Hairy Roots of Scutellaria spp. (Lamiaceae) as Promising Producers of Antiviral Flavones.

We measured and studied the growth parameters and the qualitative and quantitative composition of the flavones of hairy roots of the Scutellaria genus: S. lateriflora, S. przewalskii and S. pycnoclada. Hairy roots were obtained using wild-type Agrobacterium rhizogenes A4 by co-cultivation of explants (cotyledons) in a suspension of Agrobacterium. The presence of the rol-genes was confirmed by PCR analysis. The hairy roots of the most studied plant from the Scutellaria genus, S. baicalensis, were obtained earlier and used as a reference sample. HPLC-MS showed the predominance of four main flavones (baicalin, baicalein, wogonin and wogonoside) in the methanol extracts of the studied hairy roots. In addition to the four main flavones, the other substances which are typical to the aerial part of plants were found in all the extracts: apigenin, apigetrin, scutellarin and chrysin-7-O-ß-d-glucuronide. According to the total content of flavones, the hairy roots of the studied skullcaps form the following series: S. przewalskii (33 mg/g dry weight) > S. baicalensis (17.04 mg/g dry weight) > S. pycnoclada (12.9 mg/g dry weight) > S. lateriflora (4.57 mg/g dry weight). Therefore, the most promising producer of anti-coronavirus flavones is S. przewalskii.

A novel study approach on Scutellaria altissima L. cultivated at the Ghirardi Botanic Garden (Lombardy, Italy).

Within an Open Science project, research was carried out to describe to the public of the Ghirardi Botanic Garden (BS, Lombardy, Italy) the invisible features of plants. This work is dedicated to Scutellaria altissima L. (Lamiaceae). Micromorphological, histochemical and phytochemical investigations were conducted on the vegetative and reproductive organs to correlate the structures involved in the emission of substances and their unique productivity. This work reports volatile organic compound (VOC) profiles of leaves and flowers and the composition of essential oil (EO) obtained from aerial parts of plants cultivated in Italy that have never been described before. Three morphotypes of glandular trichomes were observed: peltate, short-stalked capitate and long-stalked capitate. Peltate trichomes were the main producers of terpenes, short-stalked capitates of polysaccharides and long-stalked capitates of terpenes and polyphenols. The leaf VOC profile showed heterogeneous composition, with non-terpene derivatives as the major chemical class (71.04%), while monoterpene hydrocarbons represented almost the totality of the flower (99.73%). The leaf presented a higher number of total (37 versus 11) and exclusive (33 versus 7) compounds. (Z)-3-Hexenol acetate was most abundant in the leaf and (E)-ß-ocimene in the flower. Four common compounds were detected: ß-pinene, ß-caryophyllene, γ-muurolene and germacrene-D. The EO contaied 21 compounds, dominated by ß-caryophyllene, linalool and hexahydrofarnesyl acetone. This research allowed us to correlate morphotypes of the secretory structures with the production of secondary metabolites, with the aim of providing the public of the Ghirardi Botanic Garden with a dedicated iconographic approach, which accounts for olfactory perception linked to S. altissima.

An Untapped Resource in the Spotlight of Medicinal Biotechnology: The Genus Scutellaria.

BACKGROUND: This review is intended to draw the attention of pharmaceutical and biotechnological communities to the untapped potential of the Scutellaria genus. Skullcaps, as they are more widely known, are found in one of the oldest materia medica in the world, that of ancient Chinese pharmacology, and their numerous wide range of medicinal bioactivities have been studied both in vivo and in vitro. For thousands of years, chemical compounds from the Scutellaria species have been safely used as antitumor, antibacterial, antiviral, anti-inflammatory, antioxidant or hepatoprotective factors. OBJECTIVE: As these effects are well known, reflected in the presence of Scutellaria plants in national pharmacopoeias, it is clear that the plant has yet enormous unexploited potential. The European pharmacological market has turned to the resources of Scutellaria only in the last two decades, and although the construction and clinical processing of a new drug is a long process, the general impression is that very few medical products in pharmacies have been inspired by the phytochemistry of skullcaps. CONCLUSION: This paper presents the current state of knowledge on the wealth of Scutellaria chemical compounds with treatment applications, its tissue culture and biotechnological achievements, especially in the context of the production of secondary metabolites.

Floral development in Scutellaria pinnatifida (Lamiaceae): the ontogenetic basis for sepal reduction.

The Scutellaria is a Labiatae genus (subfamily Scutellarioideae) with a highly specialised floral structure. The genus is characterised by a peculiar two-lobed calyx with a projecting appendage, named the scutellum. Here, we present a detailed analysis of floral development, using epi-illumination light microscopy, to clarify open questions about its floral organisation. Floral whorls appeared in an acropetal sequence, with a marked temporal overlap of petal and stamen appearance. Organ appearance in each whorl proceeded unidirectionally from the abaxial to the adaxial side. Significant developmental features included the formation of the scutellum, reduction of sepal lobes and formation of a three-lobed nectary disc. Our study revealed that both loss of organ initiation and fusion of primordia are responsible for the reduction in sepal members in Scutellaria. The nectary structure was markedly different from most other studied Lamiaceae.

[Estimating medicinal yield of Seutellaria baicalensis in Beijing-Tianjin-Hebei region based on 3S technology].

Resources survey of traditional Chinese medicine and reserves estimation are found to be the most important issues for the protection and utilization of traditional Chinese medicine resources, this paper used multi-spatial resolution remote sensing images (RS) , geographic information systems (GIS) and global positioning system (GPS) , to establish Scutellaria resources survey of 3S data platform. Combined with the traditional field survey methods, small-scale habitat types were established based on different skullcap reserve estimation model, which can estimate reserves of the wild Scutellaria in Beijing-Tianjin-Hebei region and improve the estimation accuracy. It can provide an important parameter for the fourth national survey of traditional Chinese medicine resources and traditional Chinese medicine reserves estimates based on 3S technology by multiple spatial scales model.

Diversification, biogeographic pattern, and demographic history of Taiwanese Scutellaria species inferred from nuclear and chloroplast DNA.

The ragged topography created by orogenesis generates diversified habitats for plants in Taiwan. In addition to colonization from nearby mainland China, high species diversity and endemism of plants is also present in Taiwan. Five of the seven Scutellaria species (Lamiaceae) in Taiwan, for example, are endemic to the island. Hypotheses of multiple sources or in situ radiation have arisen to explain the high endemism of Taiwanese species. In this study, phylogenetic analyses using both nuclear and chloroplast markers revealed the multiple sources of Taiwanese Scutellaria species and confirmed the rapid and recent speciation of endemic species, especially those of the "indica group" composed of S. indica, S. austrotaiwanensis, S. tashiroi, and S. playfairii. The common ancestors of the indica group colonized first in northern Taiwan and dispersed regionally southward and eastward. Climate changes during glacial/interglacial cycles led to gradual colonization and variance events in the ancestors of these species, resulting in the present distribution and genetic differentiation of extant populations. Population decline was also detected in S. indica, which might reflect a bottleneck effect from the glacials. In contrast, the recently speciated endemic members of the indica group have not had enough time to accumulate much genetic variation and are thus genetically insensitive to demographic fluctuations, but the extant lineages were spatially expanded in the coalescent process. This study integrated phylogenetic and population genetic analyses to illustrate the evolutionary history of Taiwanese Scutellaria of high endemism and may be indicative of the diversification mechanism of plants on continental islands.

Enhanced accumulation of secondary metabolites in hairy root cultures of Scutellaria lateriflora following elicitation.

Hairy root cultures of Scutellaria lateriflora were established using Agrobacterium rhizogenes A4 and produced acteoside (18.5 mg g(-1) dry wt), baicalin (14.5 mg g(-1) dry wt) and wogonoside (12 mg g(-1) dry wt). Yeast extract (50 µg ml(-1)) increased acteoside production 1.4-fold and flavone production 1.7-fold after 7 and 14 days of elicitation. Addition of Pectobacterium carotovorum lysate in the stationary phase of the hairy root culture stimulated only the accumulation of wogonin to 30 mg g(-1) dry wt. The production of wogonin in hairy roots could be associated with its role as a phytolaexin.

Development of the chromatographic fingerprint of Scutellaria barbata D. Don by GC-MS combined with Chemometrics methods.

Gas chromatography fingerprint of Scutellaria barbata D. Don (SB) from different origins was studied by gas chromatography-mass spectrometry (GC-MS) and related Chemometrics methods. The constituents of essential oil of Scutellaria barbata D. Don (SB) and its two adulterants Oldenlandia diffusa (OD) and Lobelia chinensis Lour (LCL) were analyzed and compared, 50, 36 and 38 components were identified from SB, OD and LCL, respectively, there were 16 and 18 common components between SB and OD, SB and LCL. Nine different samples collected from different producing areas of SB were studied. The relative standard deviations (RSDs) of retention time and peak area of each component were less than 0.2% and 6%, respectively. The number of common peaks was up to 52 and the un-common peak area was less than 10%. The similarity and difference among SB and its adulterants were also evaluated by correlation coefficient similarity analysis and principal component analysis, the result showed that developed fingerprint characterize the SB from different producing areas and it is useful and feasible for the discrimination of its adulterants.

Protocols for establishment of an in vitro collection of medicinal plants in the genus Scutellaria.

The study of medicinal plants has many unique challenges and special considerations. These plants are studied for their specific chemistry, or pharmacologic activity. Plants are highly sensitive to their environment and respond through changes in their chemistry. To date, one of the biggest problems in the study of medicinal plants has been the acquisition of consistent, positively identified material for chemical analysis. Successful protocols for the collection, identification, and establishment of medicinal plants species in tissue culture is invaluable for future studies. This protocol outlines methods to establish Scutellaria baicalenisis, and Scutellaria lateriflora from commercial seed sources, and collection and establishment of Scutellaria racemosa from wild populations.

[Induction and culture of hairy roots in Scutellaria viscidula and its baicalin production].

OBJECTIVE: To establish a culturing system of hairy roots of Scutellaria viscidula, and study the hairy roots growth and biosynthesis of flavonoid in the culturing system. METHOD: Hairy roots of S. viscidula were obtained from infected stem explants after infection with the disarmed Agrobacterium tumefaciens strain C58C1, elite strains were screened and growth curves were determined. The transformation of Ri T-DNA was examined through PCR and baicalin content was examined through HPLC. RESULT: Hairy roots appeared in vitro 8 days after inoculation of S. viscidula with disarmed A. tumefaciens strain C58C1. After 24 days the frequency of stems explants was up to 81%. Transformation was confirmed by the amplification of rolC genes from the hairy roots of S. viscidula. The results demonstrated that rolC genes could be expressed in hairy roots of S. viscidula. Under the 36 days suspension culture of S. viscidula hairy roots in 1/2MS medium, dry weight of hairy root increased 17.42 times, the content and baicalin increased 21.60 and 25.56 times. CONCLUSION: The establishment of the culturing system of hairy root of S. viscidula provided a foundation for further industrial production of active drug component.

[Study on rapid reproduction of Scutellaria amoena].

Tissue cultures of Scutellaria amoena were established from the young stem explants on MS + IAA 0.1 mg/L + BA 0. 5 (0.2) mg/L. Propagation cultures were established on MS + BA 0.2 (1.5) mg/L + IAA 0.1 mg/L + NAA 0.1 mg/L + 2, 4-D 0.5 (1.5) mg/L. Rooting of planlet was established on 1/2 MS + NAA 0.1 (0.2) mg/L + IBA 0.1 (0.2) mg/L + PP333 1.0 mg/L and the rate can reach 100%.

Oil formulation of entomopathogenic fungus, Beauveria bassian, against Sunn pest, Eurygaster integriceps puton (Heteroptera: Scutelleridae).

Entomopathogenic fungi with approximately 1000 known species are key regulatory factors in insect pest population. Also, these agents infect insects by direct penetration of the cuticle and thus provide the only practical means of microbial control of insects that feed on plant or animal juices as well as for the many coleopterans and orthopteran pest which have very few known viral or bacterial diseases. Beauveria bassiana is a widespread entomopathogenic fungi that infect many insect species. The aim of the current study was to determine the effect of different composition of oil in pathogenicity of the fungus. So, the fungus was grown on Potato dextrose agar (PDA) at 25 degrees C. Conidia were harvested from 14-day sporulating cultures by scraping the surface with a spatula and suspending the conidia in sterile 0.03% aqueous Tween 80. The number of conidia was determined using an improved Neubauer haemocytometer and spore concentrations of 10(4), 10(5), 10(6), 10(7), 108 and control were prepared. Two controls were done one was different concentrations of spore without oil and the other only oil. The insects were collected from the wheat farm in and maintained on wheat plants in the laboratory at 27+/-2 degrees C under a 14 h light: 10 h dark (LD 14:10) photoperiod. Assays were done with dipping methods. The results showed that there are significant differences in mortality of insects in two spore emulsions, one oil formulated and the other spores in sterile water without oil usage. Oil formulation enhances fungal virulence toward insect. Since insect cuticle especially epicuticle (lipid layer) is the primary site of establishment of mycosis, oil formulation increase the adhesion of spore to the insect cuticle through hydrophobic interaction between the spore and cuticle surface.

[Study on yield and quality of Scutellaria baicalensis from different habitats].

OBJECTIVE: To compare the economical characters, yield characters and major effective ingredient content of Scutellaria baicalensis from different habitats. METHOD: The characters were studied by field randomized block test combining analysis of variance, correlation analysis and quality analysis. RESULT: There are significant differences in yield and quality of the crude drugs from different habitats. CONCLUSION: The crude drug from Pingyi (Shandong province) is better than those from other habitats.