Differential Expression Analysis Of Floral Organ CDNA In Beet M14 Artificial Pollination

The additional chromosome of sugar beet M14 hybrization (2n=18+1) comes from Beta corolliflora Zoss which is considered to have apoximic characteristics. Previous embryological studies indicated that the reproduction pathway of M14 is of both sexual and apomictic traits. In our work, a series of crosses were established between M14 and Beta vulgaris L. (2n=18). And then RNA was extracted from flower buds. Subsequently, we used cDNA-AFLP to analyze different gene expression during flower bud development.Two hundred fifty-six selective primer combinations were used in amplifying transcript-derived fragments (TDFs). Our results show that 295 different expression fragments were observed only in the hybrid M14, not in the control-cultivated beet. Of 295 TDFs,50 TDFs were sequenced and compared their homologies by using basic local alignment search tool (BLAST). The result indicated that three fragment were similar with choline monooxygenase, S-adenosyl-L-homocysteine hydrolase and ClpC protease. Other TDFs are low homologous in the nucleic acids database. Further more, based on sequenced amplified fragments, primers with total of 50 pairs were redesigned and used for PCR amplication in the genomic DNA of M14 and cultivated beet, respectively. PCR and Southern hybridization confirmed that at least two TDFs came only from genomic DNA of the hybrid M14 and expressed during development of floral organs. This work provided a molecular basis for further analysis of genetic mechanism of M14.