, both in fasta format.
Note that contigs should be phased.
It's recommended to obtain such an assembly using hifiasm.
you can convert {prefix}.bp.hap1.p_ctg.gfa and {prefix}.bp.hap2.p_ctg.gfa generated by hifiasm to FASTA format as input, respectively.
Usage: python3 quartet.py AssemblyMapper <parameters>
-h, --help show this help message and exit
-r REFERENCE_GENOME (*Required) Reference genome file, FASTA format.
-q CONTIGS (*Required) Phased contigs file, FASTA format.
-c MIN_CONTIG_LENGTH Contigs shorter than INT (bp) will be removed, default: 50000
-l MIN_ALIGNMENT_LENGTH
The min alignment length to be select (bp), default: 10000
-i MIN_ALIGNMENT_IDENTITY
The min alignment identity to be select (%), default: 90
-p PREFIX The prefix used on generated files, default: quarTeT
-t THREADS Use number of threads, default: 1
-a {minimap2,mummer} Specify alignment program (support minimap2 and mummer), default: minimap2
--nofilter Use original sequence input, no filtering.
--keep Keep the unplaced contigs in draft genome
--plot Plot a colinearity graph for draft genome to reference alignments. (will cost more time)
--noplot Skip all ploting.
--overwrite Overwrite existing alignment file instead of reuse.
--minimapoption MINIMAPOPTION
Pass additional parameters to minimap2 program, default: -x asm5
--nucmeroption NUCMEROPTION
Pass additional parameters to nucmer program.
--deltafilteroption DELTAFILTEROPTION
Pass additional parameters to delta-filter program.Output files should be as follow:
{prefix}.draftgenome.fasta | The pseudo-chromosome-level assembly, fasta format.
{prefix}.draftgenome.agp | The structure of this assembly, AGP format.
{prefix}.draftgenome.stat | The statistic of this assembly, including total size and each chromosome's size, GC content, gap count and locations.
{prefix}.draftgenome.png | The figure draws relative length of chromosomes and gap locations for assembly.
{prefix}.contig.mapinfo | The statistic of input contigs, including total mapped and discarded size, and each contig's destination.
{prefix}.contig_map_ref.png | The alignment colinearity graph between contigs and reference genome.
{prefix}.draftgenome_map_ref.png | The alignment colinearity graph between this assembly genome and reference genome. Only available with --plot.GapFiller is a long-reads based gapfilling tool.
A gap-tied genome and corresponding long-reads are required as input, both in fasta format.
If possible, using long-reads assembled and polished contigs instead of reads may improve the quality.
GapFiller support two mode: fill and join. fill means find a segment that can be placed in the gap and link them together. join means find an evidence that the gap edge is overlaped and can be directly merged into one. For now, join mode is unstable and disabled by default. If you want to enable it, be careful and check the result manually.
Usage: python3 quartet.py GapFiller <parameters>
-h, --help show this help message and exit
-d DRAFT_GENOME (*Required) Draft genome file to be filled, FASTA format.
-g GAPCLOSER_CONTIG [GAPCLOSER_CONTIG ...]
(*Required) All contigs files (accept multiple file) used to fill gaps, FASTA format.
-f FLANKING_LEN The flanking seq length of gap used to anchor (bp), default: 5000
-l MIN_ALIGNMENT_LENGTH
The min alignment length to be select (bp), default: 1000
-i MIN_ALIGNMENT_IDENTITY
The min alignment identity to be select (%), default: 40
-m MAX_FILLING_LEN The max sequence length acceptable to fill any gaps, default: 1000000
-p PREFIX The prefix used on generated files, default: quarTeT
-t THREADS Use number of threads, default: 1
--enablejoin Enable join mode to close the gaps. (Unstable)
--joinonly Use only join mode without fill, should be used with --enablejoin.
--overwrite Overwrite existing alignment file instead of reuse.
--minimapoption MINIMAPOPTION
Pass additional parameters to minimap2 program, default: -x asm5
--noplot Skip all ploting.Output files should be as follow:
{prefix}.genome.filled.fasta | The gap-filled genome, fasta format.
{prefix}.genome.filled.modified.agp | The modified chromosome structure, AGP format.
{prefix}.genome.filled.detail | Detailed information for each gap, including gap closed and remains, total filled size and closer's ID, range, etc.
{prefix}.genome.filled.stat | The statistic of filled genome, including total size and each chromosome's size, GC content, gap count and locations.
{prefix}.genome.filled.png | The figure draws relative length of chromosomes and gap locations for assembly.TeloExplorer is a telomere identification tool.
A genome file in fasta format is required as input.
Usage: python3 quartet.py TeloExplorer <parameters>
-h, --help show this help message and exit
-i GENOME (*Required) Genome file to be identified, FASTA format.
-c {plant,animal,other}
Specify clade of this genome. Plant will search TTTAGGG, animal will search TTAGGG, other will use tidk explore's suggestion, default: other
-m MIN_REPEAT_TIMES The min repeat times to be reported, default: 100
-p PREFIX The prefix used on generated files, default: quarTeT
--noplot Skip all ploting.Output files should be as follow:
{prefix}.telo.info | The statistic of telomere, including monomer, repeat times on both end of each chromosome.
{prefix}.telo.png | The figure draws telomere location, alongside relative length of chromosomes and gap locations for assembly.CentroMiner is a centromere prediction tool.
A genome file in fasta format is required as input.
Optionally, an additional input of TE annotation (or just LTR annotation) and gene annotation in gff3 format can improve the performance.
It's recommended to obtain TE annotation using EDTA.
{genome file}.mod.EDTA.TEanno.gff3 generated by EDTA can directly feed CentroMiner, unless you have sequence ID longer than 15 characters.
Note that the sequence ID in first column should be consistent with in genome. Some tools may change sequence ID if ID is too long.
The sequence ontology in the third column should include LTR or long_terminal_repeat (EDTA default), or have LTR in the eighth column (RepeatMasker default) to be recognized.
Usage: python3 quartet.py CentroMiner <parameters>
-h, --help show this help message and exit
-i GENOME_FASTA (*Required) Genome file, FASTA format.
--TE TE TE annotation file, gff3 format.
--gene GENE gene annotation file, gff3 format.
-n MIN_PERIOD Min period to be consider as centromere repeat monomer. Default: 100
-m MAX_PERIOD Max period to be consider as centromere repeat monomer. Default: 200
-s CLUSTER_IDENTITY Min identity between TR monomers to be clustered (Cannot be smaller than 0.8). Default: 0.8
-d CLUSTER_MAX_DELTA Max period delta for TR monomers in a cluster. Default: 10
-e EVALUE E-value threholds in blast. Default: 0.00001
-g MAX_GAP Max allowed gap size between two tandem repeats to be considered as in one tandem repeat region. Default: 50000
-l MIN_LENGTH Min size of tandem repeat region to be selected as candidate. Default: 100000
-t THREADS Limit number of using threads, default: 1
-p PREFIX Prefix used by generated files. Default: quarTeT
--trf [TRF_PARAMETER ...]
Change TRF parameters: <match> <mismatch> <delta> <PM> <PI> <minscore> Default: 2 7 7 80 10 50
-r MAX_TR_LENGTH Maximum TR length (in millions) expected for trf. Default: 3
--overwrite Overwrite existing trf dat file instead of reuse.
--noplot Skip all ploting.Output files should be as follow:
Candidates/ | The folder of all centromere candidates data on each chromosome. Check the pdf line chart and vote a best candidate.
TandemRepeat/ | The folder of all tandem repeat monomers identified by trf and cluster result on each chromosome.Note that manual selection is highly recommended.
In some species, best candidate may unexpectedly fall in near telomere region or other tandem repeat region.
You can combine the gene, TR and TE line chart with Hi-C contact heatmap, pairwise colinearity etc. to better vote your candidates. Mostly the centromere region is riched in TR, surrounded by rich TE, and low in gene content, but there are also exceptions.
Yunzhi Lin, Chen Ye, Xingzhu Li, Qinyao Chen, Ying Wu, Feng Zhang, Rui Pan, Sijia Zhang, Shuxia Chen, Xu Wang, Shuo Cao, Yingzhen Wang, Yi Yue, Yongsheng Liu, Junyang Yue. quarTeT: a telomere-to-telomere toolkit for gap-free genome assembly and centromeric repeat identification. Horticulture Research 2023;10:uhad127, https://doi.org/10.1093/hr/uhad127