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。

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.

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.

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.

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.

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.

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.

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.

Cephalotane-Type Norditerpenoids from Cephalotaxus fortunei var.alpina

A chemical investigation of the 95%EtoH extract of the seeds of Cephalotaxus fortunei var.alpina led to the isolation of nine new cephalotane-type norditerpenoids,ceforalides A-I(1-9),and two known analogues(10 and 11).Compounds 1-8 belong to a rare class of A-ring-contracted cephalotane-type norditerpenoids,of which compound 8 incorporates a unique tetrasubstituted 2,5-cyclohexadienone A-ring.Compound 9 is a rare 13,14-seco-17-nor-cephalotane-type diterpenoid.Their structures were determined based on NMR,HRESIMS,ECD,and X-ray diffraction analysis.Biological tests revealed compounds 10 and 11 displayed moderate antimalarial activity.

海南粗榧愈伤组织增殖及褐变现象研究

本文通过测定海南粗榧愈伤组织的生长速率、多酚氧化酶(PPO)、过氧化物酶(POD)和多酚含量,探究外植体、培养基、抗褐变剂及激素对粗榧愈伤组织生长和褐变的影响。结果表明:叶片外植体诱导产生的愈伤组织褐变率比茎段外植体低;培养基添加0.5g·L-1聚乙烯吡咯烷酮(PVP)有良好的抗褐变效果。愈伤组织最佳增殖培养基为MS+萘乙酸(NAA)4mg·L-1+激动素(KT)0.1mg·L-1+蔗糖30g·L-1+卡拉胶6g·L-1+PVP0.5g·L-1。

粗榧中低含量黄酮类化学成分的研究

[目的]研究粗榧中低含量黄酮类化学成分。[方法]以粗榧叶子为原材料,采用多种柱色谱相结合的方法,从其乙醇提取物中分离纯化得到3个含量较低的黄酮类化合物,并运用ESI-MS和NMR技术鉴定其化学结构。[结果]鉴定得3个含量较低的黄酮类化合物分别为芹菜素5-O-β-D-葡萄糖苷(I)、芹菜素7-O-β-D-葡萄糖苷(II)和槲皮素-3-O-β-D-葡萄糖苷(III)。[结论]这3个化合物都是首次从该植物中得到。

2种种植模式下海南粗榧早期生长表现

采用全光照与遮阳(遮光率约70%)2种模式对海南粗榧(Cephalotaxus mannii)进行种植试验,其早期的生长结果表明:1)2种种植模式的海南粗榧保存率分别为96.67%和98.33%,二者不具有明显差异(P>0.05);2)2种种植模式下海南粗榧均能生长,但是遮阳环境下的生长外在表现明显更好,全光照环境下的海南粗榧叶片和顶芽容易被高温灼伤导致干枯,5—8月更为明显;3)海南粗榧在遮阳环境下的地径和树高生长均显著高于全光照环境(P<0.05);4)遮阳环境下海南粗榧的叶绿素a和叶绿素b含量均高于全光照,而类胡萝卜素的则呈现相反的结果,2种模式具有明显的差异(P<0.05)。

渗透胁迫及信号处理对海南粗榧叶绿素荧光特性的影响

为了探究甘露醇、氯化钠的渗透胁迫和脱落酸、茉莉酸甲酯、乙烯利、水杨酸等信号物质对海南粗榧(Cephalotaxus hainanensis)叶片叶绿素荧光特性的影响,本研究以海南粗榧新鲜叶片为试材,分别选用甘露醇、氯化钠、脱落酸、茉莉酸甲酯、乙烯利、水杨酸处理海南粗榧叶片,测定PSⅡ实际量子产量Y(Ⅱ)、非光化学淬灭系数(NPQ)、光化学淬灭系数(qP)等叶绿素荧光参数。结果表明:甘露醇处理和氯化钠处理在短时间内可以造成这些参数的波动,然而在24 h后,这些参数趋于平稳,大多数可以恢复到对照水平,但渗透胁迫处理后的Y(Ⅱ)值显著降低,NPQ值却显著升高。在脱落酸、茉莉酸甲酯、乙烯利、水杨酸等信号物质处理后,叶绿素荧光参数多呈现出和甘露醇处理或氯化钠处理相似的趋势,表明了渗透胁迫可能通过不同的信号转导通路影响叶绿素荧光特性,其中水杨酸信号通路可能参与了叶片实际光能转换效率适应调节。