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rotl provides an interface to the Open Tree of Life (OTL) API and allows users to query the API, retrieve parts of the Tree of Life and integrate these parts with other R packages.

The OTL API provides services to access:

  • the Tree of Life a.k.a. TOL (the synthetic tree): a single draft tree that is a combination of the OTL taxonomy and the source trees (studies)
  • the Taxonomic name resolution services a.k.a. TNRS: the methods for resolving taxonomic names to the internal identifiers used by the TOL and the GOL (the ott ids).
  • the Taxonomy a.k.a. OTT (for Open Tree Taxonomy): which represents the synthesis of the different taxonomies used as a backbone of the TOL when no studies are available.
  • the Studies containing the source trees used to build the TOL, and extracted from the scientific literature.

In rotl, each of these services correspond to functions with different prefixes:

Service rotl prefix
Tree of Life tol_
TNRS tnrs_
Taxonomy taxonomy_
Studies studies_

rotl also provides a few other functions and methods that can be used to extract relevant information from the objects returned by these functions.

Demonstration of a basic workflow

The most common use for rotl is probably to start from a list of species and get the relevant parts of the tree for these species. This is a two step process:

  1. the species names need to be matched to their ott_id (the Open Tree Taxonomy identifiers) using the Taxonomic name resolution services (TNRS)
  2. these ott_id will then be used to retrieve the relevant parts of the Tree of Life.

Step 1: Matching taxonomy to the ott_id

Let’s start by doing a search on a diverse group of taxa: a tree frog (genus Hyla), a fish (genus Salmo), a sea urchin (genus Diadema), and a nautilus (genus Nautilus).

library(rotl)
taxa <- c("Hyla", "Salmo", "Diadema", "Nautilus")
resolved_names <- tnrs_match_names(taxa)

It’s always a good idea to check that the resolved names match what you intended:

search_string unique_name approximate_match ott_id is_synonym flags number_matches
hyla Hyla FALSE 1062216 FALSE 1
salmo Salmo FALSE 982359 FALSE 1
diadema Diadema FALSE 4930522 FALSE 4
nautilus Nautilus FALSE 616358 FALSE 1

The column unique_name sometimes indicates the higher taxonomic level associated with the name. The column number_matches indicates the number of ott_id that corresponds to a given name. In this example, our search on Diadema returns 2 matches, and the one returned by default is indeed the sea urchin that we want for our query. The argument context_name allows you to limit the taxonomic scope of your search. Diadema is also the genus name of a fungus. To ensure that our search is limited to animal names, we could do:

resolved_names <- tnrs_match_names(taxa, context_name = "Animals")

If you are trying to build a tree with deeply divergent taxa that the argument context_name cannot fix, see “How to change the ott ids assigned to my taxa?” in the FAQ below.

Step 2: Getting the tree corresponding to our taxa

Now that we have the correct ott_id for our taxa, we can ask for the tree using the tol_induced_subtree() function. By default, the object returned by tol_induced_subtree is a phylo object (from the ape package), so we can plot it directly.

my_tree <- tol_induced_subtree(ott_ids = resolved_names$ott_id)
## Warning in collapse_singles(tr, show_progress): Dropping singleton nodes
## with labels: Chordata ott125642, mrcaott42ott658, Craniata (subphylum in
## Deuterostomia) ott947318, Vertebrata (subphylum in Deuterostomia) ott801601,
## Gnathostomata (superclass in phylum Chordata) ott278114, Teleostomi
## ott114656, Sarcopterygii ott458402, Dipnotetrapodomorpha ott4940726,
## Tetrapoda ott229562, Amphibia ott544595, Batrachia ott471197, Anura ott991547,
## mrcaott114ott3129, mrcaott114ott37876, mrcaott114ott18818, Neobatrachia
## ott535804, mrcaott114ott309463, mrcaott114ott177, mrcaott177ott7464,
## mrcaott177ott2199, mrcaott177ott29310, mrcaott177ott1321, Hylidae ott535782,
## mrcaott177ott17126, mrcaott177ott43017, mrcaott177ott2732, mrcaott2732ott3289,
## mrcaott3289ott40328, mrcaott3289ott173489, mrcaott3289ott110534,
## mrcaott3289ott489758, mrcaott3289ott59160, Actinopterygii ott773483,
## Actinopteri ott285821, Neopterygii ott471203, Teleostei ott212201,
## Osteoglossocephalai ott5506109, Clupeocephala ott285819, Euteleosteomorpha
## ott5517919, mrcaott274ott392, mrcaott274ott595, Protacanthopterygii ott1024043,
## mrcaott274ott3887, mrcaott3887ott9371, Salmoniformes ott216171, Salmonidae
## ott739933, mrcaott3887ott28511, Salmoninae ott936925, mrcaott31485ott79094,
## mrcaott360ott10516, Echinodermata ott451020, Eleutherozoa ott317277, Echinozoa
## ott669475, Echinoidea ott669472, mrcaott360ott26831, mrcaott360ott3985,
## mrcaott360ott367, mrcaott360ott362, Acroechinoidea ott5677511, Diadematoida
## ott631174, Diadematidae ott631173, Protostomia ott189832, mrcaott49ott6612,
## Lophotrochozoa ott155737, mrcaott56ott519, mrcaott56ott5497, Mollusca ott802117,
## mrcaott56ott1881, mrcaott1881ott102410, Cephalopoda ott7368, Nautiloidea
## ott854446, Nautilida ott854452, Nautilidae ott616361
plot(my_tree, no.margin = TRUE)
plot of chunk unnamed-chunk-3
plot of chunk unnamed-chunk-3

FAQ

How to change the ott ids assigned to my taxa?

If you realize that tnrs_match_names assigns the incorrect taxonomic group to your name (e.g., because of synonymy) and changing the context_name does not help, you can use the function inspect. This function takes the object resulting from tnrs_match_names(), and either the row number, the taxon name (you used in your search in lowercase), or the ott_id returned by the initial query.

To illustrate this, let’s re-use the previous query but this time pretending that we are interested in the fungus Diadema and not the sea urchin:

taxa <- c("Hyla", "Salmo", "Diadema", "Nautilus")
resolved_names <- tnrs_match_names(taxa)
resolved_names
##   search_string unique_name approximate_match  ott_id is_synonym flags
## 1          hyla        Hyla             FALSE 1062216      FALSE      
## 2         salmo       Salmo             FALSE  982359      FALSE      
## 3       diadema     Diadema             FALSE 4930522      FALSE      
## 4      nautilus    Nautilus             FALSE  616358      FALSE      
##   number_matches
## 1              1
## 2              1
## 3              4
## 4              1
inspect(resolved_names, taxon_name = "diadema")
##   search_string                       unique_name approximate_match  ott_id
## 1       diadema                           Diadema             FALSE 4930522
## 2       diadema                       Diademoides             FALSE 4024672
## 3       diadema Garrettia (genus in Opisthokonta)             FALSE  631176
## 4       diadema                        Hypolimnas             FALSE  643831
##   is_synonym          flags number_matches
## 1      FALSE                             4
## 2       TRUE sibling_higher              4
## 3       TRUE                             4
## 4       TRUE                             4

In our case, we want the second row in this data frame to replace the information that initially matched for Diadema. We can now use the update() function, to change to the correct taxa (the fungus not the sea urchin):

resolved_names <- update(resolved_names,
  taxon_name = "diadema",
  new_row_number = 2
)

## we could also have used the ott_id to replace this taxon:
## resolved_names <- update(resolved_names, taxon_name = "diadema",
##                          new_ott_id = 4930522)

And now our resolved_names data frame includes the taxon we want:

search_string unique_name approximate_match ott_id is_synonym flags number_matches
hyla Hyla FALSE 1062216 FALSE 1
salmo Salmo FALSE 982359 FALSE 1
diadema Diademoides FALSE 4024672 TRUE sibling_higher 4
nautilus Nautilus FALSE 616358 FALSE 1

How do I know that the taxa I’m asking for is the correct one?

The function taxonomy_taxon_info() takes ott_ids as arguments and returns taxonomic information about the taxa. This output can be passed to some helpers functions to extract the relevant information. Let’s illustrate this with our Diadema example

diadema_info <- taxonomy_taxon_info(631176)
tax_rank(diadema_info)
## $`Garrettia (genus in Opisthokonta)`
## [1] "genus"
## 
## attr(,"class")
## [1] "otl_rank" "list"
synonyms(diadema_info)
## $`Garrettia (genus in Opisthokonta)`
## [1] "Centrechinus" "Diadema"      "Diamema"     
## 
## attr(,"class")
## [1] "otl_synonyms" "list"
tax_name(diadema_info)
## $`Garrettia (genus in Opisthokonta)`
## [1] "Garrettia"
## 
## attr(,"class")
## [1] "otl_name" "list"

In some cases, it might also be useful to investigate the taxonomic tree descending from an ott_id to check that it’s the correct taxon and to determine the species included in the Open Tree Taxonomy:

diadema_tax_tree <- taxonomy_subtree(631176)
diadema_tax_tree
## $tip_label
##  [1] "Garrettia_parva_ott6356094"              
##  [2] "Garrettia_rotella_ott6356095"            
##  [3] "Diadema_savignyi_ott395692"              
##  [4] "Diadema_palmeri_ott836860"               
##  [5] "Diadema_setosum_ott631175"               
##  [6] "Diadema_paucispinum_ott312263"           
##  [7] "unclassified_Diadema_ott7669081"         
##  [8] "Diadema_africanum_ott4147369"            
##  [9] "Diadema_antillarum_scensionis_ott220009" 
## [10] "Diadema_antillarum_antillarum_ott4147370"
## [11] "Diadema_mexicanum_ott639130"             
## [12] "Diademasp.SP04-BIO_4_JGLCO_AYott7072105" 
## [13] "Diademasp.SP03-BIO_3_JGLCO_AYott7072104" 
## [14] "Diademasp.SP02-BIO_2_JGLCO_AYott7072103" 
## [15] "Diadema_sp._DSM1_ott219999"              
## [16] "Diadema_sp._DSM6_ott771059"              
## [17] "Diademasp.ACOSTI-NIOTSU3ott7072098"      
## [18] "Diademasp.LI03-BIO_JGLCO_AYott7072102"   
## [19] "Diademasp.LI02-BIO_JGLCO_AYott7072101"   
## [20] "Diademasp.LI01-BIO_JGLCO_AYott7072100"   
## [21] "Diademasp.ACOSTI-NIOTSU4ott7072099"      
## [22] "Diadema_sp._seto35_ott66618"             
## [23] "Diadema_sp._seto18_ott66623"             
## [24] "Diadema_sp._seto19_ott66624"             
## [25] "Diadema_sp._seto38_ott66625"             
## [26] "Diadema_sp._DJN9_ott66626"               
## [27] "Diademasp.CS-2014ott5502179"             
## [28] "Diadema_sp._seto17_ott587478"            
## [29] "Diadema_sp._SETO15_ott587479"            
## [30] "Diadema_sp._dsm5_ott587480"              
## [31] "Diadema_sp._DSM4_ott587481"              
## [32] "Diadema_sp._DSM3_ott587482"              
## [33] "Diadema_sp._DSM2_ott587483"              
## [34] "Diadema_sp._seto10_ott587484"            
## [35] "Diadema_sp._seto9_ott587485"             
## [36] "Diadema_sp._DSM8_ott587486"              
## [37] "Diadema_sp._DSM7_ott587487"              
## [38] "Diadema_sp._seto16_ott312262"            
## [39] "Diadema_africana_ott5502180"             
## [40] "Diadema_principeana_ott5725746"          
## [41] "Diadema_vetus_ott5725747"                
## [42] "Diadema_regnyi_ott7669077"               
## [43] "Diadema_amalthei_ott7669073"             
## [44] "Diadema_affine_ott7669072"               
## [45] "Diadema_subcomplanatum_ott7669079"       
## [46] "Diadema_ruppelii_ott7669078"             
## [47] "Diadema_calloviensis_ott7669074"         
## [48] "Diadema_megastoma_ott7669075"            
## [49] "Diadema_priscum_ott7669076"              
## [50] "Garrettia_biangulata_ott7669080"         
## [51] "Diadema_ascensionis_ott4950423"          
## [52] "Diadema_lobatum_ott4950422"              
## 
## $edge_label
## [1] "Diadema_antillarum_ott1022356"  "'Garrettia(genusinOpisthokonta"
## [3] "ott631176'"

By default, this function return all taxa (including self, and internal) descending from this ott_id but it also possible to return phylo object.

How do I get the tree for a particular taxonomic group?

If you are looking to get the tree for a particular taxonomic group, you need to first identify it by its node id or ott id, and then use the tol_subtree() function:

mono_id <- tnrs_match_names("Monotremata")
mono_tree <- tol_subtree(ott_id = ott_id(mono_id))
## Warning in collapse_singles(tr, show_progress): Dropping singleton nodes with
## labels: Tachyglossus ott16047, Tachyglossus aculeatus ott16038, Ornithorhynchus
## ott962391, Obdurodon ott3615207, Steropodon ott3615209, Teinolophos ott4119276
plot(mono_tree)
plot of chunk unnamed-chunk-8
plot of chunk unnamed-chunk-8

How do I find trees from studies focused on my favourite taxa?

The function studies_find_trees() allows the user to search for studies matching a specific criteria. The function studies_properties() returns the list of properties that can be used in the search.

furry_studies <- studies_find_studies(property = "ot:focalCladeOTTTaxonName", value = "Mammalia")
furry_ids <- furry_studies$study_ids

Now that we know the study_id, we can ask for the meta data information associated with this study:

furry_meta <- get_study_meta("pg_2550")
get_publication(furry_meta) ## The citation for the source of the study
## [1] "O'Leary, Maureen A., Marc Allard, Michael J. Novacek, Jin Meng, and John Gatesy. 2004. \"Building the mammalian sector of the tree of life: Combining different data and a discussion of divergence times for placental mammals.\" In: Cracraft J., & Donoghue M., eds. Assembling the Tree of Life. pp. 490-516. Oxford, United Kingdom, Oxford University Press."
## attr(,"DOI")
## [1] ""
get_tree_ids(furry_meta) ## This study has 10 trees associated with it
##  [1] "tree5513" "tree5515" "tree5516" "tree5517" "tree5518" "tree5519"
##  [7] "tree5520" "tree5521" "tree5522" "tree5523"
candidate_for_synth(furry_meta) ## None of these trees are yet included in the OTL
## NULL

Using get_study("pg_2550") would returns a multiPhylo object (default) with all the trees associated with this particular study, while get_study_tree("pg_2550", "tree5513") would return one of these trees.

The tree returned by the API has duplicated tip labels, how can I work around it?

You may encounter the following error message:

Error in rncl(file = file, ...) : Taxon number 39 (coded by the token Pratia
angulata) has already been encountered in this tree. Duplication of taxa in a
tree is prohibited.

This message occurs as duplicate labels are not allowed in the NEXUS format and it is stricly enforced by the part of the code used by rotl to import the trees in memory.

If you use a version of rotl more recent than 0.4.1, this should not happen by default for the function get_study_tree. If it happens with another function, please let us know.

The easiest way to work around this is to save the tree in a file, and use APE to read it in memory:

get_study_tree(
  study_id = "pg_710", tree_id = "tree1277",
  tip_label = "ott_taxon_name", file = "/tmp/tree.tre",
  file_format = "newick"
)
tr <- ape::read.tree(file = "/tmp/tree.tre")

How do I get the higher taxonomy for a given taxa?

If you encounter a taxon name you are not familiar with, it might be useful to obtain its higher taxonomy to see where it fits in the tree of life. We can combine several taxonomy methods to extract this information easily.

giant_squid <- tnrs_match_names("Architeuthis")
tax_lineage(taxonomy_taxon_info(ott_id(giant_squid), include_lineage = TRUE))
## $`564394`
##          rank               name        unique_name  ott_id
## 1      family     Architeuthidae     Architeuthidae  564393
## 2       order          Oegopsida          Oegopsida   43352
## 3  superorder     Decapodiformes     Decapodiformes  854107
## 4  infraclass       Neocoleoidea       Neocoleoidea  329546
## 5    subclass          Coleoidea          Coleoidea    7371
## 6       class        Cephalopoda        Cephalopoda    7368
## 7      phylum           Mollusca           Mollusca  802117
## 8     no rank     Lophotrochozoa     Lophotrochozoa  155737
## 9     no rank        Protostomia        Protostomia  189832
## 10    no rank          Bilateria          Bilateria  117569
## 11    no rank          Eumetazoa          Eumetazoa  641038
## 12    kingdom            Metazoa            Metazoa  691846
## 13    no rank            Holozoa            Holozoa 5246131
## 14    no rank       Opisthokonta       Opisthokonta  332573
## 15     domain          Eukaryota          Eukaryota  304358
## 16    no rank cellular organisms cellular organisms   93302
## 17    no rank               life               life  805080

Why are OTT IDs discovered with rotl missing from an induced subtree?

Some taxonomic names that can be retrieved through the taxonomic name resolution service are not part of the Open Tree’s synthesis tree. These are usually traditional higher-level taxa that have been found to be paraphyletic.

For instance, if you wanted to fetch a tree relating the three birds that go into a Turkducken as well as the pork used for stuffing, you might search for the turkey, duck, chicken, and pork genera:

turducken <- c("Meleagris", "Anas", "Gallus", "Sus")
taxa <- tnrs_match_names(turducken, context = "Animals")
## Warning in tnrs_match_names(turducken, context = "Animals"): partial argument
## match of 'context' to 'context_name'
taxa
##   search_string unique_name approximate_match ott_id is_synonym flags
## 1     meleagris   Meleagris             FALSE 446481      FALSE      
## 2          anas        Anas             FALSE 765185      FALSE      
## 3        gallus      Gallus             FALSE 153562      FALSE      
## 4           sus         Sus             FALSE 730021      FALSE      
##   number_matches
## 1              2
## 2              1
## 3              3
## 4              1

We have the OTT ids for each genus, however, if we tried to get the induced subtree from these results, we would get an error:

## Warning in collapse_singles(tr, show_progress): Dropping singleton nodes with
## labels: Mammalia ott244265, Theria (subclass in Deuterostomia) ott229558,
## Eutheria (in Deuterostomia) ott683263, mrcaott42ott3607383, mrcaott42ott3607429,
## mrcaott42ott3607455, mrcaott42ott72667, Boreoeutheria ott5334778, Laurasiatheria
## ott392223, mrcaott1548ott4697, mrcaott1548ott6790, mrcaott1548ott3607484,
## mrcaott1548ott4942380, mrcaott1548ott4942547, mrcaott1548ott3021, Artiodactyla
## ott622916, mrcaott1548ott21987, Suina ott916745, Suidae ott730008, Sauropsida
## ott639642, Sauria ott329823, mrcaott246ott2982, mrcaott246ott31216,
## mrcaott246ott3602822, mrcaott246ott4143599, mrcaott246ott3600976,
## mrcaott246ott4132107, Aves ott81461, Neognathae ott241846, Galliformes
## ott837585, mrcaott4765ott6520194, mrcaott4765ott109888, mrcaott4765ott75785,
## mrcaott4765ott104461, mrcaott4765ott151684, mrcaott4765ott54193,
## mrcaott4765ott49319, mrcaott4765ott3596087, mrcaott4765ott415487,
## mrcaott4765ott51354, mrcaott4765ott53700, mrcaott4765ott446490, Meleagridinae
## ott781250, mrcaott49310ott102705, mrcaott49310ott51349, mrcaott49310ott153554,
## mrcaott153554ott867027, Anseriformes ott241841, mrcaott30843ott714464, Anatidae
## ott765193, mrcaott30843ott75874, mrcaott30843ott962771, mrcaott30843ott145504,
## mrcaott30843ott30847, mrcaott30843ott196654, mrcaott30843ott30845

As the error message suggests, some of the taxa are not found in the synthetic tree. This occurs for 2 main reasons: either the taxa is invalid, or it is part of a group that is not monophyletic in the synthetic tree. There are two ways to get around this issue: (1) removing the taxa that are not part of the Open Tree; (2) using the complete species name.

Removing the taxa missing from the synthetic tree

To help with this situation, rotl provides a way to identify the OTT ids that are not part of the synthetic tree. The function is_in_tree() takes the output of the ott_id() function and returns a vector of logical indicating whether the taxa are part of the synthetic tree. We can then use to only keep the taxa that appear in the synthetic tree:

in_tree <- is_in_tree(ott_id(taxa))
in_tree
## Meleagris      Anas    Gallus       Sus 
##      TRUE     FALSE      TRUE      TRUE
tr <- tol_induced_subtree(ott_id(taxa)[in_tree])
## Warning in collapse_singles(tr, show_progress): Dropping singleton nodes with
## labels: Mammalia ott244265, Theria (subclass in Deuterostomia) ott229558,
## Eutheria (in Deuterostomia) ott683263, mrcaott42ott3607383, mrcaott42ott3607429,
## mrcaott42ott3607455, mrcaott42ott72667, Boreoeutheria ott5334778, Laurasiatheria
## ott392223, mrcaott1548ott4697, mrcaott1548ott6790, mrcaott1548ott3607484,
## mrcaott1548ott4942380, mrcaott1548ott4942547, mrcaott1548ott3021, Artiodactyla
## ott622916, mrcaott1548ott21987, Suina ott916745, Suidae ott730008, Sauropsida
## ott639642, Sauria ott329823, mrcaott246ott2982, mrcaott246ott31216,
## mrcaott246ott3602822, mrcaott246ott4143599, mrcaott246ott3600976,
## mrcaott246ott4132107, Aves ott81461, Neognathae ott241846, Galloanserae
## ott5839486, Galliformes ott837585, mrcaott4765ott6520194, mrcaott4765ott109888,
## mrcaott4765ott75785, mrcaott4765ott104461, mrcaott4765ott151684,
## mrcaott4765ott54193, mrcaott4765ott49319, mrcaott4765ott3596087,
## mrcaott4765ott415487, mrcaott4765ott51354, mrcaott4765ott53700,
## mrcaott4765ott446490, Meleagridinae ott781250, mrcaott49310ott102705,
## mrcaott49310ott51349, mrcaott49310ott153554, mrcaott153554ott867027

Using the full taxonomic names

The best way to avoid these problems is to specify complete species names (species being the lowest level of classification in the Open Tree taxonomy they are guaranteed to be monophyletic):

turducken_spp <- c("Meleagris gallopavo", "Anas platyrhynchos", "Gallus gallus", "Sus scrofa")
taxa <- tnrs_match_names(turducken_spp, context = "Animals")
## Warning in tnrs_match_names(turducken_spp, context = "Animals"): partial
## argument match of 'context' to 'context_name'
## Warning in collapse_singles(tr, show_progress): Dropping singleton nodes
## with labels: Mammalia ott244265, Theria (subclass in Deuterostomia)
## ott229558, Eutheria (in Deuterostomia) ott683263, mrcaott42ott3607383,
## mrcaott42ott3607429, mrcaott42ott3607455, mrcaott42ott72667, Boreoeutheria
## ott5334778, Laurasiatheria ott392223, mrcaott1548ott4697, mrcaott1548ott6790,
## mrcaott1548ott3607484, mrcaott1548ott4942380, mrcaott1548ott4942547,
## mrcaott1548ott3021, Artiodactyla ott622916, mrcaott1548ott21987, Suina
## ott916745, Suidae ott730008, Sus ott730021, Sauropsida ott639642, Sauria
## ott329823, mrcaott246ott2982, mrcaott246ott31216, mrcaott246ott3602822,
## mrcaott246ott4143599, mrcaott246ott3600976, mrcaott246ott4132107, Aves
## ott81461, Neognathae ott241846, Galliformes ott837585, mrcaott4765ott6520194,
## mrcaott4765ott109888, mrcaott4765ott75785, mrcaott4765ott104461,
## mrcaott4765ott151684, mrcaott4765ott54193, mrcaott4765ott49319,
## mrcaott4765ott3596087, mrcaott4765ott415487, mrcaott4765ott51354,
## mrcaott4765ott53700, mrcaott4765ott446490, Meleagridinae ott781250, Meleagris
## ott446481, mrcaott49310ott102705, mrcaott49310ott51349, mrcaott49310ott153554,
## mrcaott153554ott867027, Gallus ott153562, mrcaott153554ott153572,
## mrcaott153572ott240568, Anseriformes ott241841, mrcaott30843ott714464, Anatidae
## ott765193, mrcaott30843ott75874, mrcaott30843ott962771, mrcaott30843ott145504,
## mrcaott30843ott30847, mrcaott30843ott196654, mrcaott30843ott30845,
## mrcaott30845ott30850, mrcaott30850ott82420, mrcaott30850ott82414,
## mrcaott30850ott30858, mrcaott30850ott30855, mrcaott30850ott604172,
## mrcaott30850ott82410, mrcaott82410ott339494, mrcaott82410ott604175,
## mrcaott82410ott604182, mrcaott82410ott190881
plot(tr)
plot of chunk unnamed-chunk-16
plot of chunk unnamed-chunk-16