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Primer group for constructing molecular marker map of cured tobacco variety of Hunan main tobacco varieties and application of primer group

A molecular marker and primer set technology, which is applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve problems such as the lack of guidance in the identification and grading of tobacco leaves in flue-cured tobacco enterprises, and the lack of research on the identification of main plant varieties. , to achieve the effect of efficient identification and classification

Pending Publication Date: 2021-12-03
湖南省烟草公司永州市公司 +1
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, the reported studies mainly focus on the identification of a few cultivars or germplasm resources, but lack of research on the identification of most main cultivars, so the guiding significance for the identification and grading of tobacco leaves in flue-cured tobacco enterprises is not strong

Method used

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  • Primer group for constructing molecular marker map of cured tobacco variety of Hunan main tobacco varieties and application of primer group
  • Primer group for constructing molecular marker map of cured tobacco variety of Hunan main tobacco varieties and application of primer group
  • Primer group for constructing molecular marker map of cured tobacco variety of Hunan main tobacco varieties and application of primer group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] Tobacco cultivation

[0033] Tobacco seeds were sterilized with 20% NaClO for 20min, rinsed with sterile water 3-4 times, resuspended in 0.2% agar, spread on solid 1 / 2MS medium (2.4g MS salt, 3% sucrose, 0.8% agar powder, PH value 5.8, 121°C, sterilized for 15-20min), sealed with parafilm, and placed in the incubator. The culture conditions are: 25°C, 12 hours of darkness, 12 hours of light, culture for 15-20 days, and plant in pots filled with nutrient soil (nutrient soil: vermiculite = 1:1). After 10 days, it can be used for DNA extraction.

[0034] DNA extraction

[0035] (1) Weigh 0.1g~0.3g of tobacco, cut it into pieces with scissors, put it in a mortar, add appropriate amount of liquid nitrogen to grind it into powder, quickly transfer it to a 2mL centrifuge tube, add 600μL to preheat at 65°C and contain 1% CTAB extract of β-mercaptoethanol and 6 μL of 10 mg·mL -1 For RNaseA, use a vortex shaker for 1 minute, and then treat it in a 65°C water bath for 10 minut...

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Abstract

The invention discloses a primer pair for constructing a molecular marker map of a cured tobacco variety of Hunan main tobacco varieties, and the sequence of the primer pair is as shown by SEQ ID No.1-46. The primer pair can be used for identifying cured tobacco varieties of Hunan main tobacco varieties, such as Xiangyan No.3, Xiangyan No.5, Xiangyan No.6, Xiangyan No.7, Yunyan 87, Yunyan 97, Yunyan 99, Yunyan 100, Yunyan 203, Zhongyan 100 yuan, Zhongyan 100, Guangdong tobacco 9, Gui tobacco No.1, Honghua Dajinyuan, Longjiang 911, Nanjiang No.3, NC729, G80 or K326. The primer pair is of great significance in helping tobacco enterprises to quickly, accurately and efficiently identify and grade the cured tobacco.

Description

technical field [0001] The invention relates to the technical field of identification of tobacco varieties, in particular to a primer set for constructing a molecular marker map of cured tobacco leaf varieties of main tobacco varieties grown in Hunan and its application. Background technique [0002] Tobacco leaf is the foundation of the development of the tobacco industry, and its grade quality is the lifeline of the tobacco industry. Tobacco leaf grade quality status and industrial usability are ultimately reflected in the cigarette industry's demand for tobacco leaf raw materials and utilization value, which directly affects the quality of cigarettes and brand development, and affects the healthy, stable and sustainable development of the entire tobacco industry. Tobacco grading is affected by many factors, such as production factors and human factors. Production factors, such as natural factors, ecological factors, environmental conditions, etc., have a relatively obvio...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 肖钦之刘永昌胡日生李书霞邓征宇李洋洋王凯歌李泽明周立邓斌
Owner 湖南省烟草公司永州市公司
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