百合皂苷的提取、纯化及其对自由基的清除作用

本文用正交实验法对百合总皂甙的提取工艺中温度、乙醇浓度、固液比例、回流时间和提取次数5个因素进行研究,优选出简便可靠、且适合工业化生产的百合总皂甙的提取工艺。其最佳提取工艺条件是:温度为60℃,乙醇浓度为70%,固液比例为1∶6,提取时间3h,提取次数3次。以溴邻苯三酚红(BPR)为显色剂,基于Co(II)H2O2体系反应产生的羟自由基(·OH)使溴邻苯三酚红(BPR)的颜色发生变化,采用756型紫外可见分光光度计测定其吸光度的变化值,研究百合皂苷在此体系中清除羟自由基的作用,对照实验表明:百合总皂苷提取物对羟自由基的清除作用比人参皂苷强。     

百合花瓣总RNA提取方法的研究

以亚洲百合品种Pollyanna和东方百合品种Sorbonne为试材,在比较异硫氰酸胍法、SDS/酚法与CTAB-L i Cl法提取总RNA效果的基础上,针对百合组织中富含多糖的特点,在Sorbonne提取RNA中加入特殊除多糖步骤,改进了CTAB- L i Cl法.结果表明,改进CTAB- L i Cl法能有效去除多糖,提取到的RNA2 8S r RNA亮度约为18S r RNA的2倍,A2 6 0 / 2 80介于1.8～2 .0之间,A2 6 0 / 2 30为2 .0 ,Pollyanna RNA产率为36 .3μL·g- 1 ,Sor-bonne RNA产率为10 .2 μL·g- 1 ,经RT- PCR获得了特异性条带,说明用改进CTAB- L i Cl法从百合花瓣中提取到的RNA质量好、产率高、完整性强,完全适合于进一步的分子生物学研究.     

The 135-kDa actin-bundling protein from lily pollen tubes arranges F-actin into bundles with uniform polarity

 A plant 135-kDa actin-bundling protein (P-135-ABP) isolated from pollen tubes of Lilium longiflorum (Thunb.) binds stoichiometrically to F-actin filaments and bundles them in vitro (E. Yokota et al., 1998, Plant Physiol. 116: 1421–1429). To further understand the mechanism of actin-filament bundle formation by P-135-ABP, the polarity of each F-actin filament in bundles was examined using myosin subfragment 1 (S-1). Dissociation of F-actin filaments from bundles organized by P-135-ABP was induced by S-1. However, F-actin filaments that remained in a bundle and decorated by S-1 showed uniform polarity. These results indicate that P-135-ABP arranges F-actin filaments into bundles with uniform polarity and consequently plays a key role in the orientation of cytoplasmic streaming in pollen tubes. Received: 23 February 1999 / Accepted: 22 April 1999     

百合查尔酮合成酶(CHS)基因的克隆与分析

以东方百合“索邦”基因组DNA为模板进行PCR扩增,将得到的目的片段克隆至pGEM-T easy载体后进行测序.结果表明,目的序列全长1 307 bp,经BLA ST分析,与鸢尾、玉米、水稻等植物的查尔酮合成酶(CHS)基因核苷酸同源性在70%以上;与已经克隆的CHS cDNA序列相比,CHS DNA序列中包含2个外显子和1个内含子,内含子全长82 bp,符合TG-AG特征.将得到的序列提交G enB ank,序列号为DQ 471951.     

Three Previously Undescribed Chlorophenyl Glycosides from the Bulbs of Lilium regale

Three previously undescribed chlorophenyl glycosides, (2,4,6-trichloro-3-hydroxy-5-methoxyphenyl)methyl β-D-glucopyranoside ( 1 ), (2,4-dichloro-3,5-dimethoxyphenyl)methyl 6-O-β-D-glucopyranosyl-β-D-glucopyranoside ( 2 ) and 4-chloro-3-methoxy-5-methylphenyl 6-O-(6-deoxy-β-L-mannopyranosyl)-β-D-glucopyranoside ( 3 ) were obtained from Lilium regale. The absolute configurations of these new finds were elucidated by comprehensive analyses of spectroscopic data combined with acid hydrolysis derivatization. (2,4-dichloro-3,5-dimethoxyphenyl)methyl 6-O-β-D-glucopyranosyl-β-D-glucopyranoside ( 2 ) can inhibit the proliferation of lung carcinoma A549 cells with an IC₅₀ value of 29 μΜ.     

Cytological Studies of the Cytoplasmic Inheritance in Lilium regale and L. davidii

The distribution and characteristics of plastids and mitochondria in the generative and sperm cells of Lilium regale Wils. and L. davidii Duch. were described. In L. regale there were few plastids and abundant mitochondria in the newly formed generative cell. When the generative cell became free in the vegetative cytoplasm, the plastids degenerated completely within the generative cell. It was further proved by DAPI fluorescent technique that there was no organell DNA in the generative cell within the mature pollen grain or the pollen tube. However, distribution of the plastids was strictly polarizable during the division of the micmspore in L. davidii, resulting the lack of plastids in the newly formed generative cell. Data of RFLP analysis comparable between L. davidii, L. longifiorum and their interspecific hybrid have also proved the plastid inheritance in L. davidii to be of uniparental maternal transmission. Although the mitoehondria were observed both in the generative and sperm cells of L. regale and L. davidii but their DNA was decomposed in the male gametophyte stage. Therefore the mitochondda in the sperm cell could not be transmitted into the offspring. The results provided the detail, cytological evidence that organelles in the microgametophyte are incapable of genetic transmission in the two species of Lilium.     

Constituents in Easter lily flowers with medicinal activity

Easter lily (Lilium longiflorum) flowers have been used in traditional medicine for alleviating many ailments. However, the chemical basis of its bioactivity has not been investigated. We have determined bioactive components in Easter lily flowers using lipid peroxidation and cyclooxygenase enzyme inhibitory assays and found to be kaempferol (1), kaempferol glycosides (2, 3, 4, 8, 9 and 10), quercetin glycosides (5, 6 and 7), a regaloside (11), a chalcone (12) and a fatty acid fraction (13). The structures of compounds were determined by NMR, IR, UV/VIS and mass spectroscopic studies. Compound 1 showed the highest COX-1 inhibition (94.1%) followed by 3, 8 and 12 with 38.7, 30.8 and 32.4%, respectively. Only compound 1 inhibited COX-2 enzyme by 36.9% at 80 ppm. In lipid peroxidation inhibitory assay, kaempferol showed 37 and 100 % inhibitions at 1 and 10 ppm, respectively. At 10 ppm, more than 20% inhibition was observed for compounds 4, 7, 10, 11 and 12 and 53% for compound 3. The compounds reported in here are isolated for the first time from Easter lily flowers including novel compounds 10, 11 and 12. Our results suggest that kaempferol and quercetin flavonoids contributed to the anecdotal medicinal properties of Easter lily flowers.     

兰州百合精细胞表膜上存在被牛精子抗体识别的抗原决定簇

以兔抗牛精子 Ig G为一抗 ,对植物精细胞蛋白进行 Western blot分析 ,发现兰州百合 (Liliumdavidii Duch.)精细胞和生殖细胞中各有一分子量为 64k D的蛋白显示阳性反应 ;在玉米 (Zea mays)精细胞蛋白中也发现了阳性反应条带 ,其分子量为 65 k D和 2 2 k D;而兰州百合花丝、花药壁和玉米黄化苗的蛋白中均没有阳性条带。用兔抗牛精子 Ig G对兰州百合精细胞进行间接免疫荧光标记 ,结果表明在兰州百合精细胞表面 ,有兔抗牛精子 Ig G的识别位点。根据以上结果 ,作者认为植物精细胞中有与动物精子相同或相似的抗原决定簇 ,它 (们 )主要分布于精细胞表膜上 ,并为精细胞所特有     

AM真菌对百合调节激素平衡与细胞渗透性以及改善耐盐性的研究

该试验以东方百合(Lilium brownii)品种‘西伯利亚’和丛枝菌根(AM)真菌摩西斗管囊霉(Funneliformis mosseae)、变形球囊霉(Glomus versiforme)为材料,在温室盆栽条件下,设置NaCl胁迫(0、0.4%、0.8%和1.2%NaCl溶液处理)和接种AM真菌[接种摩西斗管囊霉、变形球囊霉、摩西斗管囊霉+变形球囊霉及未接种对照]双因素试验,分析各处理百合的激素平衡与细胞渗透性变化特征,以明确AM真菌提高百合耐盐性的效应,初步探索AM真菌增强百合耐盐性的作用机制。结果表明:(1)接种AM真菌能有效增加盐胁迫下百合植株的株高和生物量,显著提高百合耐盐系数,双接种处理的株高及地上、地下部分干重在1.2%NaCl胁迫下分别比未接种对照显著增加8.9%、14.5%和11.2%。(2)接种AM真菌能显著提高盐胁迫下百合植株叶片的矿质元素P、K、S含量,显著降低叶片内Na和Fe含量,双接种处理的P、K、S含量在1.2%NaCl胁迫下分别比对照提高10.9%、8.3%、13.7%,而其Na和Fe含量分别显著下降28.4%和66.4%。(3)接种AM真菌能显著提高盐胁迫下百合内源激素吲哚乙酸(IAA)和脱落酸(ABA)含量,双接种处理在1.2%NaCl胁迫下百合内源IAA、ABA含量分别是对照的1.2和1.5倍。(4)接种AM真菌能显著提高盐胁迫下百合可溶性蛋白含量,显著降低其脯氨酸含量,双接种处理在1.2%NaCl胁迫下比对照的升降幅度分别为69%和31%。(5)接种AM真菌能显著降低盐胁迫下百合叶片丙二醛和相对电导率,双接种处理在1.2%NaCl胁迫下比对照分别显著下降58.1%和9.0%。研究发现,AM真菌可以通过增强百合对养分的吸收、降低氧化胁迫造成的伤害、调节植物内源激素平衡状况与细胞渗透性来增强自身的耐盐能力,且双接种处理的效果优于单一接种。