Preliminary Analysis On The Function Of LpBCP And LpCPC Genes In Lilium Pumilum

Saline-alkali stress is an abiotic stress that restricts plant growth and development.Lilium pumilum is not only an ornamental plant,it has strong resistance to disease,cold,early,salt and alkali resistance and other resistance.Lilium pumilum is an excellent plant material in the study of saline-alkali stress.In this study,LpBCP and LpCPC genes were cloned from Lilium pumilum based on transcriptome data obtained in the laboratory.Both of them belongs to blue copper protein family.The LpBCP and LpCPC genes were bioinformatics analyzed,and the expression patterns of the genes were also analyzed.The LpBCP and LpCPC genes were transformed into yeast,tobacco and Lilium pumilum,respectively,and the transgenic strains and lines were obtained.After NaHCO3 stress treatment,the phenotypes of the transgenic yeast and transgenic plants were observed and analyzed.The results laid a foundation for further study of LpBCP and LpCPC genes in Lilium pumilum.The specific results are as follows:1.The LpBCP and LpCPC genes were cloned from Lilium pumilum.For bioinformatics analysis and gene expression pattern analysis:LpBCP and LpCPC proteins are likely to contain transmembrane protein and contain a signal peptide of hydrophobic protein,protein secondary and tertiary structure structure are some screw and curl,LpBCP protein localization may be in the cell membrane,protein LpCPC orientation may be outside the cell.The homologous sequences of LpBCP and LpCPC amino acids were compared with those of other species by NCBI,and then the phylogenetic tree analysis was carried out.The results showed that LpBCP was relatively close to Eleis guineensis,while LpCPC was more close to Solanum lycopersicum and Trifolium pratense.2.The expression characteristics of LpBCP and LpCPC genes in different organs of Lilium pumilum were analyzed.It was found that the expression of these two genes was relatively high in the bulb and root of Lilium pumilum.The expression of LpBCP and LpCPC genes in Lilium pumilum was analyzed under three kinds of abiotic stresses:NaCl,NaHCO3 and H2O2,the expression of LpBCP and LpCPC genes were increased than control.3.The yeast protein expression vectors of LpBCP and LpCPC were constructed.After the induction of galactose,the bacterial liquid was diluted in the culture medium with NaHCO3 resistance.The results showed that the recombinant yeast grew better than the control group with the increase of NaHCO3 stress concentration,indicating that LpBCP and LpCPC may be involved in the oxidative stress response of yeast cells.4.Overexpression vectors of LpBCP and LpCPC were constructed,and the recombinants were transferred into Nicotiana tabacum and Lilium pumilum,respectively.Under NaHCO3 stress,transgenic plants were more tolerant to NaHCO3 stress.Then,a series of physiological indexes including chlorophyll content,relative electrical conductivity,total soluble phenol and lignin content were further analyzed to verify the conclusion.