de Meijer Etienne P M, Bagatta Manuela, Carboni Andrea, Crucitti Paola, Moliterni V M Cristiana, Ranalli Paolo, Mandolino Giuseppe
HortaPharm B.V., 1075 VS, Amsterdam, The Netherlands.
Genetics. 2003 Jan;163(1):335-46. doi: 10.1093/genetics/163.1.335.
Four crosses were made between inbred Cannabis sativa plants with pure cannabidiol (CBD) and pure Delta-9-tetrahydrocannabinol (THC) chemotypes. All the plants belonging to the F(1)'s were analyzed by gas chromatography for cannabinoid composition and constantly found to have a mixed CBD-THC chemotype. Ten individual F(1) plants were self-fertilized, and 10 inbred F(2) offspring were collected and analyzed. In all cases, a segregation of the three chemotypes (pure CBD, mixed CBD-THC, and pure THC) fitting a 1:2:1 proportion was observed. The CBD/THC ratio was found to be significantly progeny specific and transmitted from each F(1) to the F(2)'s derived from it. A model involving one locus, B, with two alleles, B(D) and B(T), is proposed, with the two alleles being codominant. The mixed chemotypes are interpreted as due to the genotype B(D)/B(T) at the B locus, while the pure-chemotype plants are due to homozygosity at the B locus (either B(D)/B(D) or B(T)/B(T)). It is suggested that such codominance is due to the codification by the two alleles for different isoforms of the same synthase, having different specificity for the conversion of the common precursor cannabigerol into CBD or THC, respectively. The F(2) segregating groups were used in a bulk segregant analysis of the pooled DNAs for screening RAPD primers; three chemotype-associated markers are described, one of which has been transformed in a sequence-characterized amplified region (SCAR) marker and shows tight linkage to the chemotype and codominance.
将纯大麻二酚(CBD)和纯Δ-9-四氢大麻酚(THC)化学型的自交大麻植株进行了四次杂交。对所有属于F(1)代的植株进行气相色谱分析,以确定大麻素组成,结果发现它们均具有混合的CBD-THC化学型。选取10株F(1)代单株进行自花授粉,并收集和分析了10个自交F(2)代后代。在所有情况下,均观察到三种化学型(纯CBD、混合CBD-THC和纯THC)的分离符合1:2:1的比例。发现CBD/THC比值具有显著的子代特异性,并从每个F(1)代传递给由其衍生的F(2)代。提出了一个涉及一个基因座B、具有两个等位基因B(D)和B(T)的模型,这两个等位基因是共显性的。混合化学型被解释为是由于B基因座上的基因型B(D)/B(T),而纯化学型植株是由于B基因座上的纯合性(B(D)/B(D)或B(T)/B(T))。有人认为这种共显性是由于这两个等位基因对同一合酶的不同同工型进行编码,它们对共同前体大麻萜酚分别转化为CBD或THC具有不同的特异性。利用F(2)代分离群体对混合DNA进行分组分离分析,以筛选随机扩增多态性DNA(RAPD)引物;描述了三个与化学型相关的标记,其中一个已转化为序列特征性扩增区域(SCAR)标记,并显示出与化学型紧密连锁且为共显性。