Cloning of the Vacuolar H+-pyrophosphatases Gene BnVP1 from Ramie (Boehmeria nivea) and Expression Analysis Under Cadmium Stress
ZHU Shou-Jing1,2,*, SHI Wen-Juan1
1 College of Life Science and Resources and Environment, Yichun University, Yichun 336000, China; 2 Jiangxi Key Laboratory of Regulation and Control of Crop Growth and Development, Yichun 336000, China
Abstract：Cadmium is a heavy metal pollutant with high toxicity to many organisms, which enters into the environment along with industrial and agricultural activities. Vacuolar sequestration is an important mechanism to coping with cadmium stress in plants. Vacuolar H+-pyrophosphatase (V-H+-PPase), a proton pump in vacuolar membrane of plant cells, plays important roles in the transmembrane transport of inorganic ions. In this study, a V-H+-PPase gene, BnVP1 (GenBank No. MW029619) from ramie (Boehmeria nivea) was isolated by the rapid amplification of cDNA ends (RACE) method based on the previous transcriptome analysis of ramie under cadmium stress. The open reading frame sequence of BnVP1 was 2 292 bp and encoded 763 amino acids. BnVP1 was a transmembrane protein belonging to H_PPase superfamily, which contained 15 transmembrane domains. BnVP1 shared high amino acid sequence similarity (90.04%~92.27%) with typeⅠ V-H+-PPase proteins from Prunus persica (XP_007208066.1), Malus domestica (XP_008359855.1), Theobroma cacao (XP_017977384.1) and Gossypium raimondii (XP_012476518.1). The promoter sequence of BnVP1 contained several stress-related cis-acting elements. qRT-PCR showed that the expression of BnVP1 had no tissue specificity. The expression of BnVP1 in the root and leaves of ramie increased rapidly to maximum under cadmium stress, which indicated that the expression of BnVP1 was significantly induced by cadmium. The study enriches the research content of the molecular mechanism of cadmium tolerance in ramie, and provides a reference for the study of cadmium tolerance mechanism of Bnvp1 gene.
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