GABAergic phthalide dimers from Angelica sinensis (Oliv.) Diels

The methanol extract of Angelica sinensis (Oliv.) Diels roots (Dang Gui) has been shown to exhibit competitive binding to the GABAa receptor, suggesting the presence of GABAergic ligands. Chromatographic fractionation of the methanol extract led to the isolation of two GABAergic dimeric phthalides 1 and 2. Gelispirolide (1) was elucidated as a new phthalide dimer composed of a Z-ligustilide and a Z-butylidenephthalide unit on the basis of spectroscopic approaches including one- and two-dimensional NMR, HRESIMS and HRESIMS-MS. Compound 2 was identified as the known dimeric phthalide, riligustilide, by comparison of its spectroscopic data with literature values. Its dimeric linkage and stereochemistry were ascertained by a single crystal X-ray diffraction experiment. Both dimers 1 and 2 were found to be active in an in vitro GABAa receptor-binding assay with IC50 values of 29 and 24 microM, respectively.     

当归内酯对免疫抑制小鼠免疫功能的重建作用(英文)

探讨当归内酯(ASDL)对免疫抑制小鼠免疫功能的重构作用。通过小鼠腹腔注射环磷酰胺建立免疫抑制动物模型。采用免疫器官重量法和小鼠腹腔巨噬细胞吞噬鸡红细胞实验检测了ASDL对非特异性免疫功能的影响;用血清溶血素分光光度法检测了对体液免疫功能的作用;用MTT法进行了致分裂原诱导的小鼠脾淋巴细胞的增值反应实验,再用乳酸脱氢酶法测定了NK和CTL细胞活性,从而确定ASDL对小鼠细胞免疫功能的影响。结果表明:ASDL能够对免疫低下小鼠的非特性和特异性免疫功能起到一定的重构作用。但是这种效果并不是剂量依赖性的,20 mg/kg这个剂量的效果明显好于5和80 mg/kg这两个剂量。上述结果表明ASDL能够显著提高免疫低下小鼠的免疫功能。     

Structural analysis of water-soluble glucans from the root of Angelica sinensis (Oliv.) Diels

Two water-soluble glucans (designated APS-1cI and APS-1cII) were extracted from the roots of Angelica sinensis (Oliv.) Diels and further purified by anion-exchange and gel-filtration chromatography. Their molecular weights were determined to be 1.7 x 10(5) and 3.9 x 10(4)Da, respectively. The structures of the purified glucans were investigated by a combination of chemical and instrumental analysis, such as methylation analysis, periodate oxidation, GC-MS, as well as FTIR and NMR spectroscopy ((1)H, (13)C, H-H COSY, HSQC, HMBC, TOCSY and NOESY). The data obtained indicated that APS-1cI was a linear alpha-glucan composed of only (1-->6)-alpha-D-Glcp, and APS-1cII had a repeating unit consisting of (1-->4)-alpha-D-Glcp and (1-->6)-alpha-D-Glcp in a molar ratio of 4:1. Such glucans isolated from A. sinensis (Oliv.) Diels have not been previously reported.     

Somatic embryogenesis and plant regeneration of litchi (Litchi chinensis Sonn.) from leaves of mature phase trees

Embryogenic cultures were induced from leaflets from new vegetative flushes of mature ‘Brewster’ litchi trees on B5 medium containing 400 mg l⁻¹ glutamine, 200 mg l⁻¹ casein hydrolysate, 30 g l⁻¹ sucrose, 4.52 μM 2,4-D, 9.30 μM kinetin and 3 g l⁻¹ gellan gum in darkness. Embryogenic cultures consisting of proembryonic cells and masses were maintained either on semi-solid MS medium supplemented with 4.52 μM 2,4-D and 0.91 μM zeatin or as embryogenic suspension cultures in liquid medium of the same composition. Maturation of somatic embryos occurred on semi-solid MS medium with 5–20% (v/v) filter-sterilized coconut water in darkness. Recovery of plants from somatic embryos was improved with 14.4 μM GA₃ on half-strength MS medium with 0.2 g l⁻¹ activated charcoal under a 16 h photoperiod provided by cool white fluorescent lights (60–80 μmol s⁻¹ m⁻²). Plants have been successfully acclimatized in the greenhouse.     

Induction of Embryogenic Callus and Histocytological Study on Embryoid Development of Angelica sinensis (Oily.) Diels

The common calluses derived from roots of Angelica sinensis were treated with phytohormones and different kinds of organic adjuvants with various concentrations to induce embryogenic callus. The phytohormones obviously affected embryogenic callus induction and the process of globular, heartshaped, torpedo and cotyledon stages. They could emerge from both inside and surface of calluses. In the developmental process, the embryoid still kept in contact with the base of callus from which it arose.     

Callus Induction and Plantlet Regeneration from Several Organs of Angelica sinensis (Oliv.) Diels

For the culture of organs, modified B5, KU, MS, NT, 1/2 MS media were used. Callus induction and plantlet regeneration from mots, leaves, petioles, cotyledons and hypocotyls of Angelica sinensis (Oliv.) Diels have been obtained. It was found in our experiments the following results: 1. The callus induction from the rcots has arisen rapidly and increased at a high rate and with a high differentiating frequency. 2. The phytohormones play a significant role in callus induction and differentiation. 3. The suitable combination of cytokinin (6-BA) with auxins (2, 4-D, NAA) may enhance their induction and subculture, but the effect was different on the differentiation. 4. The bud was differentiated from the embryoid and the callus.     

Water-soluble polysaccharides from Angelica sinensis (Oliv.) Diels: Preparation, characterization and bioactivity

Crude water-soluble polysaccharides (ASP) were separated from Angelica sinensis (Oliv.) Diels by hot water extraction. They were fractionated into neutral and acidic polysaccharides by anion-exchange chromatography. The neutral polysaccharide (ASP1) was rich in glucose, galactose, and arabinose suggesting a mixture of glucan and arabinogalactan. The acidic polysaccharide (ASP2, ASP3) consisted mainly of galacturonic acid along with rhamnose, arabinose, and galactose indicating a pectic polysaccharide. The degree of esterification of ASP and ASP3 were 54.06% and 47.14% for the crude and purified sample, respectively. ASP3, with a molecular weight of 3.4 × 10⁴ Da determined by high-performance size-exclusion chromatography (HPSEC), was the major constituent for the crude extracts. The radioprotective effect of the pectic polysaccharide ASP3 was studied in murine models. ASP3 pretreated mice exhibited a significant decrease of apoptosis (P < 0.05, dosage of 200 mg/kg d body weight) in peripheral lymphocytes compared to the irradiated control. The results showed that ASP3 can protect leucocytes and lymphocytes of mice against radiation induced damage, which has potential radioprotective effect on acute radiation injured mice.     

Somatic Embryogenesis and Plant Regeneration from Protoplasts of Cowpea (Vigna sinensis)

Immature cotyledons of cowpea (Vigna sinensis Endlo) were used for protoplast isolation. Enzyme solution for protoplast isolation contained 40% cellulase Onozuka R-10,0.30% Macerozyme R-10 and 2% hemicellulase. The purified protoplasts were cultured in Bs,MS or KM8p liquid medium in dark (25℃) at a density of 1 × 105–5 × 105/ml. The protoplasts started cell division in 3–5 days . Sustained cell divisions resulted ill formation of cell clusters and small calli,with cell division frequency reaching 23%–28% in MS medium . Calli of 2 mm in size were transferred onto MSB (MS salts+B5 vitamins) medium with 2 mg/L 2,4-D, 0. 5mg /L BA forfurther growth. Embryogenic calli appeared on this medium. After passage to fresh medium with the same composition, the embryogenic calli were transferred into MSB liquid medium to establish suspension culture. When the suspended calli were transferred back onto MSB agar medium with 0. 1 mg /L IAA, 0.5mg/L KT, 5% mannitol (cultured in light,2000 lx,12h/d), a lot of adventitious roots formed in 7–10 days, and then somatic embryos formed from the protoplast derived calli. But only a few embryoids developed further into the cotyledonary stage ,and the others died at globular, heart-shaped, or torpeto stage . Finally, some cotyledonary embryoids germinated and developed into plantlets or shoots with leaves.     

海岛棉悬浮培养体细胞胚胎发生

取海岛棉无菌苗下胚轴作外植体,接种在含有激素２,４－Ｄ０．１ｍｇ／Ｌ和ＫＴ０．５ｍｇ／Ｌ的改良ＭＳ培养基上。经近２个月的诱导培养,获得淡黄色或淡灰色、松软、湿润、颗粒状的愈伤组织。将此类愈伤组织接入液体培养基进行悬浮培养,３～４个月后产生体细胞胚,获得各个时期的体细胞胚胎。胚性细胞悬浮系经适当的继代培养可长期保持成胚性。     

Somatic embryogenesis and plant regeneration from immature embryo cultures of onion (Allium cepa L.)

Somatic embryos were obtained and plants regenerated from immature embryos of onion following culture on embryogenic induction media. Highest rates of somatic embrogenesis resulted from 0.5- to 1.5-mm immature embryos cultured on media containing 5 mg/l of picloram. Somatic embryos formed either directly on the surface of embryos or developed from compact cultures. The production of somatic embryos was significantly affected by the addition of auxin, embryo size and cultivar. The potential of somatic embryogenic cultures for plantlet regeneration has been maintained for over 1 year in some lines. Three types of immature-embryo-derived cultures were characterized by histology. Some cultures were morphologically similar to immature-embryo-derived embryogenic cultures of other monocotyledonous species. Cultures such as these have proven to be useful target tissues in transformation studies. Received: 16 December 1997 / Revision received: 23 February 1998 / Accepted: 13 March 1998     

cDNA-AFLP比较当归早薹基因转录差异反应体系的建立

建立了利用cDNA-AFLP技术对当归早薹相关的差异表达基因进行筛选的方法,优化了影响cDNA-AFLP试验结果的关键因子,包括选择性扩增模板量、酶离子浓度、dNTP浓度。结果表明,以1μL稀释10倍的预扩增产物作为模板、1.5 mmol/LMgCl2、0.6 mmol/L dNTP,进行的选择性扩增反应获得DNA片段分子量范围较广,在100-1 000 bp长度范围中均有扩增片段,是较为理想的选择性扩增的结果。聚丙烯酰胺凝胶电泳验证了试验结果,在优化条件下利用部分引物组合可以较好地比较早薹当归在基因转录水平发生的差异。     

连作对当归药材挥发油含量的影响

采用水蒸气蒸馏法提取不同茬口当归药材挥发油,并用GC-MS法鉴定化学成分,归一化法测定相对含量,比较了不同茬口挥发油出油率及化学成分的差异.结果表明,正茬、迎茬、连作2a、连作3a栽培当归挥发油收率分别为0.95%、0.75%、0.70%、0.64%,挥发油中藁本内酯的相对含量分别为71.62%、49.04%、29.49%、59.58%,藁本内酯的异构化产物邻苯二甲酸酐和1,4-环己二烯1,2-二羧酐的相对含量分别为0.21%和4 47%、3.81%和8.55%、6.48%和10.65%、1.04%和5.33%.因此,与正茬栽培相比,连作栽培当归中挥发油的收率及其中藁本内酯的相对含量均明显降低,但挥发油的组分基本一致.     

Prolific shoot regeneration from immature embryo explants of sainfoin (Onobrychis viciifolia Scop.)

Summary Immature cotyledons and embryo axes of sainfoin were cultured on Murashige and Skoog (MS) media supplemented with various concentrations of 6-benzylaminopurine (BAP) and a-naphthaleneacetic acid (NAA) to induce adventitious shoot regeneration. The highest frequency of shoot regeneration occurred following an initial callus growth on a MS medium containing 0.5 mg/l BAP and 2 mg/l NAA. Immature embryo axes showed higher regeneration capacity than immature cotyledons, however, shoot elongation was best achieved on immature cotyledons. Regenerated shoots were excised and rooted in half strength MS medium with 1 mg/l indole-butyric acid (IBA) or 1 mg/l NAA. The rooted plantlets were finally transferred to compost.     

Immunomodulatory activity of polysaccharide isolated from Angelica sinensis

The immunomodulatory activities of an Angelica sinensis polysaccharide (AP), purified from the fresh root of A. sinensis Diels, were investigated in vitro in relation to the specificity to immune cells. AP consisted of rhamnose, arabinose, mannose, glucose, galactose with the molar ratio of 1.00:4.54:2.98:11.09:7.45. Cell proliferation results showed that proliferation of total spleen cells, macrophages and T cells were promoted by the action of AP. The treatment of AP increased the production of IL-2 and IFN-γ, while that of IL-4 was decreased. RT-PCR analysis displayed that the IL-2 and IFN-γ gene expression were enhanced but the IL-4 gene expression was decreased. Some differences in cytokines secretion pattern were also detected, the expression of IFN-γ was rapidly augmented while that of IL-2 responded later. The flow cytometry results showed that the percentage of CD4⁺T cell in total spleen cells was remarkably increased by AP, while that of CD8⁺T cell was slightly decreased. In conclusion, AP has immunomodulatory activity by regulating expression of Th1 and Th2 related cytokines. The time–effect relation of cytokines response also suggests that macrophages and natural killer cells involved in nonspecific immunity were primary activated, and helper T cell were secondarily affected by AP.     

Ultrastructural Studies of Somatic Embryogenesis in Wheat

Electron microscopic observation of somatic embryogenesis from cultured immature wheat embryos revealed the presence of a lot of small vacuoles, a large nucleus, clear nucleolus and polynucleoli. The electron density of cytoplasm was strengthened during somatic embryogenesis. Quantity and type of organelles—plastid, ribosome and mitochondrion were increased; thickened cell wall, disappeared plasmodesmata, increased organelles andstarch accumulation in the embryogenic cells. Nucleolus vacuoles, autophagic vacuoles and secretory vesicles were present in the embryogenic cells with thickened cell walls. The multicellular proembryos, globular embryoid and pear-shaped embryoid were surrounded by an envelop, but plasmodesmata existed extensively between cells of somatic embryoid. The membranous structures appeared in the plastid which underwent transformation into chloroplast in the cells of growing point in almost mature embryoid. The relation of the above-mentioned structureal changes to somatic embryogenesis is also discussed.     

当归多糖APS-3c的结构特征及体外抗肿瘤作用的研究

中国当归粗多糖经阴离子交换纤维素柱(DEAE-sephadex A-25)、凝胶柱(Sephacryl S-400及Sephadex G100)色谱分离纯化,得淡黄色粉末状多糖APS-3c.采用高效尺寸排阻色谱、红外光谱、气相色谱和糖醛酸还原等方法进行了结构的初步分析,用四甲基偶氮唑蓝比色法观察APS-3e对多种肿瘤细胞的生长抑制作用.结果表明APS-3c为均一组分,分子量为1.4×104 Da,由阿拉伯糖.鼠李糖.葡萄糖-半乳糖-甘露糖-木糖-半乳糖醛酸(6.7:6.5:6.3:4.7:1.6:1.0:13.2)构成,是首次从中国当归中分离出的新型酸性多糖.APS-3c对人白血病HL-60细胞、人结肠癌SW1116细胞的增殖有一定的抑制作用.     

Establishment of a System with High Synchronous Frequency of Somatic Embryogenesis and Embryo Seedling Formation in Camellia sinensis var. assamica

The system of high synchronous frequency of somatic embryogenesis and somatic embryo seedling formation was established by means of embryonic cell hne 1 ( CL1 ) of Camellia sinensis var. assamica Kitamura. Modified MS was used as the basic medium. Cultures of CL1 was transferred to the aqueous induced medium (0.05 mg/L 2,4-D + 0.50 mg/L 6-BA) from the maintenance medium (0.1 mg/L 2,4-D + 0.5 mg/L 6-BA) for somatic embryos induction under dark condition. 28 days later, they were cultured in the liquid differentiation medium. Various kinds of somatic embryos were obtained after another 28 days. The frequency of somatic embryos was 81.5 %. Various mesh sizes of sieves were applied to collect the somatic embryos in different developmental stages which could develop to mature stage in the aqueous growth medium ( 1/2 MS + 1.0 mg/L GA3 + 0.5 mg/L 6-BA). ABA was effective to promote the formation of highly qualified somatic embryo. The mature somatic embryos sized 20 to 70 mesh had the conversion frequency 75 %. The development of somatic embryogenesis studied under a cell suspension culture system was similar to the zygotic embryogenesis.     

当归栽培及加工技术

本文对当归的形态特征、分布区域、药用价值,特别是当归的栽培及加工技术进行了详细的论述,以期为当归的生产提供一项实用技术.