A New Hydroxyl Acetylenic Fatty Acid from the Basidiomycete Craterellus aureus (Cantharellaceae)

A new hydroxyl acetylenic acid, named (8 E , 10 R , 14 Z ) -10-hydroxy-8 , 14-octadecadien-12-ynoic acid (1), together with two known compounds (2 , 3) , were isolated from the ethanol extract of the fruiting bodies of Craterellus aureus. Their structures were established by spectroscopic methods (1D- and 2D-NMR experiments). The absolute configuration of 1 was determined to be R by computational calculation of optical rotation.     

Effect of Chemical Constituents from Syzygium cumini (Myrtaceae)on Glucose Uptake in Insulin-resistant L6 Cells

Eight known compounds isolated from the roots of Syzygium cumini were evaluated for their ability to enhance the glucose consumption in insulin-resistant L6 muscle cells induced by high concentration-insulin and glucose . All of compounds significantly enhanced the glucose consumption of insulin-resistant cells in the presence and absence of insulin. The glucose consumption was increased 17.35% and 51.11% by compound 1 ( friedelin) at a concentration of 10 μg ml^-1 without insulin and by compound 8 (5 , 7 , 3′, 4′, 5′-pentahydroxyflavone) at a concentration of 0 . 1μg ml - 1 with insulin, respectively .     

Phylogeny of Camellia sects Longipedicellata,Chrysantha and Longissima (Theaceae) Based on Sequence Data of Four Chloroplast DNA Loci

We focused on the systematic positions and relationships of three sections of Camellia subgenThea of Theaceae, i.e., sect.Longipedicellata, sect.Chrysantha (golden camellias) and sect.Longissima by using four chloroplast DNA regions (rpl16, psbA trnH, trnL F & rpl32 trnL). We sampled 28 species representing four subgenus, 11 sections in Camellia and two outgroups. Combined analyses of chloroplast DNA sequence data sets are performed with the neighbor joining, maximum parsimony and Bayesian inference methods, and the gene trees are constructed. The topologies of gene trees revealed that: 1) sect. Chrysantha is paraphyletic or polyphyletic, containing three parallel lineages and Camellia longipedicellata (type of sectLongipedicellata) nested inside; 2) all four species from Vietnam, together with C. longipedicellata, forming a well supported monophyletic clade, which implies the close relationship between sect. Longipedicellata and sect.Chrysantha; and 3) sectLongissima is not monophyletic because C.hekouensis is the sister to the rest of Camellia species. The systematic position of C.longissima and the relationship between sect.Longissima and sect.Longipedicellata are unsolved.     

Effects of Nitrogen on the Growth and Accumulation of Secondary Metabolites of Erigeron breviscapus (Compositae)

Erigeron breviscapus a medicinal plant was planted under different nitrogen levels to investigate effects of nitrogen on the growth and the accumulation of secondary metabolites, address the relationship between them in medicinal plants in the plantation. Results showed that the amount of N fertilizer was significantly positively correlated with the biomass, but negatively to contents of total caffeates and scutellarin of Ebreviscapus Moreover, increasing amount of N fertilizer did not significantly enhance yields of 2 active constituents. The biomass was negatively correlated with contents of total caffeates and scutellarin. Contents of total caffeates and scutellarin were negatively correlated with contents of N in Ebreviscapus, as predicated by the carbon/nutrient balance hypothesis. The result suggested that the nitrogen addition could increase the biomass yield, nevertheless, decrease active constituents ( flavonoids ),there could be a trade off between the growth and accumulation of secondary metabolites in medicinal plants.     

Biomass Carbon Accounting Parameters for Young and Middle Aged Plantation of Simao Pine (Pinus kesiya var.langbianensis)

Based on data collected from field surveys, biomass carbon accounting parameters including biomass conversion and expansion factor (BCEF), biomass expansion factor (BEF) and root shoot ratio (R) for Pinus kesiya var. langbianensis plantation were calculated, and relationships between the parameters and relative stand factors were studied. Main findings were as follows. (1) Mean BCEF for Pinus kesiya var. langbianensis plantation was 05483 Mg m 3(n=30, 95% confidence interval=05357-05609), lower than the IPCC default value. BCEF for Pinus kesiya var langbianensis plantation was negatively related to stand form height (FH), mean stand height (H), stand growing stock (V) and stand age(A) (P<005). BCEF was negatively related to mean diameter at breast height (D), but not statistically significant（P >005）, positively related to stand density (N), not statistically significant（P>005）. Regression equations developed for calculating BCEF with stand factors did not give satisfied estimates. (2) Mean BEF for Pinus kesiya var langbianensis plantation was 178378 (n=30, 95% confidence interval=171714-185043), higher than the IPCC default value. BEF was negatively related to D、H、FH、V and A (P <001), positively related to N (P <005). Regression equations of y=a+bx+cx2 performed well to calculating BEF with A and V as variables. Regression equation of y=a+b/x performed well to calculate BEF with N. Regression equations of y=a xb performed well to calculate BEF with FH、H and D as variables. (3) Mean R for Pinus kesiya varlangbianensis plantation was 02400 (n=30, 95% confidence interval=02194-02606), close to the IPCC default value. R was negatively related to D、H、FH、V and A (P <001), positively related to N (P <005). Regression equations of y= a+bx +cx2 performed well to calculate R with D、H、FH、V and Aas variables. Regression equation of y=a+b/x performed well to calculate R with N.     

Genetic Diversity of Wild Lycoris radiata (Amaryllidaceae) from China Revealed by SRAP

Genetic diversity of 24 wild Lycoris radiata collected from different localities in China was examined by sequence related amplified polymorphism（SRAP）. Two hundred and eighteen loci were identified with 10 SRAP primer combinations, out of which 173 were polymorphic and accounted for 7936% of total genetic diversity at species level. Observed number of alleles (na), effective number of alleles (ne), Nei′s gene diversity (h) and Shannon information index (I) were 17936, 14131, 02415 and 03664, respectively. The coefficient of gene differentiation (Gst) and gene flow (Nm) were 09547 and 00136, suggesting that most of variation occurred among different resources of Lradiata, and the genetic showed significant differentiation. UPGMA cluster analysis of the 24 resources based on Nei′s genetic distance showed two major clusters. Cluster I included 7 resources from southwest and northwest China, except for Jiangsu, Lianyungang resource（JS3）. Cluster II included the rest 17 resources from south China to east China. The subgroups exhibited related to the morphology and habit of different resources of Lradiata to some extent. There was no significant correlation between genetic diversity and longitude, altitude, latitude, annual rainfall, and annual average temperature. The results suggested that the genetic diversity of wild Lradiata was high and the possible reasons for significant genetic differentiation might due to the very low gene flow. The results of this study might be useful for guiding the exploitation and conservation of germplasm of Lradiata.     

Prokaryotic Expression and Bioinformatics Analysis of Cytosolic Malate Dehydrogenase from Camellia sinensis(Theaceae)

The complete gene of cytosolic malate dehydrogenase (cMDH) from Camellia sinensis, called Cs cMDH, was obtained by RT PCR and rapid amplification of cDNA ends (GenBank accession number GQ845406). This gene was 1 235 bp in length, encoding a protein of 332 amino acids with the putative molecular weight of 355 kD. The Ecoli Rosetta (DE3) harboring pGEX MDH was induced by 05 mmol·L 1 IPTG at 32℃ for 3 hours, and a 615 kD glutathione Stransferase (GST) fused MDH was obtained in soluble form. The results of NCBI BLAST revealed that Cs cMDH shared 88%-93% of amino acid sequence identity with other cMDH from different higher plants. According to the multiple sequence alignment based on the three dimensional structure of protein, Cs cMDH was predicted to be a dimer with thirteen β sheet and thirteen α helix of each subunit. Cs cMDH contains typical fingerprint sequence (G12AAGQIG18) as all MDHs. The amino acid D43 in Cs cMDH is conserved in all NAD MDHs. Cs cMDH also has some conserved sequence units homologous to other NAD MDHs, such as NAD+ binding sites, catalytic motif and substrate binding sites. Moreover, Cs cMDH contains six Cys which are highly conserved in all plant NAD cMDHs. Therefore, Cs cMDH was inferred to be NAD dependent cMDH. The present study may provide the fundament for the further functional characterization of Cs cMDH.     

Observations on the Formation and Ultrastructure of the Egg Membrane in the fern Ceratopteris thalictroides (Parkeriaceae)

The mature egg of ferns possesses a layer of egg membrane in the periphery. However, the fine trastructure and the formation of the egg membrane in the egg of ferns are still unclear. The present paper described the formation and the ultrastructure of the egg membrane in the egg of the fern Ceratopteris thalictroides using transmission electron microscopy (TEM). The results show that the egg membrane begins to form at the stage of the maturing egg. The egg membrane in the upper side of the egg is prominent. It is formed by attaching of osmiophilic sheets of endoplasmic reticulum to the inner surface of the plasmalemma . When mature the egg membrane in the upper side of the egg is much thick in the central region and it becomes thinner gradually towards the margin. Some osmiophilic materials are filled in the spaces between the sheets of endoplasmic reticulum . On the contrary, the egg membrane in the lower side of the egg is thin . It is composed of twoosmiophilic membranes which are associated with each other closely. The formation and the ultrastructure of the egg membrane are described for the first time . Some functions of the egg membrane are also discussed .     

Phenotypic Diversity of Acer ginnala (Aceraceae) Populations at Different Altitude

In this study, we analyzed the phenotypic variation of Acer ginnala natural populations at different altitude in Qiliyu, investigated the level of phenotypic diversity by using ANOVA analysis, correlation analysis and cluster analysis. The results showed that there were significant differences in phenotypic variation among and within populations. Coefficient of variation (CV) varied from 705% to 3812%. High phonotypic diversity (1.9253) occurred within A.ginnala populations, the mean phenotypic diversity index of populations ranged from 1.9022 to 1.9837. Phenotypic differentiation coefficient (VST) among populations (13.79%) was less than that of within populations (86.21%). The variation within populations comprised a majority of total phenotypic variation. A significant relation had occurred between phenotypic traits and soil factors. Phenotypic diversity index had a very significant correlation with soil total nitrogen (P<0.05), no significant relation with altitude. Five populations gathered into two groups by cluster analysis, which consistent with the geographic distribution of Aginnala in Qiliyu. The traits variations of natural populations at different altitude were affected mainly by micro-environmental heterogeneity of different Aginnala populations. Utilizing of variation within and among populations is important significance for genetic improvement of A.ginnala.     

Cloning and Analyses of a Dual Specific Serine􊄯Thronine Protein Kinase Gene with High Conservative and Constitutive Expression in Oryza (OsSTK)

A cytoplasmic serine&#1089839;thronine protein kinase gene (OsSTK), had been cloned from Oryza genus. It was found high conservative and constitutive expression in Oryza. OsSTK gene had two exons, separated by 114 bp short intron. The open reading frame of OsSTK gene that predicted encoded a 419 amino acids protein . The amino acid sequence of OsSTK had low identities ( less than 53% ) with any other known protein kinase . The phylogenetic tree based on the partial DNA sequences of OsSTK from different species and types of wild rice and cultivated rice, was close to the taxation system ofrice . Interestingly , OsSTK had a serine, including basic amino acids and charged amino acids abundant polypeptide with a “GDGDGDGDG”sequence at N- terminal that had not been found in any other genes. OsSTK may play dual specificity that phosphorylates both serine&#1089839;thronine and tyrosone, because the amino acids module of VIb , VIII and XI catalytic domain have both the serine&#1089839;thronine and tyrosine kinase characters .     

Altitudinal Variations in Reproductive Allocation of Bergenia purpurascens (Saxifragaceae)

The strategy of resource allocation between vegetative and reproductive functions, quantitative relationship between size and reproductive output are central aspects of plant life history. To test the tactics of resource allocation and its altitudinal trend, we examined the reproductive allocation (RA) of Bergenia purpurascens (Saxifragaceae), in six populations along a shady slope in Sejila Mountain of southeast Tibet, at an altitude gradient from 4 200 m to 4 640 m. Our results showed that (1) with increasing altitude, vegetative biomass, reproductive biomass, total aboveground biomass, flower number per plant and length of flower stalk decreased significantly, but the number of leaves did not change greatly. However, the change of RA did not show a monotonic trend when altitude increased, shifting from significantly decreasing below the tree line to slightly increasing above it; (2) vegetative biomass was positively correlated with reproductive biomass, but negatively correlated with RA in all populations, but the level of significance was different among the populations; (3) RA decreased with individual size in all populations, whereas the relationship between absolute resource allocated to reproduction and individual size was allometric; (4) reproductive allometry and a size threshold for reproduction did exist in this alpine perennial, but the obvious altitudinal trend was only found along the populations below the tree line, not above it. We then concluded the altitude could not fully explain the change of resource allocation strategy of this alpine perennial, and different effects of size and habitat on RA may result from various environmental constraints along the altitudinal gradient or genetic background. Therefore, each individual within a population will follow its own developmental trajectory shaped by its genotype and the habitats. The most innovative finding was plant adaptation and resource trade off might be sharply altered at the tree line, which is a sensitive area in alpine mountains. Further investigations are needed to better understand the relationship between the reproductive allocation and changing environmental conditions.     

Preliminary Study on Development,Germination and Desiccation Tolerance of Jatropha curcas (Euphorbiaceae)

The developmental changes in morphology and germinability of Jatropha curcas seeds, effects of temperature and light on seed germination , and changes in desiccation tolerance of mature seeds were studied in this paper. The results indicated that J. curcas seeds reached a physiological maturation at 58 days after anthesis, and that seed germination percentage reached a peak at physiological maturation, and then decreased. The optimal germination temperature was 25 to 30℃. J. curcas seeds were insensitive to dehydration at physiological maturation . There was not a notable effect of light on seed germination. Therefore, J. curcas seed was a non-photoblastic and orthodox seeds .     

Mapping and Analysis of a Yellow Mutant chlm-4 of Arabidopsis thaliana (Cruciferae)

Chlorophylls are essential for photosynthesis. Chlorophyll biosynthesis is catalyzed by a series of enzyme complexes, such as Mg protoporphyrin IX methyltransferase. A yellow mutant of Arabidopsis was isolated using an enthyl methane sulfonate (EMS) mutagenesis strategy. Chlorophyll content dramatically reduced and grana stacking was absent in the mutant. Genetic analysis indicated that the mutant was controlled by a single recessive gene. Using map based cloning strategy, the gene responsible for the mutant phenotype was mapped to a region of 114 kb between the molecular markers F13M23 and T30C3 on chromosome 4, in which the CHLM gene encoding Mg protoporphyrin IX methyltransferase was included. The mutant was proved to be an allelic mutant of CHLM gene by sequencing and allelism test and then was designated as chlm 4. Gly59 of CHLM was replaced by Glu59 in chlm 4, which indicated that Gly59 was essential for the function of Mg protoporphyrin IX methyltransferase.     

Resource Utilization Pattern Analysis in Pali Gad Watershed of Tehri,Garhwal Himalaya ( India)

The western Himalaya has had a central role as a life support system for the local people and as a source of ecological services . This long term use of these resources, including livestock grazing, fuel wood collection and a variety of other uses , has shaped its montane cultural landscape . The present paper focuses on resource utilization in the Pali Gad watershed , which consists of 25 villages situated in the lesser Himalaya . Using a satellite derived map , natural resources available in this area were assessed , and the ecosystem services provided by them and level of pressure on them were computedthrough a door to door socioeconomic survey . The study revealed a spatio-temporal variation in the resource demand and extraction at the permanent villages . Average fuel wood consumption was 1 . 12 kg&#1089839;day&#1089839;capita , fodder consumption was 3 . 69 kg&#1089839;day&#1089839;capita from lopping and 3 .25 kg/day&#1089839;capita from grass collection from forest . An attempt has been taken to evaluate the ecosystem services . The forest provides more provisional and regulatory services, whereas agriculture provides more supporting services and river bed&#1089839;water bodies serve more cultural purposes for the locals. This study contributes an example of a human shaped ecosystem in a mountainous region where an additional fragmentation of natural resources exists. An increase in resource demand has been noticed in the less productive lands of the region . This has resulted in increased resource extraction from the entire watershed may not be sustainable in the long run .     

Substratum Affects the Gametophyte Development and Reproduction of Sinopteris grevilleoides(Sinopteridaceae), an Endangered Fern Endemic to China

Sinopteris grevilleoides is an endangered fern endemic to the region of Yunnan and Sichuan in southwest China. In this study, we investigated the effects of four different culture substrata on the development of gametophyte and reproduction of this fern. Cultured on different culture substrata, the fern showed different morphology of gametophyte. In humus soil, cordate prothalli did not bear archegonia, and thus the process of sexual reproduction ofSgrevilleoides could not be finished. The substratum composed of humus soil and original soil in ratio of 1∶1 was the most suitable substratum for gametophyte development and sexual reproduction of this species. Otherwise, infant sporophytes needed extra nutrient solution to sustain its growth on improved Knop′s agar medium.     

Seed Dormancy,Germination and Storage Behavior of Magnolia wilsonii (Magnoliaceae), an Endangered Plant in China

In this paper, we measured the embryo morphology and size of Magnolia wilsonii seeds. And the effects of temperature, stratification, KNO3 and soaking on seed dormancy and germination were studied. Seeds were desiccated to different moisture content, and then stored at 4℃ and -20℃ for 100 d. The effects of desiccation and storage temperature on seed viability of M. wilsonii were analyzed. The results showed that the underdeveloped embryos of Mwilsonii seeds needed cold stratification to achieve physiological afterripening. The favorable temperature for germination of Mwilsonii seeds was 25/20℃. Cold stratification and GA3 could effectively overcome dormancy of Mwilsonii seeds. Therefore, Mwilsonii seeds have morphophysiological dormancy. About 5350% of seeds could survive in the moisture content of 539%. After 100 d storage at -20℃, all seeds in different moisture contents died. However, after 100 d storage at 4℃, 76% of seeds survived. Thus, Mwilsonii seeds exhibited intermediate seed storage behavior. The optimum storage environments was dry or moist storage at 4℃.     

Structures of Novel Sesquiterpene Ketones from Chloranthus serratus (Chloranthaceae)

Two naturally occurring new sesquiterpene ketones, neoacolamone (1) and 7α-hydroxyneoacolamone (2), were isolated from the roots of Chloranthus serratus (Thunb.) Roem. et Schult. (Japanese name: Futari-shizuka, Chloranthaceae). The structures were elucidated on the basis of their physicochemical properties and chemical reactions. Three sesquiterpene ketones, acoragermacrone (3), acolamone (4) and zederone (5), and four additional sesquiterpenes including lindenanolides were also found in the same source. The relationships of these isolates to the constituents of C. japonicus and to the sesquiterpene biogenesis in the Chloranthus species are discussed.     

Chemical Constituents of Guangdong Tu Niu Xi (Eupatorium chinense,Compositae)

One new compound, namely eupatorin A (1), was isolated from the methanolic extract of Guangdong Tu Niu Xi, the dried roots of Eupatorium chinense (Compositae). Its structure was determined to be (threo) 3 O acetyl 1 (4 hydroxy 3 methoxyphenyl) 2 [4 (3 hydroxy 1 (E) propenyl) 2, 6 dimethoxyphenoxy]propyl β D gluco pyranoside on the basis of detailed spectroscopic analysis. In addition, ten known compounds, including (threo) 3 hydroxy 1 (4 hydroxy 3 methoxyphenyl) 2 [4 (3 hydroxy 1 (E) propenyl) 2, 6 dimethoxyphenoxy]propyl β D glucopyranoside (2), ardisiacrispin A (3), ardisiacrispin B (4), euparone (5), 3 (2, 3 dihydroxy isopentyl) 4 hydroxy acetophenone (6), 12,13 dihydroxy euparin (7), gymnastone (8), N (2′ hydroxy tetracosanosyl) 2 amino 1, 3, 4 trihydroxy octadec 8 (E) ene (9), stigmasterol (10) and stigmasterol 3 O β D glucopyranoside (11) were also obtained. This was the first time that compounds 2-4 were reported from Compositae family, while 5-8 were isolated from the genus Eupatorium.     

Structures of Novel Sesquiterpene Alcohols from Chloranthus japonicus (Chloranthaceae)

Three novel sesquiterpene alcohols, shizuka-acoradienol (2), shizukafuranol (3) and shizukol-idol (4), were isolated from Chloranthus japonicus Sieb. (Japanese name: Hitori-shizuka, Chloranthaceae). Their structures were elucidated on the basis of their physicochemical properties and some chemical reactions. Furanodienone (5), scopoletin (6a) and isoscopoletin (6b) were also isolated from this plant.