Functional analysis of different promoter haplotypes of the coffee (Coffea canephora) CcDREB1D gene through genetic transformation of Nicotiana tabacum
Previous results showed that the three promoter haplotypes (HP15, HP16 and HP17) of the CcDREB1D gene (encoding the dehydration responsive element binding transcription factor) found in the drought-tolerant (HP15/HP16) and drought-sensitive (HP15/HP17) clones of Coffea canephora , diverged by severa...
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Veröffentlicht in: | Plant cell, tissue and organ culture tissue and organ culture, 2018-02, Vol.132 (2), p.279-294 |
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Zusammenfassung: | Previous results showed that the three promoter haplotypes (HP15, HP16 and HP17) of the
CcDREB1D
gene (encoding the dehydration responsive element binding transcription factor) found in the drought-tolerant (HP15/HP16) and drought-sensitive (HP15/HP17) clones of
Coffea canephora
, diverged by several single nucleotide polymorphisms and insertions/deletions. In order to compare the activities and regulation of these haplotypes in response to abiotic stresses, these sequences were cloned in front of the
uidA
and analyzed in transgenic tobacco (
Nicotiana tabacum
) for their ability to regulate the expression of this reporter gene by monitoring GUS histochemical activity under drought (mimicked by dehydration), heat shock and cold treatments. Under unstressed condition, GUS staining was mainly observed in leaf and root vascular tissues of young tobacco plants transformed by the longest sequences of
CcDREB1D
promoter haplotypes. These GUS activities were not observed in the same tissues of older plants as well as in plants transformed by shorter proximal regions, suggesting a developmentally-regulated activity of
CcDREB1D
promoters in tobacco and the existence of
cis
-regulatory elements essential for their regulation in distal regions. Under dehydration and heat shock conditions, GUS staining detected in leaf midribs and secondary veins of pHP17L-transformed plants was correlated with up-regulated expression of
uidA
reporter gene while no GUS activities were observed in pHP16L-transformed plants. However, all
CcDREB1D
promoter haplotypes were positively regulated by cold stress in transgenic tobacco. These results showed that these coffee promoters were recognized by the tobacco transcriptional machinery but were regulated in different manners in response to abiotic stress. |
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ISSN: | 0167-6857 1573-5044 |
DOI: | 10.1007/s11240-017-1328-y |