Biogeochemical Features of Platinum Accumulation in <Emphasis Type="Italic">Scutellaria baicalensis</Emphasis> Georgi (Lamiaceae)

For the first time, the platinum accumulation levels and distribution patterns in the soil and in roots, stems, leaves, flowers, and seeds of medicinal herb Scutellaria baicalensis Georgi, Lamiaceae (Baikal skullcap) have been researched using the stripping voltammetry method. The plants were collected both in their natural habitats (southwestern part of Primorskii krai, Amur oblast, and Chita oblast) and from the S. baicalensis population introduced in the Central Siberian Botanical Garden (CSBG), Siberian Branch, Russian Academy of Sciences (Novosibirsk). In the habitats of S. baicalensis, the content of platinum in soils varies from 0.001 to 0.426 g/t. The content of platinum in various parts of the plants varies from 0.001 to 0.43 g/t. The platinum bioaccumulation factor in S. baicalensis individuals collected in various habitats varies from 0.01 to 6.1 g/t.     

Pharmacognostic study of aerial parts of Baikal skullcap (<Emphasis Type="Italic">Scutellaria baicalensis</Emphasis> Georgi)

A pharmacognostic study of aerial parts of Scutellaria baicalensis Georgi (Lamiaceae family) has been carried out. A merchandising analysis has been performed to determine the basic quality parameters of S. baicalensis. A chemical composition analysis has revealed the presence of organic acids, free carbohydrates, polysaccharides, amino acids, alkaloids, flavonoids, phenolic acids, anthocyanins, tannins, photosynthetic pigments, lipids, and triterpene compounds. Ethyl acetate and ethanol fractions, extracted from the aerial parts of S. baikalensis, have shown a significant antiradical activity.     

<Emphasis Type="Italic">VpUR9</Emphasis>, a novel RING-type ubiquitin ligase gene from <Emphasis Type="Italic">Vitis pseudoreticulata</Emphasis>, is involved in powdery mildew response in transgenic <Emphasis Type="Italic">V. vinifera</Emphasis> plants

Ubiquitination plays important roles in disease resistance in plants. We report the identification and functional characterization of the RING-type ubiquitin ligase gene VpUR9 from Chinese wild Vitis pseudoreticulata accession Baihe-35-1. VpUR9, encodes 164 amino acids and possesses a RING conserved motif. It is homologously cloned from the cDNA library of the high powdery mildew (Erysiphe necator [Schw.] Burr) resistant V. pseudoreticulata accession Baihe-35-1 inoculated with E. necator. The gene is induced in response to powdery mildew and salicylic acid. VpUR9 fused with FLAG-tag controlled by 35S promoter was transformed into 15 regenerated V. vinifera L. cv. Red Globe lines via Agrobacterium tumefaciens-mediated transformation. Twelve of these lines were confirmed by Western blot of FLAG-tag. As a result, the powdery mildew-resistance of Red Globe transformed with VpUR9 was repressed. Furthermore, the expression of some disease-resistant related genes (NPR1, PR1, PR10 and PAL) of the transgenic Red Globe declined compared with wild type grapes when inoculated with powdery mildew or salicylic acid. When treated with jasmonic acid methyl ester, its PR1 gene expression decreased, while the expressions of NPR1, PR10 and PAL all increased, contrasting with the wild type grape.     

Radix Scutellariae Attenuates CUMS-Induced Depressive-Like Behavior by Promoting Neurogenesis via cAMP/PKA Pathway

Chronic mild unpredictable stress (CUMS) causes neurogenesis damage in the hippocampus and depressive-like behaviors such as cognitive impairment. Radix Scutellariae from the dry root of Scutellaria baicalensis Georgi, with the common name Baikal skullcap. In this study, we demonstrated that Radix Scutellariae (RS 500, 1000 mg/kg) notably improved the behavior of the rat, such as shortened escape latency in morris maze test, reduced immobility time in tail suspension test and in forced swimming test, as well as increased sucrose consumption in sucrose preference test. In addition, RS alleviated the damage CUMS-induced neurogenesis and the reduced levels of BrdU; DCX and NeuN, the neurons hallmark of hippocampus neurogenesis. Moreover, associated proteins in cAMP/PKA pathway were up-regulated after RS treatment. By HPLC analysis, we found that RS decoction contains four main components, including baicalin, baicalein, wogonoside and wogonin, respectively. In conclusion, RS could exert a natural antidepressant with improving depressive-like behavior via regulation of cAMP/PKA neurogenesis pathway.     

Molecular cloning,expression, and subcellular localization of a PAL gene from Citrus reticulata under iron deficiency

Phenylalanine ammonia lyase (PAL) is a specific branch point enzyme of primary and secondary metabolism. The Citrus reticulata Blanco PAL gene was cloned and designated as CrPAL1. The cDNA sequence of CrPAL1 was 2 166 bp, encoding 721 amino acid residues. Sequence alignment indicates that CrPAL1 shared a high identity with PAL genes found in other plants. Both the dominant and catalytic active sites of CrPAL1 were similar to PAL proteins observed in Petroselinum crispum. Phylogenetic tree analysis indicates that CrPAL1 was more closely related to PALs in Citrus clementina × C. reticulata and Poncirus trifoliata than to those from other plants. Subcellular localization reveals that CrPAL1-green fluorescent protein fusion protein was specifically localized in the plasma membrane. Activity of PAL as well as CrPAL1 expression increased under Fe deficiency. A similar result was noted for total phenolic content. The root exudates of C. reticulata strongly promoted reutilization of apoplastic Fe in roots. Furthermore, Fe was more desorbed from the cell wall under Fe deficiency than in sufficient Fe supply.     

Study on the chemical composition and antioxidant activity of extracts from shoot culture and regenerated plants of <Emphasis Type="Italic">Scutellaria altissima</Emphasis> L.

The flavonoid (baicalin, wogonoside, luteolin, luteolin-7-glucoside) and verbascoside contents of Scutellaria altissima in both shoot cultures, and the shoots and roots of micropropagated plants grown in the greenhouse for 12 weeks or in the field for 2 years were determined. The level of secondary metabolites was found to be strongly affected by the age and type of plant organ. A comparative analysis of S. altissima plants propagated in vitro and from seeds revealed no differences in the level of secondary metabolites when plants of the same age were studied. The antioxidant potential of methanolic extracts from shoot cultures, and the shoots and roots of S. altissima plants propagated in vitro, were evaluated using ABTS radical scavenging, FRAP metal reduction power and the lipid peroxidation test, in relation to the content of baicalin, wogonoside, verbascoside, total phenolic and total flavonoid compounds. Extracts from the roots of field-grown regenerated plants at the flowering stage were found to possess the strongest antioxidant activity. Correlation analysis revealed that the antioxidant activity of extracts correlated most closely with their total phenolic content estimated by the Folin-Ciocalteu method.     

Ecological ranges and types of rarity in the Kuznetsk Alatau of some <Emphasis Type="Italic">Saussurea</Emphasis> DC. species

An analysis of indicator values of four Saussurea DC. species, S. baicalensis (Adams) B. L. Rob., S. frolowii Ledeb., S. salicifolia (L.) DC., and S. schanginiana (Wydl.) Fisch. ex Serg., have been carried out by the method of indicator scales of soil moisture and nutrient availability and salinity in more than 1600 relevés. The size of their ecological ranges and the values of their competitiveness in phytocenoses are determined. As a result of a comparison of the locality of local populations within the ecological range and their occurrence, the vulnerability of S. baicalensis, S. frolowii, S. salicifolia, and S. schanginiana in the Kuznetsk Alatau is evaluated. The types of rarity for these species in the Kuznetsk Alatau are determined.     

Flavonoid Production in Transformed Scutellaria baicalensis Roots and Ways of Its Regulation

A root culture of skullcap (Scutellaria baicalensisGeorgi) transformed with pRi T-DNA was initiated by the inoculation of sterile seedlings with Agrobacterium rhizogenes(wild-type strain A-4). The flavonoid concentration in cultured roots comprised 5% of the root dry weight and was maintained essentially constant during a subculture. For four weeks of culturing, the weight of the roots increased by 20–30 times; when the roots were cultured for a longer time and with periodic enrichment of the nutrient medium, their weight increased 50-fold. Skullcap roots were shown to synthesize flavones characteristic of intact roots (wogonin, baicalein, and baicalin). The addition of 0.01–1 mM L-phenylalanine (a precursor of flavonoids) to the nutrient medium affected neither root growth, nor their flavonoid concentration. Root elicitation with 100 M methyl jasmonate for 72 h increased the flavonoid content per flask and per root dry weight by 1.8 and 2.3 times, respectively.     

Deep Sequencing of the Scutellaria baicalensis Georgi Transcriptome Reveals Flavonoid Biosynthetic Profiling and Organ-Specific Gene Expression

Scutellaria baicalensis Georgi has long been used in traditional medicine to treat various such widely varying diseases and has been listed in the Chinese Pharmacopeia, the Japanese Pharmacopeia, the Korean Pharmacopoeia and the European Pharmacopoeia. Flavonoids, especially wogonin, wogonoside, baicalin, and baicalein, are its main functional ingredients with various pharmacological activities. Although pharmaological studies for these flavonoid components have been well conducted, the molecular mechanism of their biosynthesis remains unclear in S. baicalensis. In this study, Illumina/Solexa deep sequencing generated more than 91 million paired-end reads and 49,507 unigenes from S. baicalensis roots, stems, leaves and flowers. More than 70% unigenes were annotated in at least one of the five public databases and 13,627 unigenes were assigned to 3,810 KEGG genes involved in 579 different pathways. 54 unigenes that encode 12 key enzymes involved in the pathway of flavonoid biosynthesis were discovered. One baicalinase and three baicalein 7-O-glucuronosyltransferases genes potentially involved in the transformation between baicalin/wogonoside and baicalein/wogonin were identified. Four candidate 6-hydroxylase genes for the formation of baicalin/baicalein and one candidate 8-O-methyltransferase gene for the biosynthesis of wogonoside/wogonin were also recognized. Our results further support the conclusion that, in S. baicalensis, 3,5,7-trihydroxyflavone was the precursor of the four above compounds. Then, the differential expression models and simple sequence repeats associated with these genes were carefully analyzed. All of these results not only enrich the gene resource but also benefit research into the molecular genetics and functional genomics in S. baicalensis.     

Genetic diversity and population structure of common walnut (<Emphasis Type="Italic">Juglans regia</Emphasis>) in China based on EST-SSRs and the nuclear gene phenylalanine ammonia-lyase (<Emphasis Type="Italic">PAL</Emphasis>)

Common walnut (Juglans regia L.) is an economically important temperate tree species valued for both its nut and wood. We investigated the genetic diversity and population structure of J. regia germplasm from 13 locations in China using 10 markers derived from expressed sequences (EST-SSR) and sequence polymorphisms within the phenylalanine ammonia-lyase (PAL) gene. Analysis of the population genetic structure based on EST-SSRs showed distinct populations in northern versus southern China that were not reflected in the spatial distribution of PAL haplotypes. High levels of population differentiation were probably the result of reproductive isolation and in southern China, hybridization with Juglans sigillata. The results indicate the possible presence of distinct evolutionary lineages of J. regia in the Qinling and Daba Mountains of China and in Yunnan province that may require ecological management if they are to be retained as in situ resources.     

Over-expression of the <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> harpin-encoding gene <Emphasis Type="Italic">hrpZm</Emphasis> confers enhanced tolerance to Phytophthora root and stem rot in transgenic soybean

Phytophthora root and stem rot (PRR) caused by Phytophthora sojae is one of the most devastating diseases reducing soybean (Glycine max) production all over the world. Harpin proteins in many plant pathogenic bacteria were confirmed to enhance disease and insect resistance in crop plants. Here, a harpin protein-encoding gene hrpZpsta from the P. syringae pv. tabaci strain Psta218 was codon-optimized (renamed hrpZm) and introduced into soybean cultivars Williams 82 and Shennong 9 by Agrobacterium-mediated transformation. Three independent transgenic lines over-expressing hrpZm were obtained and exhibited stable and enhanced tolerance to P. sojae infection in T₂–T₄ generations compared to the non-transformed (NT) and empty vector (EV)-transformed plants. Quantitative real-time PCR (qRT-PCR) analysis revealed that the expression of salicylic acid-dependent genes PR1, PR12, and PAL, jasmonic acid-dependent gene PPO, and hypersensitive response (HR)-related genes GmNPR1 and RAR was significantly up-regulated after P. sojae inoculation. Moreover, the activities of defense-related enzymes such as phenylalanine ammonia lyase (PAL), polyphenoloxidase (PPO), peroxidase, and superoxide dismutase also increased significantly in the transgenic lines compared to the NT and EV-transformed plants after inoculation. Our results suggest that over-expression of the hrpZm gene significantly enhances PRR tolerance in soybean by eliciting resistance responses mediated by multiple defense signaling pathways, thus providing an alternative approach for development of soybean varieties with improved tolerance against the soil-borne pathogen PRR.     

Engineering of <Emphasis Type="Italic">Escherichia coli</Emphasis> for the synthesis of <Emphasis Type="Italic">N</Emphasis>-hydroxycinnamoyl tryptamine and serotonin

Plants synthesize various phenol amides. Among them, hydroxycinnamoyl (HC) tryptamines and serotonins exhibit antioxidant, anti-inflammatory, and anti-atherogenic activities. We synthesized HC–tryptamines and HC–serotonin from several HCs and either tryptamine or serotonin using Escherichia coli harboring the 4CL (4-coumaroyl CoA ligase) and CaHCTT [hydroxycinnamoyl-coenzyme A:serotonin N-(hydroxycinnamoyl)transferase] genes. E. coli was engineered to synthesize N-cinnamoyl tryptamine from glucose. TDC (tryptophan decarboxylase) and PAL (phenylalanine ammonia lyase) along with 4CL and CaHCTT were introduced into E. coli and the phenylalanine biosynthetic pathway of E. coli was engineered. Using this strategy, approximately 110.6 mg/L of N-cinnamoyl tryptamine was synthesized. By feeding 100 μM serotonin into the E. coli culture, which could induce the synthesis of cinnamic acid or p-coumaric acid, more than 99 μM of N-cinnamoyl serotonin and N-(p-coumaroyl) serotonin were synthesized.     

Effect of foliar application of salicylic acid,hydrogen peroxide and a xyloglucan oligosaccharide on capsiate content and gene expression associated with capsinoids synthesis in <Emphasis Type="Italic">Capsicum annuum</Emphasis> L<Emphasis Type="Italic">.</Emphasis>

Capsinoids are non-pungent analogues of capsaicinoids in pepper (Capsicum spp). The absence of pungency, in addition to their biological activities similar to that of capsaicinoids such as anti-inflammatory, antimicrobial, and antioxidant properties, makes capsinoids an excellent option for increasing use in human and animal nutrition, as well as health and pharmaceutical industries. There are only few sources of pepper producing capsinoids, and one of them (accession 509–45-1), Capsicum annuum L., is a potential source for increasing capsinoids content using strategies as controlled elicitation during plant production in the greenhouse. In this research we evaluated the effect of weekly and one-day-before-harvest foliar applications of hydrogen peroxide, salicylic acid and a xyloglucan oligosaccharide on the concentration of capsiate in fruits of this pepper accession, as well as the gene expression of phenylalanine ammonia-lyase (pal), putative aminotransferase (pamt), capsaicin synthase (at3) and β-keto acyl synthase (kas). Results showed that the two tested concentrations of H₂O₂ significantly increased capsiate content and gene expression associated with capsaicinoids (pamt, at3 and kas) and the phenylpropanoids (pal) pathways. Plant yield was not affected using this induction strategy. Our results indicated that the pre-harvest and weekly application of hydrogen peroxide and xyloglucan oligosaccharide improved production of capsiate in C. annuum L.     

Flavone-catalyzed apoptosis in Scutellaria baicalensis

In response to mechanical damage, roots of Scutellaria baicalensis undergo cell death within 24 h. The flavone baicalein was identified as the factor regulating apoptosis in the damaged roots of S. baicalensis. Plant apoptosis is known to be triggered by oxidative damage of DNA through oxidative bursts, whereas baicalein causes apoptosis in Scutellaria cells by a copper-dependent oxidation of nuclear DNA without inducing an oxidative burst. S. baicalensis possesses an interesting system for quickly producing this apoptosis-inducing flavone in its cells. Intact Scutellaria cells contain little baicalein but store a large amount of baicalin (baicalein 7-O-β-D-glucuronide). Stress treatment of Scutellaria cells immediately initiates hydrolysis of baicalin by endogenous β-glucuronidase, and the resulting baicalein is immediately translocated to the nucleus, leading to apoptosis. Thus, S. baicalensis possesses a unique apoptosis-inducing system that is linked with metabolism of baicalin.     

A study of the intestinal microbiomes of the Lake Baikal oilfishes (Cottoidei,Comephoridae)

A comparative analysis of the intestinal microbiomes of some representatives of the Lake Baikal endemic family Comephoridae—the big oilfish Comephorus baicalensis (Pallas, 1776) and the little oilfish C. dybowski (Korotneff, 1904)—has been performed for the first time. The phylotype Akkermansia (phylum Verrucomicrobia) was detected in the C. dybowski microbiome, while Alistipes, Bacteroides, Chryseobacterium, Prevotella (Bacteroidetes), and Peptoniphilus (Firmicutes), were found in the C. baicalensis microbiome. Their presence in the intestine is associated with an elevated lipid uptake or an increase in the human and animal body weight. It is hypothesized that these microorganisms are involved in the lipid metabolism of the oilfish and determine their morphophysiological adaptations to pelagic life.     

A new species of grayling <Emphasis Type="Italic">Thymallus svetovidovi</Emphasis> sp. nova (Thymallidae) from the Yenisei basin and its position in the genus <Emphasis Type="Italic">Thymallus</Emphasis>

The Upper Yenisei grayling Thymallus svetovidovi sp. nova that inhabits the headwaters of the Yenisei River in Mongolia is described. From the other representatives of the genus Thymallus, the species differs in the elements of body coloration, dorsal fin pattern, some morphometric characters, and genetic characteristics. Besides this species, most of the Yenisei basin is inhabited by Baikal grayling T. baicalensis Dyb., and its low reaches, by the Arctic grayling T. arcticus (Pall.).     

The lignin synthesis related genes and lodging resistance of <Emphasis Type="Italic">Fagopyrum esculentum</Emphasis>

Lignin is closely related to the lodging resistance of common buckwheat (Fagopyrum esculentum Moench.). However, the characteristics of lignin synthesis related genes have not yet been reported. We investigated the lignin biosynthesis gene expression, activities of related enzymes, and accumulation of lignin monomers during branching stage, bloom stage, and milky ripe stage by real-time quantitative PCR, UVspectrophotometry, and gas chromatography-mass spectrometry in the 2^nd internode of three common buckwheat cultivars with different lodging resistance. The results showed that lignin content and the activity of phenylalanine ammonia lyase (PAL), 4-coumarate: CoA ligase (4CL), cinnamyl alcohol dehydrogenase (CAD) and peroxidase (POD) were closely related to the lodging resistance of common buckwheat. Further, we studied gene expression of cinnamate 4-hydroxylase (C4H), caffeoyl-CoA O-methyltransferase (CCoAOMT), ferulate 5-hydroxylase (F5H), cinnamoyl-CoA reductase (CCR), and caffeic acid O-methyltransferase (COMT). The lignin biosynthesis genes were divided into three classes according to their expression pattern: 1) expression firstly increasing and then descending (PAL, 4CL, CAD, C4H, CCoAOMT, F5H, and CCR), 2) expression remaining constant during maturation (C3H), and 3) expression decreasing with maturation (COMT). The present study provides preliminary insights into the expression of lignin biosynthesis genes in common buckwheat, laying a foundation for further understanding the lignin biosynthesis.