Bvm14-cam Gene Transformation And Expression In Tobacco

Apomictic monosomic addition line M14 of Beta corolliflora Zoss. was the progeny from the hybridization of Beta corolliflora Zoss. and Beta vulgaris L. in sugar beet. Constituted of the normal 18 Beta vulgaris L. chromosomes with the No.9 chromosome of Beta corolliflora Zoss., M14 was found having a chromosome transmission frequency 96.5%. Thus M14 is one of the best materials for research of apomixis.298 specifically expressed ESTs were obtained from M14 in our lab, contrasted to a control of Beta vulgaris L. All of these ESTs had been sequenced and bioinformatics analysis was also performed. BvM14-CaM gene was expressed genes obtained from 298 ESTs by the method of RACE. Bioinformatics analysis results of BvM14-CaM gene indicated that BvM14-CaM protein had a high homology with Calmodulin protein. BvM14-CaM protein is intently interrelated with adversity of plants.In this research, BvM14-CaM cDNA fragment was transformed into model plant of tobacco, in order to study the relation between BvM14-CaM gene and antireversion force of plants, as well as by detecting the differential expression of BvM14-CaM protein between un-transformed and transformed plants. We constructed a eukaryotic expression vector pBI-M14-CaM-TNOS-PcaMV35S, and then transformed it into Agrobacterium tumefaciens EHA105. Leaf disc transformation was respectively performed to tobacco, and positive transformed plants were obtained. The result of analysis of PCR, RT-PCR, Northern blot and GUS reaction showed that BvM14-CaM gene was transformed into model plant of tobacco.The stress treatments of the transformed plants had been researched in low-temperature (4℃), high-temperature (42℃), high-permeability and high-salt. Analysis of transformed plants showed that, because of the transformation of BvM14-CaM gene, adaptability of the tobacco had been obviously improved.The results of our research are such as follows:the protein of the expressed gene BvM14-CaM in M14 has a similar function with Calmodulin protein. BvM14-CaM gene plays a very important role in plant physiology adversity.