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Platycodon grandiflorum is a medicinal and edible medicinal material. Our study is aimed to explore the differences in the gene expression of P. grandiflorum in different growth years, and the expression rules of key genes in the biosynthesis of the main active substances of P. grandiflorum. Illumina Hiseq 4000 sequencing platform was used to sequence the transcriptome of P. grandiflorum in different years. Then, 59 654 unigenes were obtained through filtering, assembly, splicing and bioinformatics analysis of the sequencing data, of which 1 671 unigenes were differentially expressed between at least two samples. The results of cluster analysis showed that there was a great difference in the gene expression of P. grandiflorum from one-year-old to two/three-year-old. There were 1 128 different genes between one-and three-year old P. grandiflorum, and only 57 different genes between two-and three-year-old P. grandiflorum. KEGG enrichment results showed that the differential genes of P. grandiflorum in different years were mainly concentra-ted in the biosynthesis of sesquiterpenes and triterpenes, and the biosynthesis of terpenoid skeletons. In the triterpenoid biosynthesis-related pathways, a total of 15 unigenes were identified, involving 5 enzymes. The expression levels of ACAT, HMGR, FDFT1, SQLE decreased with the increase of the growth year of P. grandiflorum. The expression of HMGS was the highest in the one-year-old P. grandiflorum, followed by the three-year-old sample. This study provides useful data for the development of P. grandiflorum, and also provides a basis for the study of related genes in the biosynthetic pathway of platycodin.
Subject(s)
Gene Expression Profiling , Plant Roots , Platycodon/genetics , Saponins , Transcriptome , TriterpenesABSTRACT
To explore the mechanism hydroxysafflor yellow A (HSYA) biosynthesis and regulation, the effect of methyl jasmonate (MeJA) treatment on gene expression related to the biosynthesis of hydroxysafflor yellow A (HSYA) was analyzed, and expression differences in genes involved in HSYA biosynthesis in safflower of different colors was quantified. MeJA at concentrations of 0, 50, 100, and 200 μmol·L-1 was sprayed onto safflower florets to determine the optimal concentration of MeJA. Safflower was treated with 100 μmol·L-1 MeJA and florets were harvested 0, 3, 6, 12 and 24 h after treatment. The content of MeJA was determined by high performance liquid chromatography (HPLC). RNA was extracted from safflower florets treated with 100 μmol·L-1 MeJA for 6 h. The transcription of key genes involved in the biosynthesis of HSYA was quantified by qRT-PCR and differentially expressed genes were identified. The content of HSYA increased after treatment with MeJA, with 100 μmol·L-1 MeJA treatment for 6 h having the greatest effect on HSYA accumulation. qRT-PCR results showed that MeJA could significantly increase the transcription of HSYA biosynthesis genes including PAL2, PAL4, 4CL2, 4CL4, 4CL5, CHS3, CHS4 and CHI2. The content of HSYA differed between safflowers of different colors with a trend of red>orange-yellow>yellow>white. The results of qRT-PCR showed that the expression of CHS1 and CHI2 in red, orange and yellow safflower was significantly higher than that in white safflower. These results indicate that MeJA promotes the accumulation of HSYA by up-regulating the expression of genes involved in the biosynthesis of HSYA such as PAL2, PAL4, 4CL2, 4CL4, 4CL5, CHS3, CHS4 and CHI2, and the variation of HSYA content in safflower of different colors was related to a difference in the level of expression of CHS1 and CHI2.
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OBJECTIVE: To acquire the expressing information about tumor abnormal glycoprotein (TAP) in the peripheral blood of the gastric cancer patients who are treated with fluorouracil and the colorectal cancer patients treated with fluorouracil, and to discuss the factors that affect its expression and its clinical significance. METHODS: This research was conducted on the basis of 99 gastric cancer patients and colorectal cancer patients whose diseases were pathologically diagnosed from May to December, 2018, including 53 gastric cancer patients treated with fluorouracil and paclitaxel and 46 colorectal cancer patients treated with fluorouracil and oxaliplatin. It tested their TAP values, collected the basic information, the type of the cancer, the stage of the cancer, serum tumor markers, etc. of the patients as well as analyzed the differences of the expression of TAP in gastrointestinal tumors with different characteristics. RESULTS: The expression value of TAP in the peripheral blood of the gastrointestinal cancer patients is (158.11±33.63), and the expression of TAP value has no statistical differences in the expression of different sexes, ages, types of the cancer, clinical stages and serum tumor markers. CONCLUSION: The fact that the expression of TAP has no differences in the gastrointestinal cancer patients with different characteristics can't be taken as the basis of identifying or diagnosing gastrointestinal tumors which are different in character.
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Objective To obtain the transcriptome database and differentially expressed genes of Tetrastigma hemsleyanum Diels et Gilg. by Illumina HiSeq 4000; To provide important molecular information for its molecular biology research. Methods Leaves and roots of Tetrastigma hemsleyanum Diels et Gilg. were chosen as experimental materials to conduct transcriptome sequencing. Then bioinformatics analysis of gene function annotations, metabolic pathways, and microsatellites was performed on the test data. Results 24.13 Gb Clean Data were assembled. Afer assembly steps, 84 433 of T. hemsleyanum Unigene were obtained, and then they were compared in the 7 gene database, and 47 766 annotated information of Unigene was obtained. There were 27 790 annotations in the GO database. The number of differentially expressed genes in the roots, stems and leaves was 4989, of which 3511 were up-regulated and 1478 were down-regulated. The COG database obtained 16 152 homologous sequences of Unigene, which were divided into 25 categories. In the KEGG database, there were 14 511 Unigene obtained the corresponding Ko number, which could be divided into 130 branches of signal metabolism, among which the number of Unigene in the ribosome synthesis pathway was the most, with 1042, and there was only 1 Unigene in the biosynthetic pathway of isoflavones. Conclusion A large number of transcripts of the transcriptome were obtained through splicing, assembling and functional annotation of Tetrastigma hemsleyanum Diels et Gilg., which can provide genomic database resources for molecular biology research of Tetrastigma hemsleyanum Diels et Gilg.
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Objective To screen and clone the genes related to stilbene glucosides biosynthesis in Polygonum multiflorum Thunb. Methods The differentially expressed genes in the root, stem and leaf of Polygonum multiflorum which have different contents of stilbene glucosides were screened by differential display reverse transcription polymerase chain reaction (DDRT-PCR). After pMD19-T carrier was inserted into the obtained differential genes for sequencing and comparison, the gene function was analyzed. Results Fifty-one differentially expressed cDNA fragments were found. Of them, 9 were used for the identification by semi-quantitative PCR. The identification results presented 3 positive fragments, one fragment was specifically expressed in the stem and leaf of Polygon-um multiflorum Thunb., sharing high homology with glycine dehydrogenase, and 2 were specifically expressed in the root of Polygonum multiflorum Thunb., having high homology with enoyl-CoA hydratase and aminopeptidase N, respectively. Conclusion Three homologous gene sequences obtained through DDRT-PCR provide a basis for the further study of biosynthetic pathway of stilbene glucoside from Polygonum multiflorum Thunb..
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Objective:Differential expression analysis and cloning aged related genes of mouse thymus Methods:Different expressions of thymus mRNAs from 1 and 10 month old mouse were analyzed by DDRT PCR and different expression sequence tags (ESTs) were obtained One EST that represented high expressed level in one month mouse thymus was probed to screen mouse thymus cDNA library One 827 bp cDNA fragment was obtained and was extended to 1 406 bp by PCR Results:Homology analysis showed that mt22 1406 contained one 438AA coding region and showed high similarity with human elongation factor1?(EF1?) The Genbank accession number is BE241062 Conclusion:Cloning one gene related with mouse thymus aging
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Chinese materia medica (CMM) has double complexity in bioactive ingredient and its mechanism. It is difficult to explain by the modern biomedicine theory So it seriously restricts the modernization of CMM The modern CMM should have the high quality standard to meet the needs of international standard It can be guaranteed by spreading the GAP for Chinese medicinal materials and GMP for standard production The mechanism depends on using the DNA microarray to set up “the gene expression difference chart”, to study on the combination of CMM and gene expression difference chart Meanwhile, we can establish a totally new method of screening modern CMM based on the gene expression difference chart, it can really make the modernization and internationalization of CMM