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期 刊 ：植物遗传资源学报 2016年 17卷 5期

关键词：大麻素；四氢大麻酚；大麻二酚；化学型；基因型；

Keywords:cannabinoid, tetrahydrocannabinol, cannabidiol, chemotype, genotype,

摘 要 ：大麻素（Cannabinoids）是大麻植物中特有的次生代谢产物，主要成分为四氢大麻酚（Tetrahydrocannabinol, THC）和大麻二酚（Cannabidiol, CBD）。本文通过对我国不同来源地的23份大麻种质资源（共69个单株材料）中THC和CBD含量特征及其合成关键酶基因多态性进行分析，鉴定了我国大麻种质资源的大麻素化学型及基因型。结果显示，69个单株中大麻素含量差异显著，THC含量均值为0.56%，范围为0.01～2.45%；CBD含量均值为0.53%，范围为0～2.24%；根据CBD/THC含量比值，大麻资源可划分为毒品型（占44.93%，）、中间型（占20.29%）和纤维型（占34.78%）三种大麻素化学型，毒品型、中间型中分别有93.5%和71.4%的植株中THC含量>0.3%，纤维型植株中THC含量≤0.08%。三种化学型遗传位点（共显性位点B）的基因型分别为BT/BT、BT/BD和BD/BD；BT等位基因（THCAS）存在10个变异位点，氨基酸序列有4处变异，BD等位基因（CBDAS）存在4个变异位点，均为同义突变。根据THCAS和CBDAS基因多态性，设计了一个共显性复合PCR分子标记，可准确鉴定出大麻三种化学型。研究结果揭示了我国大麻种质资源中大麻素含量、化学型和基因型三者之间的关系，可为大麻素遗传研究与利用提供理论依据。

Abstract:Cannabinoids are specific secondary metabolites in the plant of Cannabis sativa L. Tetrahydrocannabinol (THC）and cannabidiol (CBD）are the main constituents of cannabinoids. To distinguish the chemotype and genotype of cannabinoids of Cannabis germplasm, 23 Cannabis germplasm resources (including 69 single plants) from different areas of China were selected to analyze content characteristics of THC and CBD, and also gene polymorphism of key enzymes during the pathway of cannabinoid biosynthesis. The results showed that the cannabinoid contents of 69 single plants were significantly different. The average content of THC was 0.56% ranging from 0.01% to 2.45%, The average content of CBD was 0.53% ranging from 0 to 2.24%. According to CBD/THC ratio, all Cannabis plants can be divided into 3 chemotypes: drug-type (accounting for 44.93%), intermediate-type (accounting for 20.29%) and fiber-type (accounting for 34.78%). The THC content of these plants, including 93.5 % plants among drug-type and 71.4% plants among intermediate-type, are more than 0.3%. The THC content of plants in fiber-type are less than 0.08%. Genotypes in drug-type, intermediate-type and fiber-type were BT/BT, BT/BD and BD/BD respectively. The sequences alignment showed that BT allele (THCAS) had 10 mutation sites in nucleotide sequences, and 4 mutation sites in the deduced amino acids. The BD allele (CBDAS) had 4 mutation sites in nucleotide sequences, but no mutation in the deduced amino acids. According to the polymorphism of THCAS and CBDAS, a codominant multiplex PCR molecular marker was designedto identify three chemotypes of cannabinoids. This study revealed that the relationship among cannabinoid contents, chemotypes and genotypes, which can provide a theoretical foundation for genetic research and utilization of cannabinoids.

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