【作者】 张莉；

【导师】 张小平；

【作者基本信息】 安徽师范大学 ， 植物学， 2003， 硕士

【摘要】 短萼黄连(Coptis chinensis var．brevisepala W．T．Wang et Hsiao)和黄连(C．chinensis Franch)隶属于毛茛科(Ranunculaceae)黄连属(Coptis Salisb)，前者为后者的变种。二者均为我国常用中药，有清热燥湿、凉血解毒的功能。但目前两种植物的野生种质资源急剧减少，野生种类已难以找到。短萼黄连和黄连同属国家三级保护植物，在中国植物红皮书中濒危级别为渐危。自20世纪70～80年代以来，一些科学工作者对黄连属植物进行了大量的研究工作，但较多的是针对黄连属植物，特别是黄连的成分化学、药材质量、药理学及临床应用等方面，但对黄连属的形态学研究，包括黄连属的解剖学和孢粉学研究，尤其在分子方面的证据几乎为零，短萼黄连与黄连的比较研究也未见报道；黄连属植物的稀危原因与机制至今了解不多，缺少对短萼黄连等的生物生态学特性在内的，有关其致濒机制的系统研究。 为此，本文从地理分布与生态环境、生长与繁殖特性、形态解剖学、种群与群落学特性及免疫血清学和同工酶等几个方面对两种黄连进行了较为详细的研究，以期探寻短萼黄连的致濒机制，并针对性地提出合理的保护对策和建议；同时为短萼黄连和黄连的分类、系统研究提供参考资料。研究结果如下： 1、短萼黄连自然分布于安徽、浙江、广西、福建及广东等省，多生于海拔600～1600m之间的山地沟边林下或山谷阴湿处，呈间断性零星或小块状分布，其模式标本采于广西全州。黄连自然分布于陕西、湖北、四川等地，多生长于海拔500～2000m之间的山地林中或山谷阴湿处，其模式标本采自于四川成口。野生黄连现已极为稀少，市场上所售黄连几乎全是栽培。两者对生态环境的要求相似，均忌高温干旱，要求气温冷凉，降雨丰富，日照少；要求土壤容重小、孔隙度大和砂粒含量高；土壤呈酸性反应，有机质含量高；养分中氮、钾丰富。 2、两种黄连均为浅根系植物，花期基本一致。从移栽于校园内的情况看，黄连始花的时间略早于短萼黄连。两种黄连每花具有雌蕊心皮8～12，每心皮具胚珠6～10枚，但由于两者的雄蕊败育及胚珠败育率均较高，加上雌雄蕊异熟，雄蕊花丝较短，雌蕊较长，而开花季节传粉昆虫因气温较低活动较少，结实率较低，这也是两种黄连的濒危原因之一。 3、短粤黄连在形态、解剖方面与黄连的结构基本一致，均为发达须根系植物，根只具有初生结构，表皮具角质层，上表皮无气孔，沿叶脉处密生指状表皮毛;下表皮分布无规则型气孔;两者花粉均为球形，具散孔，花粉外表壁及孔盖有短刺，黄连花粉孔的密度大于短曹黄连。 4、免疫血清学是植物分类学研究中有效而简便的方法之一。从两种黄连的当年生成熟叶片中提取多组分球蛋白，作为抗原，进行的抗原电泳谱带相同，均为6条带，且迁移率也相同，Rf值分别为0.013、0.081、0.200、0.338、0.513和0.656。说明两种黄连具有同源蛋白或相同的蛋白。免疫双扩散的结果显示，抗原与抗体间反应所产生的沉淀线，无论是以抗原为中心，另两个为相同或不同的抗血清，还是以抗血清为中心，另两个为相同或不同的抗原，均产生一条清晰的沉淀带，且两沉淀带相吻合，边缘光滑，说明两种黄连的抗原距离为零，二者具有相同的蛋白质。由此得出，用免疫血清学方法难以对两种黄连作出分类学上的鉴定。 5、利用同工酶电泳技术对两种黄连的三种同工酶进行了分析研究。两种黄连的过氧化物酶同工酶共有5条谱带，前三条带染色较深，后两条带较浅，两种及居群间无差别;酉旨酶同工酶共有4条谱带，其中前两条带染色较深，且种间无差别;苹果酸脱氢酶谱带最多，共有9条，且8、9两条带较粗，种间及种群间也无差异。对三种同工酶谱带的聚类分析认为，在本文研究的3种同工酶水平上，两物种及其不同居群间无差别。短尊黄连居群间的同工酶谱带无差异，说明短尊黄连的遗传多样性水平较低，居群间的遗传一致程度很高。由此认为，该物种的居群间遗传距离与地理分布差异不显著。遗传多样性较低也有可能是其稀危的因素之一。 6、根据对两物种的形态、解剖、花粉、免疫血清学、同工酶等方面的研究，确立两种黄连的分类地位，支持诚静容等将短尊黄连作为黄连一变种的处理。 7、根据短尊黄连的生物生态学特性，建议采取就地保护和迁地保护的措施对其实施保护;同时加强自然保护区的建设，保护其生存环境，制定相关的规章制度，做好宣传工作，使广大人民意识到生物多样性丧失的严重后果;严禁山民和药农大量采挖。另外应加大科学研究，以寻求解决短粤黄连的解危措施。更多还原

【Abstract】 Coptis chinensis var.brevisepala and C. chinensis which are often used as traditional Chinese medicine for removing internal heat belong to the genus Coptis Salisb from the family Ranunculaceae. The former is a variety of the latter. At present, its resources of the species are decreasing quickly. It is rather difficult to find the wild species now. C.chinensis var.brevisepala was listed as an endangered species of our country in Red Data Book. Although some workers have paid much attention to C. chinensis since 1980’, the research of C. chinensis var. brevisepala and relationship to C. chinensis are not reported. The mechanism by which the endangered situations of the two species were invoked natures is still unknown.This paper is mainly dealing with the two species regarding to population biology, growth and reproduction biology, serology and isozyme of the two species. In this study, The threatened factors and the measures for preserving the species were proposed. The results are as follows:1. C.chinensis var.brevisepala is distributed natively in the areas of Anhui, Zhejiang, Guangxi, Fujiang provinces, while C.chinensis occurs in Shanxi, Hubei and Sichuan etc. The natural distribution of C. chinensis is very limited of narrow, fragmentary areas. The population size is very small, so it is difficult to find wild individuals in some areas. Most of the populations are short of adult individuals. Every population isn’t dominant in its community. The two species grow in mountain forest or at the edge of mountain valley at the altitude of 600-1600m, with the surroundings beings moist and shady, and the soil yellow-brown acid with the pH of 5.5 ~ 7.0.2. From observation of the plants cultivated in Anhui normal university, the time C. chinensis begin to flower is earlier than that of C. chinensis var.brevisepala and the rate of pollination is very low. There are 8-12carpels in a flower, generally, of which 5-6develop into fruits and in each follicle, there are 5-6 ripe seeds. Some ovules don’t develop fully. In some flowers, the pistil is not pollinated, so the pistil didn’t develop. The pistil is higher than stamen. Earlier-flowering, cold weather and scarce of insect activities, probably caused the low set-rate of seed, which threatened the growth of the two species.3. C. chinensis var.brevisepala is identical with C. chinensis in morphological and anatomical characters. Both have fibrous root system and the primary structure is not strong. The leaf epidermis of them have cuticle. The stomatas in irregular form were distributed on the abaxial epidermis, and hairs were distributed on the adaxial along vein. The pollen grains of the two species are spheroidal, pantoporate with pore membrae spinulose. The little difference between the two species is that the bract of C.chinensis var.brevisepala has the phenomenon of special change and its sepal and petal are shorter and wider than those of C. chinensis.4. The double-diffusion serology is one of the useful and simple methods in taxonomy. The proteins extracted from the mature leaves of the two species acted as antigen. Put them in electrophoresis, then got 6 bands. The results showed that the moving rate is identical. The values of Rf are 0.013, 0.081, 0.200, 0.034, 0.513, 0.656. C. chinensis has the same protein with C. chinensis var.brevisepala. The result of the agar gel double diffusion produced one band, and the adjoining bands are in coincide. The edges of the two bands are smooth. This showed the precipitation reaction can not distinguish between the two species.5. Isozyme technology was explored to study three types of isozymes. POD produced 5 bands. The former 3 bands stained strongly, while the last two bands stained weakly. The research indicated that the two species and the populations have no difference. EST produced 4 bands, the two of which stained strogly. EST in the two species is consistent too. MDH produce 9 bands, and the 8th, 9th are wide than the others, no much difference in both, so the diversity genetic is low.更多还原