Serk And Bbm Conservative Fragment Of The Gene Was Cloned In The Sugar Beet In The M14 Strain

Beta vulgaris Line M14 was a monosomic alien addition line of Beta corolliflora in Beta vulgaris, which consisted of the normal eighteen B. vulgaris chromosomes plus B. corolliflora the ninth chromosome, and it was found to have a high chromosome transmission frequency among successive generations. Line M14 was evaluated to be a facultative apomixis material with morphology, genetics, cytology and embryology. It was predicted that the apomixis genes of Line M14 were located in the additional ninth chromosome of B. corolliflora. Line M14 was superior to those wild apomixis materials in several aspects: the size of its genome was much smaller, its genetic property was better known, and the additional ninth chromosome of B. corolliflora was comparatively independent. For these reasons, B. vulgaris Line M14 was thought to be a perfect material to clone the genes related to apomixis.At present, the expression of SERK gene has been observed in many plant species, such as Daucus carota, Arabidopsis thaliana, Zea mays, Medicago truncatula, Helian thusannuus, Poa pratensis, Oryza sativa, Cocos nucifera, Theobroma cacao and Coffea canephora; the expression of BBM gene has been observed in Brassica napus, Arabidopsis thaliana and Medicago truncatula. SERK gene was extremely correlated with the induction of somatic embryogenesis, for each single cell in which SERK gene was expressed had the ability to form a somatic embryo. Ectopic expression of BBM in Arabidopsis and Brassica led to the spontaneous formation of somatic embryos; the expression of BBM gene also induced hormone-free regeneration of explants and alterations in leaf and flower morphology. It was presumed that the apomixis of Line M14 was related with SERK and BBM gene. To valuate whether SERK and BBM gene were expressed or not in Line M14 and to isolate the specific amplified fragments of those two genes would contribute fundamentally to finding out the heredity mechanism of the apomixis of Line M14 and to eventually cloning those genes related to apomixis.In this research, the SERK conservative sequence was identified through the multiple alignment of the SERK sequences of Daucus carota, Arabidopsis thaliana and Zea mays in GenBank with the ClustalX 1.81 program. The SERK fragment was cloned in Line M14 with PCR based on the primers designed according to the SERK conservative sequence using the Primer Premier 5.0 program, and it showed 81.9% similarity to the SERK sequence of Arabidopsis, 65.5% to that of Daucus and 64.4% to that of Zea mays. The BBM conservative sequence was identified through the pairwise alignment of the BBM sequences of Arabidopsis thaliana and Brassica napus in GenBank with the ClustalX 1.81 program. The BBM fragment was cloned in Line M14 with PCR based on the primers designed according to the BBM conservative sequence with the Primer Premier 5.0 program, and it showed 86.6% similarity to the BBM sequence of Arabidopsis and 70.3% to that of Brassica. The results found in this paper strongly suggested that SERK and BBM gene be expressed in B. vulgaris Line M14.