Cloning Of Spinach SpoHIPP39 Gene And Its Stress Response

Heavy metal associated isoprenylated plant protein(HIPP),belonging to plant metal chaperone protein,plays an important role in plant cell heavy metal homeostasis and detoxification mechanism,and found that the protein may be responsive to regulatory elements in abiotic stress transcription reactions.The analysis of the response of these genes in stress can provide theoretical data for the study of plant stress tolerance.In this experiment,the full-length c DNA of Spo HIPP39 gene was obtained from the spinach transcriptome database,totaling 1747 bp,of which the largest open reading frame was 519 bp,the encoded protein was 172 amino acids,and it had HMA conserved domain and prenylated motif.The full-length c DNA of the Spo HIPP39 gene was obtained by PCR,and the Hidden Markov Model(HMM)model was established based on the amino acid sequence encoded by the gene and the known Arabidopsis HIPP protein series.The established model was used for spinach.A transcriptome database was searched to obtain 34 spinach HIPP family proteins.For the analysis of this family member,no transmembrane region and signal peptide were predicted,and the family proteins were considered to be intracellular proteins.The subcellular localization of the onion epidermis was carried out by constructing the Spo HIPP39 gene into a green fluorescent protein(GFP)plant expression vector.The subcellular localization results showed that the protein encoded by the gene was nuclear localization.The aviral stress of Spo HIPP39 gene was analyzed by q RT-PCR.The data showed that the expression of Spo HIPP39 gene was different in different organs of spinach,and the expression level was the highest in leaves.Subsequent analysis of abiotic stress in spinach Spo HIPP39 gene revealed that the expression of this gene in roots and leaves was not completely consistent.This gene indicates that the gene responds to a variety of abiotic stresses,but the expression is not exactly the same in different plant organs.It is speculated that the gene participates in various biological defenses in different organs of plants.The Spo HIPP39 gene was transformed into the model plant Arabidopsis thaliana by constructing the plant expression vector of Spo HIPP39 gene,and the Arabidopsis overexpression plants stably expressing the Spo HIPP39 gene were obtained after multiple screenings.Arabidopsis thaliana Spo HIPP39 overexpressing plants showed no significant trait changes compared with wild type.Arabidopsis thaliana Spo HIPP39 overexpressing plants had no obvious trait changes compared with wild type,and they were subjected to stress treatment to find overexpressing plants to determine the response of the gene to a variety of abiotic stresses.