Analysis of the promoter features of BpCUC2 in Betula platyphylla × Betula pendula
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CUC2 encodes a NAC domain transcription factor required for establishing boundaries around plant organ primordia. It is also associated with the initiation of the shoot apical meristem. We cloned a 1497-bp BpCUC2 promoter fragment and constructed a pBpCUC2::LUC vector for birch transgenic. A quantitative real-time polymerase chain reaction (qRT-PCR) analysis of transgenic birch buds, leaves, stems, and roots revealed that luciferase (LUC) was most highly expressed in the buds. The pBpCUC2::LUC transgenic birch plants were treated with methyl jasmonate, indoleacetic acid, abscisic acid, or gibberellic acid3 for 2 h. The subsequent qRT-PCR analysis indicated that the transgenic birch tissues responded differently to the various hormones. Our results confirmed that BpCUC2 expression is influenced by hormones. The PLACE online tool revealed that the BpCUC2 promoter sequence contains several cis-acting elements. Furthermore, an auxin response element was used to screen transcription factors in a yeast one-hybrid assay. We identified three unique cDNA sequences, with complete open reading frames containing regulatory motifs, which were related to growth and development.
KeywordsBetula platyphylla × Betula pendula Promoter expression feature Gene function Upstream regulatory gene screening
Shoot apical meristem
Betula platyphylla × Betula pendula CUC2
Synthetic dropout medium
This work was supported by the National Natural Science Foundation of China “Cloning and Functional Study on the Leaf Margin-related Genes of Betula platyphylla” Grant (Grant No. 31370660) and 111 Project (B16010). The funding source had no other involvement.
CL, HX: performed all the experiments. SW assisted with DNA sequence analysis. GL, JJ together with CL, HX and SW designed the experiments and wrote the manuscript. All authors read and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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