Isolation And Functional Analysis Of PmARF Gene In Japanese Apricot

Japanese apricot(Prunus mume Sieb.et Zucc.),originated in China,belongs to Rosaceae.The flowers of which have high ornamental value,the fruit of which was rich in nutrients.However,the phenomenon of pistil abortion widely occurs in Japanese apricot and has seriously affected the yield in production.We used ’Daqiandi’ in the ’National Field Genbank for Japanese apricot’ as experimental material.PmARF11 was isolated through Rapid Amplification of RT-PCR.Gene structural characteristics were analyzed by the bioinformatics software;quantitative real-time PCR(qRT-PCR)were carried out.The PmARF11 gene functiong was preliminaries identified by transferred it into tobacco(Nicotiana).The main research results are as follows:1 Auxin has been widely implicated in various aspects of plant growth and development,including flower development.In order to further elucidate the role of auxin during flower development,especially on the pistil development process,auxin response factors(ARFs),an important component in auxin signalling pathway,were studied in the early flower buds of Japanese apricot(Prunus mume Sieb.et Zucc).In this study,a comprehensive overview of the ARF gene family in Japanese apricot is presented,including the chromosomal locations,phylogenetic relationships,gene structures,the domain and nuclear localization analysis.Seventeen Japanese apricot genes that encode ARF proteins(PmARFs)have been identified based on the genome sequence of Japanese apricot.Comparison of the expression of some PmARF genes between perfect and imperfect flower buds in Japanese apricot suggests that PmARFs,especially the PmARF11 gene may be required for pistil development and function in Japanese apricot.These results will be useful for future functional analyses of the ARF family genes.2 The PmARF11 cDNA was obtained by using RT-PCR.The structural characteristics were analyzed by the bioinformatics software.The results showed that the PmARFll gene ORF which encoded a 711 amino acid polypeptide with a calculated molecular weight of 78.663 kD and an isoelectric point of 5.80.3 We constructed an overexpression vector of PBI121-PmARF11 and transferred it into tobacco by Agrobacterium tumefaction,obtained Kan positive tobacco plants.Analyses of GUS straining,PCR and RT-PCR showed that obtained 3 positive tobacco plants.The observer of transgenic tobacco lines and wild-type lines showed that under the same culture conditions,compared with the control,transgenic the petal color of transgenic tobacco plants became dark,the pistil became longer and the flowering became extend respectively.In addition,PmARF11 plays a major role in regulating auxin-mediated leaf longevity and the plant life cycle.In summer,PmARF11 ptotein is possible a pleiotropic developmental regulator in plant.