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Published on: April 17, 2015
The ArWRKY57-ArWRKY70-ArLEA5 module: Key regulators of drought tolerance in Anoectochilus roxburghii
Weiwei Jiang1, Aimin Lv1, Yuqiu Huang1
1National Key Laboratory for Development and Utilization of Forest Food Resources, Zhejiang A&F University, Hangzhou, 311300, China; Zhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, Zhejiang A&F University, Hangzhou, 311300, China.
Abstract:
Anoectochilus roxburghii, a valuable medicinal orchid, is severely threatened by drought stress. This study investigated its drought tolerance mechanisms focusing on late embryogenesis abundant (LEA) proteins and WRKY transcription factors (TFs). Physiological analyses showed that SLs treatment significantly reduced relative water content (RWC) and relative electrical conductivity (REC) compared to drought-stressed controls. Transcriptomic screening and qRT-PCR analyses identified ArLEA5/9 and ArWRKY57/70 as key drought-responsive genes, whose expression was upregulated under drought but suppressed by SLs. Overexpression of these genes in tobacco leaves and A. roxburghii protocorm-like bodies (PLBs) resulted in reduced water loss and enhanced drought resistance, with increased soluble proteins, proline, and decreased malondialdehyde (MDA) content. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BIFC), and yeast one-hybrid (Y1H) assays confirmed that ArWRKY57 forms a plasma membrane complex with ArWRKY70 and directly regulates ArLEA5 transcription. This study represents the first to identify candidate drought-resistant genes (ArLEA5, ArLEA9, ArWRKY57, and ArWRKY70) and characterize the ArWRKY57-ArWRKY70-ArLEA5 regulatory module as a key mechanism underlying drought tolerance in A. roxburghii, thereby facilitating molecular breeding for drought-tolerant medicinal orchids.
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