Verifying the presence and transcription of ZmLEA14A gene in Nicotiana tabacum line NiC9-1

Abstract

The function of LEA proteins in plants that has received much attention recently is the cell wall protection against abiotic stresses. Improving the expression of  LEA genes to enhance plant drought tolerance has been carried out by various genetic engineering applications for many years. In maize, the LEA genes are evenly distributed over ten chromosomes and classified into nine major groups. Among these LEA genes, ZmLEA14A belongs to 5C group and encodes for an open reading frame (ORF) of a 152-amino-acid polypeptide chain. In the current study, the ZmLEA14A gene was isolated from Tevang1, a drought-tolerant maize landrace, and transferred into the NiC9-1 tobacco line. The expression of this transgene was driven under the control of a minimised 35S promoter. The transgenic tobacco plants developed normally at all different growth stages with a survival rate of 89.2% in the laboratory condition. The ZmLEA14A gene was successfully inserted into the tobacco genome, and the presence of the transgene was verified in 7 transgenic lines by PCR. Applying the RT-PCR technique in three transgenic lines showed that the ZmLEA14A gene was currently expressed at the transcription level. In conclusion, the structure harboring the ZmLEA14A gene could be used to transfer genes to other target crops to improve the drought tolerance.