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| Cloning and Expression Analysis of Phosphomannomutase Gene in Anoectochilus roxburghii |
| LI He-Ping, LIN Jiang-Bo, HUANG Hui-Ming, ZOU Hui, DAI Yi-Min* |
| Institute of Subtropical Agriculture, Fujian Academy of Agricultural Sciences, Zhangzhou 363005, China |
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Abstract Polysaccharide is one of the main medicinal components in Anoectochilus roxburghii. To further understand the biosynthetic pathway of A. roxburghii polysaccharides, the phosphomannomutase (PMM) gene was cloned and prokaryotic expressed in this study. In addition, the bioinformatics, subcellular localization, and expression regulation were analyzed. The results showed that the ORF region of ArPMM gene was 759 bp in length, encoding 252 amino acids, and the molecular weight of fusion protein expressed by prokaryotes was 30.75 kD. Bioinformatics prediction showed that the PMM protein belonged to the haloacid dehalogenase (HAD) superfamily, and was a stable hydrophilic protein without a signal peptide and transmembrane structure. The ArPMM protein was found, for the first time, located in the nucleus by transient expression method, although the PMM was believed located in the cytoplasm in previous reports. Phylogenetic tree analysis showed that ArPMM protein was closely related to Dendrobium officinale, D. huoshanense, and Phalaenopsis equestris. The result of gene expression experiment revealed that the ArPMM gene was expressed in multiple tissues, and the highest relative expression level was detected in flowers while the lowest expression was found in stems. The ArPMM gene was up-regulated by salt stress, and it was speculated that ArPMM might catalyzing the transformation of mannose-1-phosphate to mannose-6-phosphate. Meanwhile, the ArPMM gene was also regulated by temperature, both high and low temperatures inhibited its expression, and ArPMM gene was sensitive to high-temperature treatment. This study provides basic materials for further research of ArPMM gene participating in the mannose metabolism pathway of A. roxburghii.
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Received: 01 March 2023
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Corresponding Authors:
*dymttcn@163.com
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