Anti-Influenza A Virus Effect of Hypericum perforatum L. Extract

To study the antiviral effect of Hypericum perforatum L. extract (HPE) on influenza A virus (IAV) (H1N1) in vitro and in vivo. Cytopathic effect (CPE) and neutral red (NR) dye uptake were used to examine the antiviral effect of HPE on Madin Darby Canine Kidney (MDCK) cells which were infected with IAV in vitro. HPE was effective against influenza A virus (IAV) in vitro, with a 50% effective concentration (EC50) of 40 μg/mL. The mean 50% cytotoxic concentration (CC50) in the MDCK used in these experiments was 1.5 mg/mL. Ribavirin was run in parallel with EC50 values of 5.0 μg/mL; the mean CC50 for ribavirin was 520 μg/mL. Oral gavage administrations of HPE or ribavirin to mice infected with the IAV were highly effective in preventing death, slowing the decline of arterial oxygen saturation, inhibiting lung consolidation and reducing lung virus titers. The minimum effective dose of HPE in these studies was 31.25 mg/kg/day, which was administered twice daily for 5 d beginning 4 h prior to virus exposure. Below a dosage of 2000 mg/kg/day, almost all treated mice survived, which suggests that HPE is of low toxicity. Ribavirin's minimum effective dose was 40 mg/kg/day with the LD50 determined to be 200 mg/kg/day. Delay of the initiation of either HPE or ribavirin therapy, using approximately 1/3 LD50 dose each time, could still be protective as late as 48 h after exposure to the IAV. While both agents appeared to have similar efficacy against IAV infections, HPE was considered to be less toxic and may warrant further evaluation as a possible therapy for influenza.     

Multiple shoot formation and phenotypic changes of R0 regenerants in Hypericum perforatum L.

Seedings of Hypericum perforatum L. have produced multiple shoots on RM medium supplemented with 2.22 or 4.40 μM BAP. Regenerants were evaluated for variability in several morphological characteristics. A comparison with the control showed significant differences in fresh and dry matter and height of plants, but did not effect branching and the number of glands containing dianthrones.     

Pharmacodynamic study of Hypericum perforatum L.

The effects of Hypericum perforatum L. (Hypericaceae) crude ethanol extract (A), ethyl acetate extract (B), aqueous extract (C) and infusion (I), on pentobarbital induced sleeping time, intestinal motility, and their analgesic activity, have been investigated. Extracts A and B exhibited significant stimulatory and antidepressant effects on the CNS. Both extracts prolonged sleep, increasing time up to more than 25 min. The antidepressive activity of extract A was also achieved by significant reduction of the myorelaxant activity of diazepam. Extract B exhibited strong analgesic activity reducing abdominal stretching induced by acetic acid by nearly 50 %. Extracts A, B and C exhibited spasmolytic activity, significantly reducing intestine motility.     

Chromosome number stability and mitotic activity of cryopreserved Hypericum perforatum L. meristems

Meristems of in vitro-grown Hypericum perforatum L. plants were precultured for 3, 10, or 14 days in the presence of 0.5 M mannitol, or 0.076 µM or 0.76 µM abscisic acid, in RM basal liquid culture medium supplemented with 0.5 mg/l 6-benzylaminopurine and subsequently subjected to cryopreservation by the slow freezing method. The survival rate - determined as the percentage of meristems capable of differentiating plantlets - varied between 10% and 48%. Chromosome number stability of the cryopreserved meristems was determined by chromosome counting. The mitotic index of the control did not significantly differ from that of the treated samples.     

Genetic and biochemical analysis of Hypericum perforatum L. plants regenerated after cryopreservation

Shoot-tips from in vitro cultured Hypericum perforatum L. genotypes were subjected to assessments of developmental competence, genetic stability, and biosynthetic ability to identify critical points during cryopreservation. Survival rate, chromosome number stability, alteration in VNTR sequences and hypericin content were evaluated, in plants after pre-culture, and two subsequent cryogenic steps (cryoprotection and cooling) and those recovered from cryopreserved meristems. Pre-culture and cryoprotection treatments, did not reveal any significant differences, in these studied characteristics. Genetic stability was assessed by chromosome counts and analysis of variability in the VNTR sequences. No changes in chromosome number were detected in comparison with the untreated control but minor alterations were revealed in non-coding sequences. The content of hypericin after the recovery of cryopreserved meristems remained comparable with the unfrozen control. The controlled rate freezing technique used for cryopreservation was relevant for restoration of genetic and biochemical stability in Hypericum perforatum L. shoot-tips.     

Seasonal variation in hypericin content of Hypericum perforatum L. (St. John's Wort)

Hypericin and pseudohypericin, bioactive constituents in St. John's Wort (Hypericum perforatum), have been determined in the soft tops of the plant that are most likely to be browsed by foraging livestock. In two consecutive seasons, the hypericin/pseudohypericin concentration in a broad leaf biotype varied from a winter minimum of less than 100 ppm to a summer maximum approaching 3000 ppm. In contrast the narrow leaf biotype increased from similar winter values to summer maxima approaching 5000 ppm. The latter biotype was slower in returning to low levels of hypericin/pseudohypericin.     

贯叶连翘中金丝桃素的合成与积累研究进展

金丝桃素是贯叶连翘的主要药理活性成分。本文概述了金丝桃素的化学与生物合成途径,介绍了金丝桃素在贯叶连翘个体发育过程中的积累以及利用贯叶连翘的细胞和组织培养技术生产与积累金丝桃素的研究进展。最后,指出分子生物学和电子显微镜技术的发展为深入研究金丝桃素的产生和积累提供了有利工具。     

In vitro pollen germination in Hypericum perforatum L. and Hypericum rumeliacum Boiss

In vitro pollen germination and pollen tube growth investigations are valuable tools used in identification of the effects of environmental factors and genotypic differences on pollen viability, pollen germination and tube elongation. In this study pollen viability, in vitro pollen germination capacity, abnormality ratios and tube length in germinated pollens of Hypericum perforatum L. and H. rumeliacum Boiss. were investigated. Both of these species has spheroid-shaped and tricolporate pollen grains. The diameters of Hypericum perforatum and H. rumeliacum pollens were found as 24 +/- 3 microm and 19 +/- 2 microm, respectively. Pollen viability of H. perforatum and H. rumeliacum was found as 83% and 72%, respectively. The germination percentages were found as 12.85% for H. perforatum and 64.42% for H. rumeliacurm. Tube lengths in germinated pollens of both taxa were measured approximately as 95.25 +/- 38 microm in H. perforatum and 165.92 +/- 53 microm in H. rumeliacium 4 h after inoculation. In germinated pollen grains of H. perlbratum and H. rumeliacumn abnormality percentages were determined as 13.23% and 43.97%, respectively. In germinated pollens of these two species, highly significant (P < 0.00001) differences in in vitro germination percents and abnormality percents were observed. Abnormalities such as swollen tube tip, branched tube, spiralled tube and excessive tube formation were observed in pollen tubes. The results of this study showed that there were obvious differences in pollen germinability between these two species growing under the same environmental conditions.     

Root cryopreservation to biobank medicinal plants: a case study for Hypericum perforatum L.

In this study, an effective root-based cryopreservation method was developed for Hypericum perforatum L., an important medicinal species, using in vitro plants. A systematic approach was applied to determine effective combinations of protocol steps such as preculture, osmoprotection, vitrification solution treatment, and unloading, followed by protocol optimization using a single-factor approach. The effects of root section type (root tips, middle sections, or basal sections), duration of root section culture after excision, and donor plant age were also investigated. In a wild genotype, middle and basal root sections excised from 8-wk-old plants and cryopreserved at the age of 10 d after excision showed the highest plant regrowth after cryopreservation. In the optimized protocol, root sections were precultured in 10% (w/v) sucrose for 17 h, osmoprotected with a solution composed of 17.5% (w/v) glycerol and 17.5% (w/v) sucrose for 20 min, followed by a vitrification solution of 40% (w/v) glycerol and 40% (w/v) sucrose for 30 min, and cryopreserved using aluminum foil strips (droplet-vitrification). After rewarming in preheated 25% (w/v) sucrose solution and 30-min unloading, root segments were recovered on medium supplemented with 1.0 mg L⁻¹ gibberellic acid and showed 78% plant regrowth. This cryopreservation method was successfully adapted for five elite lines of H. perforatum with a 45 to 87% regrowth rate after cryopreservation. These results suggest that root cryopreservation may be an effective method for medicinal plant conservation and should be tested with a broader range of species.

     

激素对贯叶连翘器官分化的影响

贯叶连翘 (ＨｙｐｅｒｉｃｕｍｐｅｒｆｏｒａｔｕｍＬ .)为多年生草本 ,中国民间主要用于止血、抗炎、妇科病等[1] ,欧洲民间用于治疗创伤也有相当长的历史。近年来 ,欧、美等国家和地区将其应用于抑郁症的治疗 ,取得了很好的疗效。 80年代后期 ,由于发现该植物体内含有显著抗     

贯叶连翘野生转家化前后生物学性状特征观察

野生贯叶连翘 (HypericumperforatumL .)在江苏经过了 3a的引种栽培 ,其生物学特性检测结果表明 ,栽培贯叶连翘的生物量、开花的一致性和生长的适应性等明显优于野生贯叶连翘 ;良好的繁殖方式使贯叶连翘的收获期提前了 1a ;成熟后 3个月采收的种子萌发率较高 ,不同浓度赤霉素处理能明显提高其种子的萌发率 ;贯叶连翘的黑色腺体即分泌细胞球数量与金丝桃素间也基本存在着正相关关系 ,贵州贵阳居群和陕西丹凤居群黑色腺体明显多于其他居群 ,其金丝桃素含量在所有居群中为最高 ,而甘肃武都居群黑色腺体最少 ,其金丝桃素含量在 6个居群中最低。 6个野生居群栽培后生物量相近 ,在 4个不同土壤pH的基地大田中均能良好生长 ,说明江苏地区环境条件完全适合贯叶连翘引种栽培。     

Anti-influenza A virus effect of Hypericum perforatum L. extract

To study the antiviral effect of Hypericum perforatum L. extract (HPE) on influenza A virus (IAV) (H₁N₁) in vitro and in vivo. Cytopathic effect (CPE) and neutral red (NR) dye uptake were used to examine the antiviral effect of HPE on Madin Darby Canine Kidney (MDCK) cells which were infected with IAV in vitro. HPE was effective against influenza A virus (IAV) in vitro, with a 50% effective concentration (EC₅₀) of 40 μg/mL. The mean 50% cytotoxic concentration (CC₅₀) in the MDCK used in these experiments was 1.5 mg/mL. Ribavirin was run in parallel with EC₅₀ values of 5.0 μg/mL; the mean CC₅₀ for ribavirin was 520 μg/mL. Oral gavage administrations of HPE or ribavirin to mice infected with the IAV were highly effective in preventing death, slowing the decline of arterial oxygen saturation, inhibiting lung consolidation and reducing lung virus titers. The minimum effective dose of HPE in these studies was 31.25 mg/kg/day, which was administered twice daily for 5 d beginning 4 h prior to virus exposure. Below a dosage of 2000 mg/kg/day, almost all treated mice survived, which suggests that HPE is of low toxicity. Ribavirin’s minimum effective dose was 40 mg/kg/day with the LD50 determined to be 200 mg/kg/day. Delay of the initiation of either HPE or ribavirin therapy, using approximately 1/3 LD₅₀ dose each time, could still be protective as late as 48 h after exposure to the IAV. While both agents appeared to have similar efficacy against IAV infections, HPE was considered to be less toxic and may warrant further evaluation as a possible therapy for influenza. Foundation items: One Hundred Person Project of The Chinese Academy of Sciences (2008-287); The Project of Basic Scientific Research Fund for Central Public-Welfare of Institute of Sciences (BRF070402).     

贯叶连翘总提取物对谷氨酸棒状杆菌和粪产碱杆菌的抗菌作用

报道了经光照和未经光照处理后贯叶连翘总提取物对谷氨酸棒状杆菌和粪产碱杆菌的作用,结果表明,总提取物对谷氨酸状杆菌有强烈的抑菌和杀菌作用,对粪产碱杆菌有抑菌作用,其抑菌或杀菌作用与其浓度有关,且不需光照。     

Morphoregulatory effect of plant growth regulators on Hypericum perforatum L. Seedlings

The morphogenetic response of Hypericum perforatum seedlings to different auxin and cytokinin concentrations was studied. A stimulation of the concentration-dependent rooting ability was observed under the influence of indole-3-acetic acid and indole-3-butyric acid. Rooting was not enhanced by the effects of 2,4-dichlorophenoxyacetic acid and 1-naphtaleneacetic acid. Differentiated roots were isolated and cultured in liquid media with the same combination of growth-promoting auxins. Chromosome counts in root tip cells after long-term cultivation indicated a high degree of chromosomal instability. Multiple shoot formation occurred under the influence of 6-benzylaminopurine and kinetin. Adenine and 6-(γ,γ-dimethylallylamino)-purine did not stimulate shoot differentiation. No differences in the morphogenetic response to auxins and cytokinis were detected between diploid and tetrapoloid plants.     

茉莉酸甲酯(MeJA)对贯叶连翘悬浮细胞生长和贯叶金丝桃素产量的影响

培养基中添加100 μm01·L-1MeJA后,贯叶连翘悬浮细胞生物量增加量和贯叶金丝桃素(HF)的产量分别是未经MeJA处理的1.3倍和1.73倍;培养3 d时,培养基中添加100 μmol·L-1 MeJA能提高HF产量,是未经MeJA处理的2.4倍.     

Identification and genetic analysis of the APOSPORY locus in Hypericum perforatum L.

The introduction of apomixis – seed formation without fertilization – into crop plants is a long‐held goal of breeding research, since it would allow for the ready fixation of heterozygosity. The genetic basis of apomixis, whether of the aposporous or the diplosporous type, is still only poorly understood. Hypericum perforatum (St John’s wort), a plant with a small genome and a short generation time, can be aposporous and/or parthenogenetic, and so represents an interesting model dicot for apomixis research. Here we describe a genetic analysis which first defined and then isolated a locus (designated HAPPY for H ypericum AP OSP ORY ) associated with apospory. Amplified fragment length polymorphism (AFLP) profiling was used to generate a cleaved amplified polymorphic sequence (CAPS) marker for HAPPY which co‐segregated with apospory but not with parthenogenesis, showing that these two components of apomixis are independently controlled. Apospory was inherited as a dominant simplex gene at the tetraploid level. Part of the HAPPY sequence is homologous to the Arabidopsis thaliana gene ARI7 encoding the ring finger protein ARIADNE7. This protein is predicted to be involved in various regulatory processes, including ubiquitin‐mediated protein degradation. While the aposporous and sexual alleles of the HAPPY component HpARI were co‐expressed in many parts of the plant, the gene product of the apomict’s allele is truncated. Cloning HpARI represents the first step towards the full characterization of HAPPY and the elucidation of the molecular mechanisms underlying apomixis in H. perforatum.     

Reproduction pathway analysis of several Hypericum perforatum L. somaclonal families

Flow cytometry seed screen of mature seeds originating from several in vitro regenerated Hypericum perforatum L. somaclones and their seed progenies were used to screen the ways of reproduction of 4 subsequent generations of several somaclonal families and to search for the relation between the ploidy and prevalent mode of reproduction. The prevalent reproduction pathway of diploid plants was sexual reproduction. Seed samples of plants with higher ploidy levels showed an extensive variation in the mode of reproduction: BII and BIII hybrid formation and/or aposporous pseudogamy including parthenogenetic development of a reduced embryo sac.     

Toxic Effects of Pb on Anatomy and Hypericin Content in Hypericum perforatum L.

Lead (Pb) is one of the most common heavy metal contaminants in the environment. The present study was therefore undertaken to determine the effects of Pb on structural characteristics and hypericin production in Hypericum perforatum. Mature plants were treated with contaminated soil in seven treatments (75, 150, 300, 600, 800, 1000, and 1500 mg/kg Pb in soil) with three repeats per treatment every 14 days. Maximum observed Pb content in shoot parts was observed in the treatments with 600 and 1500 mg/kg Pb. The Pb concentration in roots was higher than in shoot parts, enhanced with increasing Pb concentration in the soil. In this study, Pb treatment significantly influenced the morphology, anatomy, and hypericin content in the plant. Anatomical characteristics of leaf, stem, and root affected by Pb contamination, as well as scanning electron microscopy (SEM) studies, revealed structural changes in stomata and epicuticular waxes. Under Pb toxicity, anatomical symptoms occurred in leaves, including increase in sizes of epidermal cells, mesophyll tissue, and diameter of stems and roots, as well as amplified vascular bundles and pith area. This, therefore, indicated that metal contamination can change the chemical composition of this plant. Maximum hypericin content was observed in the treatment containing 600 mg/kg Pb in soil, which then decreased.