Alba Samples Research Articles

DNA barcodes, ISSR, RAPD and SCAR markers as potential quality control tools for molecular authentication of black and white mulberry

Morus species, especially black mulberry (Morus nigra L.), are well known for their nutritive and economic values. Black mulberry is often a subject for deliberate and undeliberate substitution with the morphologically similar white mulberry (Morus alba L.). In the current study, the genetic diversity of Morus nigra L. and Morus alba L. was assessed using different genetic profiling and fingerprinting techniques namely namely DNA barcoding, Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR). DNA barcoding was implemented using 2 plastid loci (matK, and rbcL-1) and 2 nuclear spacers (ITS and ITS2) aiming at accurate authentication of the studied species. DNA barcoding, targeting the selected loci succeeded to identify and authenticate the studies Morus species but failed to differentiate them, owing to their close genetic resemblance. Out of 52 screened RAPD primers, 27 were able to produce reproducible and clear profiles with 115 amplified fragments of which 37 were polymorphic accounting for 32.17% polymorphism. The number of amplified bands varied from 2 to 7 ranging in size from 100 to 700 bp. Moreover, 55 ISSR primers were evaluated, 108 fragments were produced, using 20 primers, of which 28 were polymorphic accounting for 25.92% polymorphism with size range of 200–800 bp. Sequence Characterized Amplified Region (SCAR) marker was developed based on reproducible banding pattern obtained by RAPD primer (OPG-03), for the purpose of accelerating the initial screening of Morus batches and enabling the preventative rejection of adulterated samples. The polymorphic band obtained was sequenced and specific primers were designed to develop a SCAR marker of approximately 558 bp exclusively for M. nigra, allowing detection of down to 1 ng, of M. nigra. The SCAR marker showed 100% success in differentiating authentic M. nigra and M. alba samples. In addition, the application of the developed SCAR marker was extended to distinguish M. nigra from its four common adulterants namely Ficus carica L., Fragaria vesca L., Ficus racemosa L. and Vitis vinifera L.. The proposed protocol was successfully implemented for the evaluation of 47 commercial M. nigra samples collected from different Egyptian locations.

Effectiveness of Essential Oils from Three Medicinal Plants Against Bayoud Disease (Fusarium oxysporum f. sp. albedinis) of Date Palm (Phoenix dactylifera L.)

Plant extracts and essential oils are promising new sources of non-toxic alternatives to chemicals substances. They are used for their antimicrobial properties against plant diseases of fungal origin, against bacterial, and those of virus origin. This study was carried out to study the antifungal activity of Artemisia herba-alba, Foeniculum vulgare and Citrus sinensis essential oils against Fusarium oxysporum f. sp. albedinis. Essential oil was extracted by steam distillation. Antifungal activity of essential oil was investigated by macro-broth method of dilution by a minimal inhibitory concentration (MIC) assay against this pathogen. The yield of essential oil obtained by steam distillation of Foeniculum vulgare samples was 2.31% greater than that obtained from Citrus sinensis sample which was 1.8%, followed by Artemisia herba alba samples 1.22%. Regarding antifungal activity, the results revealed a better inhibitory activity of Artemisia herba alba against the tested strainsat the lowest LC50 values (0.1 µl/ml). On the other hand, Foeniculum vulgare, Artemisia herba alba and Citrus sinensis essential oils show similar MICs of mycelial growth against this pathogen. The value of the MIC and CMF is greater than 50 µl/ml for the three essential oils.

Comparative Analyses of Radix Paeoniae Alba with Different Appearance Traits and from Different Geographical Origins Using HPLC Fingerprints and Chemossmetrics

In this study, Radix Paeoniae Alba with different appearance traits and from three geographical regions was analyzed using a combination of high-performance liquid chromatography fingerprints, chemometrics, and content determination. Hierarchically clustered heatmap was developed for sample visualization. Principal component analysis was used to classify 47 Radix Paeoniae Alba samples with different appearance traits and from three different geographical regions. Principal component analysis results revealed that the Radix Paeoniae Alba samples fell into three clusters, which were related to geographical location and appearance traits. In addition, orthogonal partial least squares discriminant analysis was established to predict samples classification and obtain the 14 most significant variables (chemical markers) by a variable importance in the project. The six chemical markers were gallic acid (1), oxypaeoniflorin (7), paeoniflorin (12), 1,2,3,4,6-O-pentagalloylglucose (19), benzoic acid (20), and benzoyl paeoniflorin (24). Finally, the content of gallic acid, oxypaeoniflorin, paeoniflorin, and benzoyl paeoniflorin was determined, and the total contents of the four components were class 2 > class 1 > class 3, which suggests that geographical origins and appearance traits can be used to determine the quality of Radix Paeoniae Alba.

A comparative analysis of raw and processed Radix Paeoniae Alba samples using HPLC, chemometrics, and antioxidant activity.

Radix Paeoniae Alba (RPA) and its processed products are commonly used in traditional Chinese medicine, but the chaos phenomenon among processed products often occurs. In this study, we used multiple chemometric methods to analyze raw and six different processed products of RPA based on HPLC fingerprinting. Heat map analysis was used to assess the changes in chemical composition. Principal component analysis was used for classification, and the samples were divided into four classes: class 1 (raw, wine-processed, and vinegar-processed products), class 2 (bran-processed and soil-processed products), class 3 (stir-fried products), and class 4 (coke products). Further, the orthogonal partial least squares discriminant analysis model was used to obtain chemical markers among different classes. The antioxidant property of RPA is an important factor responsible for its pharmacological effects, and so the antioxidant activity of RPA was also investigated. We measured 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and hydroxyl radical scavenging ability, and ferric reducing antioxidant power. The total antioxidant activity follows the order: coke > stir-fried > soil-processed > bran-processed > wine-processed > raw > vinegar-processed products. These results suggest that different processing methods affect the chemical composition and antioxidant power of RPA, and thus, different products of RPA should not be mixed.

First Report of Basella alba Naturally Infected with Basella Rugose Mosaic Virus in Hawaii

Basella alba, also known as Malabar spinach, is a plant belonging to the family Basellaceae. It is native to tropical Asia and Africa and introduced to Hawaii, where it is grown as a vegetable crop (Wester 1992). Basella rugose mosaic virus (BaRMV) is a potyvirus that was first isolated from B. alba and reported in December 2006 in Taiwan (Zheng et al. 2008). In December 2018, B. alba plants with leaves exhibiting mosaic and rugose symptoms were found growing in a community garden consisting of about 150 small gardens 10 m² in size on Oahu, Hawaii (21°17′05″ N, 157°49′40″ W). Five leaf samples with severe mosaic and rugose symptoms from five different B. alba plants in five different small gardens in the same community garden were tested by ELISA using potyvirus-specific antibodies (Agdia, Elkhart, IN) (Wang et al. 2019). All symptomatic leaf samples tested positive for potyvirus infection by the ELISA test. Total RNAs were extracted from these samples and tested by reverse transcription PCR (RT-PCR) using the universal primer pair CIFor and CIRev designed to amplify the cylindrical inclusion protein (CI) gene from the genomes of potyviruses (Ha et al. 2008). To identify the specific potyvirus infecting the B. alba samples, amplicons of approximately 600 bp corresponding to the targeted potyviral CI region were directly sequenced (MN075828). BLAST analysis of the 564-bp sequences showed that this virus isolate respectively shared 97 and 95% nucleotide and amino acid sequence identities with the homologous sequences of a peace lily mosaic virus (PeLMV) isolate from Vietnam (DQ851494/ABI34613). PeLMV was first reported infecting peace lily (Spathiphyllum patinii, family Araceae) in Vietnam in 2008 (Ha et al. 2008). Because the Vietnam isolate of PeLMV (DQ851494/ABI34613) respectively shared 83 and 92% nucleotide and amino acid sequence identities to an isolate of BaRMV (DQ821939/ABH10135), PeLMV is considered a strain of BaRMV (Wylie et al. 2017). However, our Hawaii isolate (MN075828) of BaRMV only shared 83 and 94% nucleotide and amino acid sequence identities, respectively, with the isolate of BaRMV (DQ821939/ABH10135). We then designed new BaRMV CI gene-specific primers B-CIF (5′-TCAGTTTTTGGTTCGACGCC-3′) and B-CIR (5′-GATTTCCGTTGCACCCGACT-3′) from the BaRMV-specific sequence (MN075828) to further characterize the viral isolate from B. alba. BLASTn analysis of the 389-bp sequence of the CI fragment amplicon (MN075829) indicated that it shared 98% nucleotide sequence identity with the same isolate from Vietnam (DQ851494), whereas BLASTx analysis showed that the CI amplicon shared 96% amino acid sequence identity with the corresponding amino acid region of the same isolate from Vietnam (ABI34613). The CI fragment amplicon (MN075829) also revealed 83 and 95% nucleotide and amino acid sequence identities, respectively, to the Taiwan isolate of BaRMV (DQ821939/ABH10135). Our sequence comparison results indicated that our Hawaii isolate was more closely related to the Vietnamese BaRMV isolate initially identified as PeLMV than to the Taiwan isolate of BaRMV. The primer set B-CIF and B-CIR was further used in RT-PCR reactions to determine infection by BaRMV in nine additional symptomatic samples of B. alba based on amplification of DNA fragments of the expected size. We have also used RT-PCR to test 20 more samples from this community garden; our results show that two asymptomatic samples were negative and 18 symptomatic samples were positive. Therefore, the disease incidence of BaRMV on B. alba was approximately 92%. To our knowledge, this is the first report of BaRMV infecting B. alba plants in the United States. Further studies to obtain information on the full-length genome sequence, insect vector(s), and geographic distribution of this BaRMV isolate are necessary to help control BaRMV infection in B. alba and other vegetables among Hawaii’s community and family gardens.

Phytostilbenoid production in white mulberry (Morus alba L.) cell culture using bioreactors and simple deglycosylation by endogenous enzymatic hydrolysis

Phytostilbenes are responsible for several biological activities of mulberry (Morus sp.), which has been widely used as a raw material in health products. This study aimed to investigate the capability of Morus alba L. cell in bioreactors to produce the major bioactive stilbenes. The cell obtained from air-driven bioreactors such as round bottom, flat bottom, and air-lift vessel shape bioreactors was collected and analyzed for the levels of mulberroside A and oxyresveratrol. The results showed that the cell culture in round bottom and air-lift vessel bioreactors had higher growth rate, as compared with the cell culture in shake flasks (1.38- and 1.41-fold, respectively). The optimized culture condition to produce mulberroside A was obtained from round bottom bioreactor culture (55.56 ± 11.41 μmol/L). Additionally, endogenous stilbenoid hydrolysis of cell from the bioreactor culture was examined. Under optimized hydrolytic conditions, mulberroside A in the cell was readily deglycosylated to give oxyresveratrol within 1 h. These results indicated that the glycoside mulberroside A in the cell is sensitive to the endogenous enzymatic hydrolysis. Interaction of the stilbenoid components with the endogenous hydrolytic enzyme triggered by cell disruption in M. alba samples was suggested to be the major cause of the alteration of the stilbenoid levels. These findings have provided a new approach to producing glycosidic compounds and corresponding aglycones in cell culture.

High-performance thin-layer chromatography/bioautography and liquid chromatography-mass spectrometry hyphenated with chemometrics for the quality assessment of Morus alba samples

Quality control is a crucial step in the production of effective and safe herbal remedies. The aim of this study was to develop a new, simple, and high throughput procedure for the quality assessment of herbal drugs using a high-performance thin-layer chromatography (HPTLC)/bioautography and UPLC–MS/MS approach combined with chemometrics. This was exemplarily shown for Morus alba L. root bark (sāng bái pí; SBP). Bioautography assays were developed for the identification of constituents with radical scavenging (DPPH assay) and antimicrobial activities (Bacillus subtilis, Escherichia coli) of 18 different M. alba samples, which was supported by UPLC–MS/MS analysis. Further, the combination of bioautography and chemometrics identified those samples with the most bioactive constituents.Plant materials collected from Serbia (11 samples) showed higher both radical scavenging and antimicrobial activities compared to samples provided from China (7 samples). Principal component analysis (PCA) confirmed the discrimination of geographically different samples and recognized their main markers responsible for differences between Serbian and Chinese samples. Most importantly for quality assessment, the combined HPTLC/bioautography and UPLC–MS/MS approach not only allowed for a fast chemical profiling of the investigated samples and their unambiguous identification, but also for the disclosure of major and minor bioactive constituents present in SBP.

Molecular cytogenetic characterization, leaf anatomy and ultrastructure of the medicinal plant Potentilla alba L.

Potentilla alba L. is a valuable medicinal plant widely used in folk and traditional medicine and particularly promising in complex treatment of thyroid pathology. Natural resources of this species are insufficient due to ever-growing use in contemporary medicine. Comprehensive investigations of different P. alba populations are essential for the successful extension of P. alba plantings. Aiming for a better understanding of karyotype structure, chromosome behaviour in meiosis and developing new diagnostic characters, we performed molecular cytogenetic characterization and leaf structure and ultrastructure analyses of two introduced P. alba samples originating from different habitats. Based on chromosome morphology, distribution of 45S/5S rDNA and DAPI-banding patterns, all chromosomes in the karyotypes were identified and the P. alba chromosomal idiogram was constructed. Our findings confirmed P. alba karyotype stability and also revealed several diagnostic characters of this species: the features of cells of upper and lower leaf epidermis, the presence of calcium oxalate druses and three types of leaf indumentum, essential for evaluation of genetic diversity in different populations, validation of raw materials and further selection progress. The meiotic abnormalities were detected probably related to low pollen activity and indicated the advantages of vegetative propagation in the development of a P. alba plantation system.

Isolation and Analysis of Extractives from White Cinquefoil (Potentilla alba L.) Grown under Different Conditions

Potentilla alba L. is used as a medicinal plant that contains a variety of biologically active substances. Natural resources are not meeting the needs of the pharmaceutical industry. We obtained the first biomass of P. alba plant-regenerants using hydroponics technique with clonal micropropagation. We have conducted a comprehensive study of the chemical composition of a vegetable raw materials P. alba, obtained in the Biotechnology Department (plant-regenerants) in comparison with raw material obtained from plants grown under field conditions (intact plants). We have established the quantitative content of the structural components in the samples: cellulose in roots and rhizomes of intact plants, 15.4%, in the roots of plant-regenerants, 4.3%, in the leaves of plant-regenerants, 2.5%; lignin in roots and rhizomes of intact plants, 40.7%, in the roots of plant-regenerants, 37.0%, and in the leaves of plant-regenerants, 36.8%. We determined the content of extractives in P. alba samples, recovered by sequential treatment with hexane, 96% and 40% ethanol solution, water and 1% aqueous sodium hydroxide solution. We identified the total content of extractives in the roots and rhizomes of intact plants at 15.3%, in the roots of plants-regenerants, 11.2%, and in the leaves of plant-regenerants, 5.1%. We characterized the chemical composition of extractives by UV-spectroscopy. Phenolic substances (λ = 279.5–280.0 nm) were dominant in the composition of the extracts. We have established the authenticity of the samples by the TLC chromatography based on the presence of flavonoids and tannins. We showed the identity of the phytochemical composition of vegetable raw materials produced by micropropagation and grown hydroponically for two months.

The suture pullout characteristics of human and porcine linea alba

There is a substantial prevalence of post-operative incisional hernia for both laparoscopic and laparotomy procedures, but there have been few attempts at quantifying abdominal wound closure methodology in the literature. One method to ascertain a more robust method of wound closure is the identification of the influence of suture placement parameters on suture pullout force. Current surgical practice involves a recommended bite depth and bite separation of 10mm, but the evidence base for this is not clear. In this paper, the suture pullout characteristics of both porcine and human linea alba were investigated to ascertain a suture placement protocol for surgical wound closure. Uniaxial suture pullout force testing on fresh frozen porcine and human linea alba samples was performed using standard materials testing machines. The influence of the number of suture loops, the bite depth and the bite separation of the sutures and the orientation of the sutures with respect to the principal fibre direction in the linea alba were assessed.Results showed a clearly identifiable relationship between pullout force of the suture, bite separation and bite depth, with low suture separation and high suture depth as optimal parameters for increasing pullout force. Resistance to pullout could be improved by as much as 290% when optimizing test conditions. Both human and porcine tissue were observed to exhibit very similar pullout force characteristics, corroborating the use of a porcine model for investigations into wound closure methodology. Orientation of suture application was also found to significantly affect the magnitude of suture pullout, with suturing applied longitudinally across a transverse defect resulting in higher pullout forces for small suture bite separations. Although further assessment in an environment more representative of in vivo conditions is required, these findings indicate that increasing the bite depth and reducing the bite separation with respect to the current surgical recommendations may reduce the risk of post-operative incisional hernia.

Molecularly imprinted solid phase extraction in a syringe filter for determination of triazine herbicides in Radix Paeoniae Alba by ultra-fast liquid chromatography

A novel, cost-effective and simple solid phase extraction (SPE) method, by using a syringe connected with a nylon membrane filter as the adsorbent container, was developed for the extraction of triazine herbicides from Radix Paeoniae Alba (RPA) samples. The selective molecularly imprinted polymers (MIPs) synthesized with the template of atrazine were employed as the adsorbents for the enrichment and purification of analytes. The extraction parameters, including the volume and type of loading solvent, the type of washing solvent and eluting solvent, were investigated. Under the optimized conditions, the final extracts were analyzed by ultra-fast liquid chromatography (UFLC). Recoveries of the developed method range from 92.4% to 107.3% with intra- and inter-day relative standard deviations (RSDs) lower than 8.2%. The calibration curve is linear in the concentration range of 0.005–2.4µgg−1 for desmetryn, atrazine and terbumeton, and 0.005–1.5µgg−1 for dimethametryn and dipropetryn, with the correlation coefficient (R2) higher than 0.9995. The limits of detection (LODs) of five triazine herbicides are in the range of 0.09–0.39ngg−1, which are lower than the maximum residue levels (MRLs) established by various official organizations. Analytical results of three real Radix Paeoniae Alba samples indicate that the proposed method is cost-effective and easy-to-use than other routine pretreatment methods.

Fingerprint analysis, multi-component quantitation, and antioxidant activity for the quality evaluation of Salvia miltiorrhiza var. alba by high-performance liquid chromatography and chemometrics.

Salvia miltiorrhiza Bge. var. alba C.Y. Wu and H.W. Li has wide prospects in clinical practice. A useful comprehensive method was developed for the quality evaluation of S. miltiorrhiza var. alba by three quantitative parameters: high-performance liquid chromatography fingerprint, ten-component contents, and antioxidant activity. The established method was validated for linearity, precision, repeatability, stability, and recovery. Principal components analysis and hierarchical clustering analysis were both used to evaluate the quality of the samples from different origins. The results showed that there were category discrepancies in quality of S. miltiorrhiza var. alba samples according to the three quantitative parameters. Multivariate linear regression was adopted to explore the relationship between components and antioxidant activity. Three constituents, namely, danshensu, rosmarinic acid, and salvianolic acid B, significantly correlated with antioxidant activity, and were successfully elucidated by the optimized multivariate linear regression model. The combined use of high-performance liquid chromatography fingerprint analysis, simultaneous multicomponent quantitative analysis, and antioxidant activity for the quality evaluation of S. miltiorrhiza var. alba is a reliable, comprehensive, and promising approach, which might provide a valuable reference for other herbal products in general to improve their quality control.

Ultra-performance liquid chromatography coupled with high-resolution quadrupole time-of-flight mass spectrometry analysis of the impact of bran-processing on the chemical profile of Radix Paeoniae Alba (Baishao)

In this study, a rapid and versatile ultra-performance liquid chromatography coupled with high-resolution quadrupole time-of-flight mass spectrometry-based chemical profiling approach was applied to evaluate chemical constitution of crude and processed Radix Paeoniae Alba (RPA) samples. A total of 44 compounds were identified, among which the contents of 9 compounds in processed samples were obviously decreased and 8 compounds were increased. Furthermore, compound 28 was not found in RPA sample after stir-frying with wheat bran. The proposed method provided a chemical basis for exploring the processed mechanism of herbal medicine.

Investigation of the Chemical Changes from Crude and Processed Paeoniae Radix Alba-Atractylodis Macrocephalae Rhizoma Herbal Pair Extracts by Using Q Exactive High-Performance Benchtop Quadrupole-Orbitrap LC-MS/MS.

The Paeoniae Radix Alba-Atractylodis Macrocephalae Rhizoma herbal pair is mainly used for regulating the functions of liver and spleen, benefiting qi, and nourishing blood. However, the bioactive compounds for the pharmacological activities of the crude and processed Paeoniae Radix Alba-Atractylodis Macrocephalae Rhizoma herbal pair extracts are still unclear to date. In the present study, Q Exactive high-performance benchtop quadrupole-Orbitrap LC-MS/MS was applied to identify the complicated components from crude and processed Paeoniae Radix Alba, crude and processed Atractylodis Macrocephalae Rhizoma, and their crude and processed herbal pair extracts. 123 and 101 compounds were identified in crude and processed Paeoniae Radix Alba samples, respectively. Meanwhile, 32 and 26 compounds were identified in crude and processed Atractylodis Macrocephalae Rhizoma samples, respectively. In the crude and processed Paeoniae Radix Alba-Atractylodis Macrocephalae Rhizoma herbal pair extracts, co-decoction could significantly change the chemical composition of Paeoniae Radix Alba and Atractylodis Macrocephalae Rhizoma in solution. The developed method may provide a scientific foundation for deeply elucidating the processing and compatibility mechanism of Paeoniae Radix Alba and Atractylodis Macrocephalae Rhizoma.

Rapid discrimination of geographical origin and evaluation of antioxidant activity of Salvia miltiorrhiza var. alba by Fourier transform near infrared spectroscopy

Radix Salvia miltiorrhiza Bge. var. alba C.Y. Wu and H.W. Li and Radix S. miltrorrhiza belong to the same genus. S. miltiorrhiza var. alba has a unique effectiveness for thromboangiitis besides therapeutical efficay of S. miltrorrhiza. It exhibits antioxidant activity (AA), while its quality and efficacy also vary with geographic locations. Therefore, a rapid and nondestructive method based on Fourier transform near infrared spectroscopy (FT-NIRS) was developed for discrimination of geographical origin and evaluation of AA of S. miltiorrhiza var. alba. The discrimination of geographical origin was achieved by using discriminant analysis and the accuracy was 100%. Partial least squares (PLS) regression was employed to establish the model for evaluation of AA by NIRS. The spectral regions were selected by interval PLS (i-PLS) method. Different pre-treated methods were compared for the spectral pre-processing. The final optimal results of PLS model showed that correlation coefficients in the calibration set (Rc) and the prediction set (Rp), root mean square error of prediction (RMSEP) and residual prediction deviation (RPD) were 0.974, 0.950, 0.163mgmL−1 and 2.66, respectively. The results demonstrated that NIRs combined with chemometric methods could be a rapid and nondestructive tool to discriminate geographical origin and evaluate AA of S. miltiorrhiza var. alba. The developed NIRS method might have a potential application to high-throughput screening of a great number of raw S. miltiorrhiza var. alba samples for AA.

DNA Barcoding as a Tool for Elucidating Species Delineation in Wide-ranging Species as Illustrated by Owls (Tytonidae and Strigidae)

The mitochondrial cytochrome c-oxidase subunit I (cox1) can serve as a fast and accurate marker for the identification of animal species, and for the discovery of new species across the tree of life. Distinguishing species using this universal molecular marker, a technique known as DNA barcoding, relies on the identifying the gap between intra- and interspecific divergence. One of the difficulties could be wide-ranging, cosmopolitan species that show large amounts of morphological variation. The barn owl Tyto alba is a case in point. It occurs worldwide and varies morphologically, leading to the recognition of many subspecies or, more recently, species. We analysed data from the cox1 gene for 31 individuals of seven subspecies, and compared this with 214 sequences from 29 other owl species. Phylogenetic analysis of the T. alba samples gives very strong support for an Old World alba-clade (three subspecies) and a New World furcata-clade (four subspecies) that are genetically equidistant. The amount of intraspecific variation within each of these clades ranges from 0.66-0.99%, but variation among these clades ranges from 5.33-6.20%. Combined these data suggest that barn owl of the Old World is indeed best considered a separate species different from that of the New World. For combined dataset, sample size of owl species (n between 1 and 21 sequences) increased with geographic range size but we did not find significant relationships between interspecific divergence and sample size or between interspecific divergence and geographic range. For 21/24 species of owls with sample sizes of n ≥4 the maximum interspecific divergences was ≤ 3.00%. However, similar to those found in barn owls, the largest amount of divergence (3.23-4.09%) was present in two other wide-ranging species (Strix nebulosa and Aegolius funereus) raising the possibility of multiple species in other wide-ranging owls as well.

UPLC-Q-TOF/MS Coupled with Multivariate Statistical Analysis as a Powerful Technique for Rapidly Exploring Potential Chemical Markers to Differentiate between Radix Paeoniae Alba and Radix Paeoniae Rubra

To explore rapidly the potential chemical markers for differentiating Radix Paeoniae Alba and Radix Paeoniae Rubra, a method is proposed based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) coupled with multivariate statistical analysis. Batches of commercial samples were analyzed by UPLC-Q-TOF/MS. The datasets of t(R)-m/z pair, ion intensities and sample codes were further processed with orthogonal partial least squared discriminant analysis (OPLS-DA) to compare holistically the difference between these two kinds of samples. Then statistics were used to generate an S-plot, in which the variables (t(R)-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components correlated to these ions should be regarded as the chemical markers. The identities of the most changed markers can be identified by comparing the mass/UV spectra and retention times with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of known compounds published in the literature. Using this proposed approach, albflorin, paeoniflorin, oxypaeoniflorin, benzoylpaeoniflorin, galloylalbiflorin and paeoniflorigenone were found to be the differentiating components for discrimination of Radix Paeoniae Alba and Radix Paeoniae Rubra. Moreover, paeoniflorin sulfonate and its isomer, isomaltopaeoniflorin sulfonate, were found to be the characteristic markers for all Radix Paeoniae Alba samples that were processed by sulfurdioxide gas fumigation. The results suggested that this newly established approach could be used to explore rapidly the potential chemical markers for herbs with similar chemical characteristics.

Exploring Potential Chemical Transformation by Chemical Profiling Approach for Rapidly Evaluating Chemical Consistency between Sun-Dried and Sulfur-Fumigated Radix Paeoniae Alba Using Ultraperformance Liquid Chromatography Coupled with Time-of-Flight Mass Spectrometry.

Ultraperformance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-QTOF/MS) based on a chemical profiling method was applied to rapidly evaluate the chemical consistency between sun-dried and sulfur-fumigated Radix Paeoniae Alba. By virtue of the high resolution, high speed of UPLC, and the accurate mass measurement of TOFMS coupled with reliable MarkerLynx software, five newly assigned monoterpene glycoside sulfonates were found and identified in sulfur-fumigated Radix Paeoniae Alba samples. This method could be applied for rapid quality evaluation of different kinds of sulfur-fumigated Radix Paeoniae Alba among commercial samples.

Development and characterization of 15 polymorphic microsatellite loci in Sonneratia alba (Lythraceae) using next-generation sequencing

A set of 15 new microsatellite loci was developed and characterized for the widespread mangrove tree species Sonneratia alba (Lythraceae) by using next-generation sequencing. Forty-eight S. alba samples from seven populations in the Indo-West Pacific region were genotyped; all loci were polymorphic, with the number of alleles ranging from three to eight. The mean observed heterozygosity per locus was 0.21 for a population from Sabah, Malaysia. No null allele or significant linkage disequilibrium was detected, indicating the robustness of the markers. Only one locus (SA103) showed deviation from Hardy–Weinberg equilibrium. As characterization of these microsatellite loci was done with samples covering most of the species’ distribution range, the markers can be applied to genetic diversity studies over the broad geographical range of the species.

Combining the use of molecular techniques and archival documentary evidence to trace the origin of Populus alba in a Central Mediterranean archipelago

The nativity of Populus alba in the Mediterranean has only been confirmed in the last decade, following the discovery of 8,000-year-old leaf imprints in Southern France. Recent evidence has even emerged from molecular studies suggesting that the species is native to some of the islands, and these populations may be relicts of a native flora that arrived there much earlier than previously thought. In view of this, samples obtained from the Central Mediterranean archipelago of Malta and other neighbouring regions were analysed to determine the native status of the Maltese populations and possibly trace their origins. All 38 samples were investigated in order to assess the genetic diversity and origin of Maltese trees. Nuclear microsatellite analysis revealed that all 28 trees sampled from the two islands of Malta belonged to one clone. Chloroplast data suggested relatedness of the Maltese clone to Italian P. alba samples. However, nuclear data suggested additional admixture through pollen from North Africa. Existing archival and palaeontological records were also examined for any supporting evidence. On considering the latter records in combination with molecular evidence, we arrived to the conclusion that arrival of this clone in Malta through human introduction in the sixteenth century is the most likely explanation, since alternative scenarios like autovegetative propagation or arrival by seed seem highly unlikely.