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Fluorescent quantitative internal reference genes and their special primers and applications under drought stress in Changshan, Haizhou

A technology of fluorescence quantification and internal reference gene, applied in the field of plant molecular biology, to achieve strong specificity, improve detection efficiency, and improve reliability

Inactive Publication Date: 2020-01-10
NANJING FORESTRY UNIV
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Problems solved by technology

There is no report on the research on genes under drought stress in Changshan, Haizhou

Method used

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  • Fluorescent quantitative internal reference genes and their special primers and applications under drought stress in Changshan, Haizhou
  • Fluorescent quantitative internal reference genes and their special primers and applications under drought stress in Changshan, Haizhou
  • Fluorescent quantitative internal reference genes and their special primers and applications under drought stress in Changshan, Haizhou

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Embodiment 1

[0030] 1. Plant tissue total RNA extraction and cDNA synthesis

[0031] Extract total RNA with EASYspin Plus Plant RNA Extraction Kit (Adelaide Biotechnology Co., Ltd.), detect RNA concentration and quality with a spectrophotometer (Mettler-Toledo Group), and use 1.5% agarose gel electrophoresis to detect total integrity. Using Beijing Quanshijin Reverse Transcription Kit (Quanshijin Biotechnology Co., Ltd.), RNA was purified and the first strand of cDNA was synthesized according to the instructions.

[0032] 2. Selection of internal reference genes and design of primers

[0033] Query the main internal reference genes in other plants according to the literature and finally design 17 internal reference genes required by this embodiment, which are Actin (SEQ ID NO.1), PP2A (SEQ ID NO.2), RPL (SEQ ID NO. .3), PK (SEQ ID NO.4), 18S (SEQ ID NO.5), RAN (SEQ ID NO.6), APT (SEQ ID NO.7), SAND (SEQ ID NO.8), PROF ( SEQ ID NO.9), MDH (SEQ ID NO.10), EF-1A (SEQ ID NO.11), UBC-E2 (SEQ...

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Abstract

The invention discloses a fluorescent quantitative internal reference gene and its special primers and applications under drought stress in Changshan, Haizhou. The fluorescent quantitative internal reference genes are Actin, AP‑2 and RAN genes, wherein the gene sequence of the Actin gene is as shown in SEQ ID NO.1 The gene sequence of the AP‑2 gene is shown in SEQ ID NO.13, and the gene sequence of the RAN gene is shown in SEQ ID NO.6. The present invention screens candidate genes with relatively stable expression from transcriptome data and internal reference genes studied by previous studies, and evaluates their stability through software, and discloses three internal reference genes suitable for the most stable expression under drought stress in Changshan, Haizhou , and based on these genes, real-time fluorescent quantitative PCR primers were designed to fill the status quo of no internal reference genes under drought stress in Changshan, Haizhou, and provide strong support for the accurate quantification of related functional genes under drought stress in Changshan, Haizhou, which can improve the stability of research repeatability, repeatability and reliability.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and specifically relates to a fluorescent quantitative internal reference gene and its primer and application under drought stress in Changshan, Haizhou. [0002] technical background [0003] Haizhou Changshan (Clerodendrum trichotomum Thunb.), also known as stinky phoenix tree, is a deciduous shrub or small tree of the Verbenaceae (Verbenaceae) genus (Clerodendrum L.). Few pests and diseases, strong adaptability. In addition, the flower shape is peculiar and beautiful, the flowering period is long, the fruit color is bright, and it does not fall for a long time in autumn and winter. It is an excellent autumn flower and fruit tree species. The existing research results show that Haizhou Changshan not only has very good salt-alkali tolerance, but also has strong early tolerance. It is an excellent garden plant with strong resistance to adverse environments. There has not been any report on...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2600/166C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 岳远征杜菊花王良桂杨秀莲施婷婷华雅洁
Owner NANJING FORESTRY UNIV
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