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Reference gene for fluorescent quantitative analysis on Cannabis satia Linn and cultivated Cannabis sativa and application thereof

An internal reference gene, real-time fluorescence quantitative technology

Active Publication Date: 2018-11-16
云南省农业科学院经济作物研究所
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Problems solved by technology

However, a large number of research results in recent years have shown that there is no internal reference gene with absolutely stable expression, and the so-called constant expression of any internal reference gene is only a "range" constant expression under certain types of cells or experimental conditions.
So far, there have been many reports on the screening and verification of reference genes in crops such as rice, corn, potato, cucumber, tomato, and apple, but there is no report on the expression analysis of genes related to cannabinoid synthesis in wild cannabis and cultivated cannabis. Appropriate internal reference genes are available

Method used

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  • Reference gene for fluorescent quantitative analysis on Cannabis satia Linn and cultivated Cannabis sativa and application thereof
  • Reference gene for fluorescent quantitative analysis on Cannabis satia Linn and cultivated Cannabis sativa and application thereof
  • Reference gene for fluorescent quantitative analysis on Cannabis satia Linn and cultivated Cannabis sativa and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The screening and stability verification of the most stably expressed internal reference genes in different parts of wild cannabis and cultivated cannabis plants at the initial fruiting stage include the following steps:

[0052] 1. Sampling of experimental materials: Sow the seeds of wild cannabis CF 1 (collected in Chifeng City, Inner Mongolia) and cultivated cannabis YM 7 (the main variety in Yunnan) in the greenhouse, and take the roots, stems, and leaves of the plants at the beginning of fruiting stage. A total of 30 samples were collected from five parts of , flower and false fruit. All samples were quickly frozen in liquid nitrogen and then stored at -80°C. Each sample of wild cannabis CF1 and cultivated cannabis YM 7 stored at -80°C was carried out according to the following steps:

[0053] 2. Total RNA extraction: RNA extraction kit (RNeasy Plant Mini Kit, Qiagen, Germany) was used to extract total RNA from the sample.

[0054] 3. Synthesis of the first stran...

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Abstract

The invention discloses a reference gene for fluorescent quantitative analysis on Cannabis satia Linn and cultivated Cannabis sativa and application thereof. The reference gene is gene PP2A or gene EF1alpha, expression stability of the gene PP2A or gene EF1alpha is the heighest in the different parts at the time of begining to bear fruit of the Cannabis satia Linn and cultivated Cannabis sativa,the problem that no reference gene in the conventional expression research on the Cannabis satia Linn and cultivated Cannabis sativa is solved, and a strong support for further gene expression research in Cannabis satia Linn and cultivated Cannabis sativa with different gene types is provided.

Description

technical field [0001] The invention relates to the field of plant molecular biology, in particular to an internal reference gene for fluorescence quantitative analysis of wild cannabis and cultivated cannabis and its application. Background technique [0002] Hemp (Cannabis sativa L.), also known as hemp and thread hemp, is an annual herb plant of the genus Cannabis in the family Cannabinaceae, with many dioecious plants. It is a traditional economic crop widely used in papermaking, textiles, fields such as food and pharmaceuticals. Cannabis mainly originated in Central Asia and is now distributed all over the world (cultivated species or wild species). my country is rich in hemp resources, among which wild hemp is distributed in Yunnan, Tibet, Xinjiang, Inner Mongolia and Northeast China, while cultivated hemp is distributed in a wider range, covering most provinces and regions in my country. Currently, it is mainly concentrated in Yunnan, Heilongjiang, Inner Mongolia, Ga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 郭蓉郭鸿彦陈璇杨明张庆滢郭孟璧许艳萍吕品
Owner 云南省农业科学院经济作物研究所
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