STMS markers related to Ascochyta blight resistance in chickpea

Chickpea (Cicer arietinum L.) is one of the important legume crops and is cultivated in large-scale throughout Türkiye as well as the world. Ascochyta blight, caused by the fungal phytopathogen Ascochyta rabiei, is the leading reason for the highest yield losses among the diseases known for chickpea. The pathogen exhibits high genetic diversity in Türkiye. Therefore, resistancy using Sequence Tagged Microsatellite Site (STMS) markers related with the genes that provide resistant against Ascochyta blight was investigated for the 205 chickpea breeding lines grown in different parts of Türkiye. The analysis for Ascochyta blight resistance was performed using Ta2, Ta146 and Ts54. It was demonstrated that Ta2, Ts54 and Ta146 were the STMS markers having distinguishable features for the detection of Ascochyta blight resistance and were shown to be used in credible fashion for the selection of resistant chickpea breeding lines.

Dried blood spots: a robust tool for malaria surveillance in countries targeting elimination

With the advancements in analytical and molecular techniques, Dried Blood Spots (DBS) are re-emerging as attractive and cost-effective alternatives for global health surveillance. The use of DBS has been well-characterized in the neonatal screening of metabolic diseases, therapeutic screening as well as in epidemiological studies for biomonitoring. Malaria is one such infectious disease where DBS use can expedite molecular surveillance for assessing drug resistance and for refining drug usage policies. In India, malaria cases have reduced significantly over the past decade but to achieve malaria elimination by 2030, country-wide DBS-based screening should be conducted to identify the presence of molecular markers of artemisinin resistance and to study parasite reservoirs in asymptomatic populations. DBS has wide applications in genomics, proteomics, and metabolomic studies concerning both host and pathogen factors. Hence, it is a comprehensive tool for malaria surveillance that can capture both host and parasite information. In this review, we elucidate the current and prospective role of DBS in malaria surveillance and its applications in studies ranging from genetic epidemiology, parasite and vector surveillance, drug development and polymorphisms to ultimately how they can pave the roadmap for countries aiming malaria elimination

Progress on basic research and clinical evaluation of novel molecular markers of liver cancer / 中华临床感染病杂志

Primary liver cancer includes three types: Hepatocellular carcinoma, intrahepatic cholangiocarcinoma, mixed hepatocellular carcinoma and cholangiocarcinoma. Among them, hepatocellular carcinoma accounts for 75% to 85%, posing a serious threat to human life and health. The screening and monitoring of high-risk populations for hepatocellular carcinoma is crucial for early detection, diagnosis, and treatment, as well as for improving the prognosis of liver cancer. Serum biomarkers play an important role in monitoring and diagnosing hepatocellular carcinoma. In recent years, new serum biomarkers such as AFP heterogeneity, abnormal prothrombin/de-γ-carboxyprothrombin, Golgi protein 73, Dickkopf-associated protein 1, aldehyde ketone reductase-AKR1B10, gypican 3, liquid biopsies and microRNAs have been recommended for screening and monitoring hepatocellular carcinoma, and some have been included as auxiliary diagnostic measures in liver cell carcinoma guidelines. This article summarizes the progress of relevant basic research and clinical evaluation of these novel biomarkers, which may provide a reference for future clinical application.

Clinical value of combined detection of thrombus precursor protein and P-selectin in the diagnosis of acute coronary syndrome / Valor clínico de la detección combinada de proteína precursora de trombo y selectina P en el diagnóstico del síndrome coronario agudo

Abstract Acute coronary syndrome (ACS), including acute myocardial infarction (AMI) and unstable angina (UA), is the most threatening and lethal form of coronary heart disease. ACS has an abrupt onset and rapid development, which may lead to fatal conditions at any time. Thus, it is never too early to detect and diagnose patients with ACS. The objective of this work was to explore the significance of the combined detection of plasma thrombus precursor protein (TpP) and serum P-selectin (Ps), in the detection and diagnosis of patients with early ACS. A total of 126 subjects were included in the study, 64 ACS patients, 30 individuals with stable angina (SA) and 32 healthy persons who were selected as the control groups. There were no differences in gender, age, ethnicity, or blood glucolipid levels among the groups. Enzyme linked immunosorbent assay (Elisa) was used to quantitatively determine the plasma levels of TpP and Ps. The levels of the two biomarkers in the case group were significantly higher than those in the control groups. Among the ACS patients, the levels of TpP and Ps were higher in AMI patients than in the UA patients. In addition, there was no significant differences in the levels of Ps between SA patients and healthy persons. In conclusion, plasma TpP and serum Ps are remarkably increased in patients with ACS. Therefore, TpP and Ps may serve as ACS indicators, and their measurement may provide a support for an early clinical identification of ACS.

Resumen El síndrome coronario agudo (SCA), que incluye el infarto agudo de miocardio (IAM) y la angina inestable (AI), es la forma más amenazante y letal de enfermedad coronaria. El SCA tiene un inicio abrupto y un desarrollo rápido, lo que puede conducir a condiciones fatales en cualquier momento. Por lo tanto, nunca es demasiado pronto para detectar y diagnosticar pacientes con SCA. El objetivo de este trabajo fue explorar la importancia de la detección combinada de la proteína precursora de trombos plasmáticos (TpP) y la selectina P sérica (Ps), en la detección y diagnóstico de pacientes con SCA precoz. Se incluyeron en el estudio un total de 126 sujetos, 64 pacientes con SCA, 30 individuos con angina estable (AE) y 32 personas sanas que fueron seleccionadas como grupos de control. No hubo diferencias en el género, la edad, el origen étnico o los niveles de glucolípidos en sangre entre los grupos. Se usó el ensayo inmunoabsorbente ligado a enzimas (Elisa) para determinar cuantitativamente los niveles plasmáticos de TpP y Ps. Los niveles de los dos biomarcadores en el grupo de casos (SCA) fueron significativamente más altos que los de los grupos de control. Entre los pacientes con SCA, los niveles de TpP y Ps fueron más altos en los pacientes con IAM que en los pacientes con AI. Además, no hubo diferencias significativas en los niveles de Ps entre pacientes con SA y personas sanas. En conclusión, la TpP plasmática y la Ps sérica están notablemente aumentadas en pacientes con SCA. Por lo tanto, TpP y Ps pueden servir como indicadores de SCA y su medición puede proporcionar un apoyo para una identificación clínica temprana de SCA.

Molecular characterization of the severe falciparum malaria with typhoid co-infection A case report

Malaria and typhoid co-infections can be a serious public health issue in tropical countries leading to incorrect diagnosis due to overlapping clinical presentations of malaria and typhoid and hence, causing a delay in implementing the appropriate treatment regimen for these concurrent infections. This study reports a case of six-year-old female child co-infected with severe malaria (Plasmodium falciparum) and typhoid (Salmonella typhi) diagnosed by rapid malaria antigen test (RMAT) and blood culture respectively. Further, analysis of the chloroquine resistance gene Pfcrt for the falciparum demonstrated the presence of K76T mutant allele in pfcrt gene with high IC50 (150nM) for chloroquine (CQ) drug. The present case highlights the significance of timely identification and treatment of co-infections and also provides information about the circulating P. falciparum clinical strains.

Prevalence of antifolate drug resistance markers in Plasmodium vivax in China / 医学前沿

The dihydrofolate reductase (dhfr) and dihydropteroate synthetase (dhps) genes of Plasmodium vivax, as antifolate resistance-associated genes were used for drug resistance surveillance. A total of 375 P. vivax isolates collected from different geographical locations in China in 2009-2019 were used to sequence Pvdhfr and Pvdhps. The majority of the isolates harbored a mutant type allele for Pvdhfr (94.5%) and Pvdhps (68.2%). The most predominant point mutations were S117T/N (77.7%) in Pvdhfr and A383G (66.8%) in Pvdhps. Amino acid changes were identified at nine residues in Pvdhfr. A quadruple-mutant haplotype at 57, 58, 61, and 117 was the most frequent (57.4%) among 16 distinct Pvdhfr haplotypes. Mutations in Pvdhps were detected at six codons, and the double-mutant A383G/A553G was the most prevalent (39.3%). Pvdhfr exhibited a higher mutation prevalence and greater diversity than Pvdhps in China. Most isolates from Yunnan carried multiple mutant haplotypes, while the majority of samples from temperate regions and Hainan Island harbored the wild type or single mutant type. This study indicated that the antifolate resistance levels of P. vivax parasites were different across China and molecular markers could be used to rapidly monitor drug resistance. Results provided evidence for updating national drug policy and treatment guidelines.

Progress of endometriosis-related ovarian cancer / 肿瘤研究与临床

As one of the common benign gynecological diseases, endometriosis (EM) can have a serious impact on the physical and mental health and quality of life of women of childbearing age. Studies have shown that the risk of ovarian cancer in patients with EM is significantly higher than that in the general population. EM is closely related to the occurrence of some types of ovarian cancer, and this kind of ovarian cancer is named endometriosis-related ovarian cancer (EAOC). The incidence rate, pathogenesis, treatment and prognosis of EAOC are summarized in order to better understand it.

Identification of the taxonomic status of Scinax nebulosus and Scinax constrictus (Scinaxinae, Anura) based on molecular markers / Identificação do status taxonômico de Scinax nebulosus e Scinax constrictus (Scinaxinae, Anura), baseado em marcadores moleculares

Abstract The validation of many anuran species is based on a strictly descriptive, morphological analysis of a small number of specimens with a limited geographic distribution. The Scinax Wagler, 1830 genus is a controversial group with many doubtful taxa and taxonomic uncertainties, due a high number of cryptic species. One example is the pair of species Scinax constrictus and Scinax nebulosus, which share a similar morphology. Scinax constrictus is restricted to the Brazilian Cerrado savanna, while S. nebulosus is widely distributed throughout northern South America. Despite the validation of many anuran species, discriminations based only on morphological traits is quite difficult due to the high conservative morphology of some groups. In this context, the present study uses mitochondrial and nuclear genes to provide a more consistent diagnosis and test the validity of S. constrictus as a distinct species from S. nebulosus, as well as evaluate the position of these taxa within the Scinax genus. The topologies obtained herein uphold the monophyletic status of Scinax based on all molecular markers assessed in this study, in all analytical approaches, with high levels of statistical support.

Resumo A validação de muitas espécies de anuros é baseada em uma análise morfológica e descritiva de um pequeno número de espécimes com uma distribuição geográfica limitada. O gênero Scinax Wagler, 1830 é um grupo controverso com muitos táxons duvidosos e incertezas taxonômicas devido ao grande número de espécies crípticas. Um exemplo são as espécies, Scinax constrictus e Scinax nebulosus, que compartilham uma morfologia similar. Scinax constrictus é restrito à savana do Cerrado brasileiro, enquanto S. nebulosus é amplamente distribuído pelo norte da América do Sul. Apesar da validação de muitas espécies de anuros, a discriminação baseada apenas em características morfológicas é bastante difícil, devido à alta morfologia conservadora de alguns grupos. Neste contexto, o presente estudo utiliza genes mitocondriais e nucleares para fornecer um diagnóstico mais consistente e para testar a validade de S. constrictus como uma espécie distinta de S. nebulosus, bem como avaliar a posição destes táxons dentro do gênero Scinax. As topologias obtidas confirmaram o status monofilético de Scinax com base em todos os marcadores moleculares, em todas as abordagens analíticas, com altos níveis de suporte estatístico.

Herramientas complementarias utilizadas en el programa de mejoramiento de arveja (Pisum sativum l.) de la Universidad Nacional de Rosario / Complementary tools utilized in the pea (Pisum sativum l.) breeding program at Universidad Nacional de Rosario

RESUMEN El mejoramiento convencional puede ser complementado mediante diferentes estrategias que incrementen la eficiencia de las metodologías y la tasa actual de aumento de los rendimientos a fin de satisfacer la demanda. El uso de marcadores moleculares con el objetivo de desarrollar mapas de ligamiento de la especie, el uso de Blup (Best Linear Unbiased Prediction) para una selección eficiente de progenitores a hibridar, el uso del cultivo in vitro para incrementar artificialmente el número de plantas F1 o el uso de fenotipificación digital para una eficiente caracterización digital que puede realizarse durante la regeneración periódica y rutinaria de accesiones en colecciones de germoplasma.

ABSTRACT Conventional breeding can be complemented by different strategies that increase the efficiency of the methodologies and the current rate of increase in yields in order to meet demand. The use of molecular markers with the aim of developing linkage maps of the species, the use of Blup (Best Linear Unbiased Prediction) for an efficient selection of progenitors to hybridize, the use of in vitro culture to artificially increase the number of F1 plants or the use of digital phenotyping for efficient digital characterization that can be performed during the periodic and routine regeneration of accessions in germplasm collections.

Mejoramiento de la calidad del fruto por la incorporación de genes de especies silvestres en el tomate (Solanum lycopersicum L.) / Fruit quality improvement through the incorporation of wild species genes in the tomato (Solanum lycopersicum L.)

RESUMEN En el mejoramiento del tomate (Solanum lycopersicum L.) se ha logrado un incremento significativo para el rendimiento y otras características productivas en un período corto de tiempo. Como consecuencia se redujo notablemente la diversidad genética. Si bien el germoplasma silvestre se ha utilizado principalmente como fuente de genes de resistencia para enfermedades y plagas, nuestro grupo inició en la década de 1990, un programa de mejoramiento genético en tomate para mejorar la calidad del fruto con especial énfasis en incrementar la vida poscosecha y también ampliar la variabilidad genética con la incorporación de estos genes al gran cultivo. Hemos desarrollado diferentes poblaciones a partir del cruzamiento interespecífico entre el cultivar argentino Caimanta de S. lycopersicum y la accesión LA0722 de S. pimpinellifolium L. Mediante la generación de cruzamientos entre estos padres selectos y el posterior avance generacional de la selección se ha tratado de dilucidar las bases genéticas que definen la calidad del fruto. Para ello se integraron al programa de mejoramiento información obtenida de datos genómicos, posgenómicos y bioinformáticos. Al mismo tiempo hemos desarrollado cuatro nuevos cultivares con características de calidad de fruto superiores al ser comparados con híbridos comerciales. Para conservar y estudiar la diversidad del cultivo también estamos desarrollado una colección de germoplasma que en la actualidad cuenta con 162 genotipos de tomate de diferentes especies y orígenes. Además, se ha iniciado la transferencia directa de plantines a huertas urbanas y periurbanas para favorecer el acceso a semillas de estos cultivares desarrollados en instituciones públicas.

ABSTRACT The genetic improvement of tomato (Solanum lycopersicum L.) has achieved an increase for yield and other agronomic traits in a short period of time. As a consequence, genetic diversity has been notably reduced. Wild germplasm has been mostly used as a source of resistance genes for diseases and pests. Our group started in the 1990' a breeding program in tomato for improving fruit quality, with special emphasis on increasing fruit shelf life and broadening the genetic variability with the incorporation of wild genes. We have developed different populations from the interspecific cross between the Argentine cultivar Caimanta of S. lycopersicum and the accession LA0722 of S. pimpinellifolium L. Through crosses between these selected parents and the subsequent generational selection advance, we attempted to elucidate the genetic bases that underlie tomato fruit quality. To do that, we use state-of-the-art technology available in the field of genetics and breeding programs, including genomic, post-genomic and bioinformatic data. At the same time, we have developed four new cultivars with improved fruit quality traits compared to commercial hybrids. To conserve and study the tomato diversity, we have developed a germplasm collection that currently contains 162 tomato genotypes from different species and origins. In addition, we have started a direct transfer of our cultivars to urban and peri-urban community orchards to facilitate them the access to genotypes that were developed in Argentine public institutions.

Large-scale advances in SSR markers with high-throughput sequencing in Euphorbia fischeriana Steud

BACKGROUND: Euphorbia fischeriana Steud is a very important medicinal herb and has significant medical value for healing cancer, edema and tuberculosis in China. The lack of molecular markers for Euphorbia fischeriana Steud is a dominant barrier to genetic research. For the purpose of developing many simple sequence repeat (SSR) molecular markers, we completed transcriptome analysis with the Illumina HiSeq 2000 platform. RESULTS: Approximately 9.1 million clean reads were acquired and then assembled into approximately 186.3 thousand nonredundant unigenes, 53,146 of which were SSR-containing unigenes. A total of 76,193 SSR loci were identified. Of these SSR loci, 28,491 were detected at the terminal position of ESTs, which made it difficult to design SSR primers for these SSR-containing sequences, and the residual SSRs were thus used to design primer pairs. Analyzing the results of these markers revealed that the mononucleotide motif A/T (44,067, 57.83% of all SSRs) was the most abundant, followed by the dinucleotide type AG/CT (9430, 12.38%). Using 100 randomly selected primer pairs, 77 primers were successfully amplified in Euphorbia fischeriana Steud, and 79 were successfully amplified in three other related species. The markers developed displayed relatively high quality and cross-species transferability. CONCLUSIONS: The large number of EST-SSRs exploited successfully in Euphorbia fischeriana Steud for the first time could provide genetic information for research on linkage maps, variety identification, genetic diversity analysis, and molecular marker-assisted breeding.

Development of Kompetitive Allele Specific PCR markers for identification of persimmon varieties using genotyping-by-sequencing

BACKGROUND: Persimmon (Diospyros kaki Thunb.) is the most widely cultivated species of the genus Diospyros. In this study, genetic diversity and variations in persimmon genotypes were investigated using single nucleotide polymorphism (SNP) markers identified by genotyping-by-sequencing (GBS) analysis. RESULTS: Ninety-five persimmon accessions grown in the Pear Research Institute, National Institute Horticultural and Herbal Science, were sequenced using the Illumina Hiseq2500 platform and polymorphic SNPs were detected to develop molecular markers. These reliable SNPs were analyzed using the Kompetitive Allele Specific PCR (KASP) assay to discriminate among persimmon genotypes. GBS generated a total of 447,495,724 trimmed reads, of which 89.7% were raw reads. After demultiplexing and sequence quality trimming, 108,876,644 clean reads were mapped to the reference transcriptome. An average of 1,146,070 genotype reads were mapped. Filtering of raw SNPs in each sample led to selection of a total of 1,725,401 high-quality SNPs. The number of homozygous and heterozygous SNPs ranged from 1,933 to 6,834 and from 846 to 5,927, respectively. CONCLUSIONS: Of the 49 SNPs selected for development of an identification system for persimmons, 15 SNPs were used in the KASP assay to analyze 32 persimmon accessions. These KASP markers discriminated among all accessions.

Cross species amplification and characterization of microsatellite markers in Leporinus friderici (Characiformes, Anostomidae) / Amplificação cruzada e caracterização de marcadores microssatélites em Leporinus friderici (Characiformes, Anostomidae)

ABSTRACT: Leporinus friderici is a migratory neotropical fish with elevated ecological and economic importance in Brazil. Microsatellite markers are highly important in population genetic studies, management, and conservation programs; however, no markers are available for this species. In this study, seven microsatellite loci, previously developed for Megaleporinus obtusidens, were successfully cross-amplified in L. friderici. Among these loci, five presented moderate to high genetic variability levels, with four to seven alleles per loci and expected heterozygosities varying from ≥ 0.574 to 1.000. These markers represent a valuable tool for the future management and ecological studies involving this species and group of neotropical fishes.

RESUMO: Leporinus friderici é um peixe neotropical migratório com elevada importância ecológica e econômica no Brasil. Os marcadores microssatélites são conhecidos por sua importância em estudos genéticos populacionais, programas de manejo e conservação, no entanto, não existem marcadores disponíveis para esta espécie. Neste estudo, sete locos microssatélites, previamente desenvolvidos para Megaleporinus obtusidens foram amplificados com sucesso em L. friderici. Dentre esses loci, cinco apresentaram variabilidade genética moderada a alta, com quatro a sete alelos por loci e heterozigosidades esperadas variando de ≥ 0,574 a 1.000. Esses marcadores representam uma ferramenta valiosa para futuros manejos e estudos ecológicos envolvendo esta espécie e este grupo de peixes neotropicais.

Research Progress of Molecular Biology Techniques in Identification of Medicinal Plants / 中国实验方剂学杂志

Medicinal plant germplasm resources are the foundation of the modern development of traditional Chinese medicine. In-depth study of medicinal plant germplasm resources is a prerequisite for cultivating fine varieties and ensuring the output and standard quality of traditional Chinese medicine（TCM）. Traditional identification methods start with appearance and are greatly affected by natural environment and human factors，with a low efficiency and accuracy of identification are generally low molecularin general. Due to such advantages as easy operation，high sensitivity，accurate results， molecular biology technology has been widely used in the related research of relevant studies for medicinal plant germplasm resources due to its advantages of easy operation，high sensitivity，accurate results，etc. It mainly involving the distinction between wild and cultivated products，researchstudy on substitutes of TCM，identification of Chinese patent medicine，good variety marker breeding，genetic diversity researchstudy，genetic map establishment and omics research，etcstudy. Among them，omics researchstudy is divided into genomics，transcriptomics，metabolomics，and proteomics due toby different analysis purposes. Genomics is divided into three sub-fields namely structural genomics，functional genomics， and comparative genomics. Eukaryotes Because eukaryotes have nuclei and organelles，so omics researchstudy also includes chloroplast genomics，mitochondrial genomics，nuclear genomics，and plastid genomics. Among them，the chloroplast genome has a simple structure，small molecular weight，and good conservation，while the mitochondrial genome has a strong variability and complex structure，the nuclear genome data isfeatures complex， data and the nucleus contains no ribosomes in nucleus，resulting in spatiotemporal differences in the translation process，even if repeated repeatedly test， the result of and the test is alsoresults remained uncertain， even after repeated tests. The molecular biology technology and omics researchstudy involved in theby current medicinal plant researchstudy still hashave shortcomings，and there iswith a large room for development，which needs and need further improvement and supplementation. This articlepaper successively introduces the characteristics and applications of cytology，molecular markers，and omics researchstudy techniques in the identification of medicinal germplasm resources，providingin order to provide a reference for subsequent identification，development and utilization of medicinal plant germplasm resources.

Cytomolecular investigations using repetitive DNA probes contribute to the identification and characterization of Characidium sp. aff. C. vidali (Teleostei: Characiformes)

Characidium sp. aff. C. vidali is a species found in coastal streams in southeastern Brazil, which has karyotypic explanatory elements as the occurrence of microstructural variations, keeping the chromosomal macrostructure of the genus. The objective of this study was to apply cytomolecular tools in the chromosomes of Characidium sp. aff. C. vidali to identify characteristics in their karyotype contributing to cytogenetic definition of this species, adding information about the evolution of the chromosomal structure of the group. The species showed 2n = 50 chromosomes and from 1 to 4 additional B microchromosomes. FISH technique showed histone H3 and H4 genes in the short arm of pair 10, and microsatellites (CA)15, (CG)15, (GA)15 and (TTA)10 clustered in the subtelomeric portions of all A chromosomes, with total accumulation by supernumerary. The telomeric probe marked terminal regions of all chromosomes, in addition to the interstitial portion of four pairs, called ITS sites, with these markings being duplicated in two pairs, hence the double-ITS classification. C-banding revealed that supernumerary chromosomes are completely heterochromatic, that ITS sites are C-banding positive, but double-ITS sites are C-banding negative. So, throughout the evolution to Characidium, genomic events are occurring and restructuring chromosomes in populations.(AU)

Characidium sp. aff. C. vidali é uma espécie encontrada em riachos costeiros do sudeste do Brasil, que apresenta elementos cariotípicos elucidativos quanto à ocorrência de variações microestruturais, conservando a macroestrutura cromossômica do gênero. O objetivo deste estudo foi aplicar ferramentas citomoleculares para identificar características no cariótipo de Characidium sp. aff. C. vidali, que contribuam para a definição citogenética desta espécie, agregando informações quanto à evolução da estruturação cromossômica do grupo. A espécie apresentou 2n = 50 cromossomos, além de 1 a 4 microcromossomos B por célula. A FISH mostrou os genes de histona H3 e H4 sintênicos no braço curto do par 10, e os microssatélites (CA)15, (CG)15, (GA)15 e (TTA)10 clusterizados nas porções subteloméricas de todos os cromossomos do complemento A, com grande acúmulo nos supranumerários. A sonda telomérica identificou marcações terminais em todos os cromossomos, além de quatro pares marcados intersticialmente, chamados de sítios ITS, e dois pares com duas marcações intersticiais, chamados de double-ITS. O bandamento C revelou que os cromossomos supranumerários são completamente heterocromáticos, que os sítios ITS são banda C positivos, mas os sítios double-ITS são banda C negativos. Então, ao longo da evolução de Characidium, eventos genômicos estão ocorrendo e reestruturando cromossomos nas populações.(AU)

Genetic Diversity Analysis of Erigeron breviscapus Based on SSR Markers / 中国实验方剂学杂志

Objective:To explore the genetic diversity and population structure of <italic>Erigeron breviscapus</italic>， so as to provide a scientific basis for its resource protection and rational utilization. Method:Twelve pairs of simple sequence repeat（SSR） primers were screened out from 243 individuals in 16 natural populations to calculate the genetic diversity parameters of <italic>E. breviscapus</italic>， which were then subjected to principal coordinate analysis and cluster analysis. Result:Twelve SSR markers generated 209 alleles， with an average of 17.417 alleles per locus. Based on 12 SSR markers and 16 populations of <italic>E. breviscapus</italic>， the observed heterozygosity （<italic>H</italic>₀） values were determined to be 0.603 and 0.613， the expected heterozygosity （<italic>H</italic>_e）to be 0.658 and 0.659， and the Shannon's information index （<italic>I</italic>） to be 1.443 and 1.446， respectively. The Wright's fixation index （<italic>F</italic>_st） was 0.123 and gene flow （<italic>N</italic>_m） was 2.077. Analysis of molecular variance （AMOVA） and genetic differentiation revealed that genetic variation within populations was the main source of total variation. The Nei's genetic distance and genetic identity coefficients were within the ranges of 0.107 （YA and XY）-0.713 （SZ and XZD） and 0.490 （SZ and XZD）-0.899 （YA and XY）， respectively. As demonstrated by the principal coordinate analysis and cluster analysis， the 16 populations of <italic>breviscapus </italic>were divided into two clusters. Conclusion:The genetic diversity of <italic>E. breviscapus</italic> was relatively high and there existed certain genetic differentiation and gene flow within and among populations. The genetic variation was mainly present within populations. All these have provided reference for subsequent study on good germplasm selection of <italic>E. breviscapus.</italic>

Molecular markers and mechanisms for stemness maintenance of liver cancer stem cells: a review / 生物工程学报

Primary liver cancer (PLC) is an aggressive tumor and prone to metastasize and recur. According to pathological features, PLC are mainly categorized into hepatocellular carcinoma, intrahepatic cholangiocarcinoma, mixed hepatocellular cholangiocarcinoma, and fibrolamelic hepatocellular carcinoma, etc. At present, surgical resection, radiotherapy and chemotherapy are still the main treatments for PLC, but the specificities are poor and the clinical effects are limited with a 5-year overall survival rate of 18%. Liver cancer stem cells (LCSCs) are a specific cell subset existing in liver cancer tissues. They harbor the capabilities of self-renewal and strong tumorigenicity, driving tumor initiation, metastasis, drug resistance and recurrence of PLC. Therefore, the identification of molecular markers and the illustration of mechanisms for stemness maintenance of LCSCs can not only reveal the molecular mechanisms of PLC tumorigenesis, but also lay a theoretical foundation for the molecular classification, prognosis evaluation and targeted therapy of PLC. The latest research showed that the combination of 5-fluorouracil and CD13 inhibitors could inhibit the proliferation of CD13+ LCSCs, thereby reducing overall tumor burden. Taken together, LCSCs could be the promising therapeutic targets of PLC in the future. This review summarizes the latest progress in molecular markers, mechanisms for stemness maintenance and targeted therapies of LCSCs.

Research progress in myosin light chain 9 in malignant tumors / 中南大学学报(医学版)

Myosin light chain 9 (MYL9) is a regulatory light chain of myosin, which plays an important role in various biological processes including cell contraction, proliferation and invasion. MYL9 expresses abnormally in several malignancies including lung cancer, breast cancer, prostate cancer, malignant melanoma and others, which is closely related to the poor prognosis, but the clinical significance for its expression varies with different types of cancer tissues. Further elucidating the molecular mechanism of MYL9 in various types of malignant tumor metastasis is of great significance for cancer prevention and treatment. At the same time, as a molecular marker and potential target, MYL9 may have great clinical value in the early diagnosis, prognosis prediction, and targeted treatment of malignant tumors.

Advancements in molecular marker technologies and their applications in diversity studies

Crop improvement is a continuous effort, since some 10,000 years ago when primitive man made the transitionfrom hunting and foraging to domestication and crop cultivation. Since then, man-made interventions havechanged the entire scenario of crop evolution, by means of genetic alterations of plants and animals made tosatisfy man’s needs. The process of domestication has led to dramatic changes in their appearance, quality andproductivity that have contributed substantially to global food security. The tremendous decline in cultivableland, freshwater, and increasing risk of biotic and abiotic stress demand immediate attention on cropimprovement to cope with the higher demand of *40% of the food by 2020. Therefore, plant genetic variationplays a key role in plant breeding for its improvement. Most of the genetic variations useful for cropimprovement have been deposited and maintained in seed gene banks across the world; they need to be broughtinto the mainstream of breeding lines. Recent advances and progress made in molecular markers have beensubstantial tools for deeper insights of genetics, and greatly complemented breeding strategies. Integration of thenext-generation sequencing (NGS) technologies with precise phenotyping, association mapping, proteome andmetabolome studies has increased the chances of finding candidate genes and their allelic variants controlling atrait of interest. Further, these functional markers (FMMs), genotype-by-sequencing and association mappingmethodologies have opened new avenues for identification of novel genetic resources (lines) that can facilitateaccelerated crop breeding programs for increased yield, high nutritional quality, and tolerance to a variety ofabiotic and biotic stresses. The details of popular molecular markers, advancement in the technologies andstrategies for crop diversity studies and their application in crop breeding programs are presented here.

Population genetics of the fish Brycon henni (Characiformes: Bryconidae) using species-specific polymorphic microsatellite loci

Introduction: The freshwater fish Brycon henni (Characiformes: Bryconidae) is endemic to Colombia and currently considered as a "least concern" species according to the International Union for Conservation of Nature (IUCN). Objective: To develop microsatellite markers to examine population genetics in B. henni. Methods: Using a low-coverage sequencing genomic library, this study developed the first set of microsatellite loci to study the population genetics of this Neotropical species. These loci were used to evaluate the genetic diversity and structure of B. henni from three sites of the Magdalena-Cauca Basin (Colombia). Results: A set of 21 polymorphic microsatellite loci was highly informative and revealed that B. henni exhibits genetic diversity (5.143-5.619 alleles/locus, observed and expected heterozygosity = 0.461-0.645 and 0.604-0.662, respectively) and is evenly genetically structured between two tributaries of the Cauca River separated by only 30 km (F'ST = 0.093, Jost's DEST = 0.311, P < 0.001) a finding that indicates these may be reproductively isolated groups. Conclusions: We reported a set of 21 polymorphic microsatellite loci that allowed the detection of genetic structure at local and regional scales. This population genetic structure, concordant with that found in eight congeners, is relevant when determining the risk categorization of B. henni, as well as management, conservation, and restocking programs for this species.

Introducción: El pez de agua dulce Brycon henni (Characiformes: Bryconidae) es una especie endémica de Colombia que actualmente está catalogada como de "menor preocupación" por la Unión Internacional para la Conservación de la Naturaleza (UICN). Objetivo: Desarrollar marcadores microsatélites para estudiar la genética poblacional de Brycon henni. Métodos: Usando una biblioteca genómica de secuenciación de baja cobertura, este estudio desarrolló el primer grupo de loci microsatélites para el estudio de la genética poblacional de esta especie neotropical. Estos loci fueron usados para evaluar la diversidad genética y estructura de B. henni en tres sitios de la cuenca Magdalena-Cauca (Colombia). Resultados: Un grupo de 21 loci polimórficos tipo microsatélite fueron altamente informativos y revelaron que B. henni exhibe diversidad genética (5.143-5.619 alelos/locus, heterocigosidad observada y esperada = 0.461-0.645 y 0.604-0.662, respectivamente) y se encuentra genéticamente estructurado entre dos tributarios del río Cauca separados únicamente por 30 km (F'ST = 0.093, Jost's DEST = 0.311, P < 0.001), un resultado que indica que puede existir aislamiento reproductivo entre dichos grupos. Conclusiones: Reportamos un grupo de 21 loci polimórficos tipo microsatélite que permitieron la detección de la estructura genética a escala local y regional. Esta estructura genética poblacional, concordante con lo que se reporta para otros ocho congéneres, es relevante al determinar la categorización de riesgo de B. henni, así como los programas de manejo, conservación y repoblamiento para esta especie.