tbtools
Apps & AutomationUse when the user asks about TBtools, TBtools-II, TBtools RPC API, TBtools CLI, or bioinformatics operations available through TBtools such as sequence manipulation, BLAST, GFF/GTF/GXF processing, expression tables, heatmaps, trees, MCScanX, DIAMOND, HMMER, MUSCLE, and IQ-TREE.
How to use this skill
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TBtools
Overview
TBtools-II is a Java-based bioinformatics toolkit that runs on Windows and macOS. Prefer TBtools' own bundled Java/RPC/CLI tools. Do not introduce Python, R, Conda, or other dependencies unless the user explicitly asks.
Honor TBTOOLS_HOME or TBTOOLS_JAR if set; fall back to the platform default.
Setup
Windows (PowerShell)
$TbtoolsHome = if ($env:TBTOOLS_HOME) { $env:TBTOOLS_HOME } else { "C:\Program Files\TBtools" }
$TbtoolsJar = if ($env:TBTOOLS_JAR) { $env:TBTOOLS_JAR } else { Join-Path $TbtoolsHome "TBtools_JRE1.6.jar" }
$env:Path = "$(Join-Path $TbtoolsHome 'bin');$env:Path"
Test-Path $TbtoolsJar
java -version
macOS / Linux (bash)
TBTOOLS_HOME="${TBTOOLS_HOME:-/Applications/TBtools-II}"
TBTOOLS_JAR="${TBTOOLS_JAR:-$(find "$TBTOOLS_HOME" -name 'TBtools*.jar' -maxdepth 4 2>/dev/null | head -1)}"
export PATH="$TBTOOLS_HOME/bin:$PATH"
[ -f "$TBTOOLS_JAR" ] && echo "jar: $TBTOOLS_JAR" || echo "TBtools jar not found — set TBTOOLS_JAR"
java -version
Use bin/ (bin\ on Windows) tools directly when the task is a standard external tool workflow:
blastn -query query.fa -db db_prefix -out results.tsv -outfmt 6
muscle -in input.fa -out aligned.fa
iqtree -s aligned.fa -m MFP -bb 1000 -nt AUTO
diamond blastp -d proteins.dmnd -q query.fa -o matches.tsv
RPC Server
Windows (PowerShell)
.\scripts\start_rpc_server.ps1 -Background
Direct form:
java -cp $TbtoolsJar biocjava.rpc.RpcServer
macOS / Linux (bash)
java -cp "$TBTOOLS_JAR" biocjava.rpc.RpcServer &
Default URLs:
| Purpose | URL |
|---|---|
| Health check | http://127.0.0.1:8765/health |
| JSON-RPC | http://127.0.0.1:8765/rpc |
rpcPort is configurable in TBtools config. rpcBindAddress is normalized to loopback only (127.0.0.1 or ::1); do not expose the RPC server on 0.0.0.0.
RPC Helper
Windows (PowerShell)
function Invoke-TBtoolsRpc {
param(
[Parameter(Mandatory=$true)][string]$Method,
[hashtable]$Params = @{},
[string]$Uri = "http://127.0.0.1:8765/rpc",
[int]$TimeoutSec = 600
)
$body = @{
jsonrpc = "2.0"
method = $Method
params = $Params
id = [guid]::NewGuid().ToString()
} | ConvertTo-Json -Depth 50
$response = Invoke-RestMethod -Method Post -Uri $Uri -ContentType "application/json" -Body $body -TimeoutSec $TimeoutSec
if ($null -ne $response.error) {
$reason = if ($response.error.data -and $response.error.data.reason) { " ($($response.error.data.reason))" } else { "" }
throw "TBtools RPC error $($response.error.code): $($response.error.message)$reason"
}
return $response.result
}
macOS / Linux (bash + curl)
tbtools_rpc() {
local method="$1"
local params="${2:-{}}"
local uri="${TBTOOLS_RPC_URI:-http://127.0.0.1:8765/rpc}"
curl -s --max-time 600 -X POST "$uri" \
-H "Content-Type: application/json" \
-d "{\"jsonrpc\":\"2.0\",\"method\":\"$method\",\"params\":$params,\"id\":\"1\"}"
}
Discovery:
# Windows
Invoke-WebRequest -UseBasicParsing http://127.0.0.1:8765/health
Invoke-TBtoolsRpc -Method system.ping
$methods = (Invoke-TBtoolsRpc -Method system.listMethods).methods
$methods | Where-Object { $_ -like "*Fasta*" }
Invoke-TBtoolsRpc -Method system.describeMethod -Params @{ method = "FastaStat.process" }
Invoke-TBtoolsRpc -Method system.toolsJson
# macOS / Linux
curl -s http://127.0.0.1:8765/health
tbtools_rpc system.ping
tbtools_rpc system.listMethods | python3 -c "import sys,json; [print(m) for m in json.load(sys.stdin)['result']['methods']]"
tbtools_rpc system.describeMethod '{"method":"FastaStat.process"}'
RPC Workflow
- Create deterministic workspace folders and pass absolute host-local file paths. RPC bodies carry paths, not file content.
- Call
system.listMethodsandsystem.describeMethodfor candidate methods. Usesystem.toolsJsonwhen many method schemas are needed. - Prefer a matching
*.validateParamsor*.validateInputbefore*.processwhen available. - Treat validation failures carefully: validate methods may return
result.ok = false/validated = falsewith structured errors instead of a JSON-RPCerror. - For long BLAST, tree, heatmap, NCBI, or large file jobs, raise client timeout and reduce concurrency.
- If JSON-RPC error
-32603haserror.data.reason = rpc pool busy, back off and retry with lower concurrency.
RPC Protocol Notes
| Topic | Contract |
|---|---|
| Request | HTTP POST /rpc, JSON-RPC 2.0 object with method, params, id |
| Batch | JSON array of request objects; response array is in order and omits notifications |
| Empty batch | HTTP 200 with body [] |
| Health | GET /health returns HTTP 200 body OK |
| Success | result object, usually at least ok: true |
| Errors | HTTP 200 with error.code, error.message, optional error.data |
Common error codes: -32700 parse error, -32600 invalid request, -32601 method not found, -32602 invalid params, -32603 internal error, -32000 timeout.
Common RPC Methods
| Area | Methods |
|---|---|
| Sequence files | FastaStat.process, FastaSeqManipulator.process, AmazingFastaExtract.process, FastaExtract.process, FastxExtract.process, FastxIndex.process, FastaIDTools.* |
| FASTA conversion | FastaToTable.process, TableToFasta.process, CdsToProtein.process, FastaMerge.process, FastaSplitByCount.process, FastaSplitAsOneSeq.process |
| Annotation | GXFFix.process, GffCdsPhase.process, GxfSeqExtract.process, GxfFilter.process, GffFeatureExtract.process, GtfFeatureExtract.process, GxfStat.process |
| Tables | TableTools.transpose, TableTools.mergeByKeyMulti, TableTools.selectRows, TableTools.selectColumnsByList, TableTools.groupAggregate, TableRowManipulator.process |
| Expression | ExpressionFpkmToTpm.process, ExpressionRpkm.process, ExpressionTpm.process, ExpressionTau.process, ExpressionCorrMatrix.process, GeneExpFilter.process, GenePairCorr.process |
| Alignment and trees | BlastCompareTwoSeqSet.process, BlastXmlToTable.process, ReciprocalBlast.process, OneStepBuildATree.process, TrimMsaSimple.process, TrimMsaGblocks.process, BlatAlign.process |
| Visualization and misc | AmazingHeatMap.process, MemeSuiteXmlToTab.process, PlantCareClassify.process, SendEmail.process, TodoList.* |
Examples
FASTA statistics (Windows PowerShell):
Invoke-TBtoolsRpc -Method FastaStat.process -Params @{
inputPath = "D:/data/genome.fa"
outputPath = "D:/data/out/genome_stat.xls"
options = @{ getLengthOnly = $false }
}
FASTA statistics (macOS / Linux bash):
tbtools_rpc FastaStat.process \
'{"inputPath":"/data/genome.fa","outputPath":"/data/out/genome_stat.xls","options":{"getLengthOnly":false}}'
Extract FASTA records with a prewritten ID list (Windows PowerShell):
Invoke-TBtoolsRpc -Method AmazingFastaExtract.process -Params @{
inputPath = "D:/data/genome.fa"
idListPath = "D:/data/ids.txt"
outputPath = "D:/data/out/extracted.fa"
options = @{
usePattern = $false
caseInsensitive = $true
dontTreatSpaceAsColSep = $false
wholeWordMatch = $false
}
}
Validate then run a tree pipeline (Windows PowerShell):
$params = @{
inputPath = "D:/data/sequences.fa"
outputPath = "D:/data/out/tree"
options = @{ ultraFastBS = $true; bbTime = 5000; model = "Auto"; threads = 2 }
}
$check = Invoke-TBtoolsRpc -Method OneStepBuildATree.validateParams -Params $params
if ($check.ok -eq $false -or $check.validated -eq $false) { $check.errors; throw "TBtools validation failed" }
Invoke-TBtoolsRpc -Method OneStepBuildATree.process -Params $params -TimeoutSec 7200
Validate then run a tree pipeline (macOS / Linux bash):
tbtools_rpc OneStepBuildATree.validateParams \
'{"inputPath":"/data/sequences.fa","outputPath":"/data/out/tree","options":{"ultraFastBS":true,"bbTime":5000,"model":"Auto","threads":2}}'
tbtools_rpc OneStepBuildATree.process \
'{"inputPath":"/data/sequences.fa","outputPath":"/data/out/tree","options":{"ultraFastBS":true,"bbTime":5000,"model":"Auto","threads":2}}'
Operating Rules
- Detect the platform first: use PowerShell helpers on Windows, bash + curl on macOS/Linux.
- Use TBtools bundled
bin/tools before suggesting external installs. - Keep stderr/error details visible for external tools and RPC errors.
- Do not assume a method exists from memory; discover it through
system.listMethods. - Do not assume
system.listMethodsreturns a raw array; the canonical result is an object withmethods. - Do not use Python helpers for normal TBtools work in this environment.
- Use forward-slash paths (
/data/...) in RPC calls even on Windows — TBtools accepts them.