Genome-wide association studies reveal the genetic basis of amino acid content variation in tea plants

Tea is one of the most popular non-alcoholic beverages in the world,and free amino acids,especially theanine,make a major contribution to the umami taste of tea.However,the genetic basis of the variation in amino acid content in tea plants remains largely unknown.Here,we measured the free amino acid content in fresh leaves of 174 tea accessions over two years using a targeted metabolomics approach and obtained genotype data via RNA sequencing.Genome-wide association studies were conducted to investigate loci affecting the content of free amino acids.A total of 69 quantitative trait loci(–log10(P-value)>5)were identified.Functional annotation revealed that branched-chain amino acid aminotransferase,glutamine synthetase,nitrate transporter,and glutamate decarboxylase might be important for amino acid metabolism.Two significant loci,glutamine synthetase(Glu1,P=3.71×10^(−4);Arg1,P=4.61×10^(−5))and branched-chain amino acid aminotransferase(Val1,P=4.67×10^(−5);I_Leu1,P=3.56×10^(−6)),were identified,respectively.Based on the genotyping result,two alleles of CsGS(CsGS-L and CsGS-H)and CsBCAT(CsBCAT-L and CsBCAT-H)were selected to perform function verification.Overexpression of CsGS-L and CsGS-H enhanced the contents of glutamate and arginine in transgenic plants,and overexpression of CsBCAT-L and CsBCAT-H promoted the accumulation of valine,isoleucine and leucine.Enzyme activity assay uncovered that SNP1054 is important for CsGS catalyzing glutamate into glutamine.Furthermore,CsGS-L and CsGS-H differentially regulated the accumulation of glutamine,and CsBCAT-L and CsBCAT-H differentially regulated the accumulation of branched-chain amino acids.In summary,the findings in our study would provide new insights into the genetic basis of amino acids contents variation in tea plants and facilitate the identification of elite genes to enhance amino acids content.

Variation and stability analysis of growth traits of poplar clones in the seedling stage in northeast China

Plant phenotypes are infl uenced by genetic and environmental factors.In this study,the growth traits of 43 one-year-old poplar clones grown at diff erent sites in northeast China(Dongling State-owned Forest Protection Center,DL;Baicheng State-owned forest farm,BC;and Cuohai Forest farm,CH)were evaluated and analyzed across clones and sites.Results show signifi cant diff erences in height and base diameter among sites and clones.Phenotypic and genetic variation coeffi cients ranged from 49.59%(BC)to 58.39%(DL)and from 49.33%(BC)to 58.06%(DL),respectively.Additive main eff ects and multiplicative interaction(AMMI)analysis showed that the eff ects of genotype,environment,and genotype×environment interaction were signifi cantly diff erent.Genetic variation was the main source of variation,accounting for 48.6%.AMMI biplot showed that clone 30 had high and stable yields at the three sites.From an evaluation of multiple traits and GGE biplot that clone 2,clone 30 and clone 25 had higher yield than the other clones at DL,CH and BC,respectively.These clones will provide material for forest regeneration in northeast China.

Graft-induced Inheritable Variation in Mungbean and Its Application in Mungbean Breeding

Graft can induce inheritable variations in the progenies of the scion plants. Seedling of mungbean ( Vigna radiata (L.) Wilczek) was grafted onto the stem of sweet potato ( Ipomoea batatas (L.) Lam.). The growth of the scion was maintained until the scion produced selfed seeds. We sowed the seeds for several generations under normal conditions. Distinct genetic variations appeared in the progenies. Similar variations did not appear in the generations of the scion sowed normally without graft. The variations seemed to be induced by the graft and they inherited steadily. For understanding the possible mechanism of the phenomenon (graft_induced inheritable variation), we analyzed the cytoplasmic and genomic DNA of the variations. The results showed that there was no restriction fragment length polymorphism (RFLP) in the cytoplasmic DNA between the original scion and the variation. However, significant difference between the scion and variation was recognized by random amplified polymorphic DNA (RAPD) analysis. In addition, there was no evidence that indicated the gene transformation from stock to scion. Our results suggest that the non_specific grafting has a pragmatic potential for plant breeding and crop improvement and, the genetic variation seems not to be caused simply by DNA transformation but most likely the stress induced mutation.

Genetic variations of wood properties and growth characters of Ko-rean pines from different provenances

Totally 26 provenance stands of 17-year-old Korean pine were selected for investigating wood properties and growth characters in Mao抏rshan Experimental Forest Farm of Northeast Forestry University in 1999. The anatomical property indexes, including tracheid length, tracheid diameter and wall-indiameter ratio, and the physical property indexes, such as growth ring width, late wood percentage and growth ring density, were measured for wood properties. Growth character indexes, including tree height and diameter at breast height, were also measured. The analytical results showed that there exited obviously dif-ference in wood property indexes between different provenances, which is suggested that wood properties are controlled by their genetic differences. The growth character indexes of Korean pines presented significant difference and they might also be controlled by their genetic differences. Most parameters of wood properties mainly varied in the direction of longitude, but the parameters of growth characters varied in the direction of latitude.

Genetic Variation of Wild and Hatchery Populations of the Pacific Oyster Crassostrea gigas Assessed by Microsatellite Markers

Microsatellite DNA technique was used to detect the genetic variation between five hatchery populations of the Pacific oyster from China and two wild populations from Japan. Seven microsatellite loci screened in this study showed high polymorphism in both hatchery and wild populations, as observed in an average number of allele per locus （19.1-29.9） and average expected heterozygosity （0.916-0.958）. No significant difference in average allelic richness or expected heterozygosity was observed between Chinese hatchery populations and Japanese wild populations. Pairwise Fsr values and heterogeneity tests of allele frequencies showed significant genetic differentiation between all populations. According to the neighbor-joining tree constructed on the basis of the Dc distance, the seven populations fell into three groups showing a clear division between hatchery and wild populations, and between the northern and southern hatchery populations. Assignment tests correctry assigned high percentages （97%-100%） of individuals to their original populations and demonstrated the feasibility of microsatellite analysis for discrimination between populations. The information obtained in this study is useful for designing suitable management guidelines and selective breeding programs for the Pacific oyster in China.

A Preliminary Study on Genetic Variation of g E Gene of an Epidemic Pseudorabies Virus Strain and Its Pathogenicity to Piglets

[Objective] This study aimed to investigate the genetic variation of g E gene of an epidemic pseudorabies virus（PRV） strain and its pathogenicity to piglets. [Method] By serial passage in Vero cells, a PRV strain was isolated from the brain tissues of stillborn fetuses delivered by sows with suspected PRV infection and preliminarily identified by PCR. g E gene of the isolated PRV strain was amplified and sequenced for phylogenetic analysis. In addition, the pathogenicity of the isolated PRV strain to 6-week-old piglets was evaluated. [Result] A PRV strain was successfully isolated and named PRV N5 B strain, which could proliferate in Vero cells and TCID50 of the 15 thgeneration virus liquid reached 10^7.125/0.1 ml. Specific bands could be amplified by PCR. g E gene in the isolated PRV strain was 1 740 bp in length. A phylogenetic tree was constructed based on full-length g E sequences, which showed that PRV N5 B strain and PRV strains isolated since 2012 were clustered into the same independent category and shared 99.7%-100% homology of nucleotide sequences. Compared with related sequences published previously, there were insertions of three consecutive bases at two loci. Animal experiments showed that intranasal inoculation of 6-week-old piglets with 2 ml of PRV N5 B strain（10^6/0.1 ml） led to a mortality rate of 100%. [Conclusion] In this study,genetic variability of g E gene in PRV N5 B isolate and its pathogenicity to piglets were analyzed, which provided a theoretical basis for the development of new vaccines to prevent and control porcine pseudorabies.

Variation of Protein Content in Tartary Buckwheat Seeds

[Objective]The aim was to select tartary buckwheat materials with high protein content.[Method]Used 35 kinds of tartary buckwheat as experimental material,and the protein content of seeds had been determined.[Result]The protein content of 35 kinds of tartary buckwheat will change in the range of 23.65-193.28 mg/g with an average of 111.85 mg/g.There was difference among different origins of tartary buckwheat.The seeds from Hezhang in Guizhou and Sichuan had highest protein content,while the seeds from Nayong had lowest protein content.[Conclusion]The study had provided theoretical basis for the further study on the genetic and variation law of protein content in different tartary buckwheat resources.

Variation of Glucose Content in Tartary Buckwheat Seeds

[Objective] This study aimed to finding out the difference in glucose content of tartary buckwheat seeds from different origins.[Method] Taking 35 kinds of tartary buckwheat from different origins as the test materials,the glucose content of seeds had been determined.[Result] The variation range of glucose content in 35 kinds of tartary buckwheat was from 0.055 6% to 0.840 2% with an average of 0.321 7%.There was a difference in glucose content of tartary buckwheat from different origins.The seeds T324 from Weining in Guizhou had the highest glucose content,while the seeds T398 also from Weining in Guizhou had the lowest glucose content.[Conclusion] This study provides a reference for the genetic variation law of glucose content in different tartary buckwheat resources.

Microsatellite Variation Within Three Populations of Inbred C57BL/6J Strain

The goal of this study was to investigate the genetic stability of the C57BL/6J （B6） inbred mouse strain maintained in different breeders. Three populations of B6, Popl and Pop2 purchased from Beijing and Pop3 purchased from Shanghai, were examined. Fifteen microsatellite loci reported to be polymorphic among inbred strains were amplified using FAM labeled primers and genotyped with ABI Prism 377 automated sequencer. Seven loci were found polymorphic, and all the loci were homozygous in all the three populations. The present study indicates that genetic variation occurs in different B6 populations although they are still inbred in each breeder. The mechanism of genetic variation is not well understood now, but it is very important to know the precise content of the B6 genome before use of this strain in research.

Progress in clinical diagnosis and treatment of colorectal cancer with rare genetic variants

Targeted therapy is crucial for advanced colorectal cancer(CRC) positive for genetic drivers. With advances in deep sequencing technology and new targeted drugs, existing standard molecular pathological detection systems and therapeutic strategies can no longer meet the requirements for careful management of patients with advanced CRC. Thus, rare genetic variations require diagnosis and targeted therapy in clinical practice. Rare gene mutations, amplifications, and rearrangements are usually associated with poor prognosis and poor response to conventional therapy. This review summarizes the clinical diagnosis and treatment of rare genetic variations, in genes including erb-b2 receptor tyrosine kinase 2(ERBB2), B-Raf proto-oncogene, serine/threonine kinase(BRAF), ALK receptor tyrosine kinase/ROS proto-oncogene 1, receptor tyrosine kinase(ALK/ROS1), neurotrophic receptor tyrosine kinases(NTRKs), ret proto-oncogene(RET), fibroblast growth factor receptor 2(FGFR2), and epidermal growth factor receptor(EGFR), to enhance understanding and identify more accurate personalized treatments for patients with rare genetic variations.

Computational and bioinformatics tools for understanding disease mechanisms

Computational methods have significantly transformed biomedical research,offering a comprehensive exploration of disease mechanisms and molecular protein functions.This article reviews a spectrum of computational tools and network analysis databases that play a crucial role in identifying potential interactions and signaling networks contributing to the onset of disease states.The utilization of protein/gene interaction and genetic variation databases,coupled with pathway analysis can facilitate the identification of potential drug targets.By bridging the gap between molecular-level information and disease understanding,this review contributes insights into the impactful utilization of computational methods,paving the way for targeted interventions and therapeutic advancements in biomedical research.

Analysis of Genetic Variation and Population Structure of Starch Synthesis-related Genes in Indica Rice Cultivars

In this study, 34 molecular markers of starch synthesis-related genes were used to evaluate the genetic variation and population structure of 87 indica rice cultivars from different countries and regions. The results showed that a total of 80 alleles were amplified using 34 primer pairs, with an average of 2.5 alleles per locus. The allele number varied from 2 to 6 among various cultivars. Shannon＇s diversity index of molecular markers varied from 0.303 to 0.796, with an average of 0.539. Polymorphism information content （PIC） varied from 0.084 to 0.658, with an average of 0.295. The genetic similarity coefficients of 87 indica rice cultivars ranged from 0.265 to 0.990, indicating significant genetic differences of starch synthesis-related genes among different cultivars, but the variation frequency of alleles varied among different cultivars. Population structure analysis showed that these 87 indica rice cultivars were divided into three categories. Genetic differences were small within the same category but great among different categories. Moreover, indica rice cultivars with simple genetic components accounted for 39.1% and those with complex genetic background accounted for 60.9%. This study may not only provide theoretical basis for genetic improvement of rice starch quality, but also lay a solid foundation for subsequent association analysis of rice quality-related traits.

Genetic Variation Analysis of 3D Gene and Molecular Detection of Porcine Kobuvirus in 2013-2014

[Objective] This study aimed to investigate the prevalence and variation of porcine kobuvirus （PKV） in suckling piglets in China. [Method] In 2013-2014, 224 feces samples from suckling piglets with diarrhea in 27 pig farms of five provinces in China were collected to detect 3D genes of PKV with RT-PCR method; the sequences and genetic variation of 29 PKV 3D genes were analyzed. [Result] Total positive rate of PKV in feces samples from suckling piglets with diarrhea was 65.18% （146/224）; total positive rate of PKV in pig farms was 85,2% （23/27）; nucleotide sequences and the deduced amino acid sequences of 29 PKV 3D genes shared 87.0%-100% and 92.7%-100% homologies with six PKV-related 3D sequences, respectively. [Conclusion] PKV infection is prevalent in suckling piglets in China; PKV 3D genes exhibit high diversity.

Genetic Variation of the ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus in East China during 2008-2010

[Objective] A total of 260 swine samples of dead or sick pigs collected from 7 provinces （municipalities） Jiangsu, Anhui, Shanghai, Shandong, Zhejiang, Fujian and Jiangxi of China during 2008-2010 were detected for porcine reproductive and respiratory syndrome virus （PRRSV）. And the ORF5 genes of some isolates were amplified and sequenced for understanding the molecule epidemiology and the genetic evolution of PRRSV in East China. [Method] Using RT-PCR method, PRRSV was detected by RT-PCR from samples. The complete ORF5 genes of 36 PRRSV positive samples was amplified, sequenced and analyzed with other 15 strains available on GenBank. [Result] PRRSV was detected in 118/260 of the clinical samples, with a positive rate was 45.4%. Sequence analysis showed that the 36 isolates of this study belonged to the North American-type PRRSV strains and were closely related to the highly pathogenic PRRSV （HP-PRRSV） with 94.6%-100% amino acid sequence identities. The sequence analysis combined with the phylogenetic analysis indicated that all these North American-type PRRSV strains in East China were further divided into five subgenotypes, subgenotype Ⅲ showed closer identity with HPPRRSV; almost all subgenotypes were found to be variable in the primary neutralizing epitope; subgenotypes Ⅲ and IV had more glycosylation sites than others. Although these 36 isolates were collected from different provinces in East China, there were no obvious relations between the distribution of PRRSV and the region. [Conclusion] The PRRSV infection was widespread and HP-PRRSV was the popular strain in East China during 2008-2010. However some different genetic characteristics appeared in the genomes, the genetic evolution was relatively stable. There exists a cross-cutting phenomenon on the genetic relationship of PRRSV isolates obtained from different provinces. Subgenotypes IV and V only appeared in some provinces, but the distribution of PRRSV did not show apparent geographical characteristics.

APOBEC3G Role on HIV Infection in Africa: A Systematic Review

The highly active antiretroviral treatment (HAART) has allowed people living with HIV to live longer with a better quality of life. However, toxicity and the emergence of drug resistance arise from HAART use. Therefore, new antiretroviral therapy is needed since no cure or vaccine is available against HIV. Virus-host interaction has been proven to be important in the last decade. Host factors such as the C-C chemokine receptor type 5 (CCR5), a receptor used by HIV to penetrate host cells, have led to the discovery of the Maraviroc, which is an antiretroviral medication used in the United States. In contrast, other factors like C-X-C Motif Chemokine Receptor 4 (CXCR4) and the Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3G (APOBEC3G), a potent host defense factor against HIV, is under investigation. APOBEC3G antiviral activity remains a possible therapeutic target against HIV. This systematic review aimed to synthesize the available evidence on the role of APOBEC3G polymorphisms and their expression on HIV infection disease progression in Africa. We used Web of Science, PubMed, Embase, and Google Scholar and searched for relevant publications in French or English reporting on APOBEC3G polymorphisms association with HIV infection in African populations from January 2009 to May 2023. The PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-analyzes) was used to process for reporting systematic review. Fifteen studies were included, of which seven were on APOBEC3G polymorphisms and eight were on APOBEC3G expression. Among the APOBEC3G polymorphisms, the most studied was H186R or rs8177832. The average of the minor allele frequency of H186R of APOBEC3G available for the studies included in this study was 0.29 with a 95% CI (0.172;0.401) and varied from 0.108 reported in Uganda to 0.47 recorded from Burkina Faso. The polymorphism H186R was not associated with HIV status in Southern Africa. However, the referent allele of H186R was protective against HIV infection in Western Central Africa, while in West Africa, it was the minor allele (G) of H186R which was protective against HIV. This review warrants a need to increase research on APOBEC3G, from its variants to its hypermutations on the continent with an essential variety of HIV-1 subtypes, to impact the research on A3G-based anti-HIV strategies.

Genetic Variation of Porcine Circovirus Type 2 Isolate 201105ZJ

[Objective] This study aimed to investigate the genetic variation of porcine circovirus type 2 （PCV2） in China. [Method] The strain was isolated from infected samples by cel passage and preliminarily identified by PCR and IFA. Ful-length genome of the isolated strain was obtained by specific amplification for homology and phylogenetic analysis. [Result] A PCV2 strain was successful y isolated and named 201105ZJ, which could proliferate in PK15 cel lines. Specific fragments could be amplified by specific PCR assay. According to results of IFA assay, specif-ic immunofluorescence was observed; the TCID50 was low （102.67）; the ful-length genome sequence of the isolated strain was 1 768 bp, sharing 94.1%-96.8% ho-mology with 13 reference strains; to be specific, the isolated strain exhibited the highest homology of 96.8% with AF055392PCV2a; the isolated strain 201105ZJ and reference strain AF055392 belonged to genotype PCV2a, exhibiting a distant genetic relationship with genotype PCV2c. [Conclusion] Characteristics of genetic variation of PCV2 isolate 201105ZJ provided theoretical basis for vaccine development, investi-gation of PCV2 pathogenesis, and prevention and control of porcine circovirus-as-sociated diseases （PCVAD） in East China.

Variation and selection analysis of Pinus koraiensis clones in northeast China

Understanding genetic variation is important for efficiently selecting excellent clones and utilizing genetic resources during tree breeding. We investigated 16 growth traits of 50 32-year-old Pinus koraiensis clones. Analyses of variance showed that all the test traits differed significantly among clones. Average height, diameter at breast height(DBH), and volume of all clones were 10.41 m, 21.30 cm,and 0.148 m3, respectively. Phenotypic coefficients of variation and repeatabilities of traits ranged from 4.37 to48.03% and from 0.013 to 0.900, respectively. There exists significant positive correlations among heights, diameter at different heights(1.3, 3.0, 5.0 m), and volumes; genetic correlation was close to phenotype correlation. Using four growth traits(height, DBH, volume, average crown width)as indices for a comprehensive evaluation, five clones(PK11, PK 19, PK 04, PK 14, and PK 28), whose traits scored in the top 10%, were selected as elite clones. For these clones,genetic gains in height, DBH, volume and crown height were 8.58, 13.02, 32.72 and 3.83%, respectively. These results provide important information for improving P.koraiensis breeding programs.

Genetic variation of Laminaria japonica (Phaeophyta) populations in China as revealed by RAPD markers

For the population genetics analysis of the naturally grown brown seaweed Laminaria japonica （Laminariales,Phaeophyta） sampled from Dalian,Yantai,Weihai,Rongcheng and Qingdao in China,ten primers were employed to produce 88 bands as revealed by randomly amplified polymorphic DNA （RAPD） markers,and all these bands were polymorphic.According to these band patterns,there were 94 distinct phenotypes occurred in 100 samples indicating the high heterozygosity of this kelp.Dalian population samples showed the highest percentage of polymorphism （71.67%）,and also the higher diversity estimated on the basis of the Shannon’s index （8.498）,suggesting that this population could be chosen as the best resource for genetic breeding.The highest diversity of Yantai population possibly resulted from the introduction of L.longissima used for interspecific cross breeding with L.japonica cultivated in China.From Dalian southwards to Qingdao,the genetic variation of the five populations became less with a decrease in latitude,possibly due to the natural selection especially of high temperature.The genetic distance （Φ ST values） of the five populations was a little significantly correlated with the geographical distance （r=0.496） at P =0.05 by Mantel’s test.Weihai,Rongcheng and Yantai populations were closely grouped genetically together by Neighbor-joining cluster analysis probably in that the dispersal of the kelp by propagules more easily occurring in the range of relatively short distance.The analysis of molecular variance （AMOVA） also demonstrated that the relatively higher variation occurred among populations （71.49%） at an extremely significant level （P 0.000 1）.All these evidence showed that there was a relatively distinct genetic differentiation among the sampled kelp populations,and L.japonica grown in China was also rather heterozygous in heredity.

Variation analyses of growth and wood properties of Larix olgensis clones in China

The growth traits (tree height, diameter at breast height, and stem straightness degree) and wood properties [wood density (WD), fiber length, fiber width, ash content (AC), lignin content, cellulose content, hemicellulose content (HEC), and holocellulose content] of 208 26-year-old Larix olgensis clones were analyzed. Except for WD and AC, there were significant differences (p < 0.01) for all traits among clones. The phenotypic coefficient of variation and repeatability of all traits were 9.34-35.33% and 0.218-0.930, respectively. Tree height and diameter at breast height showed significant positive correlation; however, the correlation coefficients among growth characteristics and wood properties were mostly not significant. Ten clones (L70, L56, L82, L90, L59, L91, L61, L92, L86, and L64) were selected as excellent clones under a selection rate of 5%, using tree height, diameter at breast height, and stem straightness degree as evaluation indexes, providing genetic gains of 28.69, 17.96, and 0.67%, respectively. Ten clones (L88, L305, L59, L66, L253, L304, L277, L298, L248, and L293) were selected as excellent clones using wood properties as an evaluation index, with a selection rate of 5%, providing genetic gains in WD, fiber length, fiber width, cellulose content, and HEC of 4.14, 3.64, 9.28, 6.77, and 9.61%, respectively. This study provides a theoretical basis for selecting excellent L. olgensis clones.

Genetic Variation in Triticum turgidum L. ssp. turgidum Landraces from China Assessed by EST-SSR Markers

It was helpful for the wheat improvement to evaluate the genetic resources of Triticum turgidum L. ssp. turgidum landraces. In this study, 68 turgidum landraces accessions, belonging to four geographic populations in China, were investigated by using EST-SSR markers. A total of 63 alleles were detected on 22 EST-SSR loci, and the number of alleles on each locus ranged from 1 to 5, with an average of 2.9. The results of the analysis of molecular variance （AMOVA） indicated that 92.5% of the total variations was attributed to the genetic variations within population, whereas only 7.5% variations among populations. Although the four populations had similar genetic diversity parameters, Sichuan population was yet distinguished from other populations when comparing the population samples in pairs. Significant correlations were detected by the statistic analysis among six genetic diversity parameters among each other. The selection difference between heterozygosty and homozygosty was also observed among different EST-SSR locus. The genetic similarity （GS） ranged from 0.18 to 0.98, with the mean of 0.72, and all accessions could be clustered into 7 groups. The dendrogram suggested that the genetic relationships among turgidum accessions evaluated by EST-SSR markers were unrelated to their geographic distributions. These results implied that turgidum landraces from China had the unique characters of genetic diversity.