A Rapid Screening Method for High Nicotine Tobacco Mutants

Abstract

The invention discloses a method for rapidly screening high-nicotine tobacco mutants, and belongs to the biotechnical field. The method comprises the following steps: 1) dibbling tobacco M2-generation seeds onto an aqueous methyl jasmonate solution; 2) 15 days later, screening single plants of which root lengths are significantly suppressed; 3) transferring the single plants screened in the step 2) to an MS culture solution, and culturing for 20 days; 4) transferring the tobacco plants in the step 3) to a provisonal planting plate containing nutrient soil, and 30 days later, transplanting to a field; 5) performing field planting and routine management; 6) determinating nicotine contents at three different time points; 7) selfing and harvesting seeds. According to the method, within 20 days, high-nicotine materials are screened from 4000 China tobacco-100 mutants, and 5 high-nicotine flue-cured tobacco materials are obtained; the highest nicotine content reaches 7.09%, which is the highest nicotine content for current flue-cured tobacco; the efficiency for screening the tobacco mutants is greatly improved, the workload for screening the high-nicotine tobacco is reduced, and the production efficiency is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for rapidly screening high-nicotine tobacco mutants. Background technique [0002] Nicotine is a major secondary metabolite of tobacco and the main addictive ingredient in cigarette products. Tobacco seeds do not contain nicotine, and the content of nicotine in tobacco seedlings and before topping is very low. After topping, the content of nicotine is very low. In the type of flue-cured tobacco, the nicotine content generally accounts for 2 to 4% of the dry weight. [0003] Flue-cured tobacco is the main raw material of cigarette products. In recent years, with the implementation of the tobacco “harm reduction and tar reduction” strategy, the tar release amount and smoke hazard index of domestic cigarettes have gradually decreased. , concentration” also decreased. In order to compensate for the lack of "aroma, strength, and concentration" of cigarettes after reducing tar, ...

AI Technical Summary

Problems solved by technology

However, because mutagenesis generally produces unfavorable traits, the probability of producing mutants with good traits is very small, so screening materials that meet the target needs from the mutagenized offspring is a lot of work.

At present, a rapid selection method for high-nicotine mutation materials has not been established. The nicotine level of each individual tobacco plant is determined by directly measuring the nicotine content after topping in the field stage. This method requires all the selected materials to be planted. Due to the tall tobacco plants, screening a mutant population of 5,000 copies (25 individual plants for each material) requires nearly 70 acres of land for planting. The huge material population needs to be planted, artificially planted, managed by conventional cultivation measures in the field, and topped. Waiting for the process is very costly, and the determination of the nicotine content of each single leaf after harvesting is also very time-consuming and expensive, and the screening efficiency is very low

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