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Nicotine converted tobacco plant color quality rapid screening method based on incubation treatment

A technology of nicotine and tobacco strains, applied in the field of chemical analysis and detection, can solve the problems of low identification efficiency, low efficiency, and difficult operation, and achieve the effects of shortening incubation time, improving work efficiency, and shortening analysis time

Active Publication Date: 2020-08-07
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is very effective but time-consuming. The incubation reaction takes 4-6 days, and it takes several hours to determine the content of nicotine and nornicotine. Therefore, it is not suitable for the screening of large-scale tobacco strain samples.
[0004] The above methods are time-consuming, inefficient, time-consuming and labor-intensive, and are not suitable for the screening of large-scale tobacco strain samples. In addition, tobacco leaves need to be hatched by adding stimulants and controlling humidity. The operation process needs to strictly control the hatching conditions. It is difficult to operate in the field environment, which leads to low identification efficiency

Method used

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  • Nicotine converted tobacco plant color quality rapid screening method based on incubation treatment
  • Nicotine converted tobacco plant color quality rapid screening method based on incubation treatment
  • Nicotine converted tobacco plant color quality rapid screening method based on incubation treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] S1. Seedling raising and sampling

[0041] Yunyan 85 tobacco seeds were sown in respective seedling trays (9×18 holes, hole diameter: 3cm×3cm) in the greenhouse, and samples were taken 45 days after sowing. For each seedling, get its largest leaf (length and width product greater than 35cm 2 ), fold the collected leaves in half along the main vein, and then fold them in half again, and use a puncher with a diameter of 6 mm to punch holes on the folded tobacco leaves for 3 times (when punching, try to make the three holes evenly distributed on the folded tobacco leaves) , collect 12 circular leaf pieces with a diameter of 6mm, and put them into a 1.5mL plastic centrifuge tube.

[0042]S2. Sample incubation: seal the leaf pieces in a container at 37°C for 10 hours to obtain hatched tobacco leaves, seal the leaf samples in a 1.5mL centrifuge tube for incubation, the moisture of the leaves is blocked in the centrifuge tube, so no additional ambient humidity.

[0043] S3,...

Embodiment 2

[0050] The method of Example 2 is consistent with that of Example 1, the only difference is that the present example uses PPNC to distinguish transformed strains from non-transformed strains. Compared with the Shi method, tobacco seedlings with a measured PPNC value greater than or equal to 1.8% are identified as transformed strains, this method can completely remove transformants without loss of non-transformers.

Embodiment 3

[0052] S1. Seedling raising and sampling

[0053] Burley tobacco TN90 seeds were sown in respective seedling trays (9×18 holes, hole diameter: 3cm×3cm) in the greenhouse, and samples were taken on the 50th day after sowing. For each seedling, get its largest leaf (length and width product greater than 35cm 2 ), fold the collected leaves in half along the main vein, and then fold them in half again, and use a puncher with a diameter of 5 mm to punch holes on the folded tobacco leaves for 3 times (when punching, try to make the three holes evenly distributed on the folded tobacco leaves) , Collect 12 circular leaf pieces with a diameter of 5mm and put them into a 1.5mL plastic centrifuge tube.

[0054] S2. Sample incubation: seal the leaf pieces in a container at 37° C. for 12 hours to obtain hatched tobacco leaves;

[0055] S3, alkaloid extraction

[0056] Methyl butyl ether (MTBE) was used as extractant, and quinoline was used as internal standard. For each seedling sample...

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Abstract

The invention discloses a nicotine converted tobacco plant color quality rapid screening method based on incubation treatment. The method comprises the steps of seedling sampling, alkaloid extraction,sample incubation, instrument analysis and data analysis. The tobacco leaves are hatched and cultured without needing to add an activator and control the humidity, and a tobacco seedling and tobaccoleaf sample is placed in a closed container to be incubated for 10-12 hours, so that the alkaloid extraction and analysis can be carried out. According to the experiment, the humidity of the incubation reaction does not need to be additionally controlled, and the activator does not need to be added, so that the reaction condition is simplified, the incubation reaction can be carried out in any temperature-controllable environment, the incubation time is greatly shortened, and the working efficiency is improved. According to the invention, thealkaloid content analysis is carried out by using agas chromatograph-mass spectrometer, and the contents of nicotine and nornicotine can be determined within 5-6 minutes, so that a PNC value is obtained, and the analysis time of a sample solution is shortened. According to the invention, the PPNC is introduced as a simplified scheme of the PNC, is easier to obtain than the PNC and is not influenced by the factors, such as standard substance purity, standard curve quality, etc.

Description

technical field [0001] The invention belongs to the technical field of chemical analysis and detection, and in particular relates to a rapid screening method for the color quality of nicotine-converted tobacco plants based on hatching treatment. Background technique [0002] In cultivated tobacco, the mutant plants that can convert most of the nicotine into nornicotine are called "transformers". The content of other carcinogens increased exponentially, and this change led to a reduction in the smoking quality of the transformed tobacco leaves and an increase in the risk of cancer. [0003] Therefore, how to identify and remove the transformed strains from the cultivated tobacco population is extremely necessary for the production of qualified tobacco leaves. At present, the method for identifying transformed strains is usually chemical analysis and detection. Ethylene, ethephon, sodium bicarbonate, etc. are used as activating agents to hatch mature tobacco leaves, so that t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/86
CPCG01N30/02G01N30/06G01N30/8606G01N30/8675G01N2030/045G01N2030/062G01N2030/862G01N33/0098G01N2030/8886A01H1/04A24B15/243G01N30/7206G01N30/8637G01N30/8679G01N2030/042
Inventor 李勇逄涛陈学军隋学艺师君丽李永平孔光辉陈萍吴玉萍
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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