Establishment and application of a SNP molecular identification system in Grifola frondosa.

Germplasm resources of edible mushrooms are essential for the breeding of varieties with improved traits. Analysis of the genetic diversity of Grifola frondosa germplasm resources and clarification of the genetic relationships among strains can provide valuable information for the selection of breeding parents. A total of 829,488 high-quality SNP loci were screened from 2,125,382 SNPs obtained by sequencing 60 G. frondose. Phylogenetic analysis, PCA, and population structure analysis based on the high-quality SNPs showed that the 60 strains could be divided into five subgroups, and the clustering results were consistent with the geographical distributions of these strains. Based on high-quality SNP loci, a core collection containing 18 representative germplasm resources was constructed, and 1,473 Kompetitive Allele-Specific PCR markers were obtained. A total of 722 SNP markers in the exonic regions were screened using KASP-genotyping experiments, and 50 candidate SNP markers and 12 core SNP markers were obtained. Genetic fingerprints of G. frondosa germplasm resources were constructed based on the selected SNP markers; these fingerprints provide an accurate, rapid, convenient, and efficient method for the identification of G. frondosa germplasm resources. The results of this study have important implications for the preservation and utilization of G. frondosa germplasm resources and the identification of varieties.

[Impact of alpha1,2-fucosyl transferase gene transfection on cancer-related gene expression profile of human ovarian cancer cell line RMG-1].

BACKGROUND & OBJECTIVE: The malignant biological behaviors are enhanced in human ovarian cancer cell line RMG-1 after transfection of alpha1,2-fucosyl transferase (alpha1,2-FT) gene. This study was to investigate the influence of alpha1,2-FT gene transfection on cancer-related gene expression profile of RMG-1 cells. METHODS: Gene expressing vector pcDNA3.1-HFT-H and empty vector pcDNA3.1 were transfected into RMG-1 cells to produce RMG-1-H and RMG-1-C cells separately. Gene expression profiles of these two cell lines were detected by gene chip assay. The acquired data were inquired on the GoMiner online database. RESULTS: After transfection, comparing with those in RMG-1-C cells, 88 differentially expressed genes were identified in RMG-1-H cells: 60 were up-regulated and 28 were down-regulated. These genes are involved in protein binding, nucleotide binding, cell proliferation, DNA-dependent regulation of transcription, signal transduction, protein amino acid phosphorylation, transcription, cell adhesion, and so on. CONCLUSION: The transfection of alpha1,2-FT gene causes the changes of gene expression profile in ovarian cancer RMG-1 cells.

[alpha1,2-fucosyltransferase gene transfection influences on biological behavior of ovarian carcinoma-derived RMG-I cells].

To explore the effect of alpha1,2-fucosyltransferase (alpha1,2-FT) gene on the biological behavior of ovarian carcinoma cells, the expression vector pcDNA3.1 -HFUT-H which containing human alpha1,2-FT gene coding region was constructed and transfected into ovarian carcinoma cell line RMG-I. alpha1,2-FT gene expression, Lewis y antigen expression, alpha1,2-FT activity, rates of cell growth and tumorogenesis in nude mice were compared before and after RMG-I being transfected. The transfection of alpha1,2-FT gene significantly increased the expression of alpha1,2-FT gene and Lewis y antigen in RMG-I cells. After the transfection, RMG-I cells showed remarkable improvement of alpha1,2-FT activity, cell proliferation, the ability of colony formation in soft agar and tumorogenesis in nude mice. The growth rates of alpha1,2-FT gene transfected RMG-I cells were obviously increased, presenting that the proportion of the cells in G2-M and S phases was obviously increased from 2.05% to 7.32% and from 23.95% to 33.27% respectively, and that of the cells in G1 phase was obviously decreased from 74.14% to 59.46%, as compared with the control group. These findings indicate that alpha1,2-FT and Lewis y antigen have the ability to enhance the proliferation and elevate the survival rates of RMG-I cells, which can promote the genesis and development of ovarian carcinoma.