Analyse scripts for "Global patterns of fish genetic diversity increase with “current” temperature"
================================================
2017
This folder contains all the scripts to reproduce all the analysis

# Table of contents

1. [Introduction](#1-introduction)
2. [Installation](#2-installation)
  1. [Prerequisite](#21-prerequisite)
  2. [Data Files](#22-data-files)
  3. [Set up](#23-set-up)
3. [Scripts Code Source](#3-scripts-code-source)
4. [Reporting bugs](#4-reporting-bugs)
5. [Running the pipeline](#5-running-the-pipeline)
  1. [Filter raw data](#51-filter-raw-data)
  2. [Georeferenced sequences alignments by species](#52-data-files)
  3. [Species sequence pairwise comparison](#53-species-sequence-pairwise-comparison)
  4. [Genetic Diversity calculation](#54-genetic-diversity-calculation)
  5. [Statistical analysis](#55-statistical-analysis)

# 1. Introduction

blablabla

# 2. Installation


## 2.1 Prerequisite
You must install the following softwares and packages :

- [JULIA Version 0.5.2](https://julialang.org/)
- [R Version 3.2.3](https://cran.r-project.org/)
    - [R-package]ggplot2 `install.packages("ggplot2")`
    - [R-package]rgeos `install.packages("rgeos")`
    - [R-package]rgdal `install.packages("rgdal")` it may require to install `libgdal-dev`
    - [R-package]nodiv `install.packages("nodiv")`
    - [R-package]raster `install.packages("raster")`
    - [R-package]lme4 `install.packages("lme4")`
    - [R-package]sp `install.packages("sp")`
    - [R-package]sjPlot `install.packages("sjPlot")`
    - [R-package]FactoMineR `install.packages("FactoMineR")`
    - [R-package]factoextra `install.packages("factoextra")`
    - [R-package]spdep `install.packages("spdep")`
    - [R-package]countrycode `install.packages("countrycode")`
- [Python Version 2.7.12](https://www.python.org/)
- [MUSCLE Version 3.8.31](https://www.drive5.com/muscle/)

## 2.2 Data Files
The included data files are :

* `02-raw_data/seqbold_data.txt`                            : Georeferenced sequences of individuals from the supergroup "actinopterygii" have been downloaded from [http://www.boldsystems.org/index.php/Public_SearchTerms?taxon=&searchMenu=records&query=actinopterygii]
* `01-infos/grid_equalarea200km`                            : Shapefile of worldmap equal area projection epsg:4326 with nested equal area grids (cell sizes of 200km)
* `01-infos/ne_110m_land`                                   : Shapefile of worldcoast from (http://www.naturalearthdata.com)
* `01-infos/equalarea_id_coords.tsv`                        : ID and left/right/top/bottom coordinates of each equal area into the shapefile grid_equalarea200km.
* `01-infos/marine_actinopterygii_species.txt`              : List of "actinopterygii" saltwater species according to (http://www.fishbase.org/)
* `01-infos/freshwater_behrman_worldcoast_data_object.RData`: R spatial object from "sp package" which is an equal area grid in Berhmann projection with worldcoast shape and presence/absence of freshwater fish species from (https://www.iucn.org/)
* `01-infos/marine_behrman_worldcoast_data_object.RData`    : ... of marine fish species.
* IL EN MANQUE...

## 2.3 Set Up
clone the project and switch to the main folder, it's your working directory

```
git clone http://gitlab.mbb.univ-montp2.fr/reservebenefit/worldmap_fish_genetic_diversity.git
cd worldmap_fish_genetic_diversity
```

Then you will need to download georeferenced sequences of actinopterygii individuals "combined TSV file" from (http://www.boldsystems.org/index.php/Public_SearchTerms?taxon=&searchMenu=records&query=actinopterygii)
Store it into the folder `02-raw_data` and rename it `seqbold_data.txt`

You're ready to run the analysis. Now follow the instructions at [Running the pipeline](#5-running-the-pipeline)


# 3. Scripts Code Source
## 3.1 [00-scripts/step1](00-scripts/step1) : filter raw data

- BASH scripts
    * [filter_raw_data.sh](00-scripts/step1/filter_raw_data.sh)       : Keep only the CO1 sequences with lat/lon information
    * [get_geonames_coordinates.sh](00-scripts/step1/get_geonames_coordinates.sh)   : Uses (http://www.geonames.org/) to find missing coordinates of individual sequences from their textual information of location.

- PYTHON scripts
    * [lat_long_DMS_DD_converter.py](00-scripts/step1/lat_long_DMS_DD_converter.py)     : Converts from DMS format to DD format the given coordinates.

## 3.2 [00-scripts/step2](00-scripts/step2) : georeferenced sequences alignments by species

- BASH scripts
    * [seq_alnt_filtered_data.sh](00-scripts/step2/seq_alnt_filtered_data.sh)     : aligns sequences from the same species with MUSCLE and creates coordinates file for each sequence.
    * [cluster_freshwater_vs_marine.sh](00-scripts/step2/cluster_freshwater_vs_marine.sh) : according to a list of marine species, moves the fasta and coords files into marine, freshwater repertories.

- PYTHON scripts
    * [fasta_coords_files_species_generator.py](00-scripts/step2/fasta_coords_files_species_generator.py) : extracts sequences and associated coordinates from the filtered data.

- R scripts
    * [equalareacoords.R](00-scripts/step2/equalareacoords.R)    : attributes at each sequence an ID of cell of the shapefile of worldmap equal area projection from its coordinates.

## 3.3 [00-scripts/step3](00-scripts/step3) : species sequence pairwise comparison

- JULIA scripts
    * [Lib_Compare_Pairwise.jl](00-scripts/step3/Lib_Compare_Pairwise.jl)      : functions to compute the Genetic Diversity value from a set of sequences.
    * [Lib_Create_Master_Matrices.jl](00-scripts/step3/Lib_Create_Master_Matrices.jl) : functions to create master data matrices that are used to compute genetic diversity.
    * [master_matrices.jl](00-scripts/step3/master_matrices.jl)            : generates master data matrices from species sequences alignments.

## 3.4 [00-scripts/step4](00-scripts/step4) : genetic Diversity calculation

- BASH scripts
    * [gdval_by_site.sh](00-scripts/step4/gdval_by_site.sh) : generates CSV files with 2 columns : cell ID and mean genetic diversity per species into the cell

- JULIA scripts
    * [equalarea_numbers.jl](00-scripts/step4/equalarea_numbers.jl)         : attributes mean genetic diversity at each equal area grid cell. Genetic diversity is calculated from master data matrices
    * [metrics_by_area_and_species.jl](00-scripts/step4/metrics_by_area_and_species.jl): generates files for statistical analysis at next step : mean genetic diversity per cell, genetic diversity per species per cell, number of individuals per species, number of species per cell, cell coordinates, cell ID...
    * [Lib_GD_summary_functions.jl](00-scripts/step4/Lib_GD_summary_functions.jl)   : functions to calculate genetic diversity at species level and cell level


## 3.5 [00-scripts/step5](00-scripts/step5) : statistical analysis
- R scripts
    * `descripteurs.R`
    * `figures.R` WORK IN PROGRESS !!!


# 4. Reporting bugs

If you're sure you've found a bug — e.g. if one of my programs crashes
with an obscur error message, or if the resulting file is missing part
of the original data, then by all means submit a bug report.

I use [GitLab's issue system](https://gitlab.com/reservebenefit/worldmap_fish_genetic_diversity/issues)
as my bug database. You can submit your bug reports there. Please be as
verbose as possible — e.g. include the command line, etc

# 5. Running the pipeline

## 5.1 Filter raw data
```
bash ./00-scripts/step1/filter_raw_data.sh
```

## 5.2 Georeferenced sequences alignments by species
```
bash ./00-scripts/step2/seq_alnt_filtered_data.sh
mkdir ./06-species_alnt_cluster/total
mkdir ./06-species_alnt_cluster/freshwater
mkdir ./06-species_alnt_cluster/marine
bash ./00-scripts/step2/cluster_freshwater_vs_marine.sh
Rscript ./00-scripts/step2/equalareacoords.R
```
## 5.3 Species sequence pairwise comparison
```
julia ./00-scripts/step3/master_matrices.jl
```

## 5.4 Genetic Diversity calculation
```
julia ./00-scripts/step4/equalarea_numbers.jl
bash ./00-scripts/step4/gdval_by_site.sh
julia ./00-scripts/step4/metrics_by_area_and_species.jl
```
## 5.5 Statistical analysis
```
Rscript ./00-scripts/step5/descripteurs.R
Rscript ./00-scripts/step5/figures.R
```