Chromosome identification and karyotype using fluorescence in situ hybridization(FISH)provides a technical platform for genome and cytogenetic studies. Brassica juncea(brown mustard, 2n = 4× = 36; genome AABB) is...Chromosome identification and karyotype using fluorescence in situ hybridization(FISH)provides a technical platform for genome and cytogenetic studies. Brassica juncea(brown mustard, 2n = 4× = 36; genome AABB) is an allopolyploid species that originated from a spontaneous hybridization of Brassica rapa and Brassica nigra and contains many valuable traits. In this study, a multicolor FISH procedure allowing the identification of all 18 chromosomal pairs was developed by two-step hybridizations with probes on the same metaphase chromosomes. The distribution patterns and chromosomal localizations of six repeat sequences(satellite repeat p Br STR, 5S r DNA, 45 S r DNA, B genome-specific repeat p BNBH35, and centromeric satellite repeats Cent Br1 and Cent Br2) on B. juncea chromosomes were characterized.Comparative karyotype analyses showed that the genome is relatively stable in comparison with its diploid progenitor species and revealed intraspecific karyotypic diversity among three accessions of B. juncea. This study provides valuable information about the genome evolution of B. juncea and a toolkit that will be helpful for chromosome identification.展开更多
Multiple BAC-FISH is a powerful tool for modern cytogenetic researching in both animals and plants.But in cotton,this technique is unavailable due to the high percentage of repetitive sequences.
Background: Various genetic technologies have been employed in the identification of genomic complexity and refinement of prognostic classification of clinically heterogeneous disease of chronic lymphocytic leukemia (...Background: Various genetic technologies have been employed in the identification of genomic complexity and refinement of prognostic classification of clinically heterogeneous disease of chronic lymphocytic leukemia (CLL). Objective: The present study of interphase cytogenetics and conventional karyotyping was undertaken to perform comprehensive analysis of CLL genetics with an approach to refine early prognostication of disease. Material & Methods: Retrospective analysis by fluorescence in situ hybridization (FISH) was carried out on total 671 patients of CLL at diagnosis between 2008 and 2015. Conventional cytogenetics studies were performed in 50 of 671 patients using CPG Oligonucleotide + IL-2 and TPA (12-O-Tetradecanyl Phorbol 13-acetate) for stimulation of lymphocytes cultures. Results: Interphase cytogenetics could detect recurrent abnormalities such as del(13q14), +12, del(17p13), del(11q22), del(6q23) in 71% of cases. The incidence of del(13q) was higher in Rai stage 0, I, II (p = 0.0005);whereas patients with ≥2 aberrations were more common in advance stage III, IV (p = 0.001). Frequency of IgH translocation was 7%. Morphology and immunophenotypic analysis revealed atypical CLL with higher frequency of t(14;19) than t(14;18). Conventional karyotype could detect abnormal karyotype in 97% of cases which displayed targeted FISH abnormalities along with additional non-targeted chromosomal abnormalities. Patients with negative FISH markers showed clonal non-recurrent numerical and structural changes. The complex karyotype was identified in 24% cases which included targeted FISH aberrations as well as non-targeted numerical and structural abnormalities like deletions, and unbalanced translocations. A significant association was observed between complex karyotype and coexistence of ≥2 FISH markers (p = 0.009) and del(11q22) &/or del(17p) (p = 0.03). Conclusion: Our data of interphase FISH with integration of conventional karyotyping revealed genomic complexity that helped identification of biological subclasses with clinical impact at diagnosis. Further, these cytogenetic subclasses along with molecular markers are likely to evolve more refined prognostic groups, which will help design risk-adapted therapies in B-CLL.展开更多
In the subtribe Maydeae, Tripsacum and Zea are closely related genera. Tripsacum is a horticultural crop widely used as pasture forage. Previous studies suggested that Tripsacum might play an important role in maize o...In the subtribe Maydeae, Tripsacum and Zea are closely related genera. Tripsacum is a horticultural crop widely used as pasture forage. Previous studies suggested that Tripsacum might play an important role in maize origin and evolution. However, our understanding of the genomics and the evolution of Tripsacum remains limited. In this study, two diploids,T. dactyloides var. meridionale(2n = 36, MR) and T. dactyloides(2n = 36, DD), and one tetraploid,T. dactyloides(2n = 72, DL) were sequenced by low-coverage genome sequencing followed by graph-based cluster analysis. The results showed that 63.23%, 59.20%, and 61.57% of the respective genome of MR, DD, and DL were repetitive DNA sequence. The proportions of different repetitive sequences varied greatly among the three species. Fluorescence in situ hybridization(FISH) analysis of mitotic metaphase chromosomes with satellite repeats as the probes showed that the FISH signal patterns of DL were more similar to that of DD than to that of MR. Comparative analysis of the repeats also showed that DL shared more common repeat families with DD than with MR. Phylogenetic analysis of internal transcribed spacer region sequences further supported the evolutionary relationship among the three species. Repetitive sequences comparison showed that Tripsacum shared more repeat families with Zea than with Coix and Sorghum. Our study sheds new light on the genomics of Tripsacum and differential speciation in the Poaceae family.展开更多
This study conducted karyotype analysis on 56 Chinese kale varieties from Guangdong and Fujian provinces using conventional chromosome analysis methods.The varieties were categorized into four groups based on their or...This study conducted karyotype analysis on 56 Chinese kale varieties from Guangdong and Fujian provinces using conventional chromosome analysis methods.The varieties were categorized into four groups based on their origin and flower color:white-flower Chinese kale originating from Guangdong(GW),yellow-flower Chinese kale originating from Guangdong(GY),white-flower Chinese kale originating from Fujian(FW),and yellow-flower Chinese kale originating from Fujian(FY).Karyotype differences among the four groups of Chinese kale were analyzed,and the evolutionary relationship between yellow-flower and white-flower Chinese kale from the two regions was inferred based on karyotype parameters.The results indicated that all Chinese kale varieties were diploid with 2n=2x=18,including a pair of satellites.The chromosome types included median-centromere(m)and sub-median-centromere(sm),and the karyotypes were 1A and 2A,a sper karyotype asymmetry index.The karyotype 1A of Chinese kale was identified for the first time.Partial least squares discriminant analysis(PLS-DA)identified nine karyotypic indicators that differentiated the four groups,and these differences were further visualized using heatmaps and box plots.Based on the evolution trends in the four groups and PLS-DA analysis,it was speculated that white-flower Chinese kale originated from Guangdong,yellow-flower Chinese kale originated from Fujian,and GY and FW were derived from the cross of GW and FY.This study provides a reference for understanding the genetic relationships between Chinese kale in Guangdong and Fujian,and offers a cytological basis for the evolution,hybridization,and phylogenetic relationships of Chinese kale.展开更多
BACKGROUND Acute myeloid leukemia(AML)is a complicated disease with uncontrolled hematopoietic precursor proliferation induced by various genetic alterations.Runt-related transcription factor-1(RUNX1)is commonly disru...BACKGROUND Acute myeloid leukemia(AML)is a complicated disease with uncontrolled hematopoietic precursor proliferation induced by various genetic alterations.Runt-related transcription factor-1(RUNX1)is commonly disrupted by chromosomal translocations in hematological malignancies.AIM To characterize RUNX1 gene rearrangements and copy number variations in newly diagnosed adult AML patients,with an emphasis on the impact of clinical and laboratory features on the outcome.METHODS Fluorescence in situ hybridization was used to test RUNX1 gene alterations in 77 newly diagnosed adult AML cases.NPM1,FLT3/ITD,FLT3/TKD,and KIT mutations were tested by PCR.Prognostic clinical and laboratory findings were studied in relation to RUNX1 alterations.RESULTS RUNX1 abnormalities were detected by fluorescence in situ hybridization in 41.6%of patients:20.8%had translocations,22.1%had amplification,and 5.2%had deletion.Translocations prevailed in AML-M2(P=0.019)with a positive expression of myeloperoxidase(P=0.031),whereas deletions dominated in M4 and M5 subtypes(P=0.008)with a positive association with CD64 expression(P=0.05).The modal chromosomal number was higher in cases having amplifications(P=0.007)and lower in those with deletions(P=0.008).RUNX1 abnormalities were associated with complex karyotypes(P<0.001)and were mutually exclusive of NPM1 mutations.After 44 months of follow-up,RUNX1 abnormalities affected neither patients’response to treatment nor overall survival.CONCLUSION RUNX1 abnormalities were mutually exclusive of NPM1 mutations.RUNX1 abnormalities affected neither patients’response to treatment nor overall survival.展开更多
In order to precisely recognize and karyotype Brassica napus L. chromosomes, C0t-1 DNA was extracted from its genomic DNA, labeled with biotin-1 1-dUTP and in situ hybridized. The hybridized locations were detected by...In order to precisely recognize and karyotype Brassica napus L. chromosomes, C0t-1 DNA was extracted from its genomic DNA, labeled with biotin-1 1-dUTP and in situ hybridized. The hybridized locations were detected by Cy3-conjugated streptavidin. Specific fluorescence in situ hybridization （FISH） signal bands were detected on all individual chromosome pairs. Each chromosome pair showed specific banding patterns. The B. napus karyotype has been constructed, for the first time, on the basis of both Cot-1 DNA FISH banding patterns and chromosome morphology.展开更多
Complex congenital disorders may be caused by multiple genetic alterations and/or environmental hazards. Diagnosis and management of these diseases are usually difficult. Robust next-generation sequencing （NGS） tech...Complex congenital disorders may be caused by multiple genetic alterations and/or environmental hazards. Diagnosis and management of these diseases are usually difficult. Robust next-generation sequencing （NGS） technologies provide unprecedented opportunities to maximize mutation detection and improve genetic counseling and clinical management. Targeted or whole exome sequencing （WES） mainly detects protein-coding DNA sequence aberrations and is the major DNA sequencing technology that is entering clinical practice （Liu et al., 2014）.展开更多
To explore an effective and reliable karyotyping method in Brassica crop plants,Cot-1 DNA was isolated from Brassica oleracea genome,labeled as probe with Biotin-Nick Translation Mix kit,in situ hybridized to mitotic ...To explore an effective and reliable karyotyping method in Brassica crop plants,Cot-1 DNA was isolated from Brassica oleracea genome,labeled as probe with Biotin-Nick Translation Mix kit,in situ hybridized to mitotic spreads,and where specific fluorescent bands showed on each chromosome pair.25S and 5S rDNA were labeled as probes with DIG-Nick Translation Mix kit and Biotin-Nick Translation Mix kit,respectively,in situ hybridized to mitotic preparations,where 25S rDNA could be detected on two chromosome pairs and 5S rDNA on only one.Cot-1 DNA contains rDNA and chromosome sites identity between Cot-1 DNA and 25S rDNA was determined by dual-colour fluores-cence in situ hybridization.All these showed that the karyo-typing technique based on a combination of rDNA and Cot-1 DNA chromosome landmarks is superior to all but one.A more exact karyotype of B.oleracea has been analyzed based on a combination of rDNA sites,Cot-1 DNA fluorescent bands,chromosome lengths and arm ratios.展开更多
Background:Chromosomal abnormalities are important causes of ventriculomegaly(VM).In mild and isolated cases of fetal VM,obstetricians rarely give clear indications for pregnancy termination.We aimed to calculate the ...Background:Chromosomal abnormalities are important causes of ventriculomegaly(VM).In mild and isolated cases of fetal VM,obstetricians rarely give clear indications for pregnancy termination.We aimed to calculate the incidence of chromosomal abnormalities and incremental yield of chromosomal microarray analysis(CMA)in VM,providing more information on genetic counseling and prognostic evaluation for fetuses with VM.Methods:The Chinese language databases Wanfang Data,China National Knowledge Infrastructure,and China Biomedical Literature Database(from January 1,1991 to April 29,2020)and English language databases PubMed,Embase,and Cochrane Library(from January 1,1945 to April 29,2020)were systematically searched for articles on fetal VM.Diagnostic criteria were based on ultrasonographic or magnetic resonance imaging(MRI)assessment of lateral ventricular atrium width:≥10 to<15 mm for mild VM,and≥15 mm for severe VM.Isolated VM was defined by the absence of structural abnormalities other than VM detected by ultrasonography or MRI.R software was used for the meta-analysis to determine the incidence of chromosomal abnormalities and incremental yield of CMA in VM,and the combined rate and 95%confidence interval(CI)were calculated.Results:Twenty-three articles involving 1635 patients were included.The incidence of chromosomal abnormalities in VM was 9%(95%CI:5%-12%)and incremental yield of CMA in VM was 11%(95%CI:7%-16%).The incidences of chromosomal abnormalities in mild,severe,isolated,and non-isolated VM were 9%(95%CI:4%-16%),5%(95%CI:1%-11%),3%(95%CI:1%-6%),and 13%(95%CI:4%-25%),respectively.Conclusions:Applying CMA in VM improved the detection rate of abnormalities.When VM is confirmed by ultrasound or MRI,obstetricians should recommend fetal karyotype analysis to exclude chromosomal abnormalities.Moreover,CMA should be recommended preferentially in pregnant women with fetal VM who are undergoing invasive prenatal diagnosis.CMA cannot completely replace chromosome karyotype analysis.展开更多
BACKGROUND The clinical manifestations of trisomy 7 mosaicism are diverse and nonspecific,so prenatal diagnosis is very difficult.CASE SUMMARY Two pregnant women with abnormal prenatal screening results were included....BACKGROUND The clinical manifestations of trisomy 7 mosaicism are diverse and nonspecific,so prenatal diagnosis is very difficult.CASE SUMMARY Two pregnant women with abnormal prenatal screening results were included.One was a 22-year-old woman(G1P0).At 31st week of gestation,ultrasound revealed that the posterior horn of the left lateral ventricle was 10 mm and the right renal pelvis had a separation of 7 mm.The other pregnant woman was 33 years old(G2P1L1A0),and her fetus was found to have a cardiac malformation at the 24th week of gestation.Copy number variation sequencing,whole-exome sequencing and karyotype analysis were carried out after amniocentesis,and both fetuses were diagnosed with trisomy 7 mosaicism.After parental counseling,one woman continued the pregnancy,and the other woman terminated the pregnancy.CONCLUSION In trisomy 7 mosaicism,the low proportion of trisomy does not lead to abortion,but can result in abnormal fetal development,which can be detected via ultrasound.Therefore,clinicians need to pay more attention to various aspects of fetal growth and development,combining with imaging,cellular,molecular genetics and other methods to perform comprehensive evaluations of fetuses to provide more reliable genetic counseling for pregnant women.展开更多
The constant evolution of pathogens poses a threat to wheat resistance against diseases,endangering food security.Developing resistant wheat varieties is the most practical approach for circumventing this problem.As a...The constant evolution of pathogens poses a threat to wheat resistance against diseases,endangering food security.Developing resistant wheat varieties is the most practical approach for circumventing this problem.As a close relative of wheat,Aegilops geniculata,particularly accession SY159,has evolved numerous beneficial traits that could be applied to improve wheat.In this study,we established the karyotype of SY159 by fluorescence in situ hybridization(FISH)using the oligonucleotide probes Oligo-pTa535 and Oligo-pSc119.2 and a complete set of wheat–Ae.geniculata accession TA2899 addition lines as a reference.Using specific-locus amplified fragment sequencing(SLAF-seq)technology,400 specific markers were established for detecting the SY159 chromosomes with efficiencies reaching 81.5%.The SY159-specific markers were used to classify the different homologous groups of SY159 against the wheat-Ae.geniculata addition lines.We used these specific markers on the 7Mg chromosome after classification,and successfully confirmed their suitability for studying the different chromosomes of SY159.This study provides a foundation for accelerating the application of SY159 in genetic breeding programs designed to improve wheat.展开更多
Scientific knowledge about the ancestral genome of core eudicot plant kingdom can potentially have profound impacts on both basic and applied research,including evolution,genetics,genomics,ecology,agriculture,forestry...Scientific knowledge about the ancestral genome of core eudicot plant kingdom can potentially have profound impacts on both basic and applied research,including evolution,genetics,genomics,ecology,agriculture,forestry,and global climate.To investigate which plant conserves best the core eudicots common ancestor genome,we compared Arcto-Tertiary relict Nyssaceae and 30 other eudicot plant families.The genomes of Davidia involucrata(a known living fossil),Camptotheca acuminata and Nyssa sinensis,one per existent genus of Nyssaceae,were performed comparative genomic analysis.We found that Nyssaceae originated from a single Nyssaceae common tetraploidization event(NCT)-autotetraploidization 28-31 Mya after the core eudicot common hexaploidization(ECH).We identified Nyssaceae orthologous and paralogous genes,determined its chromosomal evolutionary trajectory,and reconstructed the Nyssaceae most recent ancestor genome.D.involucrata genome contained the entire seven paleochromosomes and 17 ECH-generated eudicot common ancestor chromosomes and was the slowest in mutation among the analyzed 42 species of 31 plant families.Combing both its high retention of paleochromosomes and its low mutation rate,D.involucrata provides the best case in conservation of the core eudicot paleogenome.展开更多
The identification of sex chromosomes is fundamental for exploring the mechanism and evolution of sex determination.Platichthys stellatus,a species exhibiting clear sexual dimorphism and homomorphic chromosome pairs,h...The identification of sex chromosomes is fundamental for exploring the mechanism and evolution of sex determination.Platichthys stellatus,a species exhibiting clear sexual dimorphism and homomorphic chromosome pairs,has received limited research concerning its sex determination mechanisms.Clarifying the sex chromosome of P.stellatus will enhance our understanding of sex chromosome evolution in Pleuronectiformes.This study employed whole-genome resequencing to investigate the sex chromosome and sex determination system in P.stellatus.Notably,Chr23 was identified as the sex chromosome in P.stellatus,with the sex-determining region(SDR)occupying 48.1%of the chromosome and featuring an XX/XY system.Sex chromosome turnover was observed within Pleuronectiformes,with P.stellatus,Verasper variegatus,and Hippoglossus hippoglossus sharing a common ancestral karyotype.No inversions were detected within the SDR of P.stellatus,although chromosomal rearrangements between sex chromosomes and autosomes were identified.Additionally,a sex-specific marker for P.stellatus was ascertained,enabling genetic sex identification,with significant implications for improving breeding programs and aquaculture practices.展开更多
Rationale:If one of the partners is having balanced autosomal translocation,it is usually observed that the offspring inherit either normal chromosomes,balanced translocation identical to one of the parent or unbalanc...Rationale:If one of the partners is having balanced autosomal translocation,it is usually observed that the offspring inherit either normal chromosomes,balanced translocation identical to one of the parent or unbalanced chromosomal rearrangements of the same parental chromosome having translocation.Concern:A unique case presented with history of 8 miscarriages for genetic counseling.The last abortus material evaluation showed monosomy of chromosome X(Turner syndrome)in all the analyzed cells.There was a history of infertility and also repeated second trimester abortions on the paternal side.On the maternal side,there was a history of intellectual disability.Diagnose:History of repeated abnormal pregnancy outcomes.Wife’s karyotype is normal;however,husband shows translocation between chromosome 4 and 22.Intervention:Peripheral blood sample around 3 mL was collected for karyotype.Embryo biopsy was done and DNA was extracted and processed for whole exome sequencing.Outcomes:Wife’s karyotype is normal and husband has translocation between chromosome 4 and 22.Surprisingly,the entire pregnancy outcome including embryo screening has different,complete or partial aneuploidies of chromosomes other than chromosome 4 and 22.Main lesson:Though the translocation in one of the parent is balanced,we have to think beyond traditional ways for evaluating a couple with repeated pregnancy loss as we cannot predict the errors at cell division.Option of in vitro fertilization and preimplantation genetic diagnosis in couples having balanced translocations should be discussed so that early intervention can prevent the agony of pregnancy loss.展开更多
Successful crossbreeding between Oreochromis niloticus and Sarotherodon melanotheron to produce a commercial hybrid has been difficult.The karyotypes and isoenzyme of these two species and their reciprocal hybrids（O....Successful crossbreeding between Oreochromis niloticus and Sarotherodon melanotheron to produce a commercial hybrid has been difficult.The karyotypes and isoenzyme of these two species and their reciprocal hybrids（O.niloticus ♀ × S.melanotheron ♂,S.melanotheron ♀ × O.niloticus ♂,the last not included in the isoenzyme study） were investigated via metaphase chromosomes obtained from head kidney cells and electropherogram of lactate dehydrogenase（LDH） isoenzymes from the liver,kidney,white muscle,heart,and eye balls.The diploid chromosome number（2n=44） and the fundamental number（NF=50） of the four tilapia genotypes were the same.However,the karyotype of O.niloticus had three pairs of sub-metacentric（sm）,twelve pairs of sub-telocentric（st）,and seven pairs of telocentric（t） chromosomes,while S.melanotheron had one pair of metacentric（m）,two pairs of sm,12 pairs of st,and seven pairs of t chromosomes.The reciprocal hybrids both showed a mixed karyotype range between their parents：0.5 pair of m,2.5 pairs of sm,12 pairs of st,and seven pairs of t chromosomes.In view of the electropherogram of isozymes,only the LDH of the kidney showed significant clear bands,with five bands in O.niloticus,three bands in S.melanotheron,and duplicated six bands in the hybrids.The bands varied depending on their activities and mobilities.We considered that the differences in karyotype and isoenzyme were related to the genetic mechanism for post-mating isolation,and provided some additional basic genetic background of their taxonomy.展开更多
The karyotype of two Chinese species in Orius were studied in male germ cells prepared on air dried slides stained with Giemsa. It is reported that two species have 24 chromosomes in diploid and X Y sex chromoso...The karyotype of two Chinese species in Orius were studied in male germ cells prepared on air dried slides stained with Giemsa. It is reported that two species have 24 chromosomes in diploid and X Y sex chromosomal mechanism. But the beha vior of chromosomes during meiosis between two species is different. Which show in the arrangment and location of the X chromosome and autosome at the late diakinesis as well as metaphase stage respectively and so on. These characteristics may be used in ta xonomy among species of Orius .展开更多
[Objective] The research aimed to study the morphological characteristics and karyotype of Pelteobagrus fulvidraco in Dongting Lake water system.[Method] By using the conventional biological morphometry,PHA and colchi...[Objective] The research aimed to study the morphological characteristics and karyotype of Pelteobagrus fulvidraco in Dongting Lake water system.[Method] By using the conventional biological morphometry,PHA and colchicine injection method in vivo,the morphological characteristics and karyotype of P.fulvidraco in Yuanshui River and Lishui River of Dongting Lake were analyzed.[Result] In three ratio traits including standard length/head length,standard length/caudal peduncle depth,head length/snout length,P.fulvidraco of Yuanshui River and Lishui River had significant differences（P0.05）.However,the number and karyotype of their chromosomes were same.The chromosome number was 2n = 52,and the karyotype formula was 20M＋12SM＋10ST＋10T.The number of chromosome arm was 84.[Conclusion] The research result had certain theoretical guidance significance for the protection and utilization of wild P.resource in Dongting Lake water system.展开更多
Karyotype of 3 species of the genus Orius Wolff were firstly studied using the Giemsa staining and squashing slides method in male germ cell. It is reported that number of chromosome and system of sex chromosome of ha...Karyotype of 3 species of the genus Orius Wolff were firstly studied using the Giemsa staining and squashing slides method in male germ cell. It is reported that number of chromosome and system of sex chromosome of haploid chromosome complement of these species is n=11A＋X （Y） in the first meiotic stage. However, the behavior of chromosomes among the species are different distinctly, thus cytotaxonomic character can be adopted to the identification among the three species.展开更多
Karyotype analysis of Lycium ruthenicum Murr. was carried out in this study. The results showed that the chromosome number was 2n=2x=24; the arm index was 48; the ratio of the longest chromosome to the shortest one wa...Karyotype analysis of Lycium ruthenicum Murr. was carried out in this study. The results showed that the chromosome number was 2n=2x=24; the arm index was 48; the ratio of the longest chromosome to the shortest one was 1.31; the proportions of chromosomes with arm ratio higher than 2 was 0.08; the asymmetry index was 57.02; the karyotype type was 2A; and the karyotype formula was 2n=-24=20m＋4sm.展开更多
基金supported by the National Natural Science Foundation of China (Nos. 31471173, 31560302)the Inner Mongolia Science & Technology Plan (No. 2014MS0305)+2 种基金the Program for Introducing Talents to University (No. 135121)the Inner Mongolia Innovation Guide Fund (No. 5153202)the Basic and Frontier Technology Research Program of Henan Province (No. 132300410284) to GX Tan
文摘Chromosome identification and karyotype using fluorescence in situ hybridization(FISH)provides a technical platform for genome and cytogenetic studies. Brassica juncea(brown mustard, 2n = 4× = 36; genome AABB) is an allopolyploid species that originated from a spontaneous hybridization of Brassica rapa and Brassica nigra and contains many valuable traits. In this study, a multicolor FISH procedure allowing the identification of all 18 chromosomal pairs was developed by two-step hybridizations with probes on the same metaphase chromosomes. The distribution patterns and chromosomal localizations of six repeat sequences(satellite repeat p Br STR, 5S r DNA, 45 S r DNA, B genome-specific repeat p BNBH35, and centromeric satellite repeats Cent Br1 and Cent Br2) on B. juncea chromosomes were characterized.Comparative karyotype analyses showed that the genome is relatively stable in comparison with its diploid progenitor species and revealed intraspecific karyotypic diversity among three accessions of B. juncea. This study provides valuable information about the genome evolution of B. juncea and a toolkit that will be helpful for chromosome identification.
文摘Multiple BAC-FISH is a powerful tool for modern cytogenetic researching in both animals and plants.But in cotton,this technique is unavailable due to the high percentage of repetitive sequences.
文摘Background: Various genetic technologies have been employed in the identification of genomic complexity and refinement of prognostic classification of clinically heterogeneous disease of chronic lymphocytic leukemia (CLL). Objective: The present study of interphase cytogenetics and conventional karyotyping was undertaken to perform comprehensive analysis of CLL genetics with an approach to refine early prognostication of disease. Material & Methods: Retrospective analysis by fluorescence in situ hybridization (FISH) was carried out on total 671 patients of CLL at diagnosis between 2008 and 2015. Conventional cytogenetics studies were performed in 50 of 671 patients using CPG Oligonucleotide + IL-2 and TPA (12-O-Tetradecanyl Phorbol 13-acetate) for stimulation of lymphocytes cultures. Results: Interphase cytogenetics could detect recurrent abnormalities such as del(13q14), +12, del(17p13), del(11q22), del(6q23) in 71% of cases. The incidence of del(13q) was higher in Rai stage 0, I, II (p = 0.0005);whereas patients with ≥2 aberrations were more common in advance stage III, IV (p = 0.001). Frequency of IgH translocation was 7%. Morphology and immunophenotypic analysis revealed atypical CLL with higher frequency of t(14;19) than t(14;18). Conventional karyotype could detect abnormal karyotype in 97% of cases which displayed targeted FISH abnormalities along with additional non-targeted chromosomal abnormalities. Patients with negative FISH markers showed clonal non-recurrent numerical and structural changes. The complex karyotype was identified in 24% cases which included targeted FISH aberrations as well as non-targeted numerical and structural abnormalities like deletions, and unbalanced translocations. A significant association was observed between complex karyotype and coexistence of ≥2 FISH markers (p = 0.009) and del(11q22) &/or del(17p) (p = 0.03). Conclusion: Our data of interphase FISH with integration of conventional karyotyping revealed genomic complexity that helped identification of biological subclasses with clinical impact at diagnosis. Further, these cytogenetic subclasses along with molecular markers are likely to evolve more refined prognostic groups, which will help design risk-adapted therapies in B-CLL.
基金supported by the National Natural Science Foundation of China (Nos. 31471499, 91535206)
文摘In the subtribe Maydeae, Tripsacum and Zea are closely related genera. Tripsacum is a horticultural crop widely used as pasture forage. Previous studies suggested that Tripsacum might play an important role in maize origin and evolution. However, our understanding of the genomics and the evolution of Tripsacum remains limited. In this study, two diploids,T. dactyloides var. meridionale(2n = 36, MR) and T. dactyloides(2n = 36, DD), and one tetraploid,T. dactyloides(2n = 72, DL) were sequenced by low-coverage genome sequencing followed by graph-based cluster analysis. The results showed that 63.23%, 59.20%, and 61.57% of the respective genome of MR, DD, and DL were repetitive DNA sequence. The proportions of different repetitive sequences varied greatly among the three species. Fluorescence in situ hybridization(FISH) analysis of mitotic metaphase chromosomes with satellite repeats as the probes showed that the FISH signal patterns of DL were more similar to that of DD than to that of MR. Comparative analysis of the repeats also showed that DL shared more common repeat families with DD than with MR. Phylogenetic analysis of internal transcribed spacer region sequences further supported the evolutionary relationship among the three species. Repetitive sequences comparison showed that Tripsacum shared more repeat families with Zea than with Coix and Sorghum. Our study sheds new light on the genomics of Tripsacum and differential speciation in the Poaceae family.
基金funded by National Natural Science Foundation of China(32372732,32372683,32460750,32072586,31500247)Sichuan Science and Technology Program(2023ZYD0090,24NSFSC0404,2025ZNSFSC1112)+2 种基金Sichuan Innovation Team of National Modern Agricultural Industry Technology System(SCCXTD-2024-05)Key Laboratory of Storage of Agricultural Products,Ministry of Agriculture and Rural Aairs(kt202410)Chengdu Science and Technology Program(2024-YF05-02375-SN).
文摘This study conducted karyotype analysis on 56 Chinese kale varieties from Guangdong and Fujian provinces using conventional chromosome analysis methods.The varieties were categorized into four groups based on their origin and flower color:white-flower Chinese kale originating from Guangdong(GW),yellow-flower Chinese kale originating from Guangdong(GY),white-flower Chinese kale originating from Fujian(FW),and yellow-flower Chinese kale originating from Fujian(FY).Karyotype differences among the four groups of Chinese kale were analyzed,and the evolutionary relationship between yellow-flower and white-flower Chinese kale from the two regions was inferred based on karyotype parameters.The results indicated that all Chinese kale varieties were diploid with 2n=2x=18,including a pair of satellites.The chromosome types included median-centromere(m)and sub-median-centromere(sm),and the karyotypes were 1A and 2A,a sper karyotype asymmetry index.The karyotype 1A of Chinese kale was identified for the first time.Partial least squares discriminant analysis(PLS-DA)identified nine karyotypic indicators that differentiated the four groups,and these differences were further visualized using heatmaps and box plots.Based on the evolution trends in the four groups and PLS-DA analysis,it was speculated that white-flower Chinese kale originated from Guangdong,yellow-flower Chinese kale originated from Fujian,and GY and FW were derived from the cross of GW and FY.This study provides a reference for understanding the genetic relationships between Chinese kale in Guangdong and Fujian,and offers a cytological basis for the evolution,hybridization,and phylogenetic relationships of Chinese kale.
文摘BACKGROUND Acute myeloid leukemia(AML)is a complicated disease with uncontrolled hematopoietic precursor proliferation induced by various genetic alterations.Runt-related transcription factor-1(RUNX1)is commonly disrupted by chromosomal translocations in hematological malignancies.AIM To characterize RUNX1 gene rearrangements and copy number variations in newly diagnosed adult AML patients,with an emphasis on the impact of clinical and laboratory features on the outcome.METHODS Fluorescence in situ hybridization was used to test RUNX1 gene alterations in 77 newly diagnosed adult AML cases.NPM1,FLT3/ITD,FLT3/TKD,and KIT mutations were tested by PCR.Prognostic clinical and laboratory findings were studied in relation to RUNX1 alterations.RESULTS RUNX1 abnormalities were detected by fluorescence in situ hybridization in 41.6%of patients:20.8%had translocations,22.1%had amplification,and 5.2%had deletion.Translocations prevailed in AML-M2(P=0.019)with a positive expression of myeloperoxidase(P=0.031),whereas deletions dominated in M4 and M5 subtypes(P=0.008)with a positive association with CD64 expression(P=0.05).The modal chromosomal number was higher in cases having amplifications(P=0.007)and lower in those with deletions(P=0.008).RUNX1 abnormalities were associated with complex karyotypes(P<0.001)and were mutually exclusive of NPM1 mutations.After 44 months of follow-up,RUNX1 abnormalities affected neither patients’response to treatment nor overall survival.CONCLUSION RUNX1 abnormalities were mutually exclusive of NPM1 mutations.RUNX1 abnormalities affected neither patients’response to treatment nor overall survival.
文摘In order to precisely recognize and karyotype Brassica napus L. chromosomes, C0t-1 DNA was extracted from its genomic DNA, labeled with biotin-1 1-dUTP and in situ hybridized. The hybridized locations were detected by Cy3-conjugated streptavidin. Specific fluorescence in situ hybridization （FISH） signal bands were detected on all individual chromosome pairs. Each chromosome pair showed specific banding patterns. The B. napus karyotype has been constructed, for the first time, on the basis of both Cot-1 DNA FISH banding patterns and chromosome morphology.
基金supported by the grants from the National High Technology Research and Development Program of China (863 Program) (No. 2012AA02A201)the Tianjin Natural Science Foundation (No. 13JCQNJC10400)
文摘Complex congenital disorders may be caused by multiple genetic alterations and/or environmental hazards. Diagnosis and management of these diseases are usually difficult. Robust next-generation sequencing （NGS） technologies provide unprecedented opportunities to maximize mutation detection and improve genetic counseling and clinical management. Targeted or whole exome sequencing （WES） mainly detects protein-coding DNA sequence aberrations and is the major DNA sequencing technology that is entering clinical practice （Liu et al., 2014）.
基金This work was supported by the President Foundation of the Institute of Oil Crops,Chinese Academy of Agricultural Sciences(No.200301)the Topping Youth Foundation of Hubei Province(No.2005ABB028)the Chenguang Program of Wuhan City(No.20045006071-37).
文摘To explore an effective and reliable karyotyping method in Brassica crop plants,Cot-1 DNA was isolated from Brassica oleracea genome,labeled as probe with Biotin-Nick Translation Mix kit,in situ hybridized to mitotic spreads,and where specific fluorescent bands showed on each chromosome pair.25S and 5S rDNA were labeled as probes with DIG-Nick Translation Mix kit and Biotin-Nick Translation Mix kit,respectively,in situ hybridized to mitotic preparations,where 25S rDNA could be detected on two chromosome pairs and 5S rDNA on only one.Cot-1 DNA contains rDNA and chromosome sites identity between Cot-1 DNA and 25S rDNA was determined by dual-colour fluores-cence in situ hybridization.All these showed that the karyo-typing technique based on a combination of rDNA and Cot-1 DNA chromosome landmarks is superior to all but one.A more exact karyotype of B.oleracea has been analyzed based on a combination of rDNA sites,Cot-1 DNA fluorescent bands,chromosome lengths and arm ratios.
文摘Background:Chromosomal abnormalities are important causes of ventriculomegaly(VM).In mild and isolated cases of fetal VM,obstetricians rarely give clear indications for pregnancy termination.We aimed to calculate the incidence of chromosomal abnormalities and incremental yield of chromosomal microarray analysis(CMA)in VM,providing more information on genetic counseling and prognostic evaluation for fetuses with VM.Methods:The Chinese language databases Wanfang Data,China National Knowledge Infrastructure,and China Biomedical Literature Database(from January 1,1991 to April 29,2020)and English language databases PubMed,Embase,and Cochrane Library(from January 1,1945 to April 29,2020)were systematically searched for articles on fetal VM.Diagnostic criteria were based on ultrasonographic or magnetic resonance imaging(MRI)assessment of lateral ventricular atrium width:≥10 to<15 mm for mild VM,and≥15 mm for severe VM.Isolated VM was defined by the absence of structural abnormalities other than VM detected by ultrasonography or MRI.R software was used for the meta-analysis to determine the incidence of chromosomal abnormalities and incremental yield of CMA in VM,and the combined rate and 95%confidence interval(CI)were calculated.Results:Twenty-three articles involving 1635 patients were included.The incidence of chromosomal abnormalities in VM was 9%(95%CI:5%-12%)and incremental yield of CMA in VM was 11%(95%CI:7%-16%).The incidences of chromosomal abnormalities in mild,severe,isolated,and non-isolated VM were 9%(95%CI:4%-16%),5%(95%CI:1%-11%),3%(95%CI:1%-6%),and 13%(95%CI:4%-25%),respectively.Conclusions:Applying CMA in VM improved the detection rate of abnormalities.When VM is confirmed by ultrasound or MRI,obstetricians should recommend fetal karyotype analysis to exclude chromosomal abnormalities.Moreover,CMA should be recommended preferentially in pregnant women with fetal VM who are undergoing invasive prenatal diagnosis.CMA cannot completely replace chromosome karyotype analysis.
文摘BACKGROUND The clinical manifestations of trisomy 7 mosaicism are diverse and nonspecific,so prenatal diagnosis is very difficult.CASE SUMMARY Two pregnant women with abnormal prenatal screening results were included.One was a 22-year-old woman(G1P0).At 31st week of gestation,ultrasound revealed that the posterior horn of the left lateral ventricle was 10 mm and the right renal pelvis had a separation of 7 mm.The other pregnant woman was 33 years old(G2P1L1A0),and her fetus was found to have a cardiac malformation at the 24th week of gestation.Copy number variation sequencing,whole-exome sequencing and karyotype analysis were carried out after amniocentesis,and both fetuses were diagnosed with trisomy 7 mosaicism.After parental counseling,one woman continued the pregnancy,and the other woman terminated the pregnancy.CONCLUSION In trisomy 7 mosaicism,the low proportion of trisomy does not lead to abortion,but can result in abnormal fetal development,which can be detected via ultrasound.Therefore,clinicians need to pay more attention to various aspects of fetal growth and development,combining with imaging,cellular,molecular genetics and other methods to perform comprehensive evaluations of fetuses to provide more reliable genetic counseling for pregnant women.
基金funded by the National Natural Science Foundation of China(31471481)the Project of Science and Technology of Shaanxi Province of China(2021NY-081 and 2023YBNY-033)。
文摘The constant evolution of pathogens poses a threat to wheat resistance against diseases,endangering food security.Developing resistant wheat varieties is the most practical approach for circumventing this problem.As a close relative of wheat,Aegilops geniculata,particularly accession SY159,has evolved numerous beneficial traits that could be applied to improve wheat.In this study,we established the karyotype of SY159 by fluorescence in situ hybridization(FISH)using the oligonucleotide probes Oligo-pTa535 and Oligo-pSc119.2 and a complete set of wheat–Ae.geniculata accession TA2899 addition lines as a reference.Using specific-locus amplified fragment sequencing(SLAF-seq)technology,400 specific markers were established for detecting the SY159 chromosomes with efficiencies reaching 81.5%.The SY159-specific markers were used to classify the different homologous groups of SY159 against the wheat-Ae.geniculata addition lines.We used these specific markers on the 7Mg chromosome after classification,and successfully confirmed their suitability for studying the different chromosomes of SY159.This study provides a foundation for accelerating the application of SY159 in genetic breeding programs designed to improve wheat.
基金supported by the National Natural Science Foundation of China(Grant Nos.32170236,31501333,and 32000405)Natural Science Foundation of Hebei Province(Grant No.C2020209064)the Innovation and Entrepreneurship Training Program for College Students of North China University of Science and Technology(Grant No.X2019252)。
文摘Scientific knowledge about the ancestral genome of core eudicot plant kingdom can potentially have profound impacts on both basic and applied research,including evolution,genetics,genomics,ecology,agriculture,forestry,and global climate.To investigate which plant conserves best the core eudicots common ancestor genome,we compared Arcto-Tertiary relict Nyssaceae and 30 other eudicot plant families.The genomes of Davidia involucrata(a known living fossil),Camptotheca acuminata and Nyssa sinensis,one per existent genus of Nyssaceae,were performed comparative genomic analysis.We found that Nyssaceae originated from a single Nyssaceae common tetraploidization event(NCT)-autotetraploidization 28-31 Mya after the core eudicot common hexaploidization(ECH).We identified Nyssaceae orthologous and paralogous genes,determined its chromosomal evolutionary trajectory,and reconstructed the Nyssaceae most recent ancestor genome.D.involucrata genome contained the entire seven paleochromosomes and 17 ECH-generated eudicot common ancestor chromosomes and was the slowest in mutation among the analyzed 42 species of 31 plant families.Combing both its high retention of paleochromosomes and its low mutation rate,D.involucrata provides the best case in conservation of the core eudicot paleogenome.
基金supported by the National Key Research and Development Program of China(2022YFD2400100)National Natural Science Foundation of Shandong Province of China(ZR2023QC006)National Natural Science Foundation of China(32403053)。
文摘The identification of sex chromosomes is fundamental for exploring the mechanism and evolution of sex determination.Platichthys stellatus,a species exhibiting clear sexual dimorphism and homomorphic chromosome pairs,has received limited research concerning its sex determination mechanisms.Clarifying the sex chromosome of P.stellatus will enhance our understanding of sex chromosome evolution in Pleuronectiformes.This study employed whole-genome resequencing to investigate the sex chromosome and sex determination system in P.stellatus.Notably,Chr23 was identified as the sex chromosome in P.stellatus,with the sex-determining region(SDR)occupying 48.1%of the chromosome and featuring an XX/XY system.Sex chromosome turnover was observed within Pleuronectiformes,with P.stellatus,Verasper variegatus,and Hippoglossus hippoglossus sharing a common ancestral karyotype.No inversions were detected within the SDR of P.stellatus,although chromosomal rearrangements between sex chromosomes and autosomes were identified.Additionally,a sex-specific marker for P.stellatus was ascertained,enabling genetic sex identification,with significant implications for improving breeding programs and aquaculture practices.
基金support from ICMR-National Institute for Research in Reproductive Health(ICMR-NIRRH).
文摘Rationale:If one of the partners is having balanced autosomal translocation,it is usually observed that the offspring inherit either normal chromosomes,balanced translocation identical to one of the parent or unbalanced chromosomal rearrangements of the same parental chromosome having translocation.Concern:A unique case presented with history of 8 miscarriages for genetic counseling.The last abortus material evaluation showed monosomy of chromosome X(Turner syndrome)in all the analyzed cells.There was a history of infertility and also repeated second trimester abortions on the paternal side.On the maternal side,there was a history of intellectual disability.Diagnose:History of repeated abnormal pregnancy outcomes.Wife’s karyotype is normal;however,husband shows translocation between chromosome 4 and 22.Intervention:Peripheral blood sample around 3 mL was collected for karyotype.Embryo biopsy was done and DNA was extracted and processed for whole exome sequencing.Outcomes:Wife’s karyotype is normal and husband has translocation between chromosome 4 and 22.Surprisingly,the entire pregnancy outcome including embryo screening has different,complete or partial aneuploidies of chromosomes other than chromosome 4 and 22.Main lesson:Though the translocation in one of the parent is balanced,we have to think beyond traditional ways for evaluating a couple with repeated pregnancy loss as we cannot predict the errors at cell division.Option of in vitro fertilization and preimplantation genetic diagnosis in couples having balanced translocations should be discussed so that early intervention can prevent the agony of pregnancy loss.
文摘Successful crossbreeding between Oreochromis niloticus and Sarotherodon melanotheron to produce a commercial hybrid has been difficult.The karyotypes and isoenzyme of these two species and their reciprocal hybrids（O.niloticus ♀ × S.melanotheron ♂,S.melanotheron ♀ × O.niloticus ♂,the last not included in the isoenzyme study） were investigated via metaphase chromosomes obtained from head kidney cells and electropherogram of lactate dehydrogenase（LDH） isoenzymes from the liver,kidney,white muscle,heart,and eye balls.The diploid chromosome number（2n=44） and the fundamental number（NF=50） of the four tilapia genotypes were the same.However,the karyotype of O.niloticus had three pairs of sub-metacentric（sm）,twelve pairs of sub-telocentric（st）,and seven pairs of telocentric（t） chromosomes,while S.melanotheron had one pair of metacentric（m）,two pairs of sm,12 pairs of st,and seven pairs of t chromosomes.The reciprocal hybrids both showed a mixed karyotype range between their parents：0.5 pair of m,2.5 pairs of sm,12 pairs of st,and seven pairs of t chromosomes.In view of the electropherogram of isozymes,only the LDH of the kidney showed significant clear bands,with five bands in O.niloticus,three bands in S.melanotheron,and duplicated six bands in the hybrids.The bands varied depending on their activities and mobilities.We considered that the differences in karyotype and isoenzyme were related to the genetic mechanism for post-mating isolation,and provided some additional basic genetic background of their taxonomy.
基金The work was supported by National Natural Science Foundation of China( 395 70 1 0 8) for second author and Zhejiang Forestry College Science Foundation
文摘The karyotype of two Chinese species in Orius were studied in male germ cells prepared on air dried slides stained with Giemsa. It is reported that two species have 24 chromosomes in diploid and X Y sex chromosomal mechanism. But the beha vior of chromosomes during meiosis between two species is different. Which show in the arrangment and location of the X chromosome and autosome at the late diakinesis as well as metaphase stage respectively and so on. These characteristics may be used in ta xonomy among species of Orius .
基金Supported by National Natural Science Foundation of China(30972260)Science and Technology Innovation Team Plan of Hunan Higher Education(2010)~~
文摘[Objective] The research aimed to study the morphological characteristics and karyotype of Pelteobagrus fulvidraco in Dongting Lake water system.[Method] By using the conventional biological morphometry,PHA and colchicine injection method in vivo,the morphological characteristics and karyotype of P.fulvidraco in Yuanshui River and Lishui River of Dongting Lake were analyzed.[Result] In three ratio traits including standard length/head length,standard length/caudal peduncle depth,head length/snout length,P.fulvidraco of Yuanshui River and Lishui River had significant differences（P0.05）.However,the number and karyotype of their chromosomes were same.The chromosome number was 2n = 52,and the karyotype formula was 20M＋12SM＋10ST＋10T.The number of chromosome arm was 84.[Conclusion] The research result had certain theoretical guidance significance for the protection and utilization of wild P.resource in Dongting Lake water system.
基金The work was supported by Zhejiang Forestry College Science Foundation
文摘Karyotype of 3 species of the genus Orius Wolff were firstly studied using the Giemsa staining and squashing slides method in male germ cell. It is reported that number of chromosome and system of sex chromosome of haploid chromosome complement of these species is n=11A＋X （Y） in the first meiotic stage. However, the behavior of chromosomes among the species are different distinctly, thus cytotaxonomic character can be adopted to the identification among the three species.
基金Supported by National Natural Science Foundation of China(31100401)Tianjin Science and Technology Plan Project(13ZLZLZF05700)+1 种基金Fund for Special Science and Technology Correspondents of Tianjin City(15JCTPJC59500)College Students'Innovative Entrepreneurial Training Program of Tianjin City(201610061102)~~
文摘Karyotype analysis of Lycium ruthenicum Murr. was carried out in this study. The results showed that the chromosome number was 2n=2x=24; the arm index was 48; the ratio of the longest chromosome to the shortest one was 1.31; the proportions of chromosomes with arm ratio higher than 2 was 0.08; the asymmetry index was 57.02; the karyotype type was 2A; and the karyotype formula was 2n=-24=20m＋4sm.
