Bamboo Genome Sequencing Project

Bamboo Genome Sequencing Project (BGSP) is focused on the sequencing of 11 bamboo genomes. This repository is currently being developed and houses workflows and scripts that are utilized for the identification and comparative analysis of bamboo subgenomes.

Citation

Peng-Fei Ma, Yun-Long Liu, Cen Guo, Guihua Jin, Zhen-Hua Guo, Ling Mao, Yi-Zhou Yang, Liang-Zhong Niu, Yu-Jiao Wang, Lynn G. Clark, Elizabeth A. Kellogg, Zu-Chang Xu, Xia-Ying Ye, Jing-Xia Liu, Meng-Yuan Zhou, Yan Luo, Yang Yang, Douglas E. Soltis, Jeffrey L. Bennetzen, Pamela S. Soltis, De-Zhu Li. (2024). Genome assemblies of 11 bamboo species highlight diversification induced by dynamic subgenome dominance. Nature Genetics. 10.1038/s41588-024-01683-0
Yun-Long Liu, Shu-Yang Gao, Guihua Jin, Meng-yuan Zhou, Qijuan Gao, Cen Guo, Yi-Zhou Yang, Liang-Zhong Niu, Enhua Xia, Zhen-Hua Guo, Peng-Fei Ma, De-Zhu Li. BambooBase：A comprehensive database of bamboo omics and systematics. Molecular Plant. 10.1016/j.molp.2024.02.017.

Getting Started

Reconstruction of "perfect-copy" syntenic gene data set

Dependencies: JCVI v1.1.17, bedtools.
Input files: sp.cds, sp.bed, sp:SpeciesID
Script: Perfect-copy_syntenic_gene_identification.sh
Usage: bash Perfect-copy_syntenic_gene_identification.sh Alu 2 1

sp=$1
a=$2
b=$3
python -m jcvi.compara.catalog ortholog --quota=$a:$b $sp Osa --no_strip_names
python -m jcvi.compara.synteny mcscan --iter=1 $sp.bed $sp.Osa.anchors -o $sp.Osa.blocks
cat $sp.Osa.blocks|sort -k2|groupBy -g 2 -c 1,1 -o count,distinct|awk -v num=$a '{ if( $2==num) print $0}'|cut -f 1,3 |sed -e 's/,/\t/g' >$sp.block

After running for all species, use the following command to reconstruct "perfect-copy" syntenic gene data set for all species

python -m jcvi.formats.base join $sp.block >perfect-copy_syntenic_gene.txt

Reconstruction of single "perfect-copy" gene matrix and concatenated supermatrix for each syntenic block

Dependencies:
Input files: cds.fasta, pep.fasta, gene#.id
Script: syntenic_gene_matrix.nf
Usage: nextflow syntenic_gene_matrix.nf

Time tree estimation

Input files: 11bg.trees, 11bg_alignment.txt, 11bg_mcmctree.ctl
Dependencies: PAML v4.9
Usage: mcmctree 11bg_mcmctree.ctl

PhyloNet analyses

Input files: InferNetwork_MPL_430gene_r2.nex
Dependencies: PhyloNet_3.8.0
Usage: java -Xmx70g -jar $PHYLONET PhyloNet_3.8.0.jar InferNetwork_MPL_430gene_r2.nex

Gene tree topologies calculation

Dependencies: newick_utils
Input files: 11bg_430gene.tre
Script: Tree_topologies_calculation.sh
Usage: bash Tree_topologies_calculation.sh

nw_reroot 11bg_430gene.tre osa > 11bg_430gene_reroot.tre
nw_condense 11bg_430gene_reroot.tre > 11bg_430gene_reroot_condense.tre
nw_topology 11bg_430gene_reroot_condense.tre > 11bg_430gene_reroot_condense_topology.tre
nw_order 11bg_430gene_reroot_condense_topology.tre |sort|groupBy -g 1 -c 1 -o count

Subgenome dominance of hexaploid

Dependencies: R v4.1.2
Normalization of relative expression levels of the A, B, and C subgenomes

awk '{if(($2+$3+$4) >=0.5)print $1, $2/($2+$3+$4), $3/($2+$3+$4), $4/($2+$3+$4)}' ABC_111_TPM.csv >TPM_logTPM_normolized.txt

Definition of homoeologous expression bias categories for Normalization of relative expression (with M. baccifera as example).

#R version 4.1.2
data=read.csv("TPM_logTPM_normolized.txt",header=F, sep="\t",row.names = 1)
dist=as.matrix(dist(data[,1:3],method = "euclidean"))[,1:7]
write.table(t(dist),file="dist", quote=F)

for i in {9..3518};do awk '{print $"'${i}'", $1}' dist  |sort -n |head -1 ;done > dist_category.csv

Plot the ternary diagrams using the R package ggtern

#R version 4.1.2
library(ggplot2)
library(ggtern)
data=read.table("dist_category.csv",header=T,check.names=F,sep=",",quote="",dec=".")
ggtern(data=data,aes(x=A,y=B,z=C,color=Group ) ) + theme_rgbw() + geom_point(aes(fill=Group),size=1,shape=21)
ggsave("Mhu_combined_Ternary_Plot1.pdf")

Screening for positively selected genes

To address the challenge of multiple gene copies in polyploids on identifying PSGs, we used a subgenome-based approach. Using rice and two HB species as outgroups, we assembled nine datasets at the subgenome level:

(I) PWB-subA, (Osa, ((Ola, Rgu),(MbaA, (DsiA, BamA)) #1)); 
(II) PWB-subB, (Osa, ((Ola, Rgu), (MbaB, (DsiB,BamB)) #1)); 
(III) PWB-subC, (Osa, ((Ola, Rgu), (MbaC, (DsiC, BamC)) #1)); 
(IV) NWB_subB, (Osa, ((Ola, Rgu), (RhiB, (GanB, OglB)) #1)); 
(V) NWB_subC, (Osa,((Ola, Rgu), (RraC, (OglC, GanC)) #1)); 
(VI) TWB_subC, (Osa, ((Ola, Rgu), (AluC,(PedC, HcaC)) #1)); 
(VII) TWB_subD, (Osa, ((Ola, Rgu), (AluD, (PedD, HcaD)) #1));
(VIII) the subB lineage, (Osa, ((((DsiB, BamB), MbaB), ((GanB, OglB), RraB)) #1, (Rgu,Ola)));
(IX) the subC lineage, (Osa, ((Ola, Rgu), (((HcaC, PedC), AluC), ((MbaC, (DsiC, BamC)), (RraC, (OglC, GanC)))) #1)).

caculate ka/ks

Dependencies: callCodeml2

python3 single_anligment singall.tre
awk '{if($7<=0.05)print $1}' result.tsv > PWB_subA_PSG_OG

The P value was determined by a Chi-square test with a cutoff of <0.05 for positive selection

New genes identification

Dependencies: OrthoFinder v2.5.272

bash new_gene.sh

OrthoFinder was used to cluster gene families, and we classified genes into different age groups with the oldest gene representing the age of the gene

License

This project is licensed under the MIT License.

yunlongliukm/BGSP