Isolation, Characterization and Promoter Analysis of Cell Wall Invertase Gene SoCIN1 from Sugarcane (Saccharum spp.)

Abstract

Cell wall invertase (CIN) plays an important role in carbohydrate partitioning and regulation of sinksource interaction. In this study, a full length cDNA encoding CIN gene was cloned by RT-PCR and RACE-PCR from sugarcane. The open reading frame of 1731 bp encodes a protein of 577 amino acids with a predicted molecular mass of 141.04 kDa and theoretical PI of 4.67. A size of 4,537 bp DNA encoding SoCIN1 was isolated, which contains seven exons and six introns. Using genome walking approach, a promoter sequence of 974 bp of SoCIN1 was isolated, and prediction with PlantCARE and PLACE software revealed that it has several key cis-regulatory elements known to be involved in various biotic and abiotic plant stresses. Relative expression in specific tissues was done using qRT-PCR, and the results demonstrated that SoCIN1 expression level was higher in the immature leaves and maturing leaves than the matured leaves and internodes at elongation stage and processing maturing stage. The conditions of 15 % PEG and 6°C could have a similar function for inducing the expression of SoCIN gene in leaves, while 15 % PEG, 100 mM NaCl and 6°C could induce the expression of SoCIN gene in roots.

Expression analysis of Cell wall invertase under abiotic stress conditions influencing specialized metabolism in Catharanthus roseus

“…CrCWIN2 (CDS length: 1725bp; Genomic scaffold: cro_scaffold_3060381) had 7 exons and 6 introns whereas CrCWIN1 (CDS length: 1797bp; Genomic scaffold: cro_scaffold_3070386) and CrCWIN3 (CDS length: 1713bp; Genomic scaffold: cro_scaffold_3065222) were found to have 6 exons and 5 introns each. Previously characterized invertases from Agave tequilana 23 , Populus trichocarpa 24 , Sugarcane 25 and Cassava 26 have been shown to contain 6–8 exons. The genomic architecture of CrCWIN isoforms has been depicted in Fig.…”

“…These findings have been summarized in Table 1 . It is known that CWIN from Sugarcane, SoCIN1 encodes a protein 577 amino acids in length 25 and Arabidopsis thaliana CWIN with seven exons and six introns encodes 584 amino acids with mass 66.280kDa 27 , thus highlighting the molecular similarities among CWINs from C . roseus and other plants.…”

Diversity of nitrogen-fixing rhizobacteria associated with sugarcane: a comprehensive study of plant-microbe interactions for growth enhancement in Saccharum spp.

“…Real-time quantitative PCR (RT-qPCR) was conducted with SYBR Premix Ex Tap™II (TaKaRa, Japan) and the reaction was completed in RT-PCR System (Bio-Rad, USA). The total reaction volume was 20 μL, the composition of the reaction mixtures and conditions were followed by Niu et al [98]. The list of primers used is presented in Table 5.…”

Structural Properties, Production, and Commercialisation of Invertase

“…Multiple sequence alignment showed that amino acid sequences of homologous invertase contain three conserved amino acid regions, each consisting of NDPN, EC and RDP, which are highly conserved among the invertase and may constitute their catalytic domain (Hsieh et al 2006;Niu et al 2014;Yao et al 2014). These three conserved motifs are responsible for the catalytic action of invertase.…”