The internal reference gene developed based on Miscanthus transcriptome sequence and its application

A technology of transcriptome sequence and internal reference gene, which is applied to the internal reference gene developed based on Miscanthus transcriptome sequence and its application field, can solve the problems that restrict the process of gene screening and verification research, and achieve abundant available quantity, good specificity and stability sex high effect

Active Publication Date: 2020-11-27
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is still no development and verification of related internal reference genes in Miscanthus, which seriously restricts the process of subsequent gene screening and verification research

Method used

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  • The internal reference gene developed based on Miscanthus transcriptome sequence and its application
  • The internal reference gene developed based on Miscanthus transcriptome sequence and its application
  • The internal reference gene developed based on Miscanthus transcriptome sequence and its application

Examples

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Effect test

Embodiment 1

[0059] This example is the comparative application of the three internal reference genes developed based on the Miscanthus transcriptome sequence described in the present invention and nine traditional internal reference genes in the gene analysis of Miscanthus under drought stress. In order to compare the effects of developing new internal reference genes, the following seven The commonly used traditional reference genes GAPDH, 18S rRNA, ACTIN, PP2A, eIF4a, SAMS, SAMDC and the reference genes Sb09g019750 and Sb02g041180 on two closely related species sorghum were used as controls. The sequence information of common internal reference genes was obtained from the NCBI database, and primers were designed online using primer3.0 after homologous gene sequence alignment. The primer-related information is shown in Table 4.

[0060] Specifically include the following steps:

[0061] 1), material selection: select Miscanthus seeds, and sterilize the selected seeds with 75% alcohol and...

Embodiment 2

[0082] In this example, in order to verify the effect of the application of the internal reference gene, two genes, Cu / Zn SOD and CAT, were selected as the target genes. SOD and CAT are common antioxidant enzymes in plants, which can help plants remove reactive oxygen species in the body and protect cells from immune cells. Oxidative damage usually increases its expression after stress. Taking Unigene33312 and common reference gene ACTIN as examples to analyze the expression characteristics of awn SOD and CAT genes after drought stress, the specific steps are as follows:

[0083] 1), material selection: Select Miscanthus (two varieties: 0819 and 1302) seeds, and sterilize the selected seeds with 75% alcohol and 1% sodium hypochlorite. The experimental seeds were germinated in a petri dish and then transplanted into plastic pots (9cm in diameter) for sand culture, and 1 times of Hoagland nutrient solution was poured into the incubator for cultivation. Circulation, relative hum...

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Abstract

The invention discloses a reference gene developed based on a transcriptome sequence of miscanthus sinensis and application thereof. The three reference gene sequences of the invention come from the transcriptome sequence of miscanthus sinensis, and have the advantages of good specificity and high stability compared with the previous reference genes commonly used in other species. At the same time, the expression analysis of SOD and CAT genes under drought stress of miscanthus sinensis proves that the developed three reference genes have better calibration capability than other general reference genes, and provides an effective reference gene calibration tool for the analysis, screening and validation of the gene expression of miscanthus sinensis in the future. In addition, the newly developed reference gene enriches the available number of plant reference genes, as well as is applied to the related research on other andropogoneae and miscanthus plants. The reference gene provided by the invention is suitable for popularization and application in the field of reference genes.

Description

technical field [0001] The invention belongs to the field of internal reference genes, and in particular relates to an internal reference gene developed based on Miscanthus transcriptome sequence and its application. Background technique [0002] Miscanthus sinensis (Miscanthus sinensis) is a perennial rhizomatous C4 plant of the Poaceae family. It is widely used as high-quality forage grass, landscape plant ornamental grass, soil and water conservation and heavy metal pollution remediation plants. Miscanthus is considered to be one of the most potential sustainable bioenergy crops due to its high biomass yield and good combustion quality, as well as its wide distribution, strong stress resistance and suitable for planting on vast marginal land. . At present, the most widely used Miscanthus plant in the world is the triploid Miscanthus × giganteus, which is a hybrid of the diploid Miscanthus and the tetraploid Diploid, although it has higher biomass Yield, but due to its n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/13C12Q2600/158C12Q2600/166C12Q2521/107C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 聂刚钟旻依张新全杨昕颖黄琳凯马啸彭燕闫艳红
Owner SICHUAN AGRI UNIV
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