Cloning, Characterization, and Functional Analysis of EuTIL1, a Gene-Encoding Temperature-Induced Lipocalin in Eucommia ulmoides Oliv

Eucommia ulmoides Oliver is a unique tertiary relict tree species in China belonging to the family and genus. It traditional precious Chinese medicinal herb with anti-tumor, antibacterial, antioxidant, anti-inflammatory, bidirectional blood pressure regulation effects. mainly grows temperate regions of China, but due its sensitivity low-temperatures, it difficult introduce into new regions. To study role lipocalin plants. This investigation was conducted utilizing gene cloning, bioinformatics analysis, quantitative real-time polymerase chain reaction (qRT-PCR), subcellular localization, stable genetic transformation transfer EuTIL1 Nicotiana tabacum Xanthi. The wild-type transgenic vector tobacco were cold-treated, corresponding protective enzyme activity cold-induced expression levels measured analyze functions genes. In this study, full-length temperature-induced (EuTIL1) cDNA cloned from leaves using rapid amplification ends (RACE) method. sequence analysis showed that 917 bp encodes protein 188 aa residues, which member Lipocalin-2 family. Subcellular localization revealed found plasma membrane. lines expressing under control CaMV 35S promoter had increased tolerance cold compared (WT) average water loss rate plants 12.4%, conductivity at 24 h 55.11%, malondialdehyde content significantly lower than maximum soluble sugar (SS) content, superoxide dismutase (SOD) activity, peroxidase (POD) catalase after low-temperature treatment 22.03 mg/g, 726.87 U/g, 1283.94 356.84 respectively, are higher those wild-type. Meanwhile, plants, NtDREB1, NtDREB2, NtDREB4, NtCOR15a elevated condition. conclusion, our demonstrates involved cold-stress response has potential enhance providing molecular basis for introduction serving as candidate evaluating cold-tolerant