Abstract:
The gene
GeDRP1E encoding dynamin-related protein 1E in
Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of
G. elata. Bioinformatics analysis on
GeDRP1E gene was carried out by using ExPASy, ClustalW, MEGA,
etc. Positive transgenic
Arabidopsis plant and potato minituber were obtained with the genetic transformation system of
Arabidopsis and potato. The plant height and seed setting rate of transgenic
Arabidopsis, and agronomic characters, such as size, weight and starch content of potato minituber of transgenic potato were tested and analyzed. And
GeDRP1E gene function was preliminarily investigated. The results showed that the open reading frame of
GeDRP1E gene was 1 899 bp in length and 632 amino acids residues were encoded, with a relative molecular weight of 69.90 kDa and a molecular formula of C
3079H
4973N
883O
933S
19. It was predicted that the theoretical isoelectric point was 7.27, the instability coefficient was 43.34, and the average hydrophilicity index was -0.259, which was indicative of an unstable hydrophilic protein. GeDRP1E has no transmembrane structure and signal peptide, and was localized in the cytoplasm. The phylogenetic tree showed that GeDRP1E was highly homologous with DRP1E proteins of other plant species, among which GeDRP1E had the highest homology with DcDRP1E (XP_020689662.1) in
Dendrobium candidum, reaching 90.05%.
GeDRP1E plant expression vector pCambia1300-35Spro-
GeDRP1E was constructed by double digests, and
Arabidopsis complementary mutant and potato overexpression strain of
GeDRP1E gene were obtained by
Agrobacterium-mediated gene transformation. Compared with the
Arabidopsis AtDRP1E mutant, the height and seed setting rate of the
GeDRP1E complementation mutant were rescued. The minituber of
GeDRP1E overexpression potato had larger size, heavier weight and higher starch content, comparing to wild-type potato. It was preliminarily induced that
GeDRP1E was involved in mitochondrial morphology regulation, which related to the growth and development of
Arabidopsis plants and potato miniature. The research results laid a foundation for further elucidating the molecular mechanisms underlying the growth and development of
G. elata tuber development.