Functional characterization of OsHAK1 promoter in response to osmotic/drought stress by deletion analysis in transgenic rice
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The rice gene HAK1 (OsHAK1) is activated when the plant experiences drought stress. Here, a deletion analysis of the 3037 nt sequence lying upstream of the HAK1 translation initiation codon was carried out to identify which promoter region(s) are functionally important for its responsiveness to moisture stress. Four 5′ truncated sequences of the promoter (Dp1820, Dp1524, Dp1069 and Dp556) along with the intact sequence (Dp3037) were fused to GUS, and the resulting constructs transformed separately into rice. In the absence of any imposed abiotic stress, GUS activity was both ubiquitous and of similar intensity in plants carrying the Dp3037 and Dp1820 constructs, but was undetectable in those carrying either Dp1524, Dp1069 or Dp556. When the plants were subjected to moisture stress by the addition of polyethylene glycol to the culture medium, carriers of Dp3037 strongly expressed GUS, but carriers of Dp1820 did not respond, implying that a sequence(s) lying between the nucleotides − 3037 and − 1821 were responsible for osmotic stress inducibility. An in silico analysis of this key sequence revealed the presence of a number of putative cis-acting elements, including ABREs (abscisic acid-responsive) and DREs (dehydration-responsive). The 296 nt segment between the nucleotides − 1820 and − 1525 harbored both a CAAT-box and a TATA-box sequence. The Dp3037 sequence represents a potentially suitable candidate for regulating the expression of drought-responsive transgenes.
KeywordsOryza sativa Dehydration OsHAK1 GUS analysis Inducible promoter Cis-acting element
This work was funded by National Natural Science Foundation of China (Grant Nos. 31601811, 31671666 and 31871594); Zhejiang Province Outstanding Youth Fund (Grant No. LR19C130001).
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