Tissue Location And Functional Analysis Of Transgenic ChIFN-? Tobacco

Chicken interferon-gamma(ChIFN-?)is a type of cytokine which has a powerful immune regulation and broad-spectrum anti-virus,anti-bacterial,anti-parasitic.ChIFN-? is previously genetically transformed into tobacco and is found to have increased glandular trichome in the transgenic tobacco with significant insecticidal effects,and vascular bundles of stem are more developed than controls.The stable genetic inheritance of foreign genes in transgenic plants is a crucial link in the study.Multiple candidate genes associated with transgenic ChIFN-? tobacco are screened by genome-wide expression profiling microarrays in combination with bioinformatics analysis.The HD gene which significantly up-regulated is reported in the literature that its encoded HD-Zip protein is involved in the formation of vascular bundles and the differentiation of vascular bundle cells.In order to verify whether ChIFN-? can be stably inherited in transgenic tobacco,T5 transgenic ChIFN-? tobacco genome was used as the experimental material.We tested the copy number of the ChIFN-? gene using fluorescence quantitative PCR.Southern blotting was used to verify the results of fluorescent quantitative PCR.The result showed that the number of copies of ChIFN-? in the transgenic tobacco genome was 1.This indicated that ChIFN-? has genetic stability in transgenic tobacco plants.In order to verify the differentiation of transgenic tobacco stem cells,ChIFN-? tobacco leaves at different growth stages were used as experimental materials.Fluorescence in situ hybridization is used to detect the localization of ChIFN-? in leaf tissue.The results showed that ChIFN-? was located in the cytoplasm.Most of the fluorescence signals appeared in the xylem and the palisade tissue.Then,significantly up-regulated expression of HD genes was selected for histological observation.The results showed that the transgenic Arabidopsis thaliana compared with the control,the stem vascular bundle changed,xylem more developed than the control.The result indicated that this gene was associated with the differentiation of vascular fibroblasts.These studies indicated that ChIFN-? altered the xylem structure by affected the expression of HD in transgenic tobacco.The results of the study demonstrated that the screening of the genes for the differential genes in the early stages of the laboratory was correct.Combined with previous studies,it was speculated that the regulation of xylem cell differentiation by ChIFN-? which was affected the expression of HD was one of the reasons for the resistance of ChIFN-? to insects.In summary,ChIFN-? was a single copy in transgenic tobacco.The results showed that ChIFN-? was stably inherited in transgenic tobacco plants and was consistent with the Mendel's genetic law of single gene lociation.It was most needed for genetic breeding.The study provides a theoretical basis for the successful study and utilization of ChIFN-? for breeding new insect-resistant varieties.It provides research value for obtaining reversible strains of transgenic plants with homozygous genotypes in offspring screening.ChIFN-? altered the xylem structure of transgenic tobacco plants by affecting the expression of HD genes.The study provides a theoretical basis for the study of the function of ChIFN-? and reveales the insect resistance mechanism of transgenic ChIFN-? tobacco.