Identification and analysis of expressed sequence tags present in xylem tissues of kelampayan (Neolamarckia cadamba (Roxb.) Bosser)

The large-scale genomic resource for kelampayan was generated from a developing xylem cDNA library. A total of 6,622 high quality expressed sequence tags (ESTs) were generated through high-throughput 5’ EST sequencing of cDNA clones. The ESTs were analyzed and assembled to generate 4,728 xylogenesis unigenes distributed in 2,100 contigs and 2,628 singletons. About 59.3 % of the ESTs were assigned with putative identifications whereas 40.7 % of the sequences showed no significant similarity to any sequences in GenBank. Interestingly, most genes involved in lignin biosynthesis and several other cell wall biosynthesis genes were identified in the kelampayan EST database. The identified genes in this study will be candidates for functional genomics and association genetic studies in kelampayan aiming at the production of high value forests.     

团花树中一个新的环烯醚萜苷（英文）

从茜草科团花属植物团花树（Neolamarckia cadamba）枝叶的甲醇提取物中分离得到两个环烯醚萜苷,其中一个为新的环烯醚萜苷,利用现代波谱学技术鉴定了其结构为Δ^13,14-plumieride,并命名为plumieride B.     

Chemical constituents from the leaves of Zanthoxylum nitidum (Roxb.) DC

Twelve compounds were isolated from the ethyl acetate soluble portion of the 70% hydroalcoholic extract from the leaves of Zanthoxylum nitidum (Roxb.) DC. All of the compounds were determined on the basis of UV, IR, NMR and MS and comparison with literature. Among them, compounds 4, 7, 10 and 12 were first acquired from Rutaceae, compound 2 was isolated from the genus of Zanthoxylum for the first time and compounds 1, 3, 6, 9, 11 were isolated from Z. nitidum for the first time. Our present study exhibited the taxonomic relationships between Z. nitidum and other species in the same genus; those between Rutaceae and other families. Furthermore, we have summarized the chemotaxonomic significance of these compounds in Rutaceae and Zanthoxylum.     

黄梁木实时荧光定量PCR分析中内参基因的选择

为筛选黄梁木(Neolamarckia cadamba)实时定量PCR最佳内参基因, 该研究以黄梁木的根、芽、叶、花、果、皮及形成层为材料, 利用RT-qPCR技术对ACT、CAC、CYP和EF1α等21个管家基因家族43个候选内参基因进行表达量分析, 并利用geNorm、NormFinder和BestKeeper软件进行内参基因稳定性分析。geNorm的分析结果显示, UPL基因的稳定性最高(M=0.443), UBQ基因的稳定性最低(M=2.859); NormFinder的分析结果显示, UPL基因的稳定性最高(E=0.223), UBQ基因的稳定性最低(M=4.759); BestKeeper分析显示, UPL基因的标准偏差(SD=0.513)最低。研究结果表明, UPL基因作为内参基因稳定性最高, UBQ基因的稳定性最低。因此可以选择UPL基因作为黄梁木不同组织中RT-qPCR定量分析的内参基因。     

表施和混施污泥对团花根系生长的影响

林地利用被认为是污泥资源化利用的重要方式,但施用污泥对林木根系生长的影响报道较少。本研究通过根箱试验,分析表施和混施10%污泥对速生树种团花不同土层根系形态、土壤pH值和电导率动态变化及根系重金属含量的影响,并拟合土壤pH值、电导率和根系重金属含量与根长的关系。结果表明: 与不施污泥(对照)相比,混施污泥显著抑制了团花根长、根表面积和根体积增长,混施污泥120和240 d后,0～20 cm土层总根长分别为不施污泥的76.9%和67.4%;表施污泥对团花根长和根表面积的影响不显著,但显著提高了根体积。混施污泥显著提高了土壤pH值和电导率及根系重金属含量,混施污泥0～20和20～40 cm土层根系镉含量分别是不施污泥的11.5和10.0倍。线性回归拟合分析显示,不同处理0～20 cm土层的电导率与根长均呈显著负相关;表施和混施污泥根系镉含量与根长呈极显著负相关。上述结果表明,混施污泥抑制了团花根系生长,这可能是由于混施污泥提高了土壤电导率和根系镉含量所致,而表施污泥对团花根系生长的作用不明显。     

Investigations on Antioxidant,Antiproliferative and COX‐2 Inhibitory Potential of Alkaloids from Anthocephalus cadamba (Roxb.) Miq. Leaves

In the present study, an ayurvedic medicinal plant, Anthocephalus cadamba (Roxb .) Miq . commonly known as ‘Kadamb’ was explored for its potential against oxidative stress and cancer. The fractions namely AC‐4 and ACALK (alkaloid rich fraction) were isolated from A. cadamba leaves by employing two different isolation methods and evaluated for their in vitro antioxidant and antiproliferative activity. The structure of the isolated AC‐4 was characterized tentatively as dihydrocadambine by using various spectroscopic techniques such as ESI‐QTOF‐MS, ¹H‐ and ¹³C‐NMR, DEPT, COSY, HMQC, and HMBC. Results of various antioxidant assays viz. 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH), ABTS cation radical, superoxide anion radical scavenging, and plasmid nicking assay demonstrated that both the fractions viz. AC‐4 and ACALK possess ability to scavenge DPPH, ABTS radicals and effectively protected plasmid pBR322 DNA from damage caused by hydroxyl radicals. Further, when both fractions were evaluated for their potential to suppress growth of HeLa and COLO 205 cells, only ACALK fraction showed antiproliferative effects. ACALK exhibited GI₅₀ of 205.98 and 99.54 μg/ml in HeLa and COLO 205 cell lines, respectively. Results of Hoechst staining in cervical carcinoma (HeLa) cells confirmed that ACALK induced cell death in HeLa cells via apoptotic mode. Both the fractions also inhibited COX‐2 enzyme activity.     

Chemical constituents from Lycium barbarum (Solanaceae) and their chemophenetic significance

Phytochemical investigation of the fruit of Lycium barbarum L. has yielded nine alkaloids including one new compound, lycibarbarspermidine T together with eight known compounds. The structures were unambiguously determined by spectroscopic analyses and chemical methods. The chemophenetic significance of all these compounds was summarized. Among the known compounds, compounds 6 and 9 are reported from L. barbarum for the first time, whereas compounds 2–5 have previously been only found in this plant. The anti-inflammatory effects of all compounds were evaluated by measuring the NO production in RAW 264.7 cells stimulated by lipopolysaccharide (LPS). The results showed that compounds 1–3 and compounds 5–9 could inhibit the NO release of RAW 264.7 cells stimulated by LPS at 30 μM.     

Chemical constituents from the Jasminum lanceolarium Roxb.

Twelve compounds (1–12) are isolated from Jasminum lanceolarium Roxb. Among them, compounds 5, 7, 8, 11 and 12 are reported here for the first time from J. lanceolarium, and compounds 6, 9, 10 are firstly isolated from Jasminum genus. The existence of iridoids and lignanoids might be regarded as valuable chemotaxonomic markers for further classification and subdivision of the genus of Jasminum.     

两面针中新棒状花椒酰胺提取工艺的研究

本研究通过正交试验设计,对两面针中具有镇痛、抗炎和止血活性的新棒状花椒酰胺提取的不同溶剂、不同的提取时间和不同的提取温度用工业上常用的回流提取方法,进行了最佳工艺条件的选择。发现不同的提取条件,两面针的提取液中活性成分新棒状花椒酰胺的含量差别很大。在75％的乙醇、75℃温度、时间3h的条件下,两面针的提取液中新棒状花椒酰胺的含量最高,为0．76％;在水为溶剂、50℃温度、时间1h的条件下的条件下,两面针的提取液中新棒状花椒酰胺的含量最低,为0．00070％。     

Chemical constituents of Zanthoxylum schinifolium (Rutaceae)

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Phytochemical constituents from the stem barks of Goniothalamus tapis Miq

Phytochemical investigation on the stem bark of Goniothalamus tapis Miq. led to the isolation of seven known styryl-lactones, 3-acetyl-isoaltholactone (1), goniothalamin (2), isoaltholactone (3), cheliensisin A (4), 7-epi-goniofufurone (5), goniopypyrone (6), garvensintriol (7), and two steroids, stigmasterol (8) and β-sitosterol (9). The structures of all the compounds were elucidated based on the analysis of spectroscopic data and comparison with reported literature. The chemotaxonomic significance of all these compounds were summarized. Among all the styryl-lactones, compound 1 was isolated naturally from plant for the first time while compounds 4, 5 and 7 have not been previously reported in Goniothalamus tapis Miq. Compound 2 was the first styryl-lactone isolated from many species of Goniothalamus including other families such as Cucurbitaceae, Lauraceae, Stemonaceae and Apocynaceae). Compounds 3 and 6 were found in ten species of Goniothalamus where the latter were also found in other genus (Polyalthia Blume).     

Plant regeneration from shoot callus of rosewood (Dalbergia latifolia Roxb.)

In vitro propagation of trees using cell, tissue and organ culture is a fast emerging area. We report here the clonal propagation of Indian rosewood (Dalbergia latifolia Roxb.) from shoot callus cultures of 5 year old trees. Bud regeneration was obtained on MS media supplemented with BA and NAA. About 35% of the cultures showed organogenesis. Shoots measuring about 3–5 cm can be excised and rooted in White's medium supplemented with 1–2 mg/L IAA. Rooted plants were successfully established in soil.Abbreviations BA Benzyladenine - CM Coconut milk - 2,4-D 2,4 dichlorophenoxyacetic acid - IAA Indoleacetic acid - IBA Indolebutyric acid - K Kinetin - NAA Naphthaleneacetic acid - PVP-360 Polyvinyl pyrrolidone     

Chemical constituents from the stem bark of Clausena excavata Burm. f

Phytochemical study on the stem bark of Clausena excavata Burm. f. led to the isolation and identification of 11 compounds, including four prenylated coumarins (1–4), three carbazole alkaloids (5–7), a limonoid (8), a sterol (9), a depside (10) and a xanthone (11). The chemical structures of the compounds were elucidated based on NMR and mass spectroscopic data, and further supported by comparison with previously literature values. This is the first report on the isolation of compounds 10 and 11 from the genus Clausena. The chemotaxonomic significance of the isolated compounds is discussed.     

Chemical constituents from the rare mushroom Calvatia nipponica inhibit the promotion of angiogenesis in HUVECs

Calvatia species, also known as puffball mushrooms, are common sources of food worldwide. Calvatia nipponica (Agaricaceae) is one of the most rare species in the Calvatia genus. It was first validated in 2008. Due to its scarcity, C. nipponica has never been chemically investigated. Its recent discovery in Korea allowed to conduct this study, and using bioactivity-guided fractionation for antiangiogenic activity, chemical investigation of the MeOH extract of the fruiting bodies of C. nipponica led to the isolation of five alkaloids (1–5) and two phenolic compounds (6–7). This is the first study to report the chemical investigation of C. nipponica, and compound 1 was previously reported as chemically synthesized only until our report of its isolation and identification from natural sources. The structure of 1 was determined by spectroscopic analysis by 1D and 2D NMR spectra and HR-MS. All compounds (1–7) were tested for inhibition of angiogenesis using human umbilical vein endothelial cells (HUVECs). Compounds 2, 4 and 5 significantly inhibited the promotion of angiogenesis in HUVECs. Compound 5 showed the most potent inhibition via downregulation of VEGF, p38 and ERK signaling pathways. These results suggested that the rare mushroom C. nipponica might be beneficial in antiangiogenesis therapy for cancer treatment.     

Chemical constituents from Cyclocarya paliurus (Batal.) Iljinsk

A novel triterpenoid, cyclocarioside K, together with fifteen known compounds, including seven triterpenoids (2–8), five flavonoids (9–13), three phenolic derivatives (14–16), was obtained from the leaves of Cyclocarya paliurus (Batal.) Iljinsk (Juglandaceae). Their structures were elucidated by spectroscopic methods, including two-dimensional NMR experiments (HSQC DEPT and HMBC). The chemotaxonomic relationships were also discussed. As a result, 3, 4-secodammarane triterpenoids and kaempferol glycosides may represent phytochemical fingerprints for C. paliurus, and C. paliurus has close relationships with other species of Juglandaceae.     

Chemical constituents from Syringa reticulata (Bl.) Hara

A previously undescribed secoiridoid aglycone, syrinretin (1), together with a reported secoiridoid aglycone (2), three phenylethanoids (3, 4 and 6), phenolic acid (5) and two coumarins (7 and 8) were isolated from the stem bark of Syringa reticulata (Bl.) Hara (Oleaceae). The structures were elucidated from 1D and 2D NMR spectroscopic data by comparison with reported literature values. Compound 1 and compounds 2, 4–6 and 8 were identified in the genus Syringa for the first time. The chemotaxonomic significance of these compounds is also discussed.     

Antimalarial constituents from Nauclea orientalis (L.) L

Bioassay-guided fractionation of the antimalarial-active CHCl3 extract of the dried stem of Nauclea orientalis (L.) L. (Rubiaceae) has resulted in the isolation of two novel tetrahydro-beta-carboline monoterpene alkaloid glucosides, naucleaorine (= (16alpha,17beta)-3,14:15,20-tetradehydro-16-ethenyl-17-(beta-D-glucopyranosyloxy)-19alpha-methoxyoxayohimban-21-one; 1) and epimethoxynaucleaorine (2), as well as the known compounds, strictosidine lactam (= (15beta,16alpha,17beta)-19,20-didehydro-16-ethenyl-17-(beta-D-glucopyranosyloxy)oxayohimban-21-one; 3), 3,4,5-trimethoxyphenol (4), 3alpha-hydroxyurs-12-en-28-oic acid methyl ester (5), 3alpha,23-dihydroxyurs-12-en-28-oic acid (6), 3alpha,19alpha,23-trihydroxyurs-12-en-28-oic acid methyl ester (7), and oleanolic acid (8). Compounds 1, 2, 6, and 8 showed moderate in vitro activities against Plasmodium falciparum. Their structures and configurations were elucidated by spectroscopic methods including 1D- and 2D-NMR analyses.     

Characterization of microsatellite markers in the rosewood (Dalbergia monticola Bosser & R. Rabev.)

Dalbergia monticola is one of the major components of the oriental forest of Madagascar. This economically and ecologically important tree is threatened because of the dramatic decrease of the Madagascar forest. We have estimated the genetic diversity and structure of the species by studying nuclear microsatellites. We have developed eight pairs of primers to analyse 215 individuals distributed from the north to the south of the island. These markers will be useful for genetic and ecological studies of this species.     

Direct somatic embroygenesis from immature embryos of rosewood (Dalbergia latifolia Roxb.)

Direct regeneration of somatic embryos was obtained from immature zygotic embryos of Dalbergia latifolia. Immature embryos dissected from green pods 90 d after flowering gave the highest frequency of somatic embryo formation. Preculture on high 2,4-D medium for 4 weeks induced direct somatic embryogenesis, which was expressed during the second culture phase in the presence of low 2,4-D along with a high sucrose concentration. Embryos were separated and transferred to the maturation medium containing MS + 0.5–1.0 mg/L BAP, where embryos developed into plantlets. Somatic embryos failed to convert into complete plants without BAP treatment. This method of direct regeneration of somatic embryos without a callus phase has direct application for genetic manipulation studies.Abbreviations MS Murashige and Skoog (1962) medium - 2,4-D 2,4-Dichlorophenoxyacetic acid - BAP 6-benzylaminopurine - ABA Abscisic acid - KIN Kinetin