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.

云南腾冲上新统植物油丹Alseodaphne hainanensis Merr.表皮微细构造研究

研究的樟科植物化石油丹(Alseodaphne hainanensis Merr.)采于云南腾冲上新统芒棒组,它在滇西地区为首次发现。对其进行了大量的实验分析,揭示了其表皮微细构造特征。通过与现生油丹表皮构造和叶结构特征的对比分析,在宏观和微观两方面证实了该化石归属的正确性。现生Alseodaphne hainanensis Merr.分布范围的缩小显示滇西从上新世—现代气候逐渐变冷,上新世的气候比现在温暖。化石Alseodaphne hainanensis Merr.地理分布区的扩大和地质时代的延伸,为植物的发展演化和推论古气候提供了宝贵材料。

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。

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。

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.

First Record of a Male Adult of Epora hainanensis Chou & Wang(Hemiptera:Fulgoroidea:Tropiduchidae)

The male adult of Epora hainanensis Chou & Wang,1985 is reported for the first time.The female genitalia of this species is described and illustrated.The specimens examined are deposited in the Entomological Museum,Northwest A & F University(NWAFU).

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。

海南土地杆菌Pedobacter hainanensis 13-Q^T发酵生产κ-卡拉胶酶的条件优化

采用响应面法对海南土地杆菌Pedobacter hainanensis 13-QT生产κ-卡拉胶酶的发酵条件进行了优化,通过Plackett-Burman实验方法研究碳源含量、氮源含量、无机盐、初始pH值、温度等9个发酵因子对菌株发酵产κ-卡拉胶酶活力的影响。结果表明,影响产酶的显著因子是温度、蛋白胨浓度和初始pH值。根据最陡爬坡实验结果确定显著影响因子的取值范围,并采用Box-Behnken设计实验进行优化,然后应用响应面模拟预测和摇瓶发酵实验验证。结果表明,产κ-卡拉胶酶的最佳条件为κ-卡拉胶1.5g/L,蛋白胨3.9g/L,NaCl 20g/L,K2HPO41g/L,MgSO40.40g/L,CaCl20.12g/L,FeSO40.008g/L,发酵培养基初始pH值为6.9,温度30.3℃。经过优化后,发酵液中κ-卡拉胶酶酶活力达到3.387IU/mL,与优化前相比提高了5.04倍。

Neuroprotective activity of two active chemical constituents from Tinospora hainanensis

Objective:To determine the chemical structure of the new compound and investigate the protective effects of Tinosporaic acid A and B towards in-vitro neuro.Methods:The structures of two new compounds were established by analyzing its 1D and 2D NMR spectra as well as HRESIMS.Their neuroprotective effects with respect to the antioxidant properties were evaluated by radical scavenging tests and hydrogen peroxide-injured oxidative stress model in PC12 cell lines.Cell morphology of treated PC12 cells was observed by phase contrast microscopy.In-vitro MTT assay,lactate dehydrogenase activity assay and oxidative stress markers(intracellular ROS production,MDA level,and caspase-3 activity) were used to evaluate the protective effects against hydrogen peroxide induced cytotoxicity in PC12 cells.Results:The two new compounds,named Tinosporaic acid A and B,were isolated and identified from the stem bark of Tinospora hainanensis.Cell viability studies identified a representative concentration for each extract that was subsequently used to measure oxidative stress markers.Both extracts were able to reverse the oxidative damage caused by hydrogen peroxide,thus promoting PC12 cells survival.The concentration of Tinosporaic acid A and B were 86.34 μg/mL and 22.06 μg/mL respectively,which is neuroprotective for EC_(50).The results indicated that both of them significantly attenuated hydrogen peroxide-induced neurotoxicity.Conclusion:The two new compounds isolated from ethanol extracts of Tinospora hainanensis are the promising natural ones with neuroprotective activity and needed for further research.

Indole Alkaloids from the Roots of Ervatamia hainanensis

Two new indole alkaloids, named ibogamine-18-carboxylic acid, 3, 4-didehydro-7, 8-dioxo-methyl ester 1, ibogamine-18-carboxylic acid, 16, 17-didehydro-9, 17-dihydro-9-hydroxy （2-oxopropyl）-methyl ester 2, were isolated from Ervatamia hainanensis. Their structures were elucidated on the basis of spectroscopic methods.

Molecular Detection and Identification of Candidatus Ehrlichia Hainanensis,A Novel Ehrlichia Species in Rodents from Hainan Province,China

Ehrlichia(Anaplasmataceae family)are obligatory intracellular bacteria that infect humans and animals.They are hosted by mammals such as canines,bovines and wild rodents,and are vectored by ticks.In this study,we collected 121 rodent samples comprising 67 Niviventer fulvescens,27 Rattus tanezumi,24 Chiromyscus sp.,2 Rattus nitidus and 1 Leopoldamys edwardsi from Hainan province,which includes the second largest island in China.The presence and genetic diversity of Ehrlichia species was evaluated and characterized by amplification and sequencing of 16S rRNA,groEL and gltA genes.An Ehrlichia species was detected in 5 of the 67 Niviventer fulvescens samples(7.46%).The 16S rRNA,groEL and gltA genes showed the highest identity to known Ehrlichia sequences(99.20%,89.87%and 83.86%,respectively).In the phylogenetic trees they formed a cluster distinct from all other species.We propose that this species is a putative novel Ehrlichia species,which we suggest be named Candidatus Ehrlichia hainanensis.Its pathogenicity to humans remains to be further researched,and molecular surveillance in local populations is needed.

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.

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.

Effects of Different Processed Products of Ilex hainanensis Merr on Rats with Hyperlipidemia

[Objectives] This study aimed to investigate the hypolipidemic effects of different processed products of Ilex hainanensis Merr to provide experimental basis for the research on the hypolipidemic mechanism of I. hainanensis Merr. [Methods] SD rats were fed high-fat diet to establish hyperlipidemia rat models. The hyperlipidemia rats were administered with different processed products of I. hainanensis Merr by gavage to investigate the preventive and therapeutic effects of the medicinal material. [Results] The levels of total cholesterol(TC), triglyceride(TG), low density lipoprotein cholesterol(LDL-C) in the rats fed high-fat diet were significantly higher than those in of the control group(P<0.01), indicating successful modeling. Among different processing methods, the hypolipidemic effects were in the order as baking=traditional processing=frying=microwaving>drying in shade. Among different baking times, the hypolipidemic effects ranked as 180 ℃>200 ℃=220 ℃. Among different baking times, the hypolipidemic effects ranked as 20 min=40 min>30 min. The hypolipidemic effects of processed product of I. hainanensis Merr with different amounts of active ingredients were different. [Conclusions] The processed products of I. hainanensis Merr all had hypolipidemic effects, but the effects were different. This study will provide a scientific basis for expanding the resources of I. hainanensis Merr.