Abstract:
Objective To study the chemically-induced expression characteristics of Arabidopsis thaliana methyltransferase gene AtMET1 in Populus alba × P. glandulosa '84K', to lay a foundation for the establishment of the poplar methylation-induced variation system and genetic improvement of poplars.
Method The chemically-induced promoter and AtMET1 were transformed into genome of P. alba × P. glandulosa '84K' using Agrobacterium-mediated transformation. The hygromycin resistant plants were obtained and transgenic plants were identified as by traditional PCR and DNA sequencing. The chemical inducer 17- β -estradiol was used to induce expression of AtMET1 in leaves of in vitro leaves of a transgenic line for 0, 3, 6, 12, 24, 48, 96 and 144 h, and the expression of AtMET 1 gene was detected by Quantitative Real-time PCR (qRT-PCR).
Result A total of 648 hygromycin resistant buds were obtained, among them 18 hygromycin resistant plants were screened, and all of them were identified to be transgenic plants by molecular detection method. qRT-PCR showed that the expression of AtMET1 reached its highest at 3 h by17- β -estradiol treatment, then decreased at 6 h, increased at 12 h and decreased again to less than half of the expression at 12 h.
Conclusion The chemical inducer can efficiently and rapidly induce the expression of AtMET1 in transgenic poplar plant, that laid a solid foundation for the regulation mechanism of MET1 in poplars and provided new idea and method for chemical induction of genes and genetic improvement of poplars.
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