Plant Hyoscyamus Research Articles

Conservation efforts for the endangered medicinal plant Hyoscyamus boveanus (Dun.) Aschers. & Schweinf. (Endemic to Egypt): An in vitro propagation protocol

Conservation efforts for the endangered medicinal plant Hyoscyamus boveanus (Dun.) Aschers. & Schweinf. (Endemic to Egypt): An in vitro propagation protocol

A phytopharmacological review on an important indigenous medicinal plant Hyoscyamus gallagheri

Since the ancient time, medicinal plants play a central role in the day-to-day life of humankind as crucial additive agents in foods, drinks and drugs and also as components in perfumes and cosmetics. The nutritional, pharmacological, biological and toxicological properties of the medicinal plants also have extensive applications. Globally, the majority of people nowadays rely on alternative and complementary medicines as safe sources for the treatment of various ailments obtained from natural sources. Plenty of plant and marine sources are available worldwide including the Sultanate of Oman with notable medicinal values. Hyoscyamus gallagheri ( H. gallagheri ), is one of the leafy plants grown in the desert areas of the Sultanate of Oman which belongs to the family of Solanaceae. Omanis have used the selected plant species traditionally to increase hair density, hair fall, dental pain, relieve snake bite, scorpion sting pain, motion sickness, asthma and cough. This plant is considered as a new species because still now there is no evidence on phytochemicals, pharmacological and toxicological studies available of the selected species. Previously, our group from Nizwa University has conducted two studies on the pharmacological and phytochemicals of the selected plant species and the results showed that the plant extracts contain different groups of phytochemicals, which possess notable biological activity. This present review is to summarize the updated scientific information available on the selected plant species grown in Oman. Therefore, the main objective of this current review is to compare and contrast the morphology, toxicity, pharmacological activity, phytochemicals, potential uses with other species belong to this family including the selected plant species of H. gallagheri . In conclusion, the raw plant materials, its crude extracts and isolated ingredients might be used as a novel therapeutic or herbal medicine to treat various ailments.

Extraction and fractionation of subproteome from root tips of Hyoscyamus albus

Identification and quantification of different metabolites under stress, especially protein, is a vital way to understand plant adaptation mechanism. We established an efficient protein extraction method from the tiny amount (100 mg) of root tips of non-model medicinal plant Hyoscyamus albus, using bead-beating cell disruption, TRIzol extraction, and sequential chemical protein solubilization. H. albus is very well known for biosynthesized of different secondary metabolites like hyoscyamine, tropane alkaloids and scopolamine. Our method is rational for sample preparation even in small-scale proteomics of recalcitrant tissue and allows proficient, reproducible and impurity-free protein extraction. This method allows high-quality 2DE in mini-gel format (25 µg of protein/gel) for hydrophilic and hydrophobic sub-proteomes and is compatible to high-sensitive matrix-assisted laser/desorption ionization quadrupole ion trap time-of-flight (MALDI-QIT-TOF) mass spectrometry (MS). A protocol using TRIzol is more effective and reproducible to sequential chemical extraction of both hydrophilic and hydrophobic membrane proteins. We also demonstrated cell disrupted together with dithiothreitol (DTT) and polyvinylpolypyrrolidone (PVPP) is more useful to prevent polymerization of the phenolic compound than commonly used added DTT and PVPP with TRIzol reagent. Despite the unavailability of genomic sequence database, the efficacy of the protocol was also confirmed by MS/MS ion searches.
 
 J Bangladesh Agril Univ 17(4): 430–436, 2019

Enhanced production of hyoscyamine and scopolamine from genetically transformed root culture of Hyoscyamus reticulatus L. elicited by iron oxide nanoparticles

The medicinal plant Hyoscyamus reticulatus L. is a rich source of hyoscyamine and scopolamine, the tropane alkaloids. The use of hairy root cultures has focused significant attention on production of important metabolites such as stable tropane alkaloid production. Elicitation is an effective approach to induce secondary metabolite biosynthetic pathways. Hairy roots were derived from cotyledon explants inoculated with Agrobacterium rhizogenes and elicited by iron oxide nanoparticles (FeNPs) at different concentrations (0, 450, 900, 1800, and 3600 mg L−1) for different exposure times (24, 48, and 72 h). The highest hairy root fresh and dry weights were found in the medium supplemented with 900 mg L−1 FeNPs. Antioxidant enzyme activity was significantly increased in induced hairy roots compared to non-transgenic roots. The highest hyoscyamine and scopolamine production (about fivefold increase over the control) was achieved with 900 and 450 mg L−1 FeNPs at 24 and 48 h of exposure time, respectively. This is the first report of the effect of FeNP elicitor on hairy root cultures of a medicinal plant. We suggest that FeNPs could be an effective elicitor in hairy root cultures in order to increase tropane alkaloid production.

The effect of inductive photoperiod on flower formation and phytohormones level in a long day plant Hyoscyamus niger L.

The anatomical and hormonal investigations on a long day plant <i>Hyoscyamus niger</i> L. during the time of the generative photoinduction have been conducted. The plants were grown during 75 days on a short photoperiod and then they were transferred to long day conditions. The earliest anatomical symptoms of flower initiation were noticed after four long photoperiods. The inductive photoperiod causes also a general increase in the amounts of phytohormones. During the flower evocation the intensive accumulation of cytokinins and gibberellins in leaves takes place. The post-inductive period, in which the development of flower elements happens, is characterized by changing amount of phytohormones. The content of hormonal substances is subjected to the rhytmical changes related to the periods of light and darknees in the twenty four hours' cycle.

Possible Application of the Medicinal Plant Hyoscyamus albus in Phytoremediation: Excess Copper Compensates for Iron Deficiency, Depending on the Light Conditions

Seedlings of the medicinal plant Hyoscyamus albus were supplied with an excess of Cu to examine the possible application in phytoremediation. The seedlings were cultured in B5 medium supplied with basal 0.1 μM Cu and 200 μM Cu under various light conditions: short day (SD); long day (LD); and continuous light (CL). In addition, the effect of supplying 200 μM Cu under Fe deficiency was determined, in order to elucidate the interaction between Cu and Fe. Interestingly, Fe-deficiency symptoms that developed in plants grown with basic levels of Cu under LD almost disappeared when excess Cu was supplied. Plant growth mainly depended on the photo irradiation period (SD < LD~CL); and 200 μM Cu did not inhibit growth at all when Fe was available, whereas in the absence of Fe, CL caused damage to growth. Analysis of the Cu and Fe contents of the plants revealed that Cu was distributed equally in both the aerial parts and roots, whereas most of the Fe was found in the roots; under Fe deficiency, Cu accumulation in the roots apparently increased. Cu was mainly distributed in the soluble fraction, which included vacuoles and the cell-wall fraction. These results provide evidence indicating that H. albus seedlings are tolerant of Cu present in excess. Furthermore, excess Cu was able to compensate for Fe deficiency, depending on the light conditions. Continuous light inhibited this effect, probably as a result of the induction of Mn deficiency. The possible applications of this newly discovered cuprophyte are discussed.

Chromosomal localization of rDNA and DAPI bands in solanaceous medicinal plant Hyoscyamus niger L.

comprises of medicinally impor-tant species rich in tropane alkaloids. Its cultivated specieshave been subjected to genetic enhancement through chro-mosomal manipulation. Linear differentiation of chromo-somes is quite important to facilitate chromosome iden-tification, and inter-chromosomal and intra-chromosomalalterations for complementing cytogenetic analysis. There-fore, two-colour simultaneous fluorescence

Antifungal activities of hyoscyamine and scopolamine against two major rice pathogens: Magnaporthe oryzae and Rhizoctonia solani

The antifungal activities of hyoscyamine and scopolamine, major alkaloids extracted from the desert plant Hyoscyamus muticus, against two rice pathogens, Magnaporthe oryzae and Rhizoctonia solani, were studied. The minimum inhibitory concentration of hyoscyamine that resulted in distinctive inhibition (MIC50) was 1 μg/ml for both fungi. Exposure to hyoscyamine caused the leakage of electrolytes from the mycelia of both fungi. Hyoscyamine (>1 μg/ml) irreversibly delayed or inhibited conidial germination and appressorium formation in M. oryzae grown on polystyrene plates. Hyoscyamine effectively inhibited the attachment of conidia to the surface of rice (Oryza sativa) leaves and inhibited appressorium formation on the leaves. A high concentration of scopolamine (1000 μg/ml) also delayed or inhibited conidial germination in M. oryzae, but conidial germination was restored after washing the conidia with water. Antifungal activity of hyoscyamine was reduced by scopolamine. Magnaporthe oryzae infection was significantly suppressed (by >95%) in leaves of intact rice plants treated with hyoscyamine (10 μg/ml). Moreover, 10 μg hyoscyamine/ml significantly reduced the disease severity index for sheath blight to ≤0.2, when compared with the disease index of control plants (>7.0). Hyoscyamine (>20 μg/ml) completely inhibited sclerotial germination and development of R. solani by delaying the initiation, maturation, and melanization of the sclerotia. These results suggest that tropane alkaloids may be useful for controlling blast and sheath blight diseases of rice and for studying the mechanisms that regulate conidial germination in M. oryzae and sclerotial germination and development in R. solani.

Four Novel Ustilaginomycetous Anamorphic Yeast Species Isolated as Endophytes from the Medicinal Plant Hyoscyamus muticus

Four Novel Ustilaginomycetous Anamorphic Yeast Species Isolated as Endophytes from the Medicinal Plant Hyoscyamus muticus

Mechanistic Insights into the Cytochrome P450‐Mediated Oxidation and Rearrangement of Littorine in Tropane Alkaloid Biosynthesis

During the biosynthesis of certain tropane alkaloids, littorine (1) is rearranged to hyoscyamine (3). Recent evidence indicates that this isomerisation is a two-step process in which the first step is an oxidation/rearrangement to give hyoscyamine aldehyde (2). This step is catalysed by CYP80F1, a cytochrome P450 enzyme, which was recently identified from the plant Hyoscyamus niger; CYP80F1 also catalyses the hydroxylation of littorine at the 3'-position. The mechanisms of the reactions catalysed by CYP80F1 were probed with synthetic deutero and arylfluoro analogues of 1. Measurement of the primary kinetic isotope effects indicates that C3' hydrogen abstraction is the rate-limiting step for the oxidation/rearrangement of natural littorine, and for the 3'-hydroxylation reaction of the unnatural S enantiomer of littorine. The character of the intermediates in the oxidation/rearrangement and hydroxylation reaction was probed with the use of arylfluorinated analogues of (R)-littorine (natural stereoisomer) and (S)-littorine (unnatural stereoisomer) as substrates for CYP80F1. The relative conversions of ortho-, meta- and para-fluorolittorine analogues were used to obtain information on the likely intermediacy of either a benzylic radical or benzylic carbocation intermediate. The data suggest that hydroxylation takes place via a benzylic carbocation intermediate, whereas the product profile arising from rearrangement is more consistent with a benzylic radical intermediate.

Responses of fungi to tropane alkaloids produced by a medicinal plant Hyoscyamus muticus (Egyptian Henbane)

Antifungal activity of hyoscyamine (Hcy) and scopolamine (Sco) were determined by TLC-bioautography against fungi associated with H. muticus grown in Egypt, and those isolated from other plants grown in Japan. All 40 fungal strains were tolerant to Sco and sensitive to Hcy, exhibiting a growth inhibition zone around the Hcy spot on the bioautography plate. The strains were grouped into three types based on the appearance of the inhibition zone: (i) 17 strains exhibiting a clear inhibition zone, which remained clear at 8 d after incubation (type I); (ii) 22 strains exhibiting the inhibition zone with a brown circle surrounding the zone and regrowth within the inhibition zone (type II); (iii) 1 strain exhibiting the inhibition zone with no brown circle and regrowth within the inhibition zone (type III). In the type II and III strains, Hcy disappeared, and other alkaloids were found in the inhibition zones in its place. Hcy feeding experiments using Penicillium purpurogenum (type II) and Cunninghamella elegans (type III) revealed that these fungi may convert Hcy to a new alkaloid compound.

Transformation of the tropane alkaloid-producing medicinal plant Hyoscyamus muticus by particle bombardment.

We report an efficient whole plant transformation system for Hyoscyamus muticus, an important medicinal plant of the Solanaceous family. We developed a system using a plasmid carrying the nptII and gusA genes, which was delivered into leaf explants by particle bombardment. Ten percent of bombarded leaf explants formed kanamycin-resistant callus, from which putative transgenic plants were recovered. The nptII gene conferring kanamycin resistance was found to be incorporated into the genome of all transgenic plants screened. Over 50% of the kanamycin resistant plants showed strong expression of the non-selected gusA gene. The majority of transgenic plants reached maturity, could be self pollinated, and produced fertile seed. A simple and efficient whole plant transformation system for this medicinal plant is an important step in furthering our understanding of tropane alkaloid production in plants.

Genetic variation in meiotic behaviour and fertility in tetraploid Hyoscyamus muticus: correlation with diploid meiosis

Autotetraploidy was induced in 19 morphologically distinct genotypes of the medicinal solanaceous plant Hyoscyamus muticus (Egyptian henbane, 4x= 56). The multivalent association frequencies at metaphase I of meiosis ranged from 0.328 to 0.628 and were positively correlated with the chiasmate association frequency in the diploid progenitors. The frequent absence of chiasmata in one of the two chromosome arms in most of the tetraploids was attributed to interference between the arms and to pairing partner exchange in one of the chromosome arms instead of around the centromers. The bivalent frequency in the tetraploids was positively correlated with pollen fertility and seed set. The latter ranged from 7.5 to 68.5 per cent. The asymmetry of the karyotype was not reflected in the distribution of open bivalents in the diploid. The regression coefficients of metaphase I chromosome associations over fertility in the tetraploid suggested that, in the range covered, one multivalent less resulted in an increase of about 12 per cent in seed set. The regression of open bivalent frequency in the diploid progenitor over seed set in the tetraploid suggested that one open bivalent more in the diploid would result in almost 18 per cent seed set in the tetraploid. It is concluded that the selection of diploid progenitors with high frequencies of open bivalents should enhance the fertility of the artificial polyploids in H. muticus.

Meiotic Regularization, Restoration of Seed Fertility and Alkaloid Content in the Induced Autotetraploids of Hyoscyamus albus L.*

AbstractColchicine‐induced autotetraploids produced in the medicinally important solanaceous plant Hyoscyamus albus (2n = 68) revealed a general increase in pollen, stomata and seed size, but resulted in the reduction of leaf size and plant height. Also, an initial performance trial showed a decline in biomass yield but an increase in alkaloid content of 16.2 %. Thus, in terms of alkaloid yield, the tetraploids demonstrated a slight improvement over diploids for this economic product.Seed fertility of the raw colchiploids was low but a higher value (&gt; 80 %) could be obtained in advanced generations following rigid selection. High fertility was accompanied by a diploidized meiotic chromosome behaviour in the autotetraploids, with high bivalent pairing and regular anaphase separation. Bivalents were predominantly of open type in the autotetraploids as against a predominance of ring bivalents with distal chiasma localization in the parental diploids. The algebraic estimate of multivalent pairing frequency recorded even in the raw colchiploids was quite low (0.239) compared to that expected on the basis of random pairing (0.667), which further dropped to 0.107 in the C3 generation. Such reduction in multivalent frequency in autotetraploids could be brought about by a combination of reduction in crossover frequency and correction in pachytene pairing.

High bivalent frequencies in artificial autopolyploids of Hyoscyamus muticus L.

High pollen and seed fertility of induced autotetraploids of the medicinal solanaceous plant Hyoscyamus muticus (4x = 56) could, at least in part, be attributed to low quadrivalent and high bivalent frequencies at metaphase I (MI). Mean configuration frequencies per cell were 3.45 ring quadrivalents, 0.49 chain quadrivalents, 0.55 trivalents, 5.98 ring bivalents, 12.67 open (rod) bivalents, and 1.34 univalents. In the diploid, ring bivalents were most common with chiasmata predominantly distal at diakinesis–MI. Different causes can explain the low quadrivalent frequencies in the tetraploids. Using a simple algebraic model, calculations show that the most probable explanation is reduced chiasma formation in one arm per chromosome, resulting from nonmedian partner exchange and special interference patterns. This agrees best quantitatively with high open (rod) bivalent frequencies compared with the diploid, accompanied by high ring quadrivalent frequencies.Key words: Hyoscyamus, autotetraploid, seed fertility, meiotic mechanism, pairing.

Changes in the composition of in vitro translated leaf m‐RNA caused by photoperiodic flower induction ofHyoscyamus niger

Flowering of the long day plantHyoscyamus nigerL., which is strictly photoperiodically controlled, was induced by 58 h continuous white light. The RNA from the leaves was isolated from photoperiodically induced and non‐induced plants and the poly(A)‐rich RNA separated by affinity chromatography on oligo‐dT‐cellulose. The poly(A)‐rich RNA was translated in vitro in the presence of35S‐methionine using a rabbit reticulocyte lysate. Subsequent separation of the translation products by two‐dimensional polyacrylamide gel electrophoresis and fluorography allowed a comparison of the polypeptide pattern from induced and non‐induced leaf m‐RNA. The results indicate that induction of flowering is reflected by changes in the translation of several leaf polypeptides. These polypeptides were characterized by their isoelectric points and molecular masses.

A Quantitative and Cumulative Response to Photoperiodic Induction of Hyoscyamus niger, a Qualitative Long Day Plant

The photoperiodic induction of the long day plant Hyoscyamus niger has been investigated using white light and light of restricted spectral regions. Illumination with continuous white light revealed a quantitatively augmented response with both prolonged duration and increased irradiance in contrast to the otherwise qualitative photoperiodic nature of Hyoscyamus. From the three light qualities blue, far-red and red only the last was capable of inducing flowering in night breaks. Though the features of a phytochrome mechanism are apparent, complete reversibility and saturation, however, were lacking and reciprocity between duration and irradiance did not hold. Thus the long day plant reacts in a mode different from short day plants. In analogy with continuous light, a quantitative and cumulative response to subinductive pulses was found. Photosynthesis may participate in the night break response, as being tested by the incorporation of 14CO2 and CO2-free atmosphere.

Die wirkung von phytohormonen und von salicylsäure auf die blütenbildung und sproßverlängerung bei der langtagpflanze Hyoscyamus niger

The phytohormones auxin, kinetin and gibberellin A3 as well as salicylic acid have been infiltrated in various concentrations into the leaves of the qualitative long day plant Hyoscyamus niger. Only gibberellin induced flowering and stem elongation under non-inductive photoperiodic conditions. None of the substances had an inhibitory effect on photoperiodically pre-induced plants. While gibberellin of appropriate concentrations was able to induce stem elongation in all plants and increased shoot lengthening in a concentration-dependent manner, flowering was only induced up to a rate of 60 °/o. The finding that flowering without stem elongation can be induced even at low concentrations indicates that gibberel-lin-mediated flowering is independent of and not obligatorily caused by stem elongation.

Investigations on the mechanism of floral induction of the long day plant Hyoscyamus niger L. with cycloheximide

Previous investigations have shown that cycloheximide (CHI), an inhibitor of protein synthesis, strongly inhibited induction of flowering when applied to the apex of Hyoscyamus niger (H.n.), but after infi;tra into the leaves way induce flowering under short day conditions. Labelling of leaves with 14CO2 or [14C]leucine revealed that CHI inhibited protein synthesis measured as incorporation of 14C-radioactivity into TCA-insoluble material only temporarily and incompletely, but reduced the uptake of [14C]leucine to a greater 6 h after infiltration with CHI, when only 13% of [14C]leucine was incorporated compared with control plants infiltrated with water. The inhibition by CHI decreasing during the subsequent period of 72 h to the value of controls. Transport of gibberellic acid (GA3) from the leaves to the apex was also inhibited, as was demonstrated by the finding that CHI also reduced apparent stem elongation produced by infiltration of GA3 into the leaves. The results are interpreted to mean that CHI acts in flower induction by transitionally inhibiting the transport of the floral stimulus from the leaves to the apex.

Ausl�sung der Bl�tenbildung bei der Langtagpflanze Hyoscyamus niger im Kurztag durch 2-Thiouracil

Aqueous solutions of 2-thiouracil (TU) were applied selectively either to the growing point or to the leaves of the long-day plant Hyoscyamus niger in order to determine whether this antimetabolite has an effect on the synthesis of the floral stimulus in the leaves. Applications to the growing point were made by means of a small glass tube covering the shoot apex; application to the leaves was performed by vacuum infiltration. In all experiments all leaves except the three youngest fully expanded leaves and the 8-10 youngest primordia were removed before application. Plants were recorded as having initiated flowers when flower primordia were visible under a dissection microscope 5 weeks after the experiment.TU was inhibitory to photoperiodic induction by long-days of 16 hours when applied to the growing point during the second 8 hours of the daily photoperiod. A concentration of 5·10(-3) M of TU fully suppressed flowering without significant inhibition of leaf primordia increment; however, leaves developing from treated primordia had reduced leaf blades. These results are in agreement with findings already published by other investigators.However, when the leaves were infiltrated by TU, the antimetabolite did not inhibit photoperiodic induction but on the contrary initiated flowering even under short-day conditions. This effect was investigated in more detail by repeated daily infiltrations of TU-solutions in concentrations of 10(-5)-10(-2) M during the second part of an 8 hour photoperiod up to 5 following days. Even after one single infiltration of a 10(-4) M solution 18% of the treated plants were flowering; the percentage of flowering plants increased with increasing concentrations of TU and number of days of application up to approximately 80%. In no case was a flower initiation of 100% obtained. Leaves developing from primordia after infiltration of the leaves with TU have reduced and deformed leaf blades, indicating that TU is transported to the shoot apex to some extent.Some possible explanations of this inductive effect of TU were tested experimentally. Oxygen uptake of the leaves was not decreased and the respiratory quotient was not affected by TU. Photoperiodic induction is not stimulated by low concentrations of TU when applied to the growing point. Infiltration of the leaves by solutions of 2,4-dinitrophenol (10(-4) M) and sodium azide (10(-3) M) had no inductive effect under short-day conditions; a single complete defoliation (except for the 8-10 youngest primordia) is also not inductive. Under short-day conditions additional leaves remaining on the plant that were not infiltrated by TU decreased the percentage of flowering plants but did not fully suppress flower initiation.From these results it is concluded that TU does not act by inhibition of particular metabolic processes concerned in flower initiation or by inhibition of the synthesis of an inhibitor. We suggest that application of TU may lead to synthesis of a floral stimulus in the leaves under short-day conditions also.