Genetic Diversity of Cephalotaxus mannii, a Rare and Endangered Plant

Using RAPD technique, the DNA diversity of Cephalotaxus mannii Hook. f., its genetic diversity pattern, the reasons for its endangered position and conservative approaches were studied. The results show that: 1. The genetic diversity of C. mannii collected from five localities in Hainan is low, and its adaptability to environmental change is weak. 2. The differences of genetic diversity between intra- and inter-populations are great, and the major variation distributes within the population (DNA diversity is 85.1%). 3. The excessive lumbering, man-made destruction, violent typhoon, edible value of the seeds and genetic drift were the main reasons for the low-level genetic diversity of C. mannii and its endangered position. 4. The difference of the micro-environment and other random factors affecting the population should also be taken into full consideration in the study and in protection of such occasionally scattered plants. 5. Enforced measures should be taken to protect the present population, enlarge the population and lower the loss rate of its gene. Mt. Limulin should be chosen as a conservative spot because of its high genetic diversity and less destruction of the forest. Meanwhile, the protection of other populations should be enforced. 6. The differences within and between the populations are great based on different primers used. The change of proportions in polymorphic loci between the populations is more than that between the primers.

贡山三尖杉的新名称(Cephalotaxus talonensis Cheng et Feng ex S.G.Lu et X.D.Lang)及分类地位(英文)

提议贡山三尖杉的新名称Cephalotaxus talonensis Cheng et Feng ex S.G.Lu et X.D.Lang替代不合法的旧名称Cephalotaxus lanceolata K.M.Feng in Cheng et al.(晚出同名)。对比贡山三尖杉和三尖杉Cephalotaxus fortunei Hooker的模式标本和自然生长状态下的叶片形态,支持将贡山三尖杉处理为种的等级,不赞同Silba(1990)将贡山三尖杉降为三尖杉的变种[Cephalotaxus fortunei Hooker var.lanceolata(Feng)Silba]。文中提供4幅参照图片。

不同种群中海南粗榧(Cephalotaxus mannii)形态变异研究

通过变异系数和性状差异显著性检验，研究了海南粗榧在坝王岭等５个种群内和种群间不同形态变异以及形态总体变异。结果表明：①海南粗榧营养器官性状比生殖器官性状变异大；枝性状组中，分枝角度变异较小；叶性状组中，叶数量变异较大；球果、种子生物量变异都大于它们的其它性状。②在５个种群内，同一性状变异幅度基本相同。③同一性状在种群间和种群内的变异基本接近。④不同性状在种群间存在差异，有的显著，有的不显著。

海南粗榧(Cephalotaxusmannii Hook.f.)化学元素含量及变异研究

利用变异系数和差异显著性检验 ,对海南坝王岭等五个海南粗榧群体植株及其不同器官的 C、H、N、K等15种化学元素分布及变异进行研究。结果表明 :1)海南粗榧 (Cephalotaxus mannii Hook. f.)中的化学元素含量分布总体呈 Ca>K型 ,但枝、茎、根、种子等不同器官却表现为 K>Ca型 ;2 )海南粗榧群体内各化学元素含量的变异系数较大 ,但群体内和群体间基本无显著差异 ;3)海南粗榧不同群体同一器官间元素含量相对稳定 ,变异系数不太大 ,群体不同器官间则呈现显著甚至极显著的差异 。

不同种群海南粗榧(Cephalotaxus mannii)遗传多样性研究

采用垂直板型凝胶电泳技术 ,对海南粗榧 5个种群的遗传多样性进行了研究 ,结果表明 :海南粗榧遗传多样性水平低 ,多态位点比率 P=0 .3 3 ,等位基因平均数 A=1 .3 3 ,平均期望杂合度为 He=0 .1 3 5 ,观察杂合度 Ho=0 .1 3 9。

海南粗榧(Cephalotaxus mannii Hk.f.)5个种群遗传多样性及分化研究

采用垂直板型聚丙烯酰胺凝胶电泳测定了海南粗榧 5个种群遗传多样性和遗传分化程度。结果表明 :海南粗榧种群遗传多样性水平较低 ,多态位点比率 P=0 .33,等位基因平均数 A=1 .33,平均期望杂合度为 H e=0 .1 35,观察杂合度为 H o=0 .1 39。种群间遗传分化程度较低 ,基因分化系数 Gst=0 .1 2 3,种群间遗传一致度和遗传距离的均值分别为 0 .971 9、0 .0 2 88。黎母山种群与其它种群分化最大 。

海南粗榧(Cephalotaxus mannii)不同种群表型结构的数量分析

The key characters of the five different populations of Cephalotaxus mannii have been analyzed.No differentiation exists between the other four populations except the case of the Mt.Diaoluo population.

Five New Alkaloids from Cephalotaxus lanceolata and C. fortunei var. alpina

Five new alkaloids(1–5)were isolated from the leaves and twigs of Cephalotaxus lanceolata and C.fortunei var.alpina along with 24 known alkaloids.The new structures were elucidated based on spectroscopic data including 1D and 2D NMR,FTIR,UV and MS.These new alkaloids showed no cytotoxicity to HeLa,SGC-7901 gastric cancer,and A-549 lung cancer cell lines.

Two new flavonoid diglycosides from Cephalotaxus sinensis

Two new acetylated flavonoid diglycosides, scutellarein-5-O-[α-L-rhamnopyranosyl-（1 → 4）-β-D-6-O-acetylglucopyranoside]（1）, apigenin-5-O-[α-L-rhamnopyranosyl-（1→ 2）-β-D-3,6-di-O-acetylglucopyranoside] （2）, were isolated from the leaves of Cephalotaxus sinensis. Their structures were determined by chemical and spectroscopic methods including HR-FAB-MS,^1H-^1H COSY, HSQC and HMBC experiments.

Cytochalasins from Xylaria sp. CFL5, an Endophytic Fungus of Cephalotaxus fortunei

Three previously undescribed cytochalasins,named xylariasins A‒C(1‒3),together with six known ones(4‒9)were iso-lated from Xylaria sp.CFL5,an endophytic fungus of Cephalotaxus fortunei.The chemical structures of all new compounds were elucidated on the basis of extensive spectroscopic data analyses and electronic circular dichroism calculation,as well as optical rotation calculation.Biological activities of compounds 1,4‒9 were evaluated,including cytotoxic,LAG3/MHC II binding inhibition and LAG3/FGL1 binding inhibition activities.Compounds 6 and 9 possessed cytotoxicity against AGS cells at 5μM,with inhibition rates of 94%and 64%,respectively.In addition,all tested isolates,except compound 6,exhibited obvious inhibitory activity against the interaction of both LAG3/MHC II and LAG3/FGL1.Compounds 1,5,7,and 8 inhibited LAG3/MHC II with IC50 values ranging from 2.37 to 4.74μM.Meanwhile,the IC50 values of compounds 1,7,and 8 against LAG3/FGL1 were 11.78,4.39,and 7.45μM,respectively.

The Causes of Dormancy and the Changes of Endogenous Hormone Content in Cephalotaxus sinensis Seeds

In many wild species, seeds are dormancy at maturity and will not germinate even under favorable environment conditions. Dormancy is a complex trail that is determined by many factors. Some studies have shown that cold stratification and the application of gibberellic acid (GA) can break seed dormancy and promote seed germination. The present study investigated the causes of plant dormancy and the effect of cold stratification and different concentrations of exogenous GA3 in regulating Cephalotaxus sinensis seed germination. Results showed that C. Sinensis seeds have good water permeability, which suggested that seed coats were not the main cause that inhibited the seed germination. There were germination inhibitions in all parts of seeds, and the order of inhibitory effect was: testa < endosperm (embryo), which indicated that existence of germination inhibitions was the main reason causing seed dormancy. Endogenous GAs and IAA (indole-3-acetic acid) content increased, while ABA (abscisic acid) content decreased over the experiments. ZR (zeatin riboside) content decreased in the early phase of cold stratification, but rebounded by the end of the experimental period. The changes of endogenous hormone indicated that GA, IAA and ZR played a positive role in seed germination, whereas ABA was associated with seed dormancy. Besides, the relative ratio of GA/ABA, IAA/ABA and ZR/ABA may play a more important role than their absolute level during the seed development.

从种子和叶片特征论高山三尖杉(Cephalotaxus alpina(Li)L.K.Fu)的分类地位(三尖杉科)(英文)

长期以来,对高山三尖杉(Cephalotaxus alpina(Li)L.K.Fu)分类地位的争议缺少可靠证据。本文从居群采样的水平上比较了高山三尖杉和三尖杉(Cephalotaxus fortunei Hooker)的叶片和种子形态特征,两者除种子的长宽比率无显著差异外,种子长度,宽度,厚度,长厚比率,宽厚比率以及叶片长度和宽度均差异极显著。此外,通过散点图分析表明两者的种子宽厚比和叶片宽度的变异是间断而稳定的,这一特征能将高山三尖杉和三尖杉明显地区分,结果支持将高山三尖杉作为种的等级较为合适,不赞成将高山三尖杉作为三尖杉的变种或亚种的分类处理。同时,对标本和原始文献的研究认为胡秀英(1964)发表的园桃杉(Cephalotaxus fortunei Hooker var.globosa S.Y.Hu)和高山三尖杉应是同一分类群。

The Taxonomic History and Original Literatures Arrangements of Living <i>Cephalotaxus</i>(Cephalotaxaceae)

Living Cephalotaxus (plum yews) occurs mainly in East Asia and North of Indo-China peninsula. The study of original taxonomic literatures found that, in early days, the plum yews had ever been described as palnts of Taxus (Taxaceae). Since after the genus name Cephalotaxus Sieb. & Zucc. was firstly cited (from inedited and unpublished Flora Japonica) by Endlicher in 1842, most taxonomist thought which is the earliest time of the genus Cephalotaxus Sieb. & Zucc. was set. However, the genus Cephalotaxus Sieb. & Zucc. was formally effective and valid published by Sieb. & Zucc. in Flora Japonica in 1870 so that there exists some confusions in chronological order. Originally as a genus, Cephalotaxus was included in family Taxaceae and Neger (1907) separated Cephalotaxus from Taxaceae and set family Cephalotaxaceae. Later, Fu (1984) divided Cephalotaxus up into two sections of Cephalotaxus Sieb. & Zucc. sect. Pectinatae L. K. Fu and Cephalotaxus Sieb. & Zucc. sect. Cephalotaxus L. K. Fu on the bases of C. oliveri Masters differ significantly from other taxa of Cephalotaxus in morphological characteristics. Although the plum yews was a little genus of conifers, up to now, about 77 taxa have still been published by taxonomist in different countries or regions. Among them, C. tardiva Sieb. ex Endl. (1847), C. umbraculifera Sieb. ex Endl. (1847), C. sumatrana Miq. (1856) and C. celebica Warb. (1900) have been combined to Taxus, and C. argotaenia (Hance) Pilger (1903) has been combined to Amentotaxus. And at the same time, those taxa existing in Cephalotaxus courently which were accepted by most taxonomist in different countries was also cited in acticle. Lastly, the paper thought presently there still exists many debates about binary classification of Cephalotaxus and the nomenclature of this genus still particularly confusing. Based on some controversial problems, we also point out some taxonomically researching directions at creating some order out of the chaos.

Enantiomeric Cephalotaxus alkaloids from seeds of Cephalotaxus oliveri

Five undescribed alkaloids were isolated from the seeds of Cephalotaxus oliveri along with 27 known ones.The new structures were elucidated based on spectroscopic data including 1D and 2D NMR,MS and calculated ECD spectra.Among them,(+)-acetylcephalofortine C was an enantiomeric Cephalotaxine alkaloids.The performed bioassay revealed that those alkaloids were not cytotoxic against cancer cells and had no neuroprotective properties in the HEI-OC-1 cells model.

Effects of Different Treatments on Physiological Characteristics of Cephalotaxus sinensis Seeds

Cephalotaxus sinensis seeds can’t germinate even in the appropriate environment. However, numerous studies have showed that cold stratification and gibberellin acid (GA) can break the seed dormancy and promote seed germination effectively. To investigate the effect of cold stratification and different concentrations of exogenous GA3 on dormancy breaking in seeds of Cephalotaxus sinensis, we monitored germination rates and changes in soluble sugar, starch, amylase, soluble protein, free amino acid during cold stratification. The results showed that seeds stratified for 5 months germinated to 12.7%, while those disposed with 200, 400, 600 mg/L of GA3 germinated to 29.2%, 21.7%, and 18.4%, respectively. Free amino acid content was enhanced significantly, whereas soluble sugar content decreased during 45 days and then increased constantly. Additionally, the main reserves such as starch, protein decreased significantly during cold stratification, and cold stratification induced increases in the activities of α-amylase, (α + β)-amylase. The preliminary results show that the combination of GA3 and cold stratification has better effect to break seed dormancy.

篦子三尖杉在中国的地理分布及潜在分布区预测

【目的】预测不同气候情景下篦子三尖杉(Cephalotaxus oliveri Mast.)的地理分布格局,为其资源保护、合理利用与开发提供指导,也为其起源与地理演化研究奠定基础。【方法】基于100个分布点信息与19个气候因子,采用BIOCLIM模型构建当前(1970—2000年)以及未来(CO_(2)浓度倍增)篦子三尖杉的地理分布趋势,并采用主成分分析法分析影响其地理分布的主要气候因素。【结果】篦子三尖杉在中亚热带地区以低海拔分布为主。BIOCLIM模型结果显示:在当前气候条件下,篦子三尖杉最佳适生区集中在湖南、湖北、贵州和重庆;未来气候变化情境(CCM3)下,其潜在分布面积将减小,并有向西迁移的趋势;该模型受试者工作特征曲线下的面积为0.911±0.023,预测精度较高。主成分分析结果表明:温度季节性、气温年较差和昼夜温差与年温差比值是限制篦子三尖杉地理分布的主要因素,且温度因子的作用大于水分因子。【结论】随着全球气候变暖,篦子三尖杉的适生区有破碎化及丧失的风险,应重视对该物种的动态监测及迁地保护。

篦子三尖杉的生物碱成分及其抗肿瘤活性研究

为研究篦子三尖杉的化学成分,利用多种色谱分离技术和波谱手段,从其乙醇提取物中分离鉴定了11个生物碱类成分。其中博落回碱(7)、(2Z)-1,2-二氢-2-(1H-吲哚-3-亚甲基)-3H-吲哚-3-酮(8)、1-甲氧基-3-吲哚乙腈(9)、4-甲氧基-1H-吲哚-3-甲醛(10)、吲哚-3-甲醛(11)为首次从三尖杉属植物中分离得到。采用CCK-8法测试了化合物1~11对人乳腺癌细胞株MCF-7、人肝癌细胞株HepG2和人肺癌细胞株A549的体外抗增殖能力,异三尖杉酯碱(1)和三尖杉酯碱(2)对三种肿瘤细胞显示了显著的抗增殖活性。

篦子三尖杉枝叶的木脂素类化合物研究

为丰富篦子三尖杉的化学成分,采用回流提取法,硅胶、ODS、MCI、凝胶及高效液相等色谱分离手段,从篦子三尖杉乙醇提取物中分离得到了8个木脂素类化合物。鉴定为sesquipinsapol B(1)、8-hydroxypluviatolide(2)、异落叶松树脂醇(3)、2-[2-羟基-5-(3-羟丙基)-3-甲氧基苄基]-3-(4-羟基-3-甲氧基苯基)-1-丙醇(4)、trans-3-甲氧基-4,9,9′-三羟基-苯并二氧六环木脂素(5)、松脂素(6)、linderinol(7)、ligraminol E(8)。其中化合物1、2、4、5、7、8为首次从三尖杉属植物中分离得到。

4种微生物杀虫剂对点尾尺蛾幼虫的时间-剂量-死亡率模型分析

为筛选适用于防治点尾尺蛾Ourapteryx nigrociliaris的微生物杀虫剂,发展粗榧Cephalotaxus sinensis等药用植物害虫的生物防治,采用浸渍法测定了甘蓝夜蛾核型多角体病毒Mamestra brassicae nuclear polyhedrosis virus、苏云金芽胞杆菌Bacillus thuringiensis、球孢白僵菌Beauveria bassiana、金龟子绿僵菌Metarhizium anisopliae CQMa421对点尾尺蛾幼虫的致死效应,并使用时间-剂量-死亡率(TDM)模型进行模拟。结果表明,TDM模型均通过Hosmer-Lemoshow拟合异质性检验,4种微生物杀虫剂处理10 d的幼虫校正死亡率最高分别达到79.31%、86.21%、93.10%和72.41%,半致死浓度LC_(50)分别为1.81×10^(6)PIB/mL、2.71×10^(3)IU/mL、9.40×10^(6)孢子/mL以及9.86×10^(6)孢子/mL。综合考虑药剂致死率和致死速度,球孢白僵菌、苏云金芽胞杆菌和甘蓝夜蛾核型多角体病毒的杀虫效果优于金龟子绿僵菌CQMa421,更具应用潜力。本研究为点尾尺蛾的生物防治提供了参考依据。

Ethnopharmacology,chemodiversity,and bioactivity of Cephalotaxus medicinal plants

Cephalotaxus is the only genus of Cephalotaxaceae family,and its natural resources are declining due to habitat fragmentation,excessive exploitation and destruction.In many areas of China,folk herbal doctors traditionally use Cephalotaxus plants to treat innominate swollen poison,many of which are cancer.Not only among Han people,but also among minority ethnic groups,Cephalotaxus is used to treat various diseases,e.g.,cough,internal bleeding and cancer in Miao medicine,bruises,rheumatism and pain in Yao medicine,and ascariasis,hookworm disease,scrofula in She medicine,etc.Medicinal values of some Cephalotaxus species and compounds are acknowledged officially.However,there is a lack of comprehensive review summarizing the ethnomedicinal knowledge o f Cephalotaxus,relevant medicinal phytometabolites and their bioactivities.The research progresses in ethnopharmacology,chemodiversity,and bioactivities of Cephalotaxus medicinal plants are reviewed and commented here.Knowledge gaps are pinpointed and future research directions are suggested.Classic medicinal books,folk medicine books,herbal manuals and ethnomedicinal publications were reviewed for the genus Cephalotaxus(Sanjianshan in Chinese).The relevant data about ethnobotany,phytochemistry,and pharmacology were collected as comprehensively as possible from online databases including Scopus,NCBI PubMed,Bing Scholar,and China National Knowledge Infrastructure(CNKI).44 Cephalotaxus"',and the respective species name were used as keywords in database search.The obtained articles of the past six decades were collated and analyzed.Four Cephalotaxus species are listed in the official medicinal book in China.They are used as ethnomedicines by many ethnic groups such as Miao,Yao,Dong,She and Han.Inspirations are obtained from traditional applications,and Cephalotaxus phytometabolites are developed into anticancer reagents.Cephalotaxine-type alkaloids,homoerythrina-type alkaloids and homoharringtonine(HHT)are abundant in Cephalotaxus,e.g.,C.lanceolata,C.fortunei var.alpina,C.griffithii,and C.hainanensis,etc.New methods of alkaloid analysis and purification are continuously developed and applied.Diterpenoids,sesquiterpenoids,flavonoids,lignans,phenolics,and other components are also identified and isolated in various Cephalotaxus species.Alkaloids such as HHT,terpenoids and other compounds have anticancer activities against multiple types of human cancer.Cephalotaxus extracts and compounds showed anti-inflammatory and antioxidant activities,immunomodulatory activity,antimicrobial activity and nematotoxicity,antihyperglycemic effect,and bone effect,etc.Drug metabolism and pharmacokinetic studies o f Cephalotaxus are increasing.We should continue to collect and sort out folk medicinal knowledge of Cephalotaxus and associated organisms,so as to obtain new enlightenment to translate traditional tips into great therapeutic drugs.Transcriptomics,genomics,metabolomics and proteomics studies can contribute massive information for bioactivity and phytochemistry of Cephalotaxus medicinal plants.We should continue to strengthen the application of state-of-the-art technologies in more Cephalotaxus species and for more useful compounds and pharmacological activities.