Cloning Of BoFBX156 Gene Responding To Low Temperature And Its Expression Analysis In Brassica Oleracea Flower Buds

Brassica oleracea,referred to as cabbage,is a plant belonging to the genus Brassica in the family Cruciferae.The place of origin is the Mediterranean coast,because of its strong environmental adaptability,rich nutritional value and easy storage and transportation characteristics.So it can be widely planted around the world.Cabbage is the main vegetable in Europe,America and other countries.It is also one of the main leafy vegetables in China.In winter,it is easy to be affected by low temperature,which will lead to yield reduction or even crop failure.The genes encoded by the F-box family involved in the regulation of plant life activities have been shown to play an important role in regulating plant organ development and responding to stress.But at present,there are no reports about the research of F-box family genes in cabbage.In this study,"BoJF-16-1" of the cabbage was used as the material,and used RT-PCR technology to clone the length of the coding region of the cabbage BoFBX156 gene.Through the analysis of physical and chemical properties,amino acid sequence and phylogenetic tree construction of BoFBX156 gene,the theoretical basis was provided for the further study of the biological function of F-box gene in cabbage.The main results are as follows:(1)In this experiment,the flower bud of the cabbage variety“BoJF-16-1”was used as the experimental material.According to the known gene sequence information,the specific primers of the gene were designed by software.The length of the coding region of sequence of BoFBX156 gene was cloned by RT-PCR.The BoFBX156 gene sequence cloned in cabbage was 1035 bp in length,encoding 344 amino acids,and the Gen Bank accession number was XM?013846145.2.(2)The BoFBX156 gene is localized in the cytoplasm by subcellular and is a stable hydrophilic protein with a conserved domain of the F-box protein family.The homology analysis of amino acid sequences revealed that the protein encoded by the BoFBX156 gene of Brassica oleracea had the closest relationship with Brassica napus F-box protein At1g70590-like,and the homology reached 99.71%.Secondary structure analysis found that the alpha helix accounted for 50.87%,and the random coil accounted for 35.75%.(3)Real-time fluorescence quantitative PCR results show: when treated at 4? and 2?,the expression of F-box gene BoFBX156 was the highest in the cabbage "BoJF-16-1" variety,followed by the variety "BoZG-16-1",The lowest in the variety "BoCF-16-1";when treated at 0 ?,the expression of the F-box gene BoFBX156 was highest in the cabbage "BoZG-16-1" variety,and the variety "BoJF-16-1" Second,the expression level is the lowest in the variety "BoCF-16-1".With the prolongation of low temperature stress,the gene expression in "BoZG-16-1" was higher than that in "BoJF-16-1" and "BoCF-16-1" at the treatment time of 12 h.The BoFBX156 gene is expressed in the development of flower buds of different varieties,and the gene expression amount increases with the development of flower buds.The BoFBX156 gene of cabbage was the highest in the binuclear pollen stage of flower bud development,followed by the mononuclear pollen stage in flower bud development,and the lowest in the quadrant stage of flower bud development.