**4. Discussion**

Plants contend with various environmental conditions throughout their life cycle that may interfere with their development. *TaTLP* genes are members of a gene family found in multiple animals. Plants have a smaller number of functionally studied *TLPs* than animals. *TLPs* have only been discovered in a few plant species, including Arabidopsis [13], rice [7], maize [10], *Solanum lycopersicum* [11], and cotton [12]. This study found 40 genes encoding *TLP* proteins in wheat, which is higher than the numbers found in other plants: 11 in Arabidopsis, 14 in rice, and 15 in maize. This research will advance the knowledge and understanding of their functional characteristics in the future.

#### *4.1. TaTLP Genes Are Distributed Widely in the Wheat Genome*

The hexaploid wheat, created by crossing Triticum and Aegilops, is a valuable tool for studying allopolyploidization evolution [24]. An analysis of the phylogenetic tree (Figure 1) shows that the *TaTLP*s are clustered and divided into three large subfamilies: A, B, and C. The A subfamily has two groups: AI and AII. This grouping matches previous *S. lycopersicum* reports [8,10,11]. The *TLP*s within each subfamily share a high degree of homology and have evolved close to one another [25]. Interestingly, we found that *TLP*s possess the F-box domain related to plant stress resistance [13,26,27]. This finding suggests that *TLP*s in wheat are highly conserved and may have additional functions, as demonstrated in Figure 3.

### *4.2. TaTLP Genes Are Thought to Be Involved in Critical Biological and Molecular Processes*

The GO analysis revealed that *TaTLP* genes perform a wide range of biological, molecular, and cellular functions (Figure 3). Many genes are directly involved in cell wall biosynthesis, which is the first line of defense against abiotic and biotic factors [28]. The TUBBY gene family appears to be essential for wheat plant growth in both normal and stressful conditions. Trans-acting elements are required for any biological or molecular process in plants. Multiple signaling pathways regulate plant stress responses, and there is

much overlap between the gene expression patterns induced by different stresses [29–31]. Several transcription factors influence the expression of stress-related genes in plants. Several closely related transcription factors can frequently activate or repress genes via cis-acting sequences in response to specific stresses [14,32]. In our study, many hormones (ABRE, TCA, TATC-BOX, AUXRR-Core, CGTCA, and TGACG), stress, and growth-related (ARE, ACE, G-Box, LTR, CAT-Box, O2-Site, MSA-Like) cis-elements were identified in the promoter region of *TaTLP* genes (Table 3). These elements are primarily involved in drought, low-temperature, and hormone responses [33,34].

The CGTCA and TGACG motif were found in nearly all *TLP* promoters, indicating that they were associated with the jasmonate acid response in most cases. Also found in most *TLP* promoters where the enzymes ARE (associated with anaerobic reaction) and ABRE (associated with ABA response) [7,8,13,25]. Based on these results, we suggest that *TaTLP*s may play an essential role in stress responses, but this needs further experimental verification. The PPI analysis demonstrated that *TaTLP*s interact with other essential proteins, such as ATG2G20050, which play an indispensable role in signal transduction, ATP binding, metal ion binding, and protein serine phosphatase activity. Similarly, the *TLP* gene interacted with ATG2G35680, AT3G12370, AT3G10330, pBRP2, AT2G45910, ENDOL9, ATGRIP, AT2G04940, and MLO1 (Table 2 and Figure 4).

Plant *TLP* gene families have been previously studied, and it has been discovered that multiple *TLP* genes are involved in the responses of plants to biological and abiotic stresses [7,8,10–13]. According to this, *TLP* genes can be used as candidate genes in plant resistance breeding.
