Study on identification of medicinal plant Gentiana scabra Bge. in Liaoning province based on DNA barcode sequences / 国际中医中药杂志

Objective:To identify and analyze the genuine medicinal plant Gentiana scabra Bge. from 9 regions in Liaoning Province using DNA barcode technology based on the base sequence of internal transcribed spacer. Methods:DNA was extracted from the medicinal parts of 26 Gentiana scabra Bge. samples by using DNA kit extraction method. The ITS sequence was amplified through polymerase chain reaction (PCR), and then two-way sequencing was carried out. Other sources and outgroup sequences of the medicinal plant Gentiana scabra Bge. were downloaded from Genbank. After the sequencing results were spliced by using SeqMan 7.1.0 software, MEGA 7.0 software was used to analyze and compare the data, and calculate the genetic distance of K2P (Kimura 2-parameter). The phylogenetic tree was established by Neighbor-Joining (NJ) method for analysis. Results:According to the results of NJ cluster tree, all Gentiana scabra Bge. samples from different sources were clustered into one large branch, and Gentiana scabra Franch. and Gentiana triflora Pall. were clustered into one branch respectively, with obvious differences; Gentiana scabra and Gentiana manshurica Kitag. were clustered into one branch, and the genetic relationship was relatively close. In combination with the variation site and genetic distance, the base sequences of Gentiana scabra and Gentianamanshurica were very similar, and the interspecific differences were very small. Except for the intraspecific variation of only one sample collected in Liaoning Province, the base sequences of the other samples were the same, and there was no difference between " Gentiana scabra Bge. in Qingyuan" and Gentiana scabra Bge. samples from other regions in Liaoning Province. Conclusion:The DNA barcode technology of ITS sequence can be used to differentiate and identify medicinal plant Gentiana scabra Bge. and its original plants from different sources with a high success rate.

Study on DNA barcoding of Atractylodes chinensis (DC.) Koidz. herbs from Liaoning Province based on rbcL sequences / 国际中医中药杂志

Objective:To use rbcL sequences to identify the rhizomes of the Liaoning collection of Atractylodes chinensis (DC.) Koidz.; To provide a basis for ensuring the feasibility of cultivation of the native herb in Liaoning Province. Methods:A total of 30 rhizomes of Atractylodes chinensis (DC.) Koidz. were collected from 10 regions cultivated in Liaoning Province, and the total DNA was extracted. DNA barcodes were screened by PCR, and the rbcL sequences of the samples were amplified and sequenced, and the amplification and sequencing success rates were calculated. Sequence alignment was performed using MEGA 7.0 software; a systematic clustering tree was constructed using the neighbour-joining method. Results:The success rates of DNA extraction from the rhizomes of Atractylodes chinensis (DC.) Koidz. were all 93.3%, and the success rates of PCR amplification and sequencing were all 100%. Among the 30 samples of Atractylodes chinensis (DC.) Koidz. in Liaoning Province, two samples had intraspecific variation, and the rest of the base sequences of Atractylodes chinensis (DC.) Koidz. were identical. Atractylodes chinensis (DC.) Koidz. was closer to the herbs of the genus Cangzhu, a relative species of Asteraceae, and was genetically more distant from the rest of Asteraceae. The NJ tree could distinguish Atractylodes chinensis (DC.) Koidz. and its relatives. Conclusion:The quality of Atractylodes chinensis (DC.) Koidz. cultivars in Liaoning Province is basically similar, and the rbcL sequence can be used as a valid sequence fragment for the identification of Atractylodes chinensis (DC.) Koidz. DNA barcode.

Cupiennius spiders (Trechaleidae) from southern Mexico: DNA barcoding, venomics, and biological effect

Background: Members of the genus Cupiennius Simon, 1891 are categorized as wandering spiders and are part of the family Trechaleidae. The genomics and proteomics of Cupiennius spiders from North America remain uncharacterized. The present study explores for the first time molecular data from the endemic species Cupiennius chiapanensis Medina, 2006, and also presents new data for Cupiennius salei (Keyserling, 1878), both collected in southern Mexico. Methods: In total, 88 Cupiennius specimens were collected from southern Mexico and morphologically identified. DNA was extracted and the mitochondrial COI fragment was amplified. COI sequences were analyzed, and a phylogenetic tree was inferred for species from the Americas. Genetic diversity was analyzed using haplotype networks and gene distances. Venom was obtained from C. chiapanensis and C. salei by electrostimulation. The venom was separated by HPLC, visualized using SDS-PAGE, and quantified for use in toxicity bioassays in mice and insects. Results: Analysis of COI sequences from C. chiapanensis showed 94% identity with C. salei, while C. salei exhibited 94-97% identity with sequences from Central and South American conspecifics. The venom from C. chiapanensis exhibited toxic activity against crickets. Venoms from C. chiapanensis and C. salei caused death in Anastrepha obliqua flies. Analysis of venom fractions from C. salei and C. chiapanensis revealed molecular masses of a similar size as some previously reported toxins and neurotoxic components. We determined the amino acid sequences of ChiaTx1 and ChiaTx2, toxins that are reported here for the first time and which showed toxicity against mice and insects. Conclusion: Our work is the first to report COI-based DNA barcoding sequences from southern Mexican Cupiennius spiders. Compounds with toxic activity were identified in venom from both species.(AU)

Primeros registros de Echiophis brunneus (Teleosteo; Ophichthidae) "anguila pecosa" en el norte del Perú

Echiophis brunneus, comúnmente conocida como anguila pecosa, es una especie bentónica costera de la familia Ophichthidae. Su distribución se reporta para el Pacífico Oriental desde el Golfo de California (EE. UU.) hasta el Golfo de Guayaquil (Ecuador). Se reporta por primera vez la presencia de E. brunneus en el norte del Perú a partir de tres ejemplares capturados. Así mismo se registra una nueva talla máxima para la especie y se adiciona la secuencia COI a la base de datos BoldSystems. Una de las principales características para su determinación fue la presencia del diente canino grande localizado en la zona distal del vómer. Las distancias genéticas entre E. brunneus con E. punctifer y E. intertinctus fueron de 0.087±0.013 y 0.095±0.014 respectivamente. Con este trabajo se amplía la distribución geográfica de E. brunneus hasta Salaverry (08°13'28"S, 78°59'22"W), así mismo sugerimos el posible establecimiento de una población de esta especie en la costa norte del Perú.

Echiophis brunneus, commonly known as fangjaw eel, is a coastal benthic species belonging to the Ophichthidae family. Its distribution is reported to be in the Eastern Pacific from the Gulf of California (USA) to the Gulf of Guayaquil (Ecuador). In this study, we report for the first time the presence of E. brunneus based on three specimens captured in northern Peru. Additionally, a new maximum size for the species is recorded, and the first COI sequence is added to the BoldSystems database. One of the main characteristics for its determination was the presence of a large canine tooth located in the distal area of the vomer. The genetic distances between E. brunneus with E. punctifer and E. intertinctus were 0.087±0.013 and 0.095±0.014 respectively. With this work the geographical distribution of E. brunneus is extended to Salaverry (08°13'28"S, 78°59'22"W). We also suggest the possible establishment of a population of this species on the northern coast of Peru.

Specific DNA barcodes, germplasm resources, and genetic diversity of Eleutherococcus senticosus / 中国中药杂志

Eleutherococcus senticosus is one of the Dao-di herbs in northeast China. In this study, the chloroplast genomes of three E. senticosus samples from different genuine producing areas were sequenced and then used for the screening of specific DNA barcodes. The germplasm resources and genetic diversity of E. senticosus were analyzed basing on the specific DNA barcodes. The chloroplast genomes of E. senticosus from different genuine producing areas showed the total length of 156 779-156 781 bp and a typical tetrad structure. Each of the chloroplast genomes carried 132 genes, including 87 protein-coding genes, 37 tRNAs, and 8 rRNAs. The chloroplast genomes were relatively conserved. Sequence analysis of the three chloroplast genomes indicated that atpI, ndhA, ycf1, atpB-rbcL, ndhF-rpl32, petA-psbJ, psbM-psbD, and rps16-psbK can be used as specific DNA barcodes of E. senticosus. In this study, we selected atpI and atpB-rbcL which were 700-800 bp and easy to be amplified for the identification of 184 E. senticosus samples from 13 genuine producing areas. The results demonstrated that 9 and 10 genotypes were identified based on atpI and atpB-rbcL sequences, respectively. Furthermore, the two barcodes identified 23 genotypes which were named H1-H23. The haplotype with the highest proportion and widest distribution was H10, followed by H2. The haplotype diversity and nucleotide diversity were 0.94 and 1.82×10~(-3), respectively, suggesting the high genetic diversity of E. senticosus. The results of the median-joining network analysis showed that the 23 genotypes could be classified into 4 categories. H2 was the oldest haplotype, and it served as the center of the network characterized by starlike radiation, which suggested that population expansion of E. senticosus occurred in the genuine producing areas. This study lays a foundation for the research on the genetic quality and chloroplast genetic engineering of E. senticosus and further research on the genetic mechanism of its population, providing new ideas for studying the genetic evolution of E. senticosus.

Phylogeography of cliff racer (Platyceps rhodorachis Jan, 1865) from Punjab, Pakistan / Filografia de penhasco (Platyceps rhodorachis Jan, 1865) de Punjab, Paquistão

The present study reports the existence of cliff racer, Platyceps rhodorachis from the plains of Punjab, Pakistan. A total of 10 specimens were captured during the field surveys from June to September, 2018 from different sites of Punjab. Platyceps rhodorachis was identify on the basis of morphology and confirmed through COI gene sequences. The obtained DNA sequences have shown reliable and exact species identification. Newly produced DNA sequences of Platyceps rhodorachis were submitted to GenBank and accession numbers were obtained (MK936174.1, MK941839.1 and MT790210.1). N-J tree based on COI sequences of Platyceps rhodorachis clearly separated as out-group with other members of family Colubridae based on p-distance. The intra-specific genetic variation ranges from 12% to 18%. The DNA sequences of Platyceps rhodorachis kashmirensis, Platyceps rhodorachis ladacensis, Platyceps ventromaculatus, Platyceps ventromaculatus bengalensis and Platyceps ventromaculatus indusai are not available at NCBI to validate their taxonomic positions. In our recommendations, a large scale molecular based identification of Pakistan’s herpetofauna is required to report more new or subspecies from country.

O presente estudo relata a existência de um corredor de penhasco, Platyceps rhodorachis, das planícies de Punjab, Paquistão. Um total de 10 espécimes foi capturado durante os levantamentos de campo de junho a setembro de 2018 em diferentes locais de Punjab. Platyceps rhodorachis foi identificada com base na morfologia e confirmada por meio de sequências do gene COI. As sequências de DNA obtidas mostraram identificação de espécies confiável e exata. Sequências de DNA de Platyceps rhodorachis recém-produzidas foram submetidas ao GenBank e os números de acesso foram obtidos (MK936174.1, MK941839.1 e MT790210.1). Árvore N-J baseada em sequências COI de Platyceps rhodorachis claramente separadas como out-group com outros membros da família Colubridae com base na distância-p. A variação genética intraespecífica varia de 12% a 18%. As sequências de DNA de Platyceps rhodorachis kashmirensis, Platyceps rhodorachis ladacensis, Platyceps ventromaculatus, Platyceps ventromaculatus bengalensis e Platyceps ventromaculatus indusai não estão disponíveis no NCBI para validar suas posições taxonômicas. Em nossas recomendações, uma identificação de base molecular em grande escala da herpetofauna do Paquistão é necessária para relatar mais novas ou subespécies do país.

Phylogeography of cliff racer (Platyceps rhodorachis Jan, 1865) from Punjab, Pakistan

Abstract The present study reports the existence of cliff racer, Platyceps rhodorachis from the plains of Punjab, Pakistan. A total of 10 specimens were captured during the field surveys from June to September, 2018 from different sites of Punjab. Platyceps rhodorachis was identify on the basis of morphology and confirmed through COI gene sequences. The obtained DNA sequences have shown reliable and exact species identification. Newly produced DNA sequences of Platyceps rhodorachis were submitted to GenBank and accession numbers were obtained (MK936174.1, MK941839.1 and MT790210.1). N-J tree based on COI sequences of Platyceps rhodorachis clearly separated as out-group with other members of family Colubridae based on p-distance. The intra-specific genetic variation ranges from 12% to 18%. The DNA sequences of Platyceps rhodorachis kashmirensis, Platyceps rhodorachis ladacensis, Platyceps ventromaculatus, Platyceps ventromaculatus bengalensis and Platyceps ventromaculatus indusai are not available at NCBI to validate their taxonomic positions. In our recommendations, a large scale molecular based identification of Pakistans herpetofauna is required to report more new or subspecies from country.

Resumo O presente estudo relata a existência de um corredor de penhasco, Platyceps rhodorachis, das planícies de Punjab, Paquistão. Um total de 10 espécimes foi capturado durante os levantamentos de campo de junho a setembro de 2018 em diferentes locais de Punjab. Platyceps rhodorachis foi identificada com base na morfologia e confirmada por meio de sequências do gene COI. As sequências de DNA obtidas mostraram identificação de espécies confiável e exata. Sequências de DNA de Platyceps rhodorachis recém-produzidas foram submetidas ao GenBank e os números de acesso foram obtidos (MK936174.1, MK941839.1 e MT790210.1). Árvore N-J baseada em sequências COI de Platyceps rhodorachis claramente separadas como out-group com outros membros da família Colubridae com base na distância-p. A variação genética intraespecífica varia de 12% a 18%. As sequências de DNA de Platyceps rhodorachis kashmirensis, Platyceps rhodorachis ladacensis, Platyceps ventromaculatus, Platyceps ventromaculatus bengalensis e Platyceps ventromaculatus indusai não estão disponíveis no NCBI para validar suas posições taxonômicas. Em nossas recomendações, uma identificação de base molecular em grande escala da herpetofauna do Paquistão é necessária para relatar mais novas ou subespécies do país.

Taxonomía de los hongos: un rompecabezas al que le faltan muchas piezas / Fungal taxonomy: A puzzle with many missing pieces

Los hongos son organismos polifacéticos presentes en casi todos los ecosistemas de la tierra, donde establecen diversos tipos de simbiosis con otros seres vivos. A pesar de ser reconocidos por los humanos desde la antigüedad -y de la cantidad de trabajos que han profundizado sobre su biología y ecología-, aún falta mucho por conocer sobre estos organismos. Algunos de los criterios que clásicamente se han utilizado para su estudio, hoy resultan limitados y hasta cierto punto permiten un agrupamiento de los aislamientos según algunas características, pero generan confusión en su clasificación y, más aún, cuando se pretende comprender sus relaciones genealógicas. Los caracteres fenotípicos no son suficientes para identificar una especie de hongos y, menos aún, para construir una filogenia amplia o de un grupo particular. Hay grandes vacíos que hacen que los árboles generados sean inestables y fácilmente debatidos. Para los profesionales de la salud, parece que la identificación de los hongos hasta niveles inferiores como género y especie es suficiente para elegir el tratamiento más adecuado para su control, comprender la epidemiología de los cuadros clínicos asociados y reconocer los brotes y los factores determinantes de la resistencia a los antimicrobianos. No obstante, la ubicación taxonómica dentro del reino permitiría establecer relaciones filogenéticas entre los taxones fúngicos, facilitando la comprensión de su biología, su distribución en la naturaleza y la evolución de su potencial patogénico. Los avances de las técnicas de biología molecular y las ciencias de la computación en los últimos 30 años han permitido cambios importantes dirigidos a establecer los criterios para definir una especie fúngica y alcanzar una construcción filogenética más o menos estable. Sin embargo, el camino por recorrer aún es largo, y supone un trabajo mancomunado de la comunidad científica a nivel global y el apoyo a la investigación básica.

Fungi are multifaceted organisms found in almost all ecosystems on Earth, where they establish various types of symbiosis with other living beings. Despite being recognized by humans since ancient times, and the high number of works delving into their biology and ecology, much is still unknown about these organisms. Some criteria classically used for their study are nowadays limited, generating confusion in categorizing them, and even more, when trying to understand their genealogical relationships. To identify species within Fungi, phenotypic characters to date are not sufficient, and to construct a broad phylogeny or a phylogeny of a particular group, there are still gaps affecting the generated trees, making them unstable and easily debated. For health professionals, fungal identification at lower levels such as genus and species, is enough to select the most appropriate therapy for their control, understand the epidemiology of clinical pictures associated, and recognize outbreaks and antimicrobial resistance. However, the taxonomic location within the kingdom, information with apparently little relevance, can allow phylogenetic relationships to be established between fungal taxa, facilitating the understanding of their biology, distribution in nature, and pathogenic potential evolution. Advances in molecular biology and computer science techniques from the last 30 years have led to crucial changes aiming to establish the criteria to define a fungal species, allowing us to reach a kind of stable phylogenetic construction. However, there is still a long way to go, and it requires the joint work of the scientific community at a global level and support for basic research.

First record of the invasive dwarf surf clam Mulinia lateralis (Say 1822) in the eastern Pacific Ocean

Mulinia lateralis is a native bivalve from the Western Atlantic Ocean, distributed from the Gulf of Saint Lawrence in Canada to Yucatan in Mexico. Based on morphological and genetic data of specimens collected in shrimp farms, in this work, we confirm the presence of M. lateralis in the Gulf of Guayaquil, Ecuador. Presence and its consequences of this invasive bivalve in the region is discussed.

Mulinia lateralis es un bivalvo nativo de las aguas del Océano Atlántico Occidental, distribuido desde el Golfo de Saint Lawrence en Canadá hasta Yucatán en México. En este trabajo, la presencia de M. lateralis es confirmada en el Golfo de Guayaquil, Ecuador, con base en datos morfológicos y genéticos de ejemplares colectados en camaroneras. Se presenta una discusión sobre la presencia y consecuencias de este bivalvo invasor en la región.

Andrographis Echioides Molecular Characterization and Phylogenetic Study Using DNA Barcoding Technique

With the advancement in various molecular diagnostic tools, DNA Barcoding has emerged as a gold standard molecular diagnostic tool across the globe. Since ancient times, medicinal plants have been widely used in Indian Ayurvedic medicine for treating a variety of ailments. Plants of the genus Andrographis have been extensively used for treating different types of ailments. In this study, rarely studied medicinal plant species were isolated, sequenced at the genetic level and studied for their evolutionary characteristics using phylogenetic analysis. In the present study, the identity of A. echioides was confirmed by targeting different barcoding genes such as ribulose-bisphosphate carboxylase, internal transcribed spacer, RNA polymerase-beta subunit, maturase K, and photosystem II protein D1 genes using a phylogenetic approach. After successful isolation and amplification of genomic DNA, specific primers were utilised for sequencing of each barcoding gene, followed by nucleotide BLAST analysis to determine the sequence percent identity of each gene with that from other plant species. The best homologs were then utilised for conducting phylogenetic analysis which confirmed the identity of the plant as Andrographis echioides.

Ethyl Acetate Extract of Helicteres hirsuta Suppresses MCF-7 Human Breast Cancer Cell Mobility

Objective: To investigate the anticancer activity of Helicteres hirsuta (H. hirsute) extract against a breast cancer cell line- MCF7. Methods: H. hirsute was extracted in absolute methanol. Further, the crude extract was further partitioned in n-hexane, chloroform and ethyl acetate. The total phenolic and flavonoid content were determined by using spectroscopic methods with gallic acid and quercetin standard compounds, respectively. Anticancer activities of H. hirsuta extract was elucidated by MTT, wound headling, and transwell invasion assays. Results: Total phenolic compounds in H. hirsute extracts reached 22.07 � 2.54, 58.57 5.54 and 235.56 � 7.54 mg GAE/g in each n hexan, chloroform, ethyl acetate fractions, respectively. Whereas the ethyl acetate fraction showed the greatest phenolic contents with 235.56 � 7.54 mg GAE/g. Moreover, the flavonoid contents of H. hirsute extracts reached 5.76 � 0.94, 9.25 � 1.84 and 19.37 � 2.57 mg quercetin equivalent/g in each n hexan, chloroform, ethyl acetate fractions, respectively, in which, the ethyl acetate fraction also showed the highest amounts of flavonoid contents. Further, the ethyl acetate of H. hirsute significantly decreased the viability of breast cancer MCF7 cells after 48 h treatment with IC50 value of 95 � 2.54 compared to control. In addition, the ethyl acetate extract of H. hirsute suppressed the invasion and migration of MCF7 cell lines in a dose-dependent manner at non-toxic concentrations. Conclusions: The ethyl acetate of H. hirsute suppressed the growth and motility of breast cancer MCF7 cells.

Identification of Andrographis Herba and its common products using mini-barcode / 中国天然药物

Andrographis Herba, the aerial part of Andrographis paniculata (Burm. f.) Wall. ex Nees (Acanthaceae), has a wide geographic distribution and has been used for the treatment of fever, cold, inflammation, and other infectious diseases. In markets, sellers and buyers commonly inadvertently confuse with related species. In addition, most Chinese medicinal herbs are subjected to traditional processing procedures, such as steaming and boiling, before they are sold at dispensaries; therefore, it is very difficult to identify Andrographis Herba when it is processed into Chinese medicines. The identification of species and processed medicinal materials is a growing issue in the marketplace. However, conventional methods of identification have limitations, while DNA barcoding has received considerable attention as a new potential means to identify species and processed medicinal materials. In this study, 17 standard reference materials of A. paniculata, 2 standard decoctions, 27 commercial products and two adulterants were collected. Based on the ITS2 sequence, it could successfully identify A. paniculata and adulterants. Moreover, a nucleotide signature consisting of 71 bp was designed, this sequence is highly conserved and specific within A. paniculata while divergent among other species. Then, we used these new primers to amplify the nucleotide signature region from processed materials. In conclusion, the DNA barcoding method developed in the present study for authenticating A. paniculata is rapid and cost-effective. It can be used in the future to guarantee the quality of Andrographis Herba of each regulatory link for clinical use.

Molecular Identification of Tibetan Codonopsis Plants Based on ITS2 and psbA-trnH Barcode / 中国实验方剂学杂志

ObjectiveTo identify the molecular biology of various species of Tibetan Codonopsis plants based on internal transcribed spacer（ITS）2 and psbA-trnH sequence barcode technology. MethodThe genomic DNA of 28 Tibetan Codonopsis plant samples from four species （Codonopsis canescens，C. foetens subsp. nervosa，C. pilosula， and C. thalictrifolia var. mollis） were extracted，and the ITS2 and psbA-trnH sequences were amplified and sequenced. The related sequences of 81 Tibetan Codonopsis plant samples belonging to 15 species were downloaded from GenBank， and MEGA 6.0 was used for sequence comparison and mutation site analysis. The GC content and genetic distance within and between species were calculated. Additionally， phylogenetic trees were constructed by maximum likelihood （ML） method， neighbor-joining （NJ） method，and unweighted pair-group method with arithmetic means （UPGMA） . ResultAccording to the mutation site，C. canescens， C. pilosula，C. pilosula subsp. tangshen， C. pilosula var. modesta，C. bhutanica，C. clematidea，C. lanceolata，C. subglobosa and C. foetens were distinguished. In the phylogenetic trees，the optimal clustering effects for ITS2 and psbA-trnH sequences were obtained using the ML method and the UPGMA method， respectively， and 12 species were effectively clustered. ConclusionITS2 and psbA-trnH sequences have a high identification rate for species of single origin，but there are still some limitations in identifying variants and original variants. This study provides basis for the identification of affinity relationship and clinical safety of Tibetan Codonopsis plants.

Identification and Quality Evaluation of Dendrobium flexicaule and Its Related Species / 中国实验方剂学杂志

ObjectiveTo identify Dendrobium flexicaule and its related species， and analyze the differences in polysaccharide composition and D-mannose content， so as to provide theoretical basis for the accurate identification and quality control of Dendrobium medicinal materials. MethodNine samples of Dendrobium （S1-S9） were identified by DNA barcoding and infrared spectroscopy， and the contents of polysaccharides and D-mannose were determined by ultraviolet spectrophotometry （UV） and high performance liquid chromatography （HPLC）， respectively. UV detection condition was 488 nm， HPLC detection conditions were the mobile phase of 20 mmol·L-1 ammonium acetate solution-acetonitrile （81.5∶18.5） and the detection wavelength at 250 nm. ResultDNA barcoding results showed that samples S1-S3 were D. nobile， samples S4-S5 were D. officinale， sample S6 was D. huoshanense， and S7-S9 were D. flexicaule. One-dimensional infrared spectroscopy showed that only D. nobile had stable characteristics at the wavenumber of 1 570-1 467 cm-1， showing a "W" shape， while no absorption peak was found at the wavenumber of 842-740 cm-1， but the other Dendrobium samples had stable absorption peaks at the wavenumber of 842-740 cm-1. In the first derivative spectrum， at the wavenumber of 785 cm-1， D. huoshanense presented a "V" shape， while the rest of Dendrobium presented a "W" shape. At the wavenumber of 1 110 cm-1， D. flexicaule had a stable characteristic peak. In the second derivative spectrum， at the wavenumber of 1 125 cm-1， D. officinale presented an "M" shape， and the rest of Dendrobium was approximately "W" shape. The results of determination showed that the contents of polysaccharides in samples S1-S9 were 9.35%， 9.12%， 32.78%， 49.38%， 48.97%， 32.48%， 32.95%， 39.41% and 25.32%， and their contents of D-mannose were 1.39%， 0.47%， 13.57%， 3.04%， 33.85%， 23.57%， 16.64%， 17.47% and 19.49%， respectively. Among them， D. flexicaule had high polysaccharide and D-mannose contents. ConclusionBoth DNA barcoding and infrared spectroscopy can be used to identify D. flexicaule and its related species， and infrared spectroscopy is cost-effective and easy to operate. At the same time， D. flexicaule has high contents of polysaccharides and D-mannose， which can provide a scientific basis for rapid identification of D. flexicaule and its relatives， and provides a reference for its quality control， and resource development and utilization.

DNA barcoding of Gentiana crassicaulis in Lijiang and evaluation of root processing methods / 药学学报

The key factors for producing the best quality Chinese herbal medicines are high-quality germplasm, suitable cultivation area and the proper processing methods for herbal raw materials. Gentiana crassicaulis in Gentiana (Sect. Cruciata), Gentianaceae is one of the original plants of the Chinese herb Qinjiao (Gentianae Macrophyllae Radix), and its type specimen was collected in Lijiang, Yunnan. There is a long planting history of the herb in this area. In this study a sampling plot was designated in these traditional planting areas. G. crassicaulis was planted and herbal raw materials were harvested from the plot. The raw materials were prepared locally and at a pharmaceutical factory in Shanghai using processing methods such as "sweating" or "no sweating", "slicing" or "no slicing" (whole root), and "stoving" or "no stoving" (air drying). The quality of all processed samples was evaluated. In addition, molecular markers were determined for identifying cultivated and wild samples from Lijiang, Yunnan. The results are as follows: ① Samples from the sampling plot and the field are taxonomically identified as Gentiana crassicaulis. ② A total of 270 sequences of trnC-GCA-petN, atpB-rbcL, psbN, ndhB-rps7 and ycf1 were obtained, and three genotypes were determined from the cultivated samples; the type III was shared by both cultivated and wild plants. Based on the molecular markers, a DNA barcoding method to identify cultivated and wild samples of G. crassicaulis from Lijiang, Yunnan was established. ③ Total content of loganic acid and gentiopicroside in all samples was ≥ 2.5%, and above the Chinese Pharmacopoeia (2020) limit. ④ In HPLC fingerprinting, 9 common peaks were assigned and similarity between all samples was > 0.999; and ⑤ In a PCA score plot all slice samples were clustered, while whole root samples were scattered. Therefore, our studies could provide basic data for optimizing the processing method, producing best quality Gentianae Macrophyllae Radix, and evaluating the quality of different ecotype varieties and the multiple origin of herbal medicines.

Application and progress in high-throughput sequencing technology for meat adulteration detection / 生物工程学报

Adulteration in meat products is a widespread issue that could lead to serious threats to public health and religious violations. Technology that offers rapid, sensitive, accurate and reliable detection of meat species is the key to an effectual monitoring and control against meat adulteration. In recent years, high-throughput sequencing-based DNA metabarcoding technology has developed rapidly. With the characteristics of being high-throughput, highly precise and high-speed, this technology can simultaneously identify multiple species in complex samples, thus offering pronounced advantages in the surveillance of adulteration in meat and meat products. Starting with an introduction of the major developments in the high-throughput sequencing technology in the past two decades, this review provides an overview of the technical characteristics and research methods of DNA metabarcoding, summarizes the application of DNA metabarcoding technology in meat adulteration detection over the last few years, discusses the challenges of using DNA metabarcoding technology in the detection of meat adulteration, and provides future prospects on the development of this technology.

Complete chloroplast genome of Ligustrum lucidum and highly variable marker identification for Ligustrum / 中国中药杂志

Ligustri Lucidi Fructus, the sun-dried mature fruit of Ligustrum lucidum, is cool, plain, sweet, and bitter, which can be used as both food and medicine, with the effects of improving vision, blacking hair, and tonifying liver and kidney. It takes effect slowly. However, little is known about the genetic information of the medicinal plant and it is still a challenge to distinguish Ligustrum species. In this study, the complete chloroplast genome of L. lucidum was obtained by genome skimming and then compared with that of five other Ligustrum species, which had been reported. This study aims to evaluate the interspecific variation of chloroplast genome within the genus and develop molecular markers for species identification of the genus. The result showed that the chloroplast genome of L. lucidum was 162 162 bp with a circular quadripartite structure of two single-copy regions separated by a pair of inverted repeats. The Ligustrum chloroplast genomes were conserved with small interspecific difference. Comparative analysis of six Ligustrum chloroplast genomes revealed three variable regions(rbcL-accD, ycf1a, and ycf1b), and ycf1a and ycf1b can be used as the species-specific DNA barcode for Ligustrum. Phylogeny analysis provided the best resolution of Ligustrum and supported that L. lucidum was sister to L. gracile. This study clarified the genetic diversity of L. lucidum from provenance, which can serve as a reference for further analysis of pharmacological differences and breeding of excellent varieties with stable drug effects.

DNA barcoding revels first records of three rare coleopteran genera in Northern lakes of Egypt / O código de barras de DNA revela os primeiros registros de três gêneros raros de coleópteros nos lagos do norte do Egito

Abstract One aquatic coleopteran species from family Dytiscidae and two aquatic coleopteran genera from family Hydrophilidae were recorded in the summer period and represent first records in the Egyptian lakes. Beetles were collected from two northern lakes, Lake Idku and Lake Burullus. They were identified by morphological characteristics as well as the mtDNA barcoding method. A molecular phylogenetic approach was used to determine the genetic identity of the collected samples based on the mitochondrial cytochrome oxidase I (COI). Prodaticus servillianus (Dytiscidae) from Egypt showed no significant difference in the COI region and they are highly similar to P. servillianus from Madagascar. The phylogenetic analysis revealed that the other two coleopteran genera belong to family Hydrophilidae. Based on COI only, there is no clear evidence for their genetic identity at the species level. So, we defined them to the closest taxon and denoted them as Cymbiodyta type A and B. The results indicated that resolving the molecular identity of the aquatic beetles from northern lakes of Egypt need more considerations in the field of biological conservation. We concluded that utilization of COI as a barcoding region for identifying some coleopteran species is not sufficient and additional molecular markers are required to uncover the molecular taxonomy at deep levels.

Resumo Uma espécie de coleópteros aquático da família Dytiscidae e dois gêneros de coleópteros aquáticos da família Hydrophilidae foram registrados no período de verão e representam os primeiros registros nos lagos egípcios. Os besouros foram coletados em dois lagos do norte, o lago Idku e o lago Burullus, e identificados por características morfológicas e pelo método de código de barras mtDNA. Uma abordagem filogenética molecular foi usada para determinar a identidade genética das amostras coletadas com base no citocromo oxidase I mitocondrial (COI). Prodaticus servillianus (Dytiscidae) do Egito não mostrou diferença significativa na região COI e é altamente semelhante a P. servillianus de Madagascar. A análise filogenética revelou que os outros dois gêneros de coleópteros pertencem à família Hydrophilidae. Com base apenas no COI, não há evidências claras de sua identidade genética no nível da espécie. Assim, nós os agrupamos no táxon mais próximo e os denominamos Cymbiodyta tipo A e B. Os resultados indicaram que a identidade molecular dos besouros aquáticos dos lagos do norte do Egito precisa de mais considerações no campo da conservação biológica. Concluímos que a utilização de COI como região de código de barras para identificar algumas espécies de coleópteros não é suficiente, sendo necessários marcadores moleculares adicionais para descobrir a taxonomia molecular em níveis profundos.

Mutualistic interaction of aphids and ants in pepper, Capsicum annuum and Capsicum frutenscens (Solanaceae) / Interacción mutualista de pulgones y hormigas en pimiento, Capsicum annuum y Capsicum frutenscens (Solanaceae)

Abstract Introduction: Adequate biological identification is fundamental for establishing integrated pest management programs and identifying the trophic and mutualist relationships that can affect pest population dynamics. Aphids are the main pest of pepper Capsicum spp. (Solanaceae) crops in Southwestern Colombia, due to their role as vectors of viruses. However, the identification of aphid species is complex, limiting the investigations performed to address their interactions with other organisms. Ants and aphids present a facultative mutualistic relationship, that promotes the growth of hemipteran colonies, for this reason, the study of the ecological mutualistic association between aphids and ants is important. Objective: The main objective was to discriminate the aphid species present in commercial crops of Capsicum spp., and to identify the ant community that attends the aphid colonies and its effects on the size of the aphid colonies. Methods: Aphid species, and their ant mutualist, were collected from Capsicum annuum and Capsicum frutescens, in the Cauca valley, Southwestern Colombia. We used the DNA barcoding approach to identify aphid species, and the ants were identified by morphology-based taxonomy. To evaluate the effect of ant care on the size and structure of aphid colonies, generalized linear models were calculated using as the response variables the total number of aphids for each colony and the proportion of nymphs. Results: The aphid species that attack pepper crops, are: Aphis gossypii and Myzus persicae (Hemiptera: Aphididae), with A. gossypii being the species that interacts with ants (19 ant species). A. gossypii colonies attended by ants had larger sizes and more nymphs per colony, than those not attended. Conclusions: Although the aphid-ant interaction is not species-specific, it is necessary to consider its role in the propagation of viral diseases in peppers and to determine how this interaction may affect regional biological control strategies.

Resumen Introducción: La adecuada identificación biológica es fundamental para establecer programas de manejo integrado de plagas e identificar las relaciones tróficas y mutualistas que pueden afectar la dinámica poblacional de insectos plaga. Los áfidos son las principales plagas del ají Capsicum spp. (Solanaceae) en el suroccidente colombiano, debido a su rol como vectores de virus. Sin embargo, su identificación es compleja, y limita las investigaciones que intentan revelar sus interacciones con otros organismos. Las hormigas y los áfidos presentan una relación mutualista facultativa, que promueve el crecimiento de las colonias de los hemípteros, por esta razón, el estudio de la asociación ecológica y mutualista entre áfidos y hormigas es importante. Objetivo: El principal objetivo de esta investigación fue discriminar las especies de áfidos presentes en cultivos comerciales de Capsicum spp., e identificar la comunidad de hormigas que atiende las colonias de áfidos y su efecto en el tamaño de las colonias de áfidos. Métodos: Los áfidos, y las hormigas mutualistas de estos áfidos, se recolectaron de Capsicum annuum y Capsicum frutescens, en el valle del rio Cauca, en el suroccidente colombiano. Se empleó el Código de barras del ADN para identificar las especies de áfidos, y las hormigas se identificaron empleando taxonomía basada en morfología. Para evaluar el efecto que tiene el cuidado de las hormigas sobre el tamaño de las colonias de áfidos, se empleó un modelo lineal generalizado, utilizando como variables de respuesta el número total de áfidos por cada colonia y la proporción de ninfas por colonia. Resultados: Las especies de áfidos que atacan los cultivos de ají, son: Aphis gossypii y Myzus persicae (Hemiptera: Aphididae), siendo A. gossypii la especie que interactúa con hormigas (19 especies). Las colonias de A. gossypii atendidas por hormigas presentan mayor tamaño y número de ninfas, que aquellas desatendidas. Conclusiones: Aunque la interacción áfido-hormiga no es especie específica, es necesario considerar su rol en la propagación de enfermedades virales en plantas cultivadas y determinar cómo esta interacción puede afectar la implementación de estrategias de control biológico.

Molecular analysis reveals high diversity in the Hoplias malabaricus (Characiformes, Erythrinidae) species complex from different Amazonian localities

ABSTRACT DNA barcoding proposes that a fragment of DNA can be used to identify species. In fish, a fragment of cytochrome oxidase subunit I (COI) has been effective in many studies with different foci. Here we use this molecular tool to provide new insights into the cryptic diversity found in the Hoplias malabaricus species complex. Popularly known as trahira, H. malabaricus is widely distributed in South America. The clade shows molecular and cytogenetic diversity, and several studies have supported the occurrence of a complex of species. We performed molecular and karyotypic analysis of H. malabaricus individuals from eight Amazonian localities to assess the diversity present in the nominal taxon, and to clarify relationships within this group. We used 12 samples in cytogenetic analyses and found two karyomorphs: 2n = 40 (20m + 20sm) (karyomorph C) and 2n = 42 (22m + 20sm) (karyomorph A). We used 19 samples in molecular analyses with COI as a molecular marker, maximum likelihood analyses, and the Kimura-2-parameter evolutionary model with bootstrap support. We found karyomorph-related differentiation with bootstrap of 100%. However, we found high molecular diversity within karyomorph C. The observed pattern allowed us to infer the presence of cryptic diversity, reinforcing the existence of a species complex.

RESUMO O DNA barcoding propõe que um fragmento de DNA possa servir para identificar espécies. Em peixes, um fragmento do gene COI tem se mostrado eficaz em muitos estudos com focos diferentes. Nós usamos essa ferramenta molecular para fornecer novas informações sobre a diversidade críptica encontrada no complexo de espécies Hoplias malabaricus. Popularmente conhecida como traíra, H. malabaricus tem uma ampla distribuição na América do Sul. Esse clado mostra diversidade molecular e citogenética, e vários estudos dão suporte à ocorrência de um complexo de espécies. Realizamos análises molecular e cariotípica em indivíduos de H. malabaricus de oito localidades amazônicas, para acessar a diversidade no taxon nominal e elucidar as relações nesse grupo. Usamos 12 amostras em análises citogenéticas e encontramos dois cariomorfos: 2n = 40 (20m + 20sm) (cariomorfo C) e 2n = 42 (22m + 20sm) (cariomorfo A). Usamos 19 amostras em análise molecular, utilizando COI como marcador molecular, análises de máxima verossimilhança e o modelo evolutivo de Kimura-2-parâmetros com estimativa de bootstrap. Encontramos diferenciação relacionada aos cariomorfos com bootstrap de 100%. No entanto, encontramos alta diversidade molecular no cariomorfo C. O padrão observado nos permitiu inferir a presença de diversidade oculta, reforçando a existência de um complexo de espécies.