Arabidopsis CBF3 Allele Function Analysis And Regeneration Of Groundcover Plant

In our lab, with 32 core accessions of Arabidopsis, the single nucleotide polymorphism(SNP) study of CBF3 gene showed that there was one site mutation existing in the CBF3 allele of 203 AV accession, which caused terminal code ahead of 198bp compared with CBF3 of 186 AV accession (Columbia). This CBF3 allele was named CBF3-E151-. Compared with CBF3 encoding protein, CBF3-E151- encoding protein lacked C-terminal 66 amino acids. 203AV accession was more cold-resistant than 186AV accession, and SNP association analysis also showed CBF3 was related to cold-resistance. So we guess that the cold-resistant functions of CBF3-E151- might be more stronger than that of CBF3.In order to understand the functions of CBF3-E151- further. We cloned CBF3-E151- and CBF3. Structure analysis of CBF3-E151- was processed. The four plasmids harboring CBF3 or CBF3-E151' were constructed with CaMV 35S promoter or rd29a promoter respectively. The four plasmids were introduced into Arabidopsis Columbia accession (186AV accession). Transgenic plants were obtained by PCR test and kanamycin selection. Cold-tolerance test experiment showed that the transgenic plants overexpressing CBF3 or CBF3-E151' were more cold-tolerant than wild type Arabidopsis. The further study of cold-tolerance test showed the transgenic plants overexpressing CBF3-E151' was more cold-tolerant than those overexpressing CBF3.