JIA-2019-11

2508 LI Li-shu et al. Journal of Integrative Agriculture 2019, 18(11): 2505–2513 were analyzed by transgenic and WT A . thaliana after 7 d salt treatments. Each transgenic line was treated with 10 seedlings. The plant length, root length, hypocotyl length and leaf area were also calculated for plants growing for 15 d in 1/2 MS medium. 2.6. Statistical analysis All data are expressed as the mean±standard error and analyzed by SPSS 17.0 Software. Three biological replicates were used for analysis. 3. Results 3.1. Isolation and characterization of MieIF1A-b The full-length coding sequence of MieIF1A-b was 435 bp in length (Appendix A), encoding a 145-amino acid protein (Fig. 1-A; GenBank accession no.: KP676599). The MieIF1A-b protein had an estimated molecular mass of 16.39 kDa and an isoelectric point (pI) of 4.6. Protein sequence alignments showed that eIF1A family members were highly conserved with each other and eIF1A-b contained a conserved translation initiation factor ( eIF ) domain between 18–145 amino acids (Fig. 1-B). MieIF1A-b had 96, 95, and 92% amino acid sequence identity to EgeIF1A (XP_010915988.1), MeeIF1A (XP_021633493.1), and PteIF1A (XP_002302848.1) , respectively. Phylogenetic tree analysis showed that MieIF1A-b was very close to EgeIF1A (XP_010915988.1), and these genes clustered together (Fig. 1-C). 3.2. Expression analysis of the MieIF1A-b in mango qRT-PCR was used to investigate the expression of MieIF1A-b in different tissues, in different fruit development stages, under abiotic stresses and exogenous hormone treatments. MieIF1A-b was expressed in all tested tissues. A B C Translation initiation factor ( eIF ) conserved domain 0.02 100 90 90 47 52 46 62 EgeIF1A PteIF1A AteIF1A TceIF1A GheIF1A GmeIF1A OseIF1A ZmeIF1A MieIF1A-b MeeIF1A Fig. 1 Sequence analysis of eukaryotic translation initiation factor ( eIF1A ) in Mangifera indica ( MieIF1A-b ). A, nucleotide and predicted protein sequence of MieIF1A-b . B, multiple sequence alignment of MieIF1A-b with homologous proteins from other species, conserved residues are shaded in black line. C, phylogenetic analysis of plant eIF1A , from Elaeis guineensis ( EgeIF1A , XP_010915988.1); Manihot esculenta ( MeeIF1A , XP_021633493.1); Populus trichocarpa ( PteIF1A , XP_002302848.1); Arabidopsis thaliana ( AteIF1A , AAM10261.1); Theobroma cacao ( TceIF1A , XP_007049245.2); Gossypium hirsutum ( GheIF1A , XP_016740250.1); Glycine max ( GmeIF1A , XP_003548603.1); Oryza sativa ( OseIF1A , CAD91551.1); Zea mays ( ZmeIF1A , NP_001148997.2).