Isolation and characterization of sucrose phosphate synthase promoter from cotton (Gossypium hirsutum L.)

Abstract

Cotton fiber genes and promoters are of great importance in understanding fiber development mechanism as well as for improvement of fiber. Sucrose phosphate synthase gene (SPS) (insert abbreviation of gene)has found to express at higher rate in developing cotton fibers. It is an important enzyme that have major role in sucrose as well as cellulose synthesis. Upstream region of a SPS gene from cotton was retrieved through HTGS database and analyzed using bioinformatics tools. Sequence analysis identified various regulatory motifs including light, drought, heat responsiveness and MYB binding sites in the promoter. The SPS promoter was isolated from cotton genomic DNA (using what method) and fused to β-glucuronidase (GUS) gene in a plant expression vector. Transient GUS expression analysis in various cotton tissues showed that promoter was active in the fiber tissues. Full length 2 kb SPS promoter showed high expression in fibers during elongation and secondary cell wall synthesis stage. A 1.5 kb deletion fragment showed reduced expression in fibers. Our results suggest that cotton SPS promoter may be used to express genes specifically in fiber cells for improvement of cotton fiber quality traits.

Expression Analysis of Fiber Related Genes in Different Staple Length Genotypes in Cotton (G. Hirsutum L.)

“…A variable and unstable level of expression of GA-20 Oxidase was detected in all three lines at first two stages including 0 and 5 DPA but was much higher at 0 DPA in CIM-707. A large number of transcript pattern of GA-20 Oxidase were also detected in CYTO-179 as compared to other lines of G. hirsutum (Iqbal et al, 2016;Nadeem et al, 2021).…”

“…At SCW (20 DPA) phase of fiber expansion, nearly a small (negligible) transcript pattern was also observed in CIM-616. The expression level of PEPc was about to be doubled at 15 DPA stage in CIM-707 fibers than the others DPAs but the transcript level of PEPc was ultimately dropped at the same stage in both medium and short fiber lines (CYTO-179 and CIM-616 (Iqbal et al, 2016;Nadeem et al, 2021).…”

“…Cell wall expansion starts immediately after the termination of initiation phase during the long fiber development in cotton (Ruan et al, 2004). The process of fiber cells elongation starts with breakdown of hydrogen bonds between the cellulose and hemicellulose fibrils along with the overexpression of fiber elongation enhancement genes including expansin (EXP), sucrose synthases (sus), GhCaM-7, LTPs, PEPc, tubulins, and XTH, responsible for long staple length in cotton (Liu et al, 2015;Cheng et al, 2016;Iqbal et al, 2016;Balasubramanian et al, 2016). (Iqbal, 2017) reported that the activation of these long staple length-related genes at elongation stage are thought to have necessary for the opening and closing of plasmodesmata, resulting in the intake of water and solutes.…”

“…So, the fiber elongation genes especially PEPc are believed to have a positive correlation with fiber elongation. In biotechnology studies, RT-PCR analysis provided the evidences of the strong link of PEP Carboxylase with the elongation and expansion of fiber cells (David et al, 2016;Iqbal et al, 2016;Zhao et al, 2018).…”