1.Genetic diversity analysis and DNA fingerprinting of Artemisia argyi germplasm resources based on EST-SSR molecular markers.
Yu-Yang MA ; Chang-Jie CHEN ; Ming-Xing WANG ; Yan FANG ; Yu-Huan MIAO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2025;50(9):2356-2364
This study investigates the genetic diversity and evolutionary relationships of different Artemisia argyi germplasm resources to provide a basis for germplasm identification, variety selection, and resource protection. A total of 192 germplasm resources of A. argyi were studied, and EST-based simple sequence repeat(EST-SSR) primers were designed based on transcriptomic data of A. argyi. Polymerase chain reaction(PCR) amplification was performed on these resources, followed by fluorescence capillary electrophoresis to detect genetic diversity and construct DNA fingerprints. From 197 pairs of primers designed, 28 pairs with polymorphic and clear bands were selected. A total of 278 alleles were detected, with an average of 9.900 0 alleles per primer pair and an average effective number of alleles of 1.407 2. The Shannon's diversity index(I) for the A. argyi germplasm resources ranged from 0.148 1 to 0.418 0, with an average of 0.255 7. The polymorphism information content(PIC) ranged from 0.454 5 to 0.878 0, with an average of 0.766 9, showing high polymorphism. Cluster analysis divided the A. argyi germplasm resources into three major groups: Group Ⅰ contained 136 germplasm samples, Group Ⅱ contained 45, and Group Ⅲ contained 11. Principal component analysis also divided the resources into three groups, which was generally consistent with the clustering results. Mantel test results showed that the genetic variation in A. argyi populations was to some extent influenced by geographic distance, but the effect was minimal. Structure analysis showed that 190 germplasm materials had Q≥ 0.6, indicating that these germplasm materials had a relatively homogeneous genetic origin. Furthermore, 8 core primer pairs were selected from the 28 designed primers, which could distinguish various germplasm types. Using these 8 core primers, DNA fingerprints for the 192 A. argyi germplasm resources were successfully constructed. EST-SSR molecular markers can be used to study the genetic diversity and phylogenetic relationships of A. argyi, providing theoretical support for the identification and molecular-assisted breeding of A. argyi germplasm resources.
Artemisia/classification*
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Microsatellite Repeats
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Genetic Variation
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Expressed Sequence Tags
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DNA Fingerprinting
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Phylogeny
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Polymorphism, Genetic
;
DNA, Plant/genetics*
;
Genetic Markers
2.An accurate diagnostic approach for urothelial carcinomas based on novel dual methylated DNA markers in small-volume urine.
Yucai WU ; Di CAI ; Jian FAN ; Chang MENG ; Shiming HE ; Zhihua LI ; Lianghao ZHANG ; Kunlin YANG ; Aixiang WANG ; Xinfei LI ; Yicong DU ; Shengwei XIONG ; Mancheng XIA ; Tingting LI ; Lanlan DONG ; Yanqing GONG ; Liqun ZHOU ; Xuesong LI
Chinese Medical Journal 2024;137(2):232-234
3.Establishment of a standard DSS library for identification of original plants of medicinal materials in Chinese Pharmacopoeia.
Duo-Mei WANG ; Chao JIANG ; Jing-Zhe PU ; Chong HU ; Ling-Li CHEN ; Ya-Zhong ZHANG ; Yuan YUAN
China Journal of Chinese Materia Medica 2024;49(23):6249-6256
With the development of molecular pharmacognosy, the advantages of DNA molecular markers in the identification of original plants of Chinese medicinal materials are becoming increasingly significant. To compensate for the limitations of existing markers in the quality supervision of Chinese medicinal materials, our team has independently designed a new molecular marker named DNA signature sequence(DSS). This marker is a nucleotide sequence that only appears in a specific taxonomic unit, with a length of 40 bp and high identification accuracy. This article aims to screen and verify the DSS markers that can accurately identify the original plants of the medicinal materials included in the volume one of the Chinese Pharmacopoeia, establish the operating procedure for developing standard nucleotide sequences, and lay a foundation for the widespread application of polymerase chain reaction in the quality supervision of traditional Chinese medicine. Firstly, the Chloroplast Genome Information Resource(CGIR) was searched for the chloroplast genome sequences of the test samples, species of the same genus, and common background species. IdenDSS was used to obtain the DSS tags and specific identification primers of the tested species. After DNA extraction, PCR amplification, sequencing, and sequence alignments, a total of 203 DSS markers of Chinese medicinal materials were obtained for validation. The above sequences were uploaded to the Traditional Chinese Medicine Molecular Identification Platform(www.herbsdna.com), and a standard DSS library was established for identifying the original plants of medicinal materials, serving as an important tool for quality supervision of Chinese materia medica. On this basis, an operating procedure for DSS development is formed, laying a foundation for further DSS screening and application based on more diverse genome sequences.
Plants, Medicinal/classification*
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DNA, Plant/genetics*
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Drugs, Chinese Herbal
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Pharmacopoeias as Topic
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Genetic Markers
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Medicine, Chinese Traditional
4.Genetic diversity and structure of 15 full-sib families of Litopenaeus vannamei based on SSR markers.
Wenchun CHEN ; Kai PENG ; Minwei HUANG ; Jichen ZHAO ; Zhihao ZHANG ; Hui GUO ; Jinshang LIU ; Zhenxing LIU ; Huijie LU ; Wen HUANG
Chinese Journal of Biotechnology 2024;40(12):4628-4644
To clarify the genetic diversity and structure of the nucleus population of F1-generation Litopenaeus vannamei, this study utilized 15 pairs of highly polymorphic microsatellite primers to analyze the simple sequence repeat (SSR) markers and genetic diversity in 15 full-sib families of L. vannamei. A total of 112 alleles (Na) and 60.453 effective alleles (Ne) were identified among the selected 15 SSR loci, with the average polymorphic information content (PIC) of 0.648. The average Ne, observed heterozygosity (Ho), and expected heterozygosity (He) in the 15 F1 families varied from 1.925 to 2.626, 0.425 to 0.783, and 0.403 to 0.572, respectively. The 15 full-sib families were primarily clustered into three categories in the phylogenetic analysis, with the genetic distance between families ranging from 0.252 to 0.574. Additionally, the genetic differentiation coefficient (Fst) among the families varied from 0.112 to 0.278, indicating substantial genetic differentiation. Overall, this study suggested that the genetic diversity of the 15 full-sib families was moderate, providing valuable genetic insights for the subsequent breeding initiatives aimed at enhancing the tolerance of L. vannamei to high levels of soybean meal.
Penaeidae/classification*
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Microsatellite Repeats/genetics*
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Animals
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Genetic Variation
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Polymorphism, Genetic
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Phylogeny
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Alleles
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Genetic Markers
5.Identification Strategy of Biological Half Sibling Relationship.
Zheng TAN ; Guan-Ju MA ; Li-Hong FU ; Xiao-Jing ZHANG ; Qian WANG ; Guang-Ping FU ; Qing-Qing DU ; Shu-Jin LI
Journal of Forensic Medicine 2023;39(3):262-270
OBJECTIVES:
To compare the application value of the likelihood ratio (LR) method and identity by state (IBS) method in the identification involving half sibling relationships, and to provide a reference for the setting of relevant standards for identification of half sibling relationship.
METHODS:
(1) Based on the same genetic marker combinations, the reliability of computer simulation method was verified by comparing the distributions of cumulated identity by state score (CIBS) and combined full sibling index in actual cases with the distributions in simulated cases. (2) In different numbers of three genetic marker combinations, the simulation of full sibling, half sibling and unrelated individual pairs, each 1 million pairs, was obtained; the CIBS, as well as the corresponding types of cumulative LR parameters, were calculated. (3) The application value of LR method was compared with that of IBS method, by comparing the best system efficiency provided by LR method and IBS method when genetic markers in different amounts and of different types and accuracy were applied to distinguish the above three relational individual pairs. (4) According to the existing simulation data, the minimum number of genetic markers required to distinguish half siblings from the other two relationships using different types of genetic markers was estimated by curve fitting.
RESULTS:
(1) After the rank sum test, under the premise that the real relationship and the genetic marker combination tested were the same, there was no significant difference between the simulation method and the results obtained in the actual case. (2) In most cases, under the same conditions, the system effectiveness obtained by LR method was greater than that by IBS method. (3) According to the existing data, the number of genetic markers required for full-half siblings and half sibling identification could be obtained by curve fitting when the system effectiveness reached 0.95 or 0.99.
CONCLUSIONS
When distinguishing half sibling from full sibling pairs or unrelated pairs, it is recommended to give preference to the LR method, and estimate the required number of markers according to the identification types and the population data, to ensure the identification effect.
Humans
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Siblings
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Genetic Markers
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Computer Simulation
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Irritable Bowel Syndrome/genetics*
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Reproducibility of Results
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Genotype
6.Application of SNaPshot Technology in Semen-Specific cSNP Genetic Marker.
Rui-Yang TAO ; Shou-Yu WANG ; Chun-Yan YUAN ; Ruo-Cheng XIA ; Cheng-Tao LI
Journal of Forensic Medicine 2023;39(5):465-470
OBJECTIVES:
To explore the feasibility of genetic marker detection of semen-specific coding region single nucleotide polymorphism (cSNP) based on SNaPshot technology in semen stains and mixed body fluid identification.
METHODS:
Genomic DNA (gDNA) and total RNA were extracted from 16 semen stains and 11 mixtures composed of semen and venous blood, and the total RNA was reverse transcribed into complementary DNA (cDNA). The cSNP genetic markers were screened on the validated semen-specific mRNA coding genes. The cSNP multiplex detection system based on SNaPshot technology was established, and samples were genotyped by capillary electrophoresis (CE).
RESULTS:
A multiplex detection system containing 5 semen-specific cSNPs was successfully established. In 16 semen samples, except the cSNP located in the TGM4 gene showed allele loss in cDNA detection results, the gDNA and cDNA typing results of other cSNPs were highly consistent. When detecting semen-venous blood mixtures, the results of cSNP typing detected were consistent with the genotype of semen donor and were not interfered by the genotype of venous blood donor.
CONCLUSIONS
The method of semen-specific cSNPs detection by SNaPshot technology method can be applied to the genotyping of semen (stains) and provide information for determining the origin of semen in mixed body fluids (stains).
Genetic Markers
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Semen
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Polymorphism, Single Nucleotide
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DNA, Complementary/genetics*
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Body Fluids
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RNA, Messenger/genetics*
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DNA
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Saliva
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Forensic Genetics/methods*
7.Application Progress of Massively Parallel Sequencing Technology in STR Genetic Marker Detection.
Rui-Yang TAO ; Xin-Yu DONG ; An-Qi CHEN ; Ye-Hui LÜ ; Su-Hua ZHANG ; Cheng-Tao LI
Journal of Forensic Medicine 2022;38(2):267-279
In recent years, more and more forensic genetics laboratories have begun to apply massively parallel sequencing (MPS) technology, that is, next-generation sequencing (NGS) technology, to detect common forensic genetic markers, including short tandem repeat (STR), single nucleotide polymorphism (SNP), the control region or whole genome of mitochondrial DNA (mtDNA), as well as messenger RNA (mRNA), etc., for forensic practice, such as individual identification, kinship analysis, ancestry inference and body fluid identification. As the most widely used genetic marker in forensic genetics, STR is currently mainly detected by capillary electrophoresis (CE) platform. Compared with CE platform, MPS technology has the advantages of simultaneous detection of a large number of genetic markers, massively parallel detection of samples, the polymorphism of sequence detected by NGS makes STR have the advantages of higher resolution and system efficiency. However, MPS technology is expensive, there is no uniform standard so far, and there are problems such as how to integrate MPS-STR data with the existing CE-STR database. This review summarizes the current status of the application of MPS technology in the detection of STR genetic markers in forensic genetics, puts forward the main problems that need to be solved urgently, and prospects the application prospect of this technology in forensic genetics.
DNA Fingerprinting/methods*
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Forensic Genetics/methods*
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Genetic Markers
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High-Throughput Nucleotide Sequencing/methods*
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Microsatellite Repeats/genetics*
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Polymorphism, Single Nucleotide
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Sequence Analysis, DNA
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Technology
8.Advance in research on genetic polymorphisms associated with athletic ability.
Bonan DONG ; Tingting ZHANG ; Qiuyan LI ; Songbin FU
Chinese Journal of Medical Genetics 2022;39(4):438-441
Genetic factors play a key role in human athletic ability, and endurance quality and explosive power quality are the important components of athletic ability. In this review, we aimed to reveal the biological genetic mechanism of human athletic ability at the molecular level through summarizing the relationship between genetic variants and human athletic ability, including endurance quality related genetic markers angiotensin converting enzyme (ACE) gene, creatine kinase MM (CKMM) gene and explosive power quality related genetic markers alpha actinin 3 (ACTN3) gene, angiotensinogen (AGT) gene and interleukin6 (IL6) gene. Meanwhile, we also summarized the distribution of allele frequencies among various populations.
Actinin/genetics*
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Athletic Performance
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Gene Frequency
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Genetic Markers
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Genotype
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Humans
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Polymorphism, Genetic
10.Forensic Application of Next Generation Sequencing Technology in the Typing of Y Chromosome Genetic Markers.
Yue YANG ; Rui Yang TAO ; Min LI ; Huan YU ; Li Qin CHEN ; Ya Li WANG ; Cheng Tao LI
Journal of Forensic Medicine 2021;37(1):91-98
The paternal inheritance characteristics of Y chromosome have been widely used in the forensic genetics field to detect the genetic markers in the non-recombining block, and used in the studies such as, genetic relationship identification, mixed stain detection, pedigree screen and ethnicity determination. At present, capillary electrophoresis is still the most common detection technology. The commercial detection kits and data analysis and processing system based on this technology are very mature. However, the disadvantages of traditional detection technology have gradually appeared with the rapid growth of bio-information amount, which promotes the renewal of forensic DNA typing technology. In recent years, next generation sequencing (NGS) technology has developed rapidly. This technology has been applied to various fields including forensic genetics and has provided new techniques for the detection of Y chromosome genetic markers. This article describes the current situation and application prospects of the NGS technology in forensic Y chromosome genetic markers detection in order to provide new ideas for future judicial practice.
Chromosomes, Human, Y/genetics*
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DNA Fingerprinting
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Forensic Genetics
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Genetic Markers
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High-Throughput Nucleotide Sequencing
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Humans
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Microsatellite Repeats
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Technology
;
Y Chromosome

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