LEAFY COTYLEDON2 gene expression and auxin treatment in relation to embryogenic capacity of Arabidopsis somatic cells

Abstract

The expression pattern of the LEC2 gene during somatic embryogenesis (SE) in Arabidopsis explants (immature zygotic embryos) induced in vitro was followed, using real-time quantitative PCR (qRT-PCR). The analysis revealed differential expression of LEC2 transcripts within a 30 days time course of somatic embryo development. A significant auxin-dependent upregulation of the LEC2 gene was found to be associated with the induction phase of SE. In contrast to embryogenic culture the level of LEC2 expression was noticeably lower in non-embryogenic callus of Col-0 and hormonal mutants (cbp20 and axr4-1) with low SE-efficiency. The study with 35S::LEC2-GR transgenic plants showed that overexpression of LEC2 can compensate for the auxin requirement, and that transgenic explants formed somatic embryos when cultured in vitro under auxin-free conditions. However, unlike in auxin-induced SE, intense callus formation preceded the embryogenic response triggered via LEC2 overexpression, suggesting an indirect pathway of morphogenesis. Moreover, a negative interaction between auxin treatment and LEC2 overexpression in terms of SE efficiency was observed, as transgenic explants cultured on auxin medium displayed a significantly reduced level of embryogenic potential. The study provides further experimental evidence that in the determination of the embryogenic response in Arabidopsis somatic cells, a close link exists between auxin and the LEC2 activity.