滇南农家大麻品种中大麻素化学型及基因型研究

以1个滇南农家大麻品种群体为研究对象,通过化学分析及同源克隆方法,研究了21个单株中2种主要大麻素——四氢大麻酚(THC)和大麻二酚(CBD)的化学型和基因型,以揭示大麻素含量、化学型以及基因型三者之间的关系,为工业大麻新品种选育提供理论依据。研究表明:(1)化学检测结果显示,21个单株均含有THC,THC含量在0.07%~1.35%之间,其中7个单株仅含THC,5个单株含THC和微量CBD,9个单株同时含有THC和CBD,CBD含量范围为0~0.58%。(2)CBD/THC比值显示,该群体仅存在毒品型和中间型2种化学型,且中间型大麻中THC和CBD含量显著正相关。(3)基因扩增及测序分析结果显示,该群体为基因型杂合群体,群体内THCA合成酶基因存在5个变异位点,CBDA合成酶基因存在2个变异位点,但变异位点和THC及CBD的含量无直接关系。(4)群体内单株的基因型和化学型完全对应,且THCA合成酶基因及CBDA合成酶基因可作为分子标记来鉴定单株化学型。     

大麻种质资源中大麻素化学型及基因型鉴定与评价

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(THC,tetrahydrocannabinol)和大麻二酚(CBD,cannabidiol)。本研究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%)3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%。3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3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。     

大麻和大麻受体与免疫应答

大麻类物质通过与免疫细胞表面抑制性G蛋白偶联受体CB1和CB2结合 ,调节免疫细胞的功能和细胞因子的产生。大麻受体可根据组织分布不同分为中枢型和外周型两类 ,前者主要分布在脑组织中而后者主要分布在免疫细胞表面。绝大多数大麻类物质具有结合两种类型受体的能力 ,而且内源性大麻可以快速通过血脑屏障 ,提示其可能是神经 免疫轴中的活跃递质。     

DNA条形码技术在毒品原植物大麻鉴定中的应用

准确鉴定毒品原植物大麻的种属及品种具有重要的理论和实践意义。为了探讨DNA条形码技术用于毒品原植物大麻种属鉴定及品种鉴定的可行性,该研究以60份大麻原植物(分别采自内蒙、黑龙江、陕西延安、陕西榆林4个地区的栽培大麻雌雄各6株及新疆玛纳斯地区的野生大麻雌雄各6株)为材料,通过从其叶片中提取的DNA为模版,利用核糖体DNA基因间隔区的通用引物ITS2和叶绿体DNA的通用引物psbAtrnH进行PCR扩增,对扩增片段进行双向测序,将测序结果进行人工矫正和比对。结果显示:所有大麻样本的ITS2扩增片段序列没有变异完全一致,但psbA-trnH扩增片段变异较大共检测出8种cpDNA单倍型,用MEGE5.1软件计算种间遗传距离,并构建NJ系统聚类树可以有效把这五个地区的大麻样本区别开来,因此证明DNA条形码技术在毒品原植物大麻的种属鉴定方面具有可行性,但其用于大麻的种属鉴定的准确性、可靠性及在其来源地鉴定及品种鉴定中的可能性还有待进一步深入地研究。     

天然C19—二萜生物碱的核磁共振谱（Ⅱ）

本文将迄今为止已发表的230余个天然C_(19)二萜生物碱,按其结构和核磁共振波谱特征分为8个类型,着重叙述了每个类型的结构和核磁共振波谱,并对某些立体构型与~(13)C-NMR化学位移的关系进行了讨论。最后以表格的形式报导了每个天然C_(19)-二萜生物碱的结构及其主要植物来源,并将其中170余个生物碱的~(13)C-NMR化学位移数据分门别类,整理成表,有助于C_(19)-二萜生物碱的结构鉴定。这是本文的第二部分,包括二氧亚甲基型,氨缩醛型,C-18甲基型。C_(18)型不饱和型和内脂型等六个类型的结构特征及其核磁共振波谱。     

ISSR分子标记检测大麻遗传多样性的初步分析

目的:利用ISSR分子标记技术初步检测和分析中国云南和内蒙地区毒品原植物大麻的遗传多样性。方法:用CTAR法提取大麻基因组DNA,设计10个ISSR引物,扩增产物采用6%中性聚丙烯酰胺凝胶电泳-硝酸银染色法检测,根据出现的条带数目和片段大小等分析大麻的多样性。结果:从10个ISSR引物中筛选出的4个引物用于2个地区的大麻基因组DNA扩增,PCR产物可以检测到51条重复性较好、带型清晰的DNA片段,其多态性总体比率为78.43%。云南地区和内蒙地区大麻样品可分别获得43和33条带,其中多态性条带分别为33条(76.74%)和21条(63.64%)。结论:ISSR分子标记技术揭示了大麻具有较高的遗传多样性,对于鉴别犯罪现场大麻检材的产地及种属来源具有一定的价值。     

花生籽仁外观和营养品质特征及食用型花生育种利用分析

花生籽仁的外观品质和营养品质是评价食用花生品种的重要指标。本研究对285个不同类型或来源的花生种质资源进行了籽仁外观和营养品质性状的检测和分析,旨在为食用型花生品种选育提供依据。按照植物学类型分析了各类型的8个外观性状和5个营养品质性状的分布。结果表明,我国花生种质资源包含丰富的外观性状变异类型,能够满足不同加工用途对原料的需求,但总体上高油酸、高蛋白和低脂肪资源类型偏少。借鉴美国等烤制花生仁和糖果花生的外观和营养品质特征及我国煮食和烤制花生的生产消费市场,提出了食用型品种选育亲本的策略和备选品种类型,烤制和糖果用途花生育种亲本选配应以普通型、中间型或珍珠豆型中粒农家品种或育成品种为骨干亲本,进一步改良高油酸、高蛋白和低脂肪等品质性状。     

天然C_(19)-二萜生物碱的核磁共振谱(Ⅰ)

本文将迄今为止已发表的230余个天然C_(19)-二萜生物碱按其结构和核磁共振波谱特征分为8个类型,着重叙述了每个类型的结构和核磁共振波谱,并对某些立体构型与~(13)C-NMR化学位移的关系进行了讨论。最后以表格的形式报导了每个天然C_(19)-二萜生物碱的结构及其主要的植物来源,并将其中170余个生物碱的~(13)C-NMR化学位移数据分门别类,整理成表,以便于对相似化合物的~(13)C-NMR进行比较,这样对于分析鉴定C_(19)-二萜生物碱将大有帮助。第一部分包含两个主要的类型——乌头碱型和牛扁碱型,其它类型待第二部分进行讨论。     

中国部分地方黄牛品种线粒体D-loop区的遗传变异与进化分析

测定了13个黄牛品种125个个体的线粒体D-loop区段的全序列,包括12个中国地方黄牛品种的123个个体和德国黄牛2个个体,并进行了分析。结果显示,共检测到93个变异位点,57个单倍型,平均核苷酸差异(average number ofnucleotide differences,k)为22.708,核苷酸多样度(nucleotide diversity,π)为0.0251±0.00479,单倍型多样度(haplotypediversity,Hd)为0.888±0.026,表明我国黄牛品种遗传多样性非常丰富。构建的Neighbor-Joining进化树显示这13个品种主要分成两大类型:普通牛和瘤牛;新发现的特殊类型Ⅲ只有一个西藏阿沛甲咂牛的个体,它与牦牛D-loop序列最相近,证明西藏地区的黄牛与牦牛之间存在基因渗入现象。普通牛和瘤牛在日喀则驼峰牛中占的比例分别是64.3%和35.7%,在阿沛甲咂牛中占的比例分别是50.0%和50.0%,证明了西藏的黄牛也有瘤牛类型。云南牛品种的单倍型非常丰富证明了云南在中国黄牛起源上的重要地位;在27个中国黄牛品种中(本研究11个品种以及GenBank上的16个品种)找到了中国瘤牛的核心单倍型i1,并且对它进行了讨论。同时证明了西藏瘤牛独立于中国瘤牛核心类群的特殊性。     

水稻抗冷性与不同生长阶段的关系

从国际水稻研究所种质库取得50个水稻品种,其中包括粳型品种12个,釉稻18个,爪哇型品种10个,釉×粳杂种3个,以及野生稻7个。在芽期、苗期、移栽期、孕穗期、开花期,进行了耐冷性的测定。结果指出,在不同的水稻类型中,其耐冷性的次序为:粳稻>爪哇型>秈稻>野生稻。从本实验所采用的有限量的品种来看,在苗期粳型品种的耐冷性比其它类型好;在开花期某些爪哇型品种比粳型品种好;在芽期和苗期之间,以及在芽期和开花期之间,其耐冷性有明显的正相关。在其它时期之间未找到这种相关性。这指出,在特殊地区的低温发生时期的一定生育阶段中,进行筛选是必要的。     

A chemotaxonomic analysis of cannabinoid variation in Cannabis (Cannabaceae)

Cannabinoids are important chemotaxonomic markers unique to Cannabis. Previous studies show that a plant's dry-weight ratio of Δ(9)-tetrahydrocannabinol (THC) to cannabidiol (CBD) can be assigned to one of three chemotypes and that alleles B(D) and B(T) encode alloenzymes that catalyze the conversion of cannabigerol to CBD and THC, respectively. In the present study, the frequencies of B(D) and B(T) in sample populations of 157 Cannabis accessions were determined from CBD and THC banding patterns, visualized by starch gel electrophoresis. Gas chromatography was used to quantify cannabinoid levels in 96 of the same accessions. The data were interpreted with respect to previous analyses of genetic and morphological variation in the same germplasm collection. Two biotypes (infraspecific taxa of unassigned rank) of C. sativa and four biotypes of C. indica were recognized. Mean THC levels and the frequency of B(T) were significantly higher in C. indica than C. sativa. The proportion of high THC/CBD chemotype plants in most accessions assigned to C. sativa was <25% and in most accessions assigned to C. indica was >25%. Plants with relatively high levels of tetrahydrocannabivarin (THCV) and/or cannabidivarin (CBDV) were common only in C. indica. This study supports a two-species concept of Cannabis.     

The inheritance of chemical phenotype in Cannabis sativa L

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.     

大麻植物中大麻素成分研究进展

大麻（Cannabis sativa）是一种古老的栽培植物, 它既是一种毒品原植物, 又是一种极具开发利用价值的经济作物。大麻素是大麻植物中特有的含有烷基和单萜分子结构的一类次生代谢产物, 目前已分离出70多种, 其中包含使人致幻成瘾的四氢大麻酚（THC）。该文就大麻植物中几种主要的大麻素成分： 四氢大麻酚、大麻二酚（CBD）和大麻环萜酚（CBC）的存在特征、含量变化、生物合成途径、各关键酶及其基因、遗传方式等方面的研究进行概括和归纳, 并展望了当前大麻素的主要研究方向, 对加快我国大麻素的相关研究及大麻育种具有参考意义。     

Flower power: floral reversion as a viable alternative to nodal micropropagation in Cannabis sativa.

In Vitro Cellular & Developmental Biology - Plant - The legalization of recreational and medicinal Cannabis sativa L. has been gaining global momentum, therefore increasing interest in Cannabis...     

The endocannabinoid system and prospects her stimulation in clinical medicine

The history of use by the man of plants of a sort Cannabis totals more than 4000 years. The people have begun to use Cannabis in stone century. On Taiwan archaeologist the rests of utensils made with application of stalks Cannabis more of 10000 years back were found. Cannabis (Cannabis sativa, Cannabis sowing, named also "Indian") - cultural plant, which has set of applications. From it received fibres for hemp of ropes and make a fabric similar on linen. Its stalks went on manufacture glossy of a paper and building of plates. Her sabadilla were used for graziery a bird; oil from sabadilla Cannabis offered as fuel instead of diesel. The greatest popularity Cannabis has received as raw material for reception of products (marijuana, hashish etc.), causing at the man psychotropic--first of all psychomymetic--effects, that at their regular application can result in formation of dependence. Besides the attempts were undertaken to use preparations Cannabis in the medical purposes: at migraine, spasmes, vomiting, pains of a different origin etc.     

Expanding the laticifer knowledge in Cannabaceae: distribution,morphology, origin,and latex composition

Protoplasma - Cannabaceae is a known family because of the production of cannabinoids in laticifers and glandular trichomes of Cannabis sativa. Laticifers are latex-secreting structures, which in...     

THE STRUCTURE OF GRANA IN FLOWERING PLANTS

A model of granum structure, consisting of multiple helical frets wound around each cylindrical granum, is proposed as a representation of granum structure for flowering plants in general. This model was originally formulated for grana in the mesophyll plastids of Zea mays and was subsequently extended to grana of Phaseolus vulgaris by additional studies. The model is now shown to be applicable also to grana in the plastids of Cannabis sativa, Elodea canadensis, Nicotiana rustica, Pisum sativum, and Spinacia oleracea. This sample of seven angiosperms includes genera commonly studied by other workers in their ultrastructural investigations of plastid structure.     

Back to the roots: protocol for the photoautotrophic micropropagation of medicinal Cannabis

Plant Cell, Tissue and Organ Culture (PCTOC) - The aim of this protocol was to develop an alternative in vitro propagation system for Cannabis sativa L. by mimicking nursery-based vegetative...     

Effect of Cannabis sativa L. root,leaf and inflorescence ethanol extracts on the chemotrophic response of entomopathogenic nematodes

Plant and Soil - Soils represent the natural habitat of entomopathogenic nematodes (EPNs). When moving in soil, EPNs are oriented to follow a chemical signal (chemotaxis). Cannabis sativa L. is...