Research Article
Molecular Cloning, Subcellular Localization and Expression Analysis of a BeCWINV1 Gene in Bamboo (Bambusa emeiensis)
Author Correspondence author
Tree Genetics and Molecular Breeding, 2022, Vol. 12, No. 5 doi: 10.5376/tgmb.2022.12.0005
Received: 08 Apr., 2022 Accepted: 15 Apr., 2022 Published: 20 May, 2022
Feng J.C., Li T.J., Yang Q.Q., Wang B.W., and Cao Y., 2022, Molecular cloning, subcellular localization and expression analysis of a BeCWINV1 gene in bamboo (Bambusa emeiensis), Tree Genetics and Molecular Breeding, 12(5): 1-10 (doi: 10.5376/tgmb.2022.12.0005)
Cell wall invertase (CWINV) is one of the key enzymes in sucrose degradation, which plays a key role in assimilate partitioning, the regulation of sink strength, and plant development. In order to explore the subcellular localization and expression level of CWINV gene in bamboo speices, a BeCWINV1 gene was cloned from the shoots of B. emeiensis based on the transcriptome database. The BeCWINV1 was 1 758 bp in the length of open reading frame, encoding a predicted protein of 585 amino acid. Sequence alignment and phylogenetic analysis showed that BeCWINV1 contains a cell-wall invertase active site (WECPD) motif, and shared higher identity with green bamboo Boβfruct1, rice CIN2 and maize INCW2, all of them belonging to a CWINV-specific cluster with other known CWINVs. Transient expression of the BeCWINV1-GFP fusion protein in the onion epidermis cells suggested that BeCWINV1 was located in the cell wall, indicating its role on sucrose hydrolysis in extracellular space. qRT-PCR analysis showed that the expression of BeCWINV1 was significantly higher in the stem than other tissues tested. The extra application of sucrose had no significant effect on the expression of BeCWINV1, but its expression was activated when the sugar supply was limited. These results suggested that BeCWINV1 might be involved in phloem unloading of B. emeiensis stem, and might also have an essential role in maintaining intracellular sugar concentration and sink activity under stress.