---
title: "Update of a treestructure object with new sequences"
author: "Vinicius Franceschi and Fabricia F Nascimento"
date: "`r Sys.Date()`"
output: 
  bookdown::html_vignette2:
  #rmarkdown::html_vignette
  #bookdown::pdf_book:
    toc: TRUE
pkgdown:
  as_is: true
fontsize: 12pt
vignette: >
  %\VignetteIndexEntry{Update of a treestructure object with new sequences}
  %\VignetteEngine{knitr::rmarkdown}
  %\VignetteEncoding{UTF-8}
---

```{r setup, include = FALSE}
knitr::opts_chunk$set(
  collapse = TRUE,
  comment = "#>",
  fig.width = 7,
  fig.height = 11,
  warning = FALSE, 
  message = FALSE
)
```

# Introduction

In this tutorial, we will exemplify how to update a previous `treestructure` 
object with new sequences using a down sampled version of the [Ebola dated tree](https://github.com/ebov/space-time/blob/master/Data/Makona_1610_cds_ig.GLM.MCC.tree), which is publicly available.



First, we will load all the R packages that we will use in this tutorial.
```{r message=FALSE}
library(ape)
library(treestructure)
library(phangorn)
```

Now we can read the down-sampled time-tree for Ebola. In this pruned tree, we
have 1,310 tips.

```{r}
pruned_tree <- readRDS( system.file('Ebola_down_sampled_tree.rds',
                                    package='treestructure') )

```


## Assign clusters using node support

Now we will assign clusters using the posterior probability node support to the 
Ebola  down-sampled phylogenetic tree:

```{r eval=FALSE}
trestruct_res <- trestruct(pruned_tree, 
                           minCladeSize = 30, 
                           nodeSupportValues = TRUE, 
                           nodeSupportThreshold = 95,
                           level = 0.01)
```

In the above example, the `trestruct` function took 32 seconds to run on a macOS M2. 
Here, we will load the results.

```{r}
trestruct_res <- readRDS( system.file('downsampled_tree_struc.rds',
                                      package='treestructure') )

plot(trestruct_res, use_ggtree = T) + ggtree::geom_tippoint()
```

The `treestructure` analyses resulted in 4 clusters.



## Update a previous treestrucuture object with new sequences 

To update the previous `treestructure` object with new sequences, we will now use
the maximum likelihood [Ebola tree](https://github.com/ebov/space-time/blob/master/Data/Makona_1610_genomes_2016-06-23.ml.tree).

Note that this new tree must be rooted, but does not need to be time-scaled or 
binary.

```{r}
#Note that this tree has more sequences than the previous tree used in this
#tutorial.
new_tree <- ape::read.nexus( system.file('Makona_1610_genomes_2016-06-23.ml.tree',
                                         package='treestructure') )

#now we can root the tree using mid-point rooting for illustration
ml_rooted_tree <- phangorn::midpoint(new_tree)

#now we need to remove the quotes from the tip names (to avoid an error with 
#treestructure function)
ml_rooted_tree$tip.label <- unlist(lapply(ml_rooted_tree$tip.label, 
                                          function (x) gsub("'", "", x)))

```

And without the need to re-estimate a timetree or re-run `trestruct` from scratch, 
we are now able to add the new sequences to the existing `treestructure` object:

```{r}
trestruct_add_tips <- addtips(trst = trestruct_res, tre = ml_rooted_tree)

plot(trestruct_add_tips, use_ggtree = T) + ggtree::geom_tippoint()
```


If you would like to compare the sequence names that comprise each cluster in
each tree, you can do:

```{r}

#compare sequences in cluster 1 from trestruct_res object and the 
#trestruct_add_tips object

tree1_cluster1 <- trestruct_res$clusterSets$`1`
tree2_cluster1 <- trestruct_add_tips$clusterSets$`1`

length(tree1_cluster1)
length(tree2_cluster1)

```

Note that the length of tree1_cluster1 and tree2_cluster1 is different.
That is because we _added_ tips from the ML tree, _ml_rooted_tree_, to the 
`treestructure` object, _trestruct_res_.

You can also see that all elements in tree1_cluster1 is contained in tree2_cluster1

```{r}

sum(tree1_cluster1 %in% tree2_cluster1)

```
