Objective Regulatory network of HcWRKY71 in response to salt stress in kenaf was studied, and proteins interacting with the transcription factor identified.

Method Roots, stems, leaves, and phloem of Hibiscus cannabinus L. were used to construct a yeast two-hybrid cDNA library using the Gateway method. Proteins interacting with HcWRKY71 were screened, sanger-sequenced, and validated by means of one-to-one rotation.

Result The constructed yeast library had a capacity of 1.2×10⁷ CFU, a recombination rate of 100%, and an average insertion fragment length of over 1,000bp. Screened by the yeast two-hybrid, 48 positive clones with 25 different proteins were obtained with the unknown protein sequences deleted. In them, the MYB transcription factor 81, triple helix transcription factor ASIL2, and BAHD acyltransferase protein were postulated to be involved with the salt stress response of the plant.

Conclusion The large-capacity, wide-coverage, and high-quality yeast hybrid library for kenaf established by this study could become a useful tool for research. The selection of potential proteins that might interact with the transcription factor HcWRKY71 would aid future studies in elucidating the molecular mechanism of the plant under salt stress.