Identification And Analysis Of Specific Proteins Of Flowering Stage Of Sugar Beet Unit

Sugar beet hybrid M14 (2n=18+1) was produced by the backcross progenies between B. corolliflora and B. vulgaris. It has sexual and apomictic characteristics. In our previous work, differentially expressed protein spots of the floral organs after 1 h of flowering stage were compared by proteomic analysis between M14 (2n=18+1) and cultivated beet (2n=18) using 2D-PAGE. For the present work, two specific protein spots from M14 floral organs (M14F1h-1 and M14F1h-2) were purified and analyzed by mass spectrometry (MS) technique. Results show that M14F1h-1 contains 175 amino acids, and the "M" is the first amino acid whose protein score confidence level is in line with the requirements of data. The results are satisfactory. Although M14Flh-2 contains 231 amino acids, the first amino acid is not the "M" whose protein score confidence level does not meet the requirements of the data very much. Consequently, the results are less than ideal.The amino acid sequence of M14Flh-1 has a very high similarity and homology with ribulose 1,5-bisphosphate carboxylase or its small subunit. Based on the amino acid sequence of the mass spectrometry (24 M14Flh-1 primers,36 M14Flh-2 primers), the results of M14F1h-1-12 is consistent with the mass spectrometry results after amplified to the 3 ’end using RACE technology. In the 36 primers of M14Flh-2, since the amplification results of the M14F1h-2-3’s primers show out the dispersion state, the results are abandoned. The 5’RACE primers were designed by the 3 ’sequence of M14F1h-1-12, and were amplified the 5 ’sequence by SMART RACE. The full-length cDNA(m14f1h-1-12,854bp) was obtained that expresses the M14Flh-1. Then the full-length cDNA was analyzed. Results of translation are consistent with the result of mass spectrometry. Results of this cDNA were analyzed in the NCBI database. Its has a very high similarity and homology with ribulose 1,5-bisphosphate carboxylase or its small subunit. This work provides the evidence of differentially expressed proteins to further explain the reason that the progenies of M14 have the genetic phenomenon with different chromosome numbers.