ETE构建、绘制进化树

ETE能做什么

A Python framework for construction, analysis and visualization of trees.

安装和使用

• conda安装 # Install Minconda (you can ignore this step if you already have Anaconda/Miniconda) wget http://repo.continuum.io/miniconda/Miniconda-latest-Linux-x86_64.sh -O Miniconda-latest-Linux-x86_64.sh bash Miniconda-latest-Linux-x86_64.sh -b -p ~/anaconda/ export PATH=~/anaconda/bin:$PATH; # Install ETE conda install -c etetoolkit ete3 ete3_external_apps # Check installation ete3 version ete3 build check
• github源码安装 wget https://github.com/etetoolkit/ete/archive/master.zip -O ete3.20160719.zip unzip ete3.20160719.zip python setup.py install yum install python-six.noarch ete3 upgrade-external-tools

ETE运行

• 输入序列
  • 输入序列为标准的fasta格式文件，名字可以为任意形式
  • 如果需要在序列名字中区分物种信息，fasta序列名需满足SpeciesCode_SequenceName例如HUMAN_p53 = HUMAN, p53。 可以通过参数--spname-delimiter指定使用其它字符作为分隔符。
• ete预先定义了多个流程用以完成从原始fasta序列到后续进化树生成的各个步骤。 运行以下命令可以列出系统自带的流程及其解释 ete3 build workflows genetree
• 使用既定流程最简单运行 -w指定所用的流程，-a指定输入序列，--tools-dir指定安装的外部程序的路径 ete3 build -w standard_fasttree -a diTPS.prot.fa -o standard_fasttree --tools-dir /root/.etetoolkit/ext_apps-latest/
• 自己定制流程
  • 获取可以定制的各部分命令 ete3 build apps
  • 选择预定义好的模块，如tree builders: phyml_default_bootstrap, aligners: mafft_einsi, model testers: pmodeltest_full_slow, alg cleaners:trimal_gappyout.
  • 流程定制模板: 顺序为aligner-trimmer-model_tester-builder
    • totally 4 parts included as stated above, multiple sequence alignment, trimming MSA results, select best model, use appropriate softwares to build tree.
    • - represents command separator
    • none represents skipping related operations
  • 基于我们的选择定制的流程mafft_einsi-trimal_gappyout-pmodeltest_full_slow-phyml_default_bootstrap ete3 build -w mafft_einsi-trimal_gappyout-pmodeltest_full_slow-phyml_default_bootstrap -a diTPS.prot.fa -o custom_phymltree
  • -w可以接受多个流程(空格分开)，进而得到不同的比对工具、处理方式和建树工具 输出的多个结果，可以通过ete3 compare比较这些结果的吻合度, 比如Robinson-Foulds距离等。 ete3 compare -r newtree1.nwq -t "tree2.nw tree3.nw tree4.nw" --unrooted # Tree file can be got using find find custom_phymltree -name *.nw
  • 定制不同的分析模块
    • 查看已有分析模块的定义 ete3 build show phyml_default [phyml_default] _desc = Phyml tree using +G+I+F, 4 classes and aLRT branch supports. Default models JTT/GTR _app = phyml _aa_model = JTT -nt_model = GTR --pinv = e --alpha = e --nclasses = 4 -o = tlr -f = m --bootstrap = -2
    • 修改部分定义获得新的模块 ete3 build show phyml_default >customized.config ## 修改后的customized.config [phyml_bootstrap_100] _desc = Phyml tree using +G+I+F, 4 classes and aLRT branch supports. Default models JTT/GTR _app = phyml _aa_model = JTT -nt_model = GTR --pinv = e --alpha = e --nclasses = 4 -o = tlr -f = m --bootstrap = 100 [trimal_auto] _desc = trimal alignment cleaning using auto algorithm _app = trimal -automated1 = ## 使用新定义的模块 ete3 build -a diTPS.prot.fa --clearall -o phyml_bootstrap_100 -w mafft_einsi-trimal_auto-none-phyml_bootstrap_100 -c customized.cfg --cpu 5
• 氨基酸比对指导核苷酸比对的进化树构建 (要求氨基酸序列与核苷酸序列名字一一对应，核苷酸序列可以含有终止密码子，最终获得的核苷酸比对序列存储在*.used_alg.fa文件中。) ete3 build -a diTPS.prot.fa -n diTPS.nucl.fa -o aa2nt -w standard_fasttree --clearall --nt-switch-threshold 0.9 -C 20
• 使用预先比对好的序列, 使用none代替aligner ete3 build -a diTPS.prot.aln.fa -w none-none-none-fasttree -o manual_alg --clearall
• 设置树的根节点 from ete3 import Tree tree = Tree('tree.nw') root = 'one_node_name' tree.set_outgroup(root) #use mid-point as root mid = tree.get_midpoint_outgroup() tree.set_outgroup(mid) tree.write('tree.rooted.nw') tree.render('tree.rooted.pdf')

问题解决

• ETE: cannot connect to X server 如果程序运行出现错误ETE: cannot connect to X server则安装Xvfb， 并运行 xvfb-run ete3取代ete3, 后面的代码不变。 yum install xorg-x11-server-Xvfb.x86_64 xvfb-run ete3 build -w standard_fasttree -a diTPS.prot.fa -o standard_fasttree
• ETE: cannot connect to X server (solve in python script or jupyter ref)
  • Install xvfbwrapper using pip install xvfbwrapper

  # Add the following 4 lines at the beginning of python code # or the first cell in Jupyter from xvfbwrapper import Xvfb vdisplay = Xvfb() vdisplay.start() # launch stuff inside virtual display here # other python codes here # Add this line at the end of python code # or the last cell in Jupyter vdisplay.stop()

• External applications directory are not found 指定ETE使用的工具的安装路径；一般发生在普通用户使用根用户编译的ETE时。 --tools-dir /root/.etetoolkit/ext_apps-latest/

Tree annotation

# A virtual X-server XVFB is used in case you do not have X-server
from xvfbwrapper import Xvfb

vdisplay = Xvfb()
vdisplay.start()

# launch stuff inside virtual display here

#vdisplay.stop()

from ete3 import Tree, faces, TreeStyle, NodeStyle
from ete3 import ClusterTree, RectFace, AttrFace, ProfileFace, TextFace
from ete3.treeview.faces import add_face_to_node
import pandas as pd
import numpy as np
import colorsys

The most simple way of showing a tree.

t = Tree()
t.populate(7,names_library=['A','B','C','D','E','F','G'])
## %%liline is used for showing plots in ipythonnotebook.
## t.render(file_name="tree.pdf") # will save tree into pdf file
t.render(file_name="%%inline")

Get the randomly generated tree in newick format and save to a string variable which can be read using Tree() function.

t_str = t.write(outfile=None, format=0)
t_str

'(((B:1,A:1)1:1,(G:1,F:1)1:1)1:1,(E:1,(D:1,C:1)1:1)1:1);'

t = Tree(t_str)
ts = TreeStyle()
ts.show_leaf_name = True
ts.show_branch_length = True
ts.show_branch_support = True
t.render(file_name="%%inline", tree_style=ts)

Get the randomly generated tree in newick format and save to file which can also be read using Tree() function.

t.write(outfile="tree.nw", format=0)
t = Tree("tree.nw")
ts.mode = "c"
ts.arc_start = -180 # 0 degrees = 3 o'clock
ts.arc_span = 180
t.render(file_name="%%inline", w=500, tree_style=ts)

设置根节点、叶节点和中间节点的属性

ts = TreeStyle()
ts.show_leaf_name = True
ts.show_branch_length = True
ts.show_branch_support = True

# Draws nodes as small red spheres of diameter equal to 10 pixels
for n in t.traverse():  # Traverse each node and set attribute for each type of nodes
    if n.is_leaf(): # Decide if leaf node
        nstyle = NodeStyle()
        nstyle["shape"] = "sphere"
        nstyle["size"] = 10
        nstyle["fgcolor"] = "darkred"
        n.set_style(nstyle)
    else:
        nstyle = NodeStyle()
        nstyle["shape"] = "square"
        nstyle["size"] = 15
        nstyle["fgcolor"] = "orange"
        n.set_style(nstyle)        

t.img_style["size"] = 30
t.img_style["fgcolor"] = "blue"

t.render(file_name="%%inline", w=500, tree_style=ts)

修改节点的名字

t = Tree(t_str)

nameMap = {'A': 'American', 'B': 'Britain', 'C':'China', 
           'D':'Dutch', 'E':'Egypt','F':'France','G':'German'}

ts = TreeStyle()
ts.show_leaf_name = False
ts.show_branch_length = True
ts.show_branch_support = True

# Creates my own layout function. I will use all previously created
# faces and will set different node styles depending on the type of
# node.
def mylayout(node):   
    # If node is a leaf, add the nodes name and a its scientific
    # name
    if node.is_leaf():
        # We can also create faces on the fly
        newName = nameMap.get(node.name, node.name)
        newNameFace = faces.TextFace(newName)
        faces.add_face_to_node(newNameFace, node, column=1, aligned=True)

        # Sets the style of leaf nodes
        node.img_style["size"] = 12
        node.img_style["shape"] = "sphere"
        node.img_style["fgcolor"] = "blue"
    #If node is an internal node
    else:
        # Sets the style of internal nodes
        node.img_style["size"] = 1
        node.img_style["shape"] = "circle"
        node.img_style["fgcolor"] = "darkred"

ts.layout_fn = mylayout

t.img_style["size"] = 30
t.img_style["fgcolor"] = "black"

t.render(file_name="%%inline", w=600, tree_style=ts)

不同的节点设置不同的背景色，树枝的颜色

t = Tree(t_str)

nameMap = {'A': 'American', 'B': 'Britain', 'C':'China', 
           'D':'Dutch', 'E':'Egypt','F':'France','G':'German'}

colorMap = { 'American': '#ACFFFF',
             'Britain': '#ACACFF',
             'China': '#ACACAC',
             'Dutch': '#59ACAC',
             'Egypt': '#5959AC',
             'France': '#595959',
             'German': '#065959'}

ts = TreeStyle()
ts.show_leaf_name = False
ts.show_branch_length = True
ts.show_branch_support = True

# Creates my own layout function. I will use all previously created
# faces and will set different node styles depending on the type of
# node.
def mylayout(node):   
    #Change branch color
    node.img_style["hz_line_color"] = 'orange'  # change horizontal branch color 
    node.img_style["vt_line_color"] = 'red' # Change vertical branch color

    # If node is a leaf, add the nodes name and a its scientific
    # name
    if node.is_leaf():
        # We can also create faces on the fly
        newName = nameMap.get(node.name, node.name)
        newNameFace = faces.TextFace(newName)
        faces.add_face_to_node(newNameFace, node, column=1, aligned=True)

        # Sets the style of leaf nodes
        node.img_style["size"] = 12
        node.img_style["shape"] = "sphere"
        node.img_style["fgcolor"] = "blue"
        node.img_style["bgcolor"] = colorMap[newName]

        node.img_style["hz_line_color"] = 'blue'  # change branch color 
    #If node is an internal node
    else:
        # Sets the style of internal nodes
        node.img_style["size"] = 1
        node.img_style["shape"] = "circle"
        node.img_style["fgcolor"] = "darkred"

ts.layout_fn = mylayout

t.img_style["size"] = 30
t.img_style["fgcolor"] = "black"

t.render(file_name="%%inline",tree_style=ts)

树+热图（自定义颜色+列名字）

自定义热图函数，同时该函数也支持替换或新增节点的名字。

nameFace = AttrFace("name", fsize=12) #Set leaf node attribute

def setup_heatmap(tree, tree_style, header, center_value=0.0, nameMap ={}, nameLabel = '',
                  color_up=0.7, color_down=0.2, color_center="white"):
    DEFAULT_COLOR_SATURATION = 0.5
    BASE_LIGHTNESS = 0.7
    def gradient_color(value, max_value, saturation=0.5, hue=0.1):    
        def rgb2hex(rgb):
            return '#%02x%02x%02x' % rgb
        def hls2hex(h, l, s):
            return rgb2hex( tuple(map(lambda x: int(x*255), 
                          colorsys.hls_to_rgb(h, l, s))))

        lightness = 1 - (value * BASE_LIGHTNESS) / max_value
        return hls2hex(hue, lightness, DEFAULT_COLOR_SATURATION)

    # Calculate max gradient value from the ClusterTree matrix
    maxv = abs(center_value - tree.arraytable._matrix_max)
    minv = abs(center_value - tree.arraytable._matrix_min)
    if center_value <= tree.arraytable._matrix_min:
        MAX_VALUE = minv + maxv
    else:
        MAX_VALUE = max(maxv, minv)

    # Add heatmap colors to tree
    cols_add_before_heat = 0
    if nameMap:
        cols_add_before_heat = 1
    for lf in tree:
        if nameMap:
            longNameFace = faces.TextFace(nameMap.get(lf.name, lf.name))
            lf.add_face(longNameFace, column=0, position="aligned")

        for i, value in enumerate(getattr(lf, "profile", [])):
            if value > center_value:
                color = gradient_color(abs(center_value - value), MAX_VALUE, 
                                       hue=color_up)
            elif value < center_value:
                color = gradient_color(abs(center_value - value), MAX_VALUE, 
                                       hue=color_down)
            else:
                color = color_center
            lf.add_face(RectFace(20, 20, color, color), position="aligned", 
                        column=i+cols_add_before_heat)
            # Uncomment to add numeric values to the matrix
            #lf.add_face(TextFace("%0.2f "%value, fsize=5), position="aligned", column=i)
        lf.add_face(nameFace, column=i+cols_add_before_heat+1, position="aligned")

    if nameMap and nameLabel:
        nameF = TextFace(nameLabel, fsize=7)
        #nameF.rotation = -90
        tree_style.aligned_header.add_face(nameF, column=0)
    # Add header 
    for i, name in enumerate(header):
        nameF = TextFace(name, fsize=7)
        nameF.rotation = -90
        tree_style.aligned_header.add_face(nameF, column=i+cols_add_before_heat)
 #-------------END setup_heatmap----------------------------------------------

读入矩阵 （可把文后的测试矩阵存储到文件中读入）

矩阵需满足三个条件：

• 矩阵为TAB键分割，第一行是每列的名字
• 矩阵每一行第一列为行名字，与树的节点对应
• 矩阵可以存储与一个文件中，也可以是如下的字符串

data = pd.read_table("matrix", header=0, index_col=0)
data.index.name = "#Names"  #修改第一行的名字使其符合ETE的要求
data_mat = data.to_csv(None, sep="t", float_format="%.2f")
header = list(data.columns.values)  #获取列的名字用于标记

data

data_mat

'#Namestcol1tcol2tcol3tcol4tcol5tcol6tcol7nAt-1.23t-0.81t1.79t0.78t-0.42t-0.69t0.58nBt-1.76t-0.94t1.16t0.36t0.41t-0.35t1.12nCt-2.19t0.13t0.65t-0.51t0.52t1.04t0.36nDt-1.22t-0.98t0.79t-0.76t-0.29t1.54t0.93nEt-1.47t-0.83t0.85t0.07t-0.81t1.53t0.65nFt-1.04t-1.11t0.87t-0.14t-0.80t1.74t0.48nGt-1.57t-1.17t1.29t0.23t-0.20t1.17t0.26n'

header

['col1', 'col2', 'col3', 'col4', 'col5', 'col6', 'col7']

调用函数绘制热图

t = ClusterTree(t_str, data_mat)

ts = TreeStyle()
ts.show_leaf_name = False
ts.show_branch_length = True
ts.show_branch_support = True

setup_heatmap(t, ts, header, center_value=0, color_up=0.9, color_down=0.3, color_center="white")

t.render(file_name="%%inline", tree_style=ts)

绘制热图时修改Layout

def mylayout_only(node):   
    #Change branch color
    node.img_style["hz_line_color"] = 'orange'  # change horizontal branch color 
    node.img_style["vt_line_color"] = 'red' # Change vertical branch color

    # If node is a leaf, add the nodes name and a its scientific
    # name
    if node.is_leaf():
        newName = nameMap.get(node.name)
        node.img_style["size"] = 12
        node.img_style["shape"] = "sphere"
        node.img_style["fgcolor"] = "blue"
        node.img_style["bgcolor"] = colorMap[newName]        
        node.img_style["hz_line_color"] = 'blue'  # change branch color 
    #If node is an internal node
    else:
        # Sets the style of internal nodes
        node.img_style["size"] = 1
        node.img_style["shape"] = "circle"
        node.img_style["fgcolor"] = "darkred"

t = ClusterTree(t_str, data_mat)

ts = TreeStyle()
ts.show_leaf_name = False
ts.show_branch_length = True
ts.show_branch_support = True
ts.layout_fn = mylayout_only

setup_heatmap(t, ts, header, center_value=0, color_up=0.9, color_down=0.3, 
              color_center="white", nameMap=nameMap, nameLabel="Full")

t.render(file_name="%%inline", tree_style=ts)

测试矩阵

## 矩阵为TAB键分割
## 矩阵每一行第一列为行名字，与树的节点对应
## 矩阵可以存储于一个文件中，也可以是如下的字符串
matrix = """
#Namestcol1tcol2tcol3tcol4tcol5tcol6tcol7
At-1.23t-0.81t1.79t0.78t-0.42t-0.69t0.58
Bt-1.76t-0.94t1.16t0.36t0.41t-0.35t1.12
Ct-2.19t0.13t0.65t-0.51t0.52t1.04t0.36
Dt-1.22t-0.98t0.79t-0.76t-0.29t1.54t0.93
Et-1.47t-0.83t0.85t0.07t-0.81t1.53t0.65
Ft-1.04t-1.11t0.87t-0.14t-0.80t1.74t0.48
Gt-1.57t-1.17t1.29t0.23t-0.20t1.17t0.26
"""

产生颜色的辅助函数，给定一个列表，这个函数会自动返回一个字典包含每个字段对应的颜色。

def hex2rgb(hexcolor):
    return [(hexcolor>>16) & 0xff, (hexcolor>>8) & 0xff, hexcolor & 0xff]

def rgb2hex(rgbcolor):
    r, g, b = rgbcolor
    rgb = hex((r << 16) + (g << 8) +b)[2:].upper()
    zero = '0'* (6-len(rgb))
    return '#'+zero+rgb
#----------------------------------
def generateColor(labelL):
    labelL = list(set(labelL))
    labelL.sort()
    colorD = {}

    r = 255
    g = 255
    b = 255
    len_label = int(len(labelL) / 3 + 1)
    step = int(250 / len_label)

    cnt = 1
    for labels in labelL:
        if cnt % 3 == 1:
            r = r - step 
        elif cnt % 3 == 2:
            g = g -step
        else:
            b = b - step
        cnt += 1
        color = rgb2hex([r, g, b])
        colorD[labels] = color
    return colorD

clan_colorD = generateColor(nameMap.values())
clan_colorD

{'American': '#ACFFFF',
 'Britain': '#ACACFF',
 'China': '#ACACAC',
 'Dutch': '#59ACAC',
 'Egypt': '#5959AC',
 'France': '#595959',
 'German': '#065959'}

带有Support value的Newick树，供测试不同的属性使用

nw = """
(((Dre:0.008339,Dme:0.300613)1.000000:0.596401,
(Cfa:0.640858,Hsa:0.753230)1.000000:0.182035)1.000000:0.106234,
((Dre:0.271621,Cfa:0.046042)1.000000:0.953250,
(Hsa:0.061813,Mms:0.110769)1.000000:0.204419)1.000000:0.973467);

关闭Virtual X-server

vdisplay.stop()

Ipython notebook for easy usage

https://github.com/Tong-Chen/notebook/blob/master/ETE.ipynb

Reference

• ETE tutorial http://etetoolkit.org/docs/latest/tutorial/index.html
• ETE googlegroup https://groups.google.com/d/topic/etetoolkit/pXr4B71Ozt0

Blog link

原文链接 http://blog.genesino.com/2016/07/ete/