TRANSFORMATION OF TOBACCO WITH EJLFY FROM LOQUAT AND MOLECULAR IDENTIFICATION OF TRANSGENIC PLANTS

H.W. Song, Y.X. Liu, S.Q. Lin, G.B. Hu
The gene ejLFY related to floral formation from loquat (Eriobotrya japonica Lindl.) was transformed to tobacco by the method of leaf disk mediated by Agrobacterium tumefaciens. A total of 46 Kan-resistant tobacco plantlets regenerated from leaf explants via organogenesis were obtained. With PCR technique, each tobacco plant was analyzed for the genes, ejLFY, 35SP, nptII and chvA, and 6 transformed plants were identified preliminarily. Three of them were further analyzed using T-DNA flanking sequence method. The result verified two transgenic tobacco plants, in which a 359 bp unknown fragment linked to the right border of T-DNA. BLAST search results showed the fragment had above 96% homology with the genomic or cDNA sequences from ten plant species such as Arabidopsis, rice (Oryza sativa) and citrus. These suggested T-DNA sequence containing the target gene ejLFY had been integrated into the two tobacco plants. Besides, it was also known from T-DNA flanking sequence that ejLFY was single-copied and the inserted sites of T-DNA sequence were similar in different transgenic tobacco plants. Finally, RT-PCR assay exhibited that the exogenous ejLFY and nptII had expressed on the level of mRNA in the transgenic tobacco plants. However, the transgenic tobacco plants did not flower early. Possible reasons were discussed.
Song, H.W., Liu, Y.X., Lin, S.Q. and Hu, G.B. (2007). TRANSFORMATION OF TOBACCO WITH EJLFY FROM LOQUAT AND MOLECULAR IDENTIFICATION OF TRANSGENIC PLANTS. Acta Hortic. 750, 233-240
DOI: 10.17660/ActaHortic.2007.750.36
https://doi.org/10.17660/ActaHortic.2007.750.36
Eriobotrya japonica, ejLFY gene of loquat, transgenic tobacco, PCR detection, T-DNA flanking sequence analysis, RT-PCR
English

Acta Horticulturae