| Lily (Lilium spp.) is considered as one of the five world-famous cut flowers,provided with so many good characters of big petals, rich color, beautiful flowerarchitecture and good implied wishes for lovers, and loved by people all over the world.However, many existing problems on lily (e.g. flower architecture simplification, singlepetal, too many pollens etc.) are expected to be solved in the practice. So how to create andculture new varieties of lily with improved flower architectures (such as no pollen, multi-petals and so on) is always the hotspot scientists focus on.In this study,14homologous genes related to flower development and subordinate to CGroup, named AGAMOUS (AG) genes, were respectively cloned from7lily cultivars of3strains, and analyzed by bio-software. Furthermore, expression characteristics of AG genewere studied and antisense plant expression vectors of AG gene were constructed. Themain results are as follows:1. A pair of degenerate primers was designed according to the reported AG genesoriginated from lily and other materials, and14genes were cloned by RT-PCR method.The new-cloned genes had high homology with the reported AG genes in the Genbank,which initially inferred that all new-cloned genes related to flower development belongedto C family.2. Based on analysis of amino acid sequences inferred from14nucleotide sequencesand their structures, the result showed that every amino acid sequence contained thehighly-conserved area (M region), and also had both AG I and AG II conserved regionswhich were regarded as one of the most important traits to indent C family gene.14newgenes cloned in this study were classified and their identifications were further confirmed.3. Expression characteristics of AG gene were tested by using semi-quantitativeRT-PCR detection method. The result indicated that AG gene only expressed in the stamenand pistil tissues, which coincided with the classic flower development theory of ABCmodel; that expression level of AG gene existed difference at the different developmentstages in the same organ.4. Based on analysis of the homology between new-cloned AG genes and reported ones cloned from closely related plants, the result demonstrated that the genes cloned in thisstudy had high homology with AG genes in Genbank cloned from lily genus, reach to91percent.5. A pair of new specific primers was designed and synthesized according to theintegrated analysis of sequencing results and vector structure; furthermore, appropriaterestriction sites were introduced. Antisense plant expression vector pCLtAG1withLtAG1gene and pCLtAG2with LtAG2promoted by CHSA were successfully constructed.It laid the foundation for their future genetic transformation. |
