Pattern formation and cell interactions in epidermal development of Anemarrhena asphodeloides (Liliaceae)

In the leaf epidermis of the monocotyledon Anemarrhena asphodeloides Bunge a characteristic pattern of stomata and papilla cells is developed. The ontogeny of both kinds of specialized cells is initiated by a formative mitosis yielding one daughter cell with aberrant structure and competence. It is shown that these distinctive cell types affect the fate of neighbouring epidermal cells. The final epidermal pattern is due to random initial events, as well as cell lineage, and cell-to-cell interactions during development.     

Intestinal Peyer’s patch-immunomodulating glucomannans from rhizomes of Anemarrhena asphodeloides Bunge

During screening for intestinal Peyer’s patch-immunomodulating polysaccharides from plant resources including medicinal herbs, a potent modulating activity was observed in a crude polysaccharide fraction (AS-1) from the rhizome of Anemarrhena asphodeloides Bunge. Oral administration of AS-1 (100 mg/kg/day) to aged BALB/c mice enhanced productions of IL-10, IFN-γ and IL-6 from Peyer’s patch immunocompetent cells, and its oral administration to ovalbumin (OVA)-fed B10.A mice led to significant suppression on induction of OVA-specific IgE in systemic immune system. Further fractionation of the polysaccharides in the crude polysaccharide fraction, AS-1, yielded 4 polysaccharide fractions that were potently active, and contained glucomannans. Treatment of these polysaccharide fractions with endo-β-d-(1 → 4)-mannanase significantly decreased their activities. Mannanase digestion of the active glucomannan gave both long and short hexosyl-oligosaccharides, whereas konjac glucomannan, which was inactive, released short oligosaccharides. Structural analysis indicates that the long oligosaccharides from the active glucomannan contain mannanase-resistant complex structure comprising β-d-Man and β-d-Glc.     

Karyotype Analysis of Reineckia carnea (Andr.) Kunth (Liliaceae)

Karyotype analysis for the species Reineckia carnea (Andr.) Kunth of the monotypic genus Reineckia Kunth is given for the first time. The number of chromosomes in root-tip cell was found to be 38, which is in accord with those reported by most of the previous authors^{[5,7,8,9,11,12,]}. The somatic complement shows a slight variation in size, i.e., the 2, 3, 5, 6, 7th pairs of the chromosomes have submedian constrictions, while the other pairs have median centromeres. The karyotype is therefore a rather symmetrical one, and according to the chromosomal terminology defined by Levan et al^[4], the karyotype formula of the species is 2n=38=28 m+10 sm. In spite of the presence of two nucleoli in the telophase as observed by the authors and Noguchi^[8] as well, the two corresponding Sat-chromosomes have not been found. Photomicrograph of the chromosome complement and idiogram are given in Fig. 1 and 2 respectively.     

Investigation on Embryology of Anemarrhena asphodeloides

Anemarrhena asphodeloides Bunge is the only species of Anemarrhena in Liliaceae, which possesses three stamens. The flowers in this species have following features: (1) Crystalliferous cells are present in the perianth and the filament. (2) Epidermal cells of filaments and the inner perianth appears verruciform. (3) In longitudinal section, a number of the multicellular hairs were found in the apex of the inner perianth. The above characteristics of Anemarrhena are possibly important and differ from those of the other genera in Liliaceae. The main aim of the present paper is to deal with the female gametophyte and embryogenesis in Anemarrhena. The development of embryo sac is similar to that of Ornithogalum (Tilton et al., 1981), belonging to the Polygonum type, but there is a short embryo sac haustorium at the antipodal end. Before fertilization the two polar muclei fuse into a secondary nucleus. The filiform apparatus was found in the synergid. The early development of proembryo in Anemarrhena is similar to that of Najas (Hu, 1982). After fertilization the zygote has a short stage of dormancy. When the endosperm has 12-16 free nuclei, the first division of the zygote takes place, forming an apical cell and a basal cell. Then the apical cell undergoes transversal divisions 2 or 3 times, forming a line of three to four cells. The basal cell usually does not further divide. The endosperm formation in Anemarrhena is the Helobial type. The small chalazal chamber is usually ephemeral and 2-4-nucleate, while the large micropylar one may be a multi-nucleate before wall formation.     

知母中甾体皂苷类成分的研究(英文)

从中药知母(Anemarrhena asphodeloides Bge.)的70%乙醇提取物中分离得到6个化合物,经波谱学方法和与已知样品对照鉴定为:3-O-β-D-glucopyranosyl-(1→2)-β-D-galactopyranosyl-(3β,5β)-pregn-16(17)-ene-20-one(1)、timosaponin AIII(2)、timosaponin BIII(3)、22-hydroxy-5β-furost-3β,15α-diol-3-O-β-D-glucopyranosyl(1→2)-β-D-galactopyranoside(4)、timosaponin G(5)、β-daucosterol(6)。其中化合物1为新天然产物,命名为知母孕甾A,是知母中首次发现的C21甾体类化合物,经HR ESI-MS、1D NMR、2D NMR确定其结构,并首次对其核磁数据进行归属。     

知母资源化学及药理研究进展

对中药知母的化学成分、积累动态及药理研究概况进行了综述.     

Karyotype variation inAllium commutatum (Liliaceae s. lato)

Chromosome numbers (2×, 3×, 4×), B-chromosomes, and variation among marker chromosomes inAllium commutatum Guss. was studied. These different chromosomal types exhibit distinct distribution patterns and a centre of variation has been located in the central Aegean area. A common origin ofA. commutatum, the Aegean endemicA. bourgeaui Rech. fil.,A. ampeloprasum L. and the cultivatedA. porrum L. is discussed.     

A Study on Karyotype Differentiation of Allium fasciculatum (Liliaceae)

Karyotypes of three local populations of Allium fasciculatum Rendle were analyzed. The karyotype of the population from Dagze of Xizang was 2n=2x＝20=4m+10sm +4T+2t(2SAT), the one from Xiangcheng of Sichuan was 2n=2x＝20＝10sm+6t(2SAT)+4T, and the one from Lixian of Siehuan was 2n＝2x=20＝6m+10sm+2t (2SAT)+2T. Although there were differences in karyotypes among the three populations, they were very similar in morphological characters, except that plants in the Dagze population were lower. The karyotype of the Lixian population is considered to be a primitive one that has evolved into the more asymmetrical karyotypes of the Dagze population and Xiangcheng population by structural aberration. In the genus Allium, only four species, A. fasciculaturn, A. decipiens Fisch., A. kujukense Vved. and A. cheloturm Wendelbo. have so far been found to have the basic chromosome number of x＝10. According to karyotype characters and the distribution pattern of these four species, the basic number x = 10 may have different origins. At least that of A. fasciculatum is different from that of the other three species and might have originated the latest. A. fasciculatum was similar to A. hookeri Thwaites (the latter has x＝11) in karyotypical and morphological characters. Therefore, they are considered to have originated from a common extinct ancestor that had the basic number of x = 10.ey words Allium;A．fasciculatum Rendle;Karyotype differentiation     

Isolation and anti-oomycete activity of nyasol from Anemarrhena asphodeloides rhizomes

The methanol extract of Anemarrhena asphodeloides rhizomes exhibited strong antifungal activity against the plant pathogenic fungi Magnaphothe grisea, Rhizoctonia solani, and the plant pathogenic oomycete Phytophthora capsici. The antifungal substance isolated from the rhizomes of A. asphodeloides was identified to be nyasol, (Z)-1,3-bis(4-hydroxyphenyl)-1,4-pentadiene by NMR and mass spectral analysis. Nyasol effectively inhibited the mycelial growth of Colletotrichum orbiculare, P. capsici, Pythium ultimum, R. solani, and Cladosporium cucumerinum in a range of 1-50 mug/ml, but did not affect the growth of bacteria and yeast. In a greenhouse test, treatment with the antifungal compound nyasol was significantly effective in suppressing the Phytophthora blight on pepper plants.     

知母分蘖一步法快速繁殖体系的初步构建

以知母分蘖为试材,对知母的一步法快速繁殖体系进行了初步研究。结果表明,知母分蘖直接再生植株的最佳培养基是MS+KT 1 mg/L+NAA 0.5 mg/L;知母分蘖愈伤组织诱导和再生的最佳培养基是MS+KT 2 mg/L+NAA 0.5 mg/L。本实验构建了知母分蘖的一步法快速繁殖体系,为知母试管苗的工厂化生产提供了理论和技术基础。     

The Ultrastructural Aspect of Tapetum and Ubisch Bodies in Anemarrhena asphodeloides

From ontogeny of tapetum in Anemarrhena asphodeloides, the ultrastructnral features of tapetal cells are as follows: 1. The profuse rough endoplasmic reticula are often closely associated with lipid bodies and vesicles, and linking each other into compound organelles. This is one of the striking features in Anemarrhena tapetal cell. 2. After meiosis of the micro- spore mother cell, the tapetal cytoplasm contains a large number of vesicles, in which the electron opaque substances are accumulated. Then they fuse to form a large zone of storage material similar to lipid bodies. Before accumulation of opaque material, these vesicles in the tapetal cytoplasm are larger than those in elaioplast (see Plate II, Fig. 2). 3. During stage of pollen maturation the tapetal cytoplasm becomes disorganized and the cells are almost occupied by the elaioplasts at various degree of development. On the basis of the report of Dickinson (1973), the formation of a pollen coatings of Lilium is different from that of Raphanus. The osmiophilic bodies in the former have originated from membrane lamellae or membranous system of plastid, and those in the latter are formed from the plastid vescles. It is intereting to note that the mode of origin of the plastid osmiophilic bodies in Anemarrhena is rather similar to that of Raphanus than to Lilium. About the origin of the pro-Ubisch bodies in tapetal cytoplasm of Anemarrhena studies revealed that a large number of the medium electron dense bodies appear in the tapetal cytoplasm. This is the first sign of the formation of the pro-Ubisch bodies and its character is very similar to spherosome in many respects. From many ultrasections, it can be seen that the ER profile is closely associated with the pro-Ubisch bodies. Thus we can conclude that the proubisch bodies of Anemarrhena are derived from rough endoplasmic reticulum. Although Heslop-Harrison et al. (1969) has considered that the compound Ubisch bodies do not occur in Lilium, there are prominent aggregation of Ubisch bodies in Anemarrhena, same as reported in Oxalis (Cariel, 1967), Silene (Heslop-Harrison, 1963a) and Helleborus (Echlin et al., 1968). After investigation on certain angiosperm in 1972, Gupta and Nanda have reported that the peritapetal membrane belonging to tapetum of secretory type lies against the inner tang- ential wall; in the plasmodial type of tapetum, it is formed on the outer tangential wall. But in some species of Poaceae and Solanaceae, the peritapetal membrane is formed on both sides of the tapetal cells (Banerjee, 1967; Reznickov & Willemse, 1980). In the secretory tapetum of Anemarrhena, the peritapetal membrane, which do not comply with the conclusion of Gupta & Nanta (1972), is formed on outer tangential wall.     

KT和NAA对知母愈伤组织再分化的影响

以知母愈伤组织为材料,采用单因子试验方法,探讨不同浓度KT和NAA对其再分化的影响。结果表明:知母愈伤组织生根的最佳培养基为MS+KT 2 mg/L;愈伤组织再生芽的最佳培养基为MS+KT 2 mg/L+NAA 1 mg/L;愈伤组织再生苗长高的最佳培养基为MS+KT 2 mg/L+NAA 0.5 mg/L;愈伤组织增殖的最佳培养基为MS+KT 2 mg/L+NAA 1 mg/L。     

正交试验法优选超声提取知母皂甙的研究

用正交实验法对知母(Anemarrhena asphodeloides Bge.)中总皂甙的超声提取工艺进行了研究,采用正交表L9(34),以提取剂的体积分数、料液比(mA∶VB)、提取时间以及提取次数为因素,以知母总皂甙元的含量为指标,用紫外分光光度计法进行检测,所得提取液经大孔树脂纯化,得到的最佳提取参数和结果为:以φ(乙醇)=75%的水溶液为提取剂、提取料液比mA∶VB=1∶8、提取次数2次、每次30min,提取率92.8%,纯度91.0%。结果证明了对知母而言,超声提取法是一种提取效率高、操作简便、省时的方法。     

Effect of timosaponin A-III,from Anemarrhenae asphodeloides Bunge (Liliaceae), on calcium mobilization in vascular endothelial and smooth muscle cells and on vascular tension

The effects of timosaponin A-III (TA-III), from Rhizoma Anemarrhenae, on Ca(2+) mobilization in vascular endothelial cells and smooth muscle cells and on vascular tension have been explored. TA-III increased intracellular Ca(2+) concentrations ([Ca(2+)](i)) in endothelials cells at a concentration larger than 5 microM with an EC(50) of 15 microM, and increased [Ca(2+)](i) in smooth muscle cells at a concentration larger than 1 microM with an EC(50) of 8 microM. Within 5 min, the [Ca(2+)](i) signal was composed of a gradual rise, and the speed of rising depended on the concentration of TA-III. The [Ca(2+)](i) signal was abolished by removing extracellular Ca(2+) and was recovered after reintroduction of Ca(2+). The TA-III-induced [Ca(2+)](i) increases in smooth muscle cells were partly inhibited by 10 microM nifedipine or 50 microM La(3+), but was insensitive to 10 microM verapamil and diltiazem. TA-III (10-100 microM) inhibited 0.3 microM phenylephrine-induced vascular contraction, which was abolished by pretreatment with 100 microM N(omega)-nitro-L-arginine (L-NNA) or by denuding the aorta. TA-III also increased [Ca(2+)](i) in renal tubular cells with an EC(50) of 8 microM. Collectively, the results show for the first time that TA-III causes [Ca(2+)](i) increases in the vascular system. TA-III acted by causing Ca(2+) influx without releasing intracellular Ca(2+). TA-III induced relaxation of phenylephrine-induced vascular contraction via inducing release of nitric oxide from endothelial cells.     

Karyotype studies in thirty‐two species of Lilium (Liliaceae) from China1

Fifty‐two populations representing thirty‐two taxa of Lilium L. from China are karyologically analyzed. The results showed that all populations have the same basic chromosome number x=12, and all species are diploid except Lilium tigrinum, which is triploid with 2n=36. In addition, one population of L. sulphureum is aneuploid with 2n=23. The karyotype evolution in the genus Lilium is mainly in terms of alterations of the fine structure of the chromosomes, not in ploidy or basic chromosome number variation. When combining previous studies and our results, we consider the major driving forces for evolution of Lilium to be an increase in karyotypic asymmetry, and unequal translocation and formation of secondary constrictions. The existence of intercalary satellites on the two largest pairs of chromosomes is correlated with the geographic distribution pattern of Lilium across the world. The Hengduan Mountains is a modern diversity and differentiation center of Lilium and it also could be the center of origin for this genus. In addition, some taxonomic conclusions were verified on the subgenus level. Among the species investigated, the chromosome number and karyotypes of 13 taxa were documented for the first time.     

化学诱变剂EMS对知母种子萌发的影响

以知母（Anemarrhena asphodeloides）种子为材料,分别采用1%、2%、4%、6%浓度的EMS处理6、8、12和24 h,在恒温培养箱内进行种子萌发实验,研究不同浓度的化学诱变剂甲基磺酸乙酯（ethyl methane sulfonate,EMS）处理对知母种子萌发的影响,筛选适宜的诱变浓度和诱变时间。结果表明：随EMS浓度的增加和处理时间的延长,知母种子发芽率呈现逐渐降低的趋势,以相对发芽率达到半致死浓度为标准,6%EMS浸种12和24 h可作为EMS诱导知母建立突变体库的适宜条件。     

The Biology of Allium monanthum (Liliaceae) I. Polyploid Complex and Variations in Karyotype

Abstract Plants of Allium monanthum Maxim., whose gender expression are usually dioecious, but rarely hermaphrodite or gynomonoecious, proved to constitute a polyploid complex, consisting of diploid, triploid, and tetraploid individuals. The basic chromosome complement of this species consists of seven metacentric or submetacentric chromosomes and one acrocentric, the latter possessing a satellite on the short arm. Thus, the karyotype formula is expressed as 7V+11 (x=8). The diploid plants (2n = 16) were confined to central Honshu, Japan. Typical female plants possessed the standard karyotype, whereas male plants were heterozygous for two kinds of translocations. The 3x plants (2n=24) are somewhat widely distributed in the areas from the Kanto to Hokuriku district in Honshu. All female triploid plants possessed the standard karyotype. The geographical distribution of 4x plants (2n=32) which express mostly a female phenotype occurred nearly throughout the whole areas investigated; they are geographically isolated from the 2x plants. A majority of 4x plants had the standard karyotype. The remaining tetraploids were of the aberrant type, 4x/51, which has five acrocentric chromosomes, and two aneuploids 4x+1 and 4x-1. Both 3x and 4x forms seem to be of autopolyploid origin. Three kinds of aberrant nucleolar chromosomes with an extra satellite or an inseried secondary constriction were found in the heterozygotes for translocations of 2x plants and also in some plants of the 3x form. These aberrant plants usually form their own homogeneous populations, but were somewhat scattered throughout the range in their distribution. Thus, these individuals are considered to have perpetuated these types of chromosome aberrations which originated in the remote past.     

Identification of nyasol and structurally related compounds as the active principles from Anemarrhena asphodeloides against respiratory syncytial virus (RSV)

Three known phenolic compounds, (-)-(R)-nyasol (= 4,4'-(1Z,3R)-Penta-1,4-diene-1,3-diyldiphenol; 1), its derivative 2, and broussonin A (3)--isolated from the rhizomes of Anemarrhena asphodeloides--were for the first time identified as the active principles capable of efficient respiratory-syncytial-virus (RSV) inhibition. The IC50 values of 1-3 against the RSV-A2 strain, propagated in HEp-2 cells, were determined, their activities being higher than that of the standard antiviral drug ribavirin (IC50 = 1.15 microM). In addition, the known, but inactive, compound 'trans-N-(para-coumaroyl)tyramine' (= (2E)-3-(4-hydroxyphenyl)-N-[2-(4-hydroxyphenyl)ethyl]prop-2-enamide; 4) was isolated from this plant for the first time.     

KT和2,4-D对知母愈伤组织再分化的影响

本文探讨KT和2,4-D组合对知母愈伤组织再分化的影响。结果表明：知母愈伤组织再生芽和促进再生苗长高以及愈伤组织增殖的最佳培养基均为MS + KT 4.0 mg/L。单独使用KT或KT与2,4-D组合均可抑制知母愈伤组织生根。     

The apoptotic effect of sarsasapogenin from Anemarrhena asphodeloides on HepG2 human hepatoma cells

Sarsasapogenin, a kind of mainly effective components of Anemarrhena asphodeloides Bunge (Liliaceae) has the effects of being anti-diabetes and improving memory. However, there are few reports focusing on its anti-tumor effects. In this study, the sarsasapogenin was extracted from rhizomes of A. asphodeloides Bunge and applied to inhibit HepG2 human hepatoma cells. MTT assay showed that sarsasapogenin induced a distinct dose- and time-dependent diminution of cell viability with IC(50) of 42.4+/-1.0microg/ml for 48h. Furthermore, sarsasapogenin-induced apoptosis of HepG2 cells was verified by Hoechst 33258 staining, electron microscopy, DNA fragmentation and PI staining. Flow cytometry analysis showed that sarsasapogenin-induced cell apoptosis was through arrest of cell cycle in G(2)/M phase. Hence we proposed that sarsasapogenin could be used as an anti-liver cancer drug for future studies.