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Fluorescence quantitative internal reference gene under drought stress in haizhou Changshan Mountain, special primer and application thereof

A fluorescent quantitative and drought stress technology, applied in the field of fluorescent quantitative internal reference genes and their primers and applications, to achieve the effects of improving detection efficiency, strong specificity, and improving reliability

Inactive Publication Date: 2019-01-15
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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  • Fluorescence quantitative internal reference gene under drought stress in haizhou Changshan Mountain, special primer and application thereof
  • Fluorescence quantitative internal reference gene under drought stress in haizhou Changshan Mountain, special primer and application thereof
  • Fluorescence quantitative internal reference gene under drought stress in haizhou Changshan Mountain, special primer and application thereof

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Embodiment 1

[0029] 1. Extraction of total RNA from plant tissue and synthesis of cDNA

[0030] Total RNA was extracted with EASYspin Plus Plant RNA Extraction Kit (Adelai Biotechnology Co., Ltd.), and the RNA concentration and quality were detected by spectrophotometer (Mettler-Toledo Group), and detected by 1.5% agarose gel electrophoresis total integrity. Purify RNA and synthesize the first strand of cDNA using Beijing Quanshijin Reverse Transcription Kit (Quanshijin Biotechnology Co., Ltd.) according to the instructions.

[0031] 2. Selection of internal reference genes and design of primers

[0032] The main reference genes in other plants were searched according to the literature, and 17 reference genes required in this example were finally designed, namely 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-E...

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Abstract

A fluorescence quantitative internal reference gene under the drought stress of Haizhou Changshan Mountain, a special primer and the application thereof are disclosed, wherein the fluorescence quantitative internal reference gene is Actin, AP-2 and RAN gene, wherein that gene sequence of the Actin gene is as shown in SEQ ID NO. 1, AP-The gene sequence of RAN gene is shown in SEQ ID NO. 6 and the gene sequence of RAN gene is shown in SEQ ID NO. 13. The invention screens out the candidate gene with stable expression from the transcriptome data and the internal reference gene studied by the predecessor, its stability is evaluated by software, 3 most stable internal reference gene suitable for expression under drought stress in Haizhou Changshan mountain are disclose, Based on these genes, real-time fluorescence quantitative PCR primers were designed, which filled the situation that there were no internal reference genes under drought stress in Haizhou Changshan Mountain, and provided a strong support for the precise quantification of related functional genes under drought stress in Haizhou Changshan Mountain, and could improve the stability, repeatability and reliability of the research.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and particularly relates to a fluorescent quantitative internal reference gene, primers and applications thereof under drought stress in Changshan, Haizhou. technical background [0002] Haizhou Changshan (Clerodendrum trichotomum Thunb.), also known as the stinking sycamore, 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 the autumn and winter are long-lasting. It is an excellent autumn flower and fruit viewing tree species. The existing research results show that Haizhou Changshan not only has excellent salt and alkali tolerance, but also has strong early tolerance, and is an excellent garden plant with strong resistance to adverse environments. So far, the research on genes under droug...

Claims

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Application Information

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Patent Type & Authority Applications(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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