Cloning, Expression and Functional Analysis of GhrbcMT
Zhang Mengna, Shen Li, Nnaemeka Ekene Vitalis, Sun Yuqiang, Ke Liping*
The plant rbcMT gene probably encoded bifunctional enzyme as ribulose-l,5-bisphosphate carboxylase/ oxygenase large subunit (LS) N-methyltransferase, which catalyzes methylation of the ε-amino group of lys-14 in the LS of ribulose-l,5-bisphosphate carboxylase/oxygenase (Rubisco). The rbcMT gene was identified in peas, wheat, tobacco, possibly involved in plant growth and development, the function of GhrbcMT gene has not been reported. In this study, total RNA was extracted from G. hirsutum cv. C312, and cDNA was synthesized by reverse transcription. The full-length cDNA of GhrbcMT was isolated by utilizing PCR and analyzed by bioinformatics. The GhrbcMT gene was detected by real-time PCR (qRT-PCR) for gene expression pattern analysis. GhrbcMT was interferenced by virus induced gene silence (VIGS) technique in C312. The leaf chlorophyll of wild-type and GhrbcMT-silenced cotton plants was extracted by ethanol soaking. The results showed that there were multiple potential phosphorylation and glycosylation sites in GhrbcMT protein. Loop structure accounted for 56.94% which indicated the conformation of GhrbcMT was flexible. The GhrbcMT gene was specifically expressed in cotton leaves. The GhrbcMTi cotton plants significantly grew slowly, became wrinkled and sterile with GhrbcMT decreased expression. The GhrbcMT gene had significant effect on cotton growth, development and fertility. This study focused on exploring the function of GhrbcMT, which provides a reference for further exploring the mechanism of the growth and development even sterility in plants.
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