7 results about "Quantitative PCR analysis" patented technology

The invention belongs to the technical field of biology, and particularly relates to application of hepcidin expression quantity in larimichthys crocea liver as larimichthys crocea visceral white-spot disease resistance evaluation index and a construction method. The hepcidin expression quantity in the larimichthys crocea liver is applied as the larimichthys crocea visceral white-spot disease resistance evaluation index. The construction method of the resistance evaluation index comprises the following steps: taking healthy small yellow croaker intraperitoneal injection pseudomonas proteinus strain bacterial liquid, and determining a half lethal dose in 96 hours; the method comprises the following steps: injecting a half lethal dose of bacterial liquid of pseudomonas proteinus strains into the abdominal cavity of healthy small yellow croaker for 96 hours, and after infection, taking different tissues to carry out hepcidin gene expression fluorescent quantitative PCR analysis; a regression model of gene expression quantity and bacterium loading quantity is established through fluorescence quantification, and the relationship between hepcidin expression quantity and disease resistance is constructed. Accurate disease-resistant phenotype information is provided for development of breeding of visceral white-spot disease-resistant improved varieties of the small yellow croakers, the accuracy of disease-resistant breeding is promoted, early detection of diseases is realized, and prevention and treatment of the diseases are effectively guided.

This invention discloses a method for delaying leaf senescence in Paspalum notatum under salt stress. The method includes the following steps: plant material culture and salt stress treatment; H2O2 content determination; DAB staining; chlorophyll content and Fv / Fm value determination; lipidomics data determination and analysis; endogenous hormone level analysis; real-time quantitative PCR analysis of gene expression levels; and transcriptomics data analysis. This invention discovers that root-derived H2O2 in Paspalum notatum under salt-driven conditions delays leaf senescence by participating in the regulation of jasmonic acid. The role of root-derived H2O2 was studied using diphenylthioiodine (DPI), a specific inhibitor of NADPH oxidase. Furthermore, the influence of the presence or absence of root-derived H2O2 on the expression of genes related to jasmonic acid synthesis precursors was investigated at the gene expression level. Mass spectrometry-based omics technology and data analysis are the first key technologies for implementing this project. Lipidomics, as one of the most important branches of metabolomics, can perform qualitative and quantitative analysis at the level of individual lipid species.

This invention discloses the artificial synthesis of the disease-resistant gene NLP27 and its application, belonging to the field of plant genetic engineering technology. This invention selects a conserved 27-amino acid fragment from Verticillium dahliae NLP1, modifies its nucleotide sequence, and synthesizes the NLP27 gene, which is then overexpressed in potato plants through genetic transformation. Inoculation analysis shows that transgenic plants with the NLP27 gene exhibit high resistance to potato late blight and scab. Quantitative PCR analysis shows that excessively high NLP27 transcript levels significantly inhibit potato plant growth. After screening for expression levels and disease resistance, three transgenic lines with moderate NLP27 expression levels (such as PT1, PT6, and PT7) were obtained. These lines significantly enhanced resistance to potato late blight and scab without sacrificing yield.

The invention relates to the technical field of plantago asiatic L. gene engineering, and particularly provides a plantago asiatic L. real-time fluorescent quantitative PCR (Polymerase Chain Reaction) reference gene and application thereof. According to the method, tender leaves, old leaves, stems, flowers and roots of plantago asiatica are selected as experimental materials, candidate reference genes are screened through a plantago asiatica transcriptome, and then the gene expression stability is analyzed through real-time fluorescent quantitative PCR analysis, dissolution curve analysis, amplification efficiency analysis and various software (geNorm, NormFinder, BestKeeper, delta Ct and RefFinder). Reference genes, namely eIF and Cycl genes, suitable for gene quantification of different tissue parts of plantago asiatica are screened out, and the sequences of the eIF and Cycl genes are shown as SEQ ID NO.1 and SEQ ID NO.2 respectively. The reference gene with stable expression is obtained, is suitable for analyzing the expression quantity of the gene at different parts of the plantago asiatica, makes up for the current situation that the plantago asiatica lacks the reference gene, and provides a basis for the gene expression quantity and function analysis in the plantago asiatica.

The present application belongs to the technical field of plant genetic engineering, and particularly relates to an internal reference gene in papaya fluorescent quantitative PCR analysis and a screening method and application thereof. The present application relates to 18S rRNA, Actin, TUB, GAPDH and EF-1 alpha 5 internal reference genes, and the internal reference genes can be applied to the research on gene expression in different varieties, different periods and different tissues of papaya, and provide more accurate gene expression quantification standards for subsequent experimental research on papaya.

This invention discloses a barley stripe disease resistance gene. Rdg2b This invention relates to the field of bioengineering technology and its applications. It describes the cloning of [specific species / organisms] from Ganbei No. 2 barley. Rdg2b The complete CDS segment of the gene was extracted, and an overexpression vector was constructed. Agrobacterium-mediated transformation was then performed to obtain overexpression-positive lines. Infection experiments with *Strombus striatum* showed that, compared to the wild type, [the expression was positive]. Rdg2b Overexpression lines showed significantly enhanced disease resistance. Real-time quantitative PCR analysis revealed that overexpression lines exhibited... Rdg2b Gene expression was significantly upregulated, confirming Rdg2b This invention positively regulates barley disease resistance. It provides important genetic resources for molecular breeding of barley for disease resistance.

The invention discloses reference genes for real-time fluorescent quantitative PCR detection of different tissues of rhizoma bletillae in different growth periods, and belongs to the technical field of plant molecular biology, the reference genes are BsGAPDH and BsEIF, the nucleotide sequence of the BsGAPDH is as shown in SEQ ID NO.1, and the nucleotide sequence of the BsEIF is as shown in SEQ ID NO.2. Three different tissues of rhizoma bletillae in seven growth periods are used as experimental materials, and the reference genes are used for real-time fluorescent quantitative PCR detection of different tissues of rhizoma bletillae in seven growth periods. The reference genes BsGAPDH and BsEIF with stable expression are obtained through fluorescent quantitative PCR (polymerase chain reaction) analysis and gene expression stability analysis by various technologies, and the two genes are stably expressed in different tissues in different growth periods of rhizoma bletillae and can be used as reference genes for researching gene expression in different tissues in different growth periods of rhizoma bletillae. The method can lay a foundation for functional research of genes related to accumulation of traditional Chinese medicinal components of rhizoma bletillae, and has relatively good practical value and scientific research application value.