Biomics

The features of expression of the uidA reporter gene under the control of tissue-specific promoters of the apetala3 (ap3) and rpt2a genes of Arabidopsis thaliana L. in the transgenic callus culture of Nicotiana tabacum were studied. The starting material for obtaining callus culture was previously created monoinsertion homozygous (T2) transgenic tobacco plants obtained by agrobacterial transformation. Callus cultures were obtained from two types of explants (stem, leaf) and were cultivated on three cultural media that differed in the ratio and amount of plant hormones auxin (NAA) and cytokinin (kinetin). In the studied callus lines, variability was revealed both in the enzymatic activity of β-glucuronidase and in the accumulation of mRNA. No reliably significant differences in the expression of the uidA gene were detected between different media for each type of explant and between types of explant, which suggests the insensitivity of these promoters to changes in the hormonal composition of nutrient media. The level of expression of the uidA gene under the control of the tissue-specific promoters of the ap3 and rpt2a genes of A.thaliana in callus lines was lower than the level of expression of the uidA gene under the control of the 35S CaMV promoter in the control callus line. The level of uidA reporter gene transcript accumulation did not correlate with the level of β-glucuronidase enzymatic activity. Histochemical analysis of transgenic calli revealed that in all lines, different color variants were present with different frequencies (fully colored calli, mosaic colored and not colored at all), which indicates epigenetic changes associated with the loss of reporter gene expression in vitro. In general, the results of the analysis showed that the promoters of the ap3 and rpt2a genes of A. thaliana are active in tobacco callus culture and can be effective as regulatory elements in synthetic promoters.  

Arabidopsis thaliana ap3 gene promoter, Arabidopsis thaliana rpt2a gene promoter, callus, gene expression, Nicotiana tabacum

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