The invention discloses an automatic analysis method of biodiversity. The method comprises the following specific analysis steps: step 1, using 24 pairs of primers containing sequences matched to an illumina sequencing chip, 8-base index sequences, sequencing primer sequences, difference sequences and universal amplification primer sequences, obtaining a library of on-machine sequencing through PCR reaction, and selecting a corresponding sequencing mode; step 2, converting stopping data into a fastq format by bcl2fastq software, and adding a parameter ''-create-fastq-for-index-reads''; step 3,using a Phred algorithm to intercept low-quality sequences through a filtering software BBDuk software package; and step 4, using a script in a QIIME package for secondary filtering on Tags sequencesobtained by splicing. According to the method, a project cycle is compressed, a database construction method, a data filtering method and software are improved, analysis deviations brought by chimeras and sequencing errors are effectively decreased, processes from data production to filtering and cleantags forming can be uniformity carried out without the need for distinguishing of each project even for microbial diversity projects, and processing time is saved.