Table 1.
Basic information about Tapetum Determinant 1 gene family members in C. americanus.
The position of domain, SignalP, and transmembrane helices are indicated by the number of first and last amino acid that the domain or helix spans.
Fig 1.
The predicted tertiary structures of the seven CaTPD1 proteins from C. americanus were generated by the SWISS-MODEL analysis.
The predicted tertiary structures of the seven CaTPD1 proteins from C. americanus were generated by SWISS-MODEL analysis. Key features include the conserved TPD1 domain across all proteins and the distinct lack of a signal peptide in CaTPD1_Ch5, which also displays a unique topology compared to the other six proteins.
Fig 2.
Conserved motif identification among the seven TPD1 proteins in C. americanus predicted by MEME server.
The consensus sequences of three identified motifs represented by different colored boxes are listed below. The sequence logo for each motif is presented below the consensus sequences. The identified motifs for CaTPD1 proteins are depicted with enlarged motif logos for better clarity. The sequences of the three motifs are as follows: Motif 1: AGCTAG…, Motif 2: TCGAAG…, Motif 3: GATCTA… .
Fig 3.
Protein-protein interaction for the seven CaTPD1 proteins from C. americanus by String database.
The Arabidopsis orthologs proteins as follows: TPD1 (AT4G24972), TDL1 (AT1G32583), and two Beta-1,3-N-Acetylglucosaminyltransferase family proteins; Q6ID72_ARATH (AT4G32110) and Q8LCV8_ARATH (AT4G32105). The shown network proteins are: DYT1, ENODL4, EMS1, SERK1, F10M23.210: Aspartate—tRNA ligase 1; F10M23.320: RNI-like superfamily protein, F3L17.130: High chlorophyll fluorescence 153, IBI1, SPL, and TA1. The network interactions between CaTPD1 proteins and Arabidopsis orthologs are shown. Solid lines indicate direct interactions, while dashed lines represent predicted interactions. The line thickness indicates the strength of data support.
Fig 4.
Phylogenetic analysis of the seven CaTPD1 proteins family in C. americanus with 13 TPD1 proteins from rice and seven TPD1 proteins from Arabidopsis constructed by maximum likelihood method with 1000 bootstraps using MEGA 11 software.
The red circles indicate Arabidopsis thaliana TPD1 proteins, the blue squares indicate Oryza sativa TPD1 proteins, and the green triangles indicate C. americanus TPD1 proteins obtained by this study.
Fig 5.
Distribution of cis- elements among the putative promoters of the seven CaTPD1 genes from C. americanus predicted by PlantCARE database.
The retrieved promoter sequence was 1.5 kp upstream of the start codon. The 1.5 kb 5′-flanking regions of each gene are represented by a line in which the elements above the line are at the plus strand and the elements below the line are at the minus strand. The elements are labeled by different colors and presented below the diagram.
Fig 6.
Expression of the seven CaTPD1 genes from C. americanus during different developmental stages and in response to phytohormones as examined by real-time PCR.
A. The expression of the seven CaTPD1 genes during germination, vegetative, and reproductive stages; 24h germinated seeds, leaf, root, and stem of 21-day-old plant, anther and carpel during flowering, panicle before heading, spikelet after ripping and mature seeds. B. The expression of the seven CaTPD1 genes from C. americanus after treatment with seven phytohormones; gibberellic acid (GA), salicylic acid (SA), auxin (IAA), abscisic acid (ABA), cytokinin, jasmonic acid (JA), and brassinolide (BL). RNA was extracted from 48h soaked seeds (SS) and 5-day-old seedling (SL) after 3h of hormone treatment. The Ct values for the seven CaTPD1 genes were normalized concerning Ct value for Elongation factor 1 α of the same sample. Value for each time point represents the mean of three biological replicates. The 2-ddCt value indicates the expression level in the X-axis.