**3. Results**

#### *3.1. Identification and Analysis of TaTLPs Genes*

In the current study, 40 *TaTLP* proteins from wheat were retrieved using the Ensemble Plants (http://plants.ensemble.org/Triticum\_aestivum) (accessed on 11 March 2022) database. The genes were named based on their chromosomal position from *TaTLPq1* to *TaTLP40* (Table 1). Among these, *TaTLP2, TaTLP4, TaTLP5, TALP13, TaTLP16,* and *TaTLP35* genes were located in the *Extracellular* region, *TaTLP14* and *TaTLP24* in the mitochondrial region, and *TaTLP23* was located in the chloroplast, while the remaining 31 *TaTLPs* genes were found in the nucleus (Table 1). More details regarding *TaTLPs* were also recorded, including Locus ID, Proteins, and Molecular weight.

**Table 1.** The gene features of the wheat TUBBY gene family.


CDS: Coding Sequence, MW: Molecular Weight, SL: Sub Cellular Location, EC: Extracellular, N: Nuclear, PM: Plasma membrane, C: Chloroplast.

#### *3.2. Phylogenetic Analysis of TaTLPs*

We used the Neighbor-Joining method to construct a phylogenetic tree that included *Triticum aestivum, A. thaliana*, and *O. sativa TLPs* to investigate their phylogenetic relationship (Figure 1). The results showed that 40 *TaTLPs*, 15 *OsTLPs,* and 14 *AtTLPs* were clustered and further divided into three families, namely, A, B, and C. Furthermore, Family A was divided into two subfamilies, Family AI and Family AII. Family AI was the largest family containing most *TLPs* including 18 *TaTLP*s, (*TaTLP1, TaTLP6, TaTLP7, TaTLP9, TaTLP11, TaTLP16, TaTLP17, TaTLP20, TaTLP22, TaTLP25, TaTLP27, TaTLP28, TaTLP29, TaTLP30, TaTLP31, TaTLP38, TaTLP39, TaTLP40*), six *OsTLPs*, and three *AtTLPs.* The subfamily AII contained 14 *TLPs* including nine *TaTLPs* (*TaTLP2, TaTLP3, TaTLP4, TaTLP5, TaTLP13, TaTLP14, TaTLP18, TaTLP19, TaTLP21),* four *OsTLPs*, and one *AtTLP.* Family B was the second-largest family, containing 10 *TaTLP*s including (*TaTLP8, TaTLP10, TaTLP12, TaTLP15, TaTLP23, TaTLP24, TaTLP26, TaTLP33, TaTLP35, TaTLP37)*, four Os*TLP*s, and five At*TLP*s. Family C was the smallest, containing three *TaTLPs,* including (*TaTLP32, TaTLP34, TaTLP36,)* one *OsTLP,* and two *AtTLPs*. The results confirmed that the evolutionary relationships of *A. thaliana, O. sativa,* and *Triticum aestivum* are closer.

**Figure 1.** *TLPs* protein phylogeny in three plants: *T. aestivum, A. thaliana*, and *O. sativa*. MEGA 7 was used to generate the phylogenetic tree using the following parameters: Bootstrap D 1000 replicates, Neighbor-Joining method, Poisson correction. All group members are divided into four groups, each represented by an assorted color. Different labels are used to identify members of various species.

#### *3.3. Conserved Motif Analysis of TaTLPs-Genes*

A total of 10 conserved motifs were discovered using the MEME online server, and they were found to be appropriate for explaining the *TaTLPs'* structure (Figure 2). Among the 40 *TaTLP*s genes, *TaTLP7, TaTLP9, TaTLP22, TaTLP24,* and *TaTLP39* contained more than seven *TLP*s motifs.

**Figure 2.** The *TaTLP*s genes' conserved motifs.
