Cloning of SjCA gene and its expression analysis on upland cottons

Abstract

Acquisition of salt-tolerant genes from exogenous plants to improve cotton salt resistance has always been a hotspot of research on cotton salt resistance. However, the information regarding the method of conversion of living cotton pollens by portable gene gun technology is still little. Complete sequence information of the Carbonic Anhydrase (CA) gene was obtained from NCBI database, and its full ORF length sequence was cloned by RT-PCR technology. After the construction of pBI121-CA::GFP fusion expression vector, we used cotton pollens from upland cotton varieties Y-2067, ZA-23, and GZ-2, which have weaker autofluorescence, to conduct the research on the transient expression of cotton pollens in vivo via particle bombardment (gene gun technology). The results indicated that green fluorescence enhancement of the three kinds of cotton pollens was realized after the CA gene transformation, meaning that the CA gene expression level was increased. Besides, the salt tolerance germination ability of transgenic T1 seeds was also improved. Our research initially established a transient expression system of cotton pollen in vivo via particle bombardment technology, laying the theoretical foundation for further research of cotton genetic transformation and creation of cotton salt-tolerant germplasm.