# Composition Diversity and Expression Specificity of the _Tps_ Gene Family among 24 _Ficus_ Species
## 2 Results  
### Characteristics of the TPS Genes in the Receptive Phase of Male Figs of 24 Ficus Species  
After RNA-sequencing, we obtained 41.52-56.03 million raw reads from the ostiole bract transcriptomes of 24 _Ficus_ species. For different species, we achieved 40.17-55.09 million clean reads after adapter clipping and quality control. Using the Trinity program, the clean reads of each species were assembled into 34,454-90,120 transcripts. The high-quality transcripts of the 24 species were subjected to cluster and assembly analyses, resulting in coding of 18,344-68,322 unigenes, N50 lengths of 877-1970 bp, and GC contents of 43.77-52.24%. All assembly statistics are summarized in Table S1. Using blast scanning, a total of 248 _TPS_ genes with two conserved N-terminal and C-terminal domains (PF01397 and PF03936) were identified from the unigenes of the 24 ostiole bract transcriptomes (Table S2). Among the genes, 180 _TPSs_ were found to have complete coding sequences, and 68 were found to have incomplete coding regions (Table S2). All _TPS_ genes containing a typical Terpene_Synth_C (or Terpene_cyclase_plant_C1) conserved domain were confirmed after revalidation by the NCBI Online Batch CD-Search. At the protein level, those complete _TPS_ genes showed an obvious sequence length polymorphism ranging from 405 aa in _F. variolosa_ to 850 aa in _F. microcarpa_ (Table S2). Different numbers of _TPS_ genes were found to be expressed during the receptive phase of male figs. Among the 24 _Ficus_ species. _F. altissima_ showed the most _TPS_ genes (19), while _F. fistulosa_ and _F. montana_ showed the fewest (3) (Table 1). \begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline
**Subgenus** & **Section** & **Subsection** & **Species** & **Name Abbreviations** & _TPS No._ & **Site** & **Latitude (N)** & **Longitude (B)** \\ \hline \multirow{6}{*}{Ficus} & & _F. chartauca_ & ficha & 8 & China—Guangdong & 8.776 & 99.724 \\  & & _F. fidus_ & fital & 14 & China—Guangdong & 8.776 & 99.724 \\  & _F. grassunivaloides_ & fgroo & 3 & Thailand—Ncarthiwat & 5.799 & 101.762 \\  & & _F. hitta_ & fihir & 16 & China—SCBG & 23.171 & 113.349 \\  & & _F. lampokensis_ & filian & 11 & China—Guangdong & 24.227 & 112.006 \\  & & _F. rinficial_ & artaousis & firuf & 4 & Taiwan & 21.962 & 120.811 \\  & & _F. rinficial_ & firii & 13 & China—SCBG & 23.180 & 112.537 \\ \cline{2-8}  & & _F. auli_ & fabe & 12 & China—Guangdong & 23.636 & 113.780 \\  & & _F. erreta_ & var. bechevngan_ & firee & 11 & China—Guangdong & 23.765 & 113.915 \\  & & _F. fernosa_ & fior & 6 & China—Guangdong & 23.623 & 113.811 \\  & & _F. heteromorpha_ & fihir & 15 & China—Guangdong & 24.918 & 113.033 \\  & & _F. isclumpada_ & fisc & 11 & Thailand—Chiang Mai & 18.504 & 98.665 \\  & & _F. rundurata_ & fignon & 13 & China—Guangdong & 24.252 & 112.006 \\  & & _F. pyriformis_ & fipyr & 12 & Thailand—Chiang Mai & 18.504 & 98.665 \\  & & _F. urriolosa_ & fivar & 13 & China—Guangdong & 23.624 & 113.797 \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Twenty-four _Ficus_ species used in this study. A total of 10 motifs were extracted from the 248 _TPS_ genes, but three (DDXXD, DTE, and RXR) showed the highest degree of conservation. Generally, these _TPS_ genes contained DDXXD and DTE motifs at the C-terminus (Figure 1A). \begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline  
Figure 1: Sequence characteristic, phylogeny, subfamily classification, and expression profile of 248 _FTPS_ genes. (**A**) Three conserved motif logos of the 248 _Ficus_ genes, showing DDXXD, DTE, and RXR motif, respectively. (**B**) A maximum likelihood tree based on amino acid sequences of 248 _FTPS_ genes and 33 _A. thaliana_ genes, showing that these _FTPSs_ are classified into five subfamilies and 33 orthologous genes (_A. thaliana_ genes are colored in red). (**C**) Expression heatmap of 248 _FTPS_ genes across 24 species. * Genes with positive sites.