Nicotine quantitative conversion rate and gene expression based cherry-red tobacco seed screening method

A nicotine conversion rate and gene expression technology, applied in biochemical equipment and methods, seed and rhizome treatment, measuring devices, etc., can solve the problems of nicotine-free quantitative conversion rate of flue-cured cinnabar seeds and other problems, and improve nicotine conversion The effect of preventing tobacco leaves from being moldy and rotten, and promoting the yellowing of young leaves

Inactive Publication Date: 2020-02-28
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no method for screening flue-cured tobacco c

Method used

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  • Nicotine quantitative conversion rate and gene expression based cherry-red tobacco seed screening method
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  • Nicotine quantitative conversion rate and gene expression based cherry-red tobacco seed screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Screening flue-cured cinnabar seeds with a nicotine conversion rate of 60-75%:

[0045] S100: Raise seedlings with Yunyan 87 cinnabar tobacco seeds with a nicotine conversion rate of 45-60%.

[0046]S200: After the tobacco plant grows in the large seedling tray for 30 to 40 days and grows to a height of 30 to 40 cm from the emergence of the seedling, select the tobacco leaves that are in good condition and 24 to 26 cm in length from the second to fourth leaves at the bottom of the tobacco plant as the sampling leaves. Among them, fertilization was stopped 10 days before sampling.

[0047] S300: Immerse the sampled tobacco leaves in an equal volume mixed solution of 1.2 g / L ethephon and 5 mg / L abscisic acid for 10 s, collect fresh tobacco leaves by punching method for gene expression detection, and place the remaining tobacco leaves at 35°C Dry in an oven until there is no free moisture on the surface.

[0048] S400: The pre-treated surface moisture-free tobacco leaves...

Embodiment 2

[0058] Screening flue-cured cinnabar seeds with a nicotine conversion rate of 70-85%:

[0059] S100: Raise seedlings with Yunyan 97 cinnabar tobacco seeds with a nicotine conversion rate of 55-70%.

[0060] S200: After the tobacco plant grows in the large seedling tray for 30-40 days from the emergence of the seedling and grows to a height of 30-40 cm, select the tobacco leaves that are in good condition and 22-24 cm in length from the 2nd to 4th leaves at the bottom of the tobacco plant as sampling leaves, Among them, fertilization was stopped 5 days before sampling, and nitrogen fertilization was stopped 15 days before sampling.

[0061] S300: Immerse the sampled tobacco leaves in an equal volume mixed solution of 1.0g / L ethephon and 7mg / L abscisic acid for 15s, collect fresh tobacco leaves by punching method for gene expression detection, and place the remaining tobacco leaves at 35°C Dry in an oven until there is no free moisture on the surface.

[0062] S400: Dry the pr...

experiment example 1

[0072] Starting from 25 days after emergence, the tobacco leaves in Example 1 were treated with an equal volume mixed solution of 1.2g / L ethephon, 5mg / L abscisic acid, 1.2g / L ethephon and 5mg / L abscisic acid every 5 days. , according to the method of the present invention to detect the nicotine conversion rate, until 40 days after emergence. After 100 days of emergence, the standard conversion rate of nicotine was directly detected without the aforementioned solution treatment. The positive rates of nicotine conversion are statistically compared in Tables 3, 4, and 5.

[0073] Table 3 The nicotine conversion rate of tobacco leaves treated with ethephon solution at different times

[0074]

[0075] Note: "+" in the table indicates that the nicotine conversion rate of tobacco leaves is greater than or equal to 3.5%; "-" indicates that the nicotine conversion rate of tobacco leaves is less than 3.5%.

[0076] Table 4 The nicotine conversion rate of tobacco leaves treated wit...

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Abstract

The invention discloses a nicotine quantitative conversion rate and gene expression based cherry-red tobacco seed screening method. The method includes: selecting seeds from a cherry-red tobacco seedbank, and performing seedling cultivation; after seedling emergence, when seedlings grow to 30-40cm in 30-40d, sampling from bottom 2th-4th tobacco leaves in good conditions; soaking samples in ethephon and abscisic acid mixed solution, perforating, and drying remaining tobacco leaves after perforation until no moisture is left on surfaces; drying the moisture-free tobacco leaves to a constant weight; chemically analyzing nicotine and nornicotine contents of the tobacco leaves, and calculating the nicotine conversion rate; detecting perforated tobacco leaf CYP82E4 gene expression; according tothe nicotine conversion rate and CYP82E4 gene expression, screening out tobacco plants according with a nicotine quantitative conversion rate to cultivate, and removing tobacco plants not accordant with the conversion rate; bagging the screened tobacco plants until maturity is achieved, and reserving seeds to obtain cherry-red tobacco seeds in the nicotine quantitative conversion rate. The methodhas advantages of stability, reliability, high screening speed, high accuracy, controllable seed nicotine conversion rate and the like.

Description

technical field [0001] The invention belongs to the technical field of pre-screening of tobacco seeds, and in particular relates to a method for screening cinnabar tobacco seeds based on quantitative conversion rate of nicotine and gene expression, which is stable, reliable, easy to operate, has fast screening speed, high accuracy and controllable seed nicotine conversion rate. method. Background technique [0002] Cinnabar tobacco refers to the cinnabar color of traditional Chinese medicine in the appearance of tobacco leaves after curing, commonly known as "cinnabar" tobacco leaves. Although cinnabar tobacco is rare, it exists objectively in the history of tobacco production. For example, because the color of cinnabar tobacco is similar to that of cherries in the last century, it is called "Cherry-Red" tobacco. [0003] Conventional tobacco nicotine is mainly composed of nicotine (chemical name is nicotine), nornicotine (desmethylnicotine), mesmin, anatine, cotinine, anat...

Claims

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Application Information

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IPC IPC(8): A01C1/00C12Q1/6895G01N30/02
CPCA01C1/00C12Q1/6895C12Q2600/13C12Q2600/158G01N30/02
Inventor 邹聪明宗吉建隋学艺李勇胡彬彬赵高坤陈颐宋中邦何鲜姜永雷顾华国
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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