Sisal hemp reference gene ACT/GAPDH and application thereof

Abstract

The invention provides a sisal hemp gene sequences ACT and GAPDH which are used as sisal hemp reference genes, further provides specific primers of the sisal hemp ACT and GAPDH as reference genes, establishes a qRT-PCR (Quantitative Reverse Transcription-Polymerase Chain Reaction) method based on an SYBR Green fluorescent dye technique, and provides stable and reliable reference genes and technique methods for gene expression analysis under adverse situations such as tobacco phytophthora infection, low temperature stress, salt stress and injury treatment on sisal hemp through real-time fluorescent quantitative PCR.

Description

technical field [0001] According to an existing nucleic acid sequence, the present invention screens out an internal reference gene stably expressed in sisal under adverse conditions such as Phytophthora nicotiana infection, low temperature stress, salt stress and wound treatment, and belongs to the technical field of plant molecular biology. Background technique [0002] Sisal is one of the unique economic crops in the tropics of our country. Sisal is not only the main tropical fiber raw material, the sisal stem core is the main raw material for brewing tequila, and the sisal hemp residue contains high saponin, which can be used for pharmaceuticals , sisal is also an important biomass energy, and in recent years, as a model plant for sedum metabolism, sisal has very important theoretical and economic value. [0003] With the rapid development of sequencing technology and bioinformatics, the use of omics to study gene expression profiles under a certain stress has become a r...

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Problems solved by technology

[0003] With the rapid development of sequencing technology and bioinformatics, the use of omics to study gene expression profiles under a certain stress has become a routine method for molecular biology research, but omics is only a preliminary analysis of gene expression under a certain stress However, the results need to be verified by fluorescent quantitative RT-PCR; in addition, the spatio-temporal expression of genes has become one of the main contents of gene function research, and the above studies require relatively stable expressed genes as internal reference genes; With the in-depth development of hemp molecular biology, some functional genes have been successfully isolated. At present, there are many nucleic acid sequences of agave hemp in NCBI public data, but there are few internal reference gene sequences, only actin1 (232bp) and 18S rRNA (186bp) Because the expression level of internal reference genes may change under different conditions, different tissues, different periods and different varieties, this will lead to unstable research results and reduce the reliability of the results. Therefore, the screening of sisal is different. Stable expression of internal reference genes under adverse conditions is the premise of sisal gene function research; this not only improves the current situation of few internal reference genes in sisal, but also provides a guarantee for the reliability of the research results of sisal gene expression levels

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