This study aimed to investigate the signaling pathway of LpNAC14,a NAC transcription factor in Lilium pumilum,in response to salt stress,to analyze the protein interaction,so as to provide a basis for the further study of the functions of the related genes.RNA collected from the leaves and roots of L.pumilum under salt stress was used to construct a yeast two-hybrid cDNA library using the Gateway method.Two LpNAC14 gene fragments without self-activating activity were used to construct bait vec-tors,and then they were co-transformed into the library plasmids to screen for interaction proteins.The screened results were subjected to yeast validation and GO enrichment analysis,and one of the transcription factors,LpDi19-2,was selected for BiFC validation with LpNAC14.The yeast library was constructed with a library capacity of 1.12×107 CFU,with a recombination rate of 100%,and the average length of the in-serted fragments was greater than 1 000 bp.Nineteen proteins potentially interacting with LpNAC14 were preliminarily screened by library transformation.BiFC verification test showed that LpDi19-2 interacted with LpNAC14 in vivo.It is concluded that the quality of the constructed yeast library of L.pumilum un-der salt stress is high,which can meet the screening criteria,so that the screening results are reliable.This study provides a new way to explore the mechanism of LpNAC14 in response to salt stress.
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