Cloning and Ectopic Expression of <i>ScYCF</i>1 Gene from <i>Saccharomyces cerevisiae</i> in Cotton

Abstract

Yeast cadmium factor 1 (YCF1), is a member of the ATP-binding cassette (ABC) transporter family. To explore the functions of YCF1 of Saccharomyces cerevisiae (ScYCF1) in the cotton, ScYCF1 was cloned from Saccharomyces cerevisiae As2.375, with the full-length of 4548 bp. The bioinformatics analysis revealed that the largest component of ScYCF1 protein is leucine (12%). ScYCF1 is alkaline and positive charged, stable, and hydrophilic protein. The predictive secondary structure is mainly composed of α-helix areas, random coils and β-sheets. We constructed the pBI121-ScYCF1:GFP infusion expression vector and verified it by enzyme ingestion. The transient expression results of cotton pollen showed that the green fluorescence phenomenon of three kinds of upland cotton pollen significantly increased after transforming ScYCF1. The salt sensitive material upland cotton CCRI12 was transformed in vivo simultaneously, and the germination ability of trans-ScYCF1-gene T0 seeds was much better than the acceptor material CCRI12 under the stress of 100 mM NaCl saline solution. According to the gene nucleotide sequences, four pairs of primers were designed for molecular detection of T0 generation, and the sequencing results of PCR products of four specific primers evidence that the transgene is successful. Salt tolerance analysis of leaf discs of identified transgenic cotton showed that the chlorophyll content of leaf discs of transgenic cotton was higher than the content of the control cotton under salt stress. ScYCF1 gene was cloned and introduced into cotton, showing that ScYCF1 plays an important role in improving the salt tolerance of cotton.