Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the...Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization (M-FISH) for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.展开更多
Standard FISH protocols using fluorochrome-labeled oligonucleotide probes have been successfully applied for in situ detection.However,optimized protocols of FISH for specific eukaryotes in marine environments are oft...Standard FISH protocols using fluorochrome-labeled oligonucleotide probes have been successfully applied for in situ detection.However,optimized protocols of FISH for specific eukaryotes in marine environments are often not developed.This study optimized the conditions of fluorescence in situ hybridization (FISH) by using two polar isolated microalgae.The modified conditions were as follows:(1) 10 mg·mL^(-1) lysozyme solution pretreatment at 37℃for 30 min;(2) the hybridization buffer including 20%formamide;(3) the hybridization condition was 47℃for 6 h.The cells enumerated by FISH were compared with those enumerated by flow cytometry(FCM) and DAPI to confirm the cell loss and hybridization efficiency.The optimized protocol was also successfully applied to Arctic Ocean samples,which were found to be dominated by Micromonas sp.The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples.展开更多
Objective: The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis a...Objective: The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis and prognosis. Methods: To observe MRP-1/C9mRNA expression, tissue microarray (TMA) containing 54 lung cancers and 10 normal lung tissues was prepared and Fluorescence in situ hybridization was used. Results: The positive rate of MRP-1/CD9 expression was 48.1% in lung cancer, lower than that of normal lung tissues. The statistical difference was significant (P〈0.05). Its protein expression had no relationship with the patients' ages, sex and the macroscopic type of tumor, but had relationships with the histological type, clinical stage, differentiated degree and metastasis. The expression in non-small cell lung cancer (NSCLC) was higher than that in small cell lung cancer (SCLC); in well-moderately differentiated group was higher than that in poorly differentiated group; Earlier period group (I+II) was higher than in later period group (Ⅲ+Ⅳ); and in group without lymphoid metastasis was higher than in patients with lymphoid metastasis. Conclusion: The progression of the lung cancer maybe related with the descended MRP-1/Cd9 expression, which may be useful in evaluating the prognosis of cancer patients.展开更多
A molecular biology method, fluorescent in situ hybridization(FISH), in which the pre-treatment was improved in allusion to the media of the constructed wetlands(CW), e.g. the soil and the grit, was used to invest...A molecular biology method, fluorescent in situ hybridization(FISH), in which the pre-treatment was improved in allusion to the media of the constructed wetlands(CW), e.g. the soil and the grit, was used to investigate the vertical distribution characteristics of ammonia-oxidizing bacteria(AOB) quantity and the relation with oxidation-reduction potential(ORP) in the Typha latifolia constructed wetlands under three different Ioadings in summer from May to September. Results showed that the quantity of the AOB decreased in the Typha latifolia CW with the increase of vertical depth. However, the AOB quantity was 2-4 times the quantity of the control in the root area. Additionally, ORP in the rhizosphere was found to be higher than other areas, which showed that Typha latifolia CW was in an aerobic state in summer when using simulated non-point sewage at the rural area of Taihu Lake in China and small town combined sewage.展开更多
Telomeres form the ends of eukaryotic chromosomes and serve as protective caps that keep chromosomes structure independency and completeness. The first plant telomere DNA was isolated from Arabidopsis thaliana and was...Telomeres form the ends of eukaryotic chromosomes and serve as protective caps that keep chromosomes structure independency and completeness. The first plant telomere DNA was isolated from Arabidopsis thaliana and was shown to have tandemly repeated sequence 5-TTTAGGG-3: The Arabidopsis-type telomere has been found in many plants, but several reports indicate that this sequence is absent in some plants. Up to now, no research has been conducted on the telomere of cotton. In this paper, the Arabidopsis-type telomere sequence was amplified and cloned using the primers designed based on the fragment containing telomere sequence in an Arabidopsis bacterial artificial chromosome (BAC). Fluorescence in situ hybridization (FISH) with cotton metaphase chromosomes using the Arabidopsis-type telomere sequence as probes indicated that the signals were located at all chromosome ends of seven diploid and two tetraploid cotton species with different signal intensities among chromosome complements of different cotton species, even between long and short arms of the same chromosome. To identify the signals of FISH, the genome DNA of Xinhai 7, a cultivar of Gossypium barbadense, digested by BAL-31 nuclease was introduced in this study. The result of BAL-31 digestion indicated that the hybridization signals of FISH represent the outermost DNA sequence of each cotton chromosomes. So we first proved that the telomeric repeats of cotton cross-hybridize with that of Arabidopsis. The results of terminal restriction fragment (TRF) showed significant variation in telomere length among cotton species. The telomere length of cultivated cotton was close to 20 kb and was larger than those of wild cotton species whose telomere length rahged from 6 to 20 kb.展开更多
Objective To identify the deletions in Duchenne/Becker muscular dystrophy (DMD/ BMD) by using fluorescence in situ hybridization (FISH)Methods The exon-specific cosmid DNA probes (representing 18 exons) were used to p...Objective To identify the deletions in Duchenne/Becker muscular dystrophy (DMD/ BMD) by using fluorescence in situ hybridization (FISH)Methods The exon-specific cosmid DNA probes (representing 18 exons) were used to perform one-color FISH on metaphase and interphase preparations. The peripheral blood samples from 9 normal people (4 males and 5 females) and 5 females from independent deletion DMD/BMD families, as well as 2 amniotic fluid specimens and 2 chorionic villus samples (CVS) from normal pregnant females were analyzed. Results 72%-100% of peripheral blood lymphocyte metaphases or interphases, 60% -70% of amniocyte interphases, and 95 - 99% of chorionic villus cell interphases showed expected signals. One suspected female was identified as deletion carriers and two were excluded.Conclusion FISH in combination with other available techniques allows efficient screening of DMD/BMD deletion carriers, which also lay the ground work for prenatal diagnosis for potential fetal carriers.展开更多
It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of p...It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of primers were designed for amplification to verify this linkage of two genes in a kelp cultivar of Saccharina japonica,one of species in Ochrophyta.This result supplemented the previous report that 5S rDNA was unlinked to 25S rDNA in this kelp.In order to simultaneously visualize this unlinkage of two genes,dual-color fluorescence in situ hybridization(FISH)technique was applied to the cytogenetics of S.japonica.Dual-color FISH images showed that two and four hybridization signals were present in the kelp gametophyte and sporophyte,respectively,metaphase nuclei hybridized simultaneously with the labeled probes of 18S rDNA and 5S rDNA.Both haploid and diploid karyotypes in decreasing length of chromosomes showed that 18S-5.8S-25S rDNA was localized at the interstitial region of Chromosome 23,whereas 5S rDNA resided at the sub-telomeric region of Chromosome 27.These karyotypes suggested that the kelp nuclear genome had only one locus of each rRNA gene,and their loci on different chromosomes indicated the physical unlinkage of 5S rDNA to 18S-5.8S-25S rDNA in this kelp.Therefore,dual-color FISH seems to be a powerful technique for the discrimination and pairing of chromosomes featured in both small size and nearly identical shape in S.japonica.展开更多
In order to explore researches about the chromosome karyotype analysis and fluorescence in situ hybridization(FISH) technology in China,using the bibliometric method,taking " fluorescence in situ hybridization(FI...In order to explore researches about the chromosome karyotype analysis and fluorescence in situ hybridization(FISH) technology in China,using the bibliometric method,taking " fluorescence in situ hybridization(FISH) " and " chromosome" as key words,this paper made a statistical analysis on the literature published in China National Knowledge Infrastructure(CNKI) during 2002-2016.The results indicated that the number of papers published in 2002 was the smallest(37),while the number of papers published in 2012 was the largest(125).In terms of the distribution of organizations of authors,in 1201 papers,11 organizations published papers ≥15,accounting for 21.65%.In terms of distribution of papers published by different periodicals,11 periodicals published papers ≥10,accounting for 17.65%.In terms of the papers supported by foundation projects,in all papers searched,377 papers were supported by foundation projects,accounting for 31.39%.In terms of the distribution of doctoral and master's dissertations,259 papers were master's dissertations,accounting for 21.57%;92 papers were doctoral dissertations,accounting for 7.66%.展开更多
The karyotype of the primary wheat wheatgrass alien addition line TAI 27 was 2 n=44 in which all of the chromosomes were metacentric and submetacentric.However,in the progeny of TAI 27 a pair of chromosomes had become...The karyotype of the primary wheat wheatgrass alien addition line TAI 27 was 2 n=44 in which all of the chromosomes were metacentric and submetacentric.However,in the progeny of TAI 27 a pair of chromosomes had become small chromosomes in the two morphologically different plants.Fluorescence in situ hybridization(FISH)technique was used to analyze the two different plants.The observations indicate that a pair of small chromosomes in one variation line are from wheatgrass.In another variation line,a pair of small chromosomes are also from wheatgrass,while another pair of wheatgrass chromosomes have substituted the wheat chromosomes.TAI 27 and its variant lines showed a high level of resistance to barley yellow dwarf virus(BYDV).The possible explanation for such a variation and the potential use of the variant lines were discussed briefly.展开更多
It has been reported that endosperm undergoes programmed cell death (PCD) during maize kernel development.Both bz1 (bronze ) and bz2 are anthocyanin biosynthetic genes,and related to development of aleuronic la...It has been reported that endosperm undergoes programmed cell death (PCD) during maize kernel development.Both bz1 (bronze ) and bz2 are anthocyanin biosynthetic genes,and related to development of aleuronic layer of maize seeds.Tyramide signal amplification fluorescence in situ hybridization (TSA FISH) is a novel and high sensitive FISH technique,which is suitable for routine application in plant cytogenetic research.Using this technique,we physically mapped the bz1 gene onto the short arm of chromosome 9 and the long arm of chromosome 1;the percentage distances from centromere to hybridization site were 40.2,75.4 respectively,and the bz2 onto the long arm of chromosome 1 and the short arm of chromosome 5;the percentage distances from centromere to hybridization site were 21.6,15.3 separately.The TSA FISH techniques of small low copy DNA sequences for plants are discussed.展开更多
Toxic cyanobacterial blooms constitute a threat to human safety because Microcystis sp. releases microcystins during growth, and particularly during cell death. Therefore, analysis of toxic and nontoxic Microcystis in...Toxic cyanobacterial blooms constitute a threat to human safety because Microcystis sp. releases microcystins during growth, and particularly during cell death. Therefore, analysis of toxic and nontoxic Microcystis in natural communities is required in order to assess and predict bloom dynamics and toxin production by these organisms. In this study, an analysis combining fluorescence in situ hybridization (FISH) with flow cytometry (FCM) was used to discriminate between toxic and nontoxic Microcystis and also to quantify the percentage of toxic Microcystis present in blooms. The results demonstrate that the combination of FISH and flow cytometry is a useful approach for studying the ecology of Microcystis toxin production and for providing an early warning for toxic Microcystis blooms.展开更多
The localization of 18S ribosomal RNA genes (rDNA) by fluorescence in situ hybridization (FISH) had been performed for some species of Paeonla. However, the pattern of 18S rDNA loci among populations Is Indistinct...The localization of 18S ribosomal RNA genes (rDNA) by fluorescence in situ hybridization (FISH) had been performed for some species of Paeonla. However, the pattern of 18S rDNA loci among populations Is Indistinct. In the present study, we localized 18S rDNA loci on meiotic or mitotic chromosomes of six populations of Paeonla obovata Maxim. (Paeonlaceae). Different numbers of rDNA loci were found with different diploid (2n=10) populations, namely eight (Lushl and Mt. JIuhua populations), 10 (Mt. Talbal population), and seven (Mt. Guandl population), whereas tetraplold (2n=20) populations were all found with 16 loci. Aii rDNA loci were mapped near teiomeres of mitotic chromosomes and there was no chromosome with two loci. The present results show that molecular cytological polymorphlsm exists among P. obovata diploid populations, Indicating that structural variations occurred frequently during the evolutionary history of this species, accompanied with differentiation among populations.展开更多
Objective To compare the roles of fluorescence in situ hybridization (FISH) and karyotype analysis on chorionic villus in spontaneous abortion. Methods A total of 201 cases were included in this study and were rando...Objective To compare the roles of fluorescence in situ hybridization (FISH) and karyotype analysis on chorionic villus in spontaneous abortion. Methods A total of 201 cases were included in this study and were randomly divided into 2 groups by the same gestational age. The villi from 100 cases (group A) were cultured and used for karyotype analysis, while the uncultured villi from 101 women (group B) were used for FISH. A case was randomly selected from pregnant patients of 6-11 weeks at each gestational week in each group, 6 cases from each group and a total of 12 cases were analyzed by FISH and karyotype at the same time. Results The successful karyotype analysis rate was 66. 0%, and the abnormality karyotype rate was 30.3%; FISH success analysis rate was 100%, and the abnormality rate was 46. 5%; there were significant differences between FISH and karyotype analysis (P=0.036). Conclusion There were obvious differences between the two techniques. FISH was more successfully analyzed, and was used to more precisely determine fetal chromo- some number. It is an effective way to determine abnormal chromosome by integrating FISH and karyotype analysis in spontaneous abortion.展开更多
Jellyfish blooms have increased worldwide, and the outbreaks of jellyfish population not only affect the food web structures via voracious predation but also play an important role in the dynamics of nutrients and oxy...Jellyfish blooms have increased worldwide, and the outbreaks of jellyfish population not only affect the food web structures via voracious predation but also play an important role in the dynamics of nutrients and oxygen in planktonic food webs. However, it remains unclear whether specific carbon compounds released through jellyfish metabolic processes have the potential to shape bacterial community composition. Therefore, in this study, we aimed to investigate the compositional succession of the bacterioplankton community in response to the dissolved organic matter (DOM) released by the live Scyphomedusae Cyanea lamarckii and Chrysaora hysoscella collected from Helgoland Roads of the North Sea. The bacterial community was significantly stimulated by the DOM released form live jellyfish and different dominant phylotypes were observed for these two Scyphomedusae species. Furthermore, the bacterial community structures in the different DOM sources, jellyfish-incubated media, Kabeltonne seawater, and artificial seawater (DOM-free) were significantly different, as revealed by automated ribosomal intergenic spacer analysis fingerprints. Catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) revealed a rapid species-specific shift in bacterial community composition. Gammaproteobacteria dominated the community instead of the Bacteroidetes community for C. lamarckii, whereas Gammaproteobacteria and Bacteroidetes dominated the community for C. hysoscella. The significant differences in the bacterial community composition and succession indicate that the components of the DOM released by jellyfish might differ with jellyfish species.展开更多
Performance of biological phosphorus removal in the oxic-settling-anaerobic(OSA) process was investigated. Cell staining and fluorescent in situ hybridization(FISH) were used to analyze characteristics and microbial c...Performance of biological phosphorus removal in the oxic-settling-anaerobic(OSA) process was investigated. Cell staining and fluorescent in situ hybridization(FISH) were used to analyze characteristics and microbial community of sludge. Experimental results showed that phosphorus removal efficiency was near 60% and the amount of biological phosphorus accumulation in aerobic sludge of the OSA system was up to 26.9 mg/g. Biological phosphorus removal efficiency was partially inhibited by carbon sources in the continuous OSA system. Contrasted to the OSA system,biological phosphorus removal efficiency was enhanced by 14% and the average total phosphorus(TP) contents of aerobic sludge were increased by 0.36 mg/g when sufficient carbon sources were supplied in batch experiments. Staining methods indicated that about 35% of microorganisms had typical characteristics of phosphorus accumulating organisms(PAOs) . FISH analysis demonstrated that PAOMIX-binding bacteria were predominant microbial communities in the OSA system,which accounted for around 28% of total bacteria.展开更多
基金Acknowledgements This study was supported by the National Science Foundation (No. 30630067);the State Key Basic Research Grant of China (No. 2004CB518705); the Program for Changjiang Scholars and Innovative Research Team in University (No. IRT0416).
文摘Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization (M-FISH) for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.
基金supported by the National Natural Science Foundation of China(Grants No.40806073,40876097)the Polar Science Strategic Research Foundation of China and the Youth Foundation for Marine Science of SOA (Grants No.2008128)
文摘Standard FISH protocols using fluorochrome-labeled oligonucleotide probes have been successfully applied for in situ detection.However,optimized protocols of FISH for specific eukaryotes in marine environments are often not developed.This study optimized the conditions of fluorescence in situ hybridization (FISH) by using two polar isolated microalgae.The modified conditions were as follows:(1) 10 mg·mL^(-1) lysozyme solution pretreatment at 37℃for 30 min;(2) the hybridization buffer including 20%formamide;(3) the hybridization condition was 47℃for 6 h.The cells enumerated by FISH were compared with those enumerated by flow cytometry(FCM) and DAPI to confirm the cell loss and hybridization efficiency.The optimized protocol was also successfully applied to Arctic Ocean samples,which were found to be dominated by Micromonas sp.The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples.
基金This work was supported by a grant from Tianjin Science and Technology Committee (No. 033804211)
文摘Objective: The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis and prognosis. Methods: To observe MRP-1/C9mRNA expression, tissue microarray (TMA) containing 54 lung cancers and 10 normal lung tissues was prepared and Fluorescence in situ hybridization was used. Results: The positive rate of MRP-1/CD9 expression was 48.1% in lung cancer, lower than that of normal lung tissues. The statistical difference was significant (P〈0.05). Its protein expression had no relationship with the patients' ages, sex and the macroscopic type of tumor, but had relationships with the histological type, clinical stage, differentiated degree and metastasis. The expression in non-small cell lung cancer (NSCLC) was higher than that in small cell lung cancer (SCLC); in well-moderately differentiated group was higher than that in poorly differentiated group; Earlier period group (I+II) was higher than in later period group (Ⅲ+Ⅳ); and in group without lymphoid metastasis was higher than in patients with lymphoid metastasis. Conclusion: The progression of the lung cancer maybe related with the descended MRP-1/Cd9 expression, which may be useful in evaluating the prognosis of cancer patients.
文摘A molecular biology method, fluorescent in situ hybridization(FISH), in which the pre-treatment was improved in allusion to the media of the constructed wetlands(CW), e.g. the soil and the grit, was used to investigate the vertical distribution characteristics of ammonia-oxidizing bacteria(AOB) quantity and the relation with oxidation-reduction potential(ORP) in the Typha latifolia constructed wetlands under three different Ioadings in summer from May to September. Results showed that the quantity of the AOB decreased in the Typha latifolia CW with the increase of vertical depth. However, the AOB quantity was 2-4 times the quantity of the control in the root area. Additionally, ORP in the rhizosphere was found to be higher than other areas, which showed that Typha latifolia CW was in an aerobic state in summer when using simulated non-point sewage at the rural area of Taihu Lake in China and small town combined sewage.
基金the National Natural Science Foundation of China (30170501)
文摘Telomeres form the ends of eukaryotic chromosomes and serve as protective caps that keep chromosomes structure independency and completeness. The first plant telomere DNA was isolated from Arabidopsis thaliana and was shown to have tandemly repeated sequence 5-TTTAGGG-3: The Arabidopsis-type telomere has been found in many plants, but several reports indicate that this sequence is absent in some plants. Up to now, no research has been conducted on the telomere of cotton. In this paper, the Arabidopsis-type telomere sequence was amplified and cloned using the primers designed based on the fragment containing telomere sequence in an Arabidopsis bacterial artificial chromosome (BAC). Fluorescence in situ hybridization (FISH) with cotton metaphase chromosomes using the Arabidopsis-type telomere sequence as probes indicated that the signals were located at all chromosome ends of seven diploid and two tetraploid cotton species with different signal intensities among chromosome complements of different cotton species, even between long and short arms of the same chromosome. To identify the signals of FISH, the genome DNA of Xinhai 7, a cultivar of Gossypium barbadense, digested by BAL-31 nuclease was introduced in this study. The result of BAL-31 digestion indicated that the hybridization signals of FISH represent the outermost DNA sequence of each cotton chromosomes. So we first proved that the telomeric repeats of cotton cross-hybridize with that of Arabidopsis. The results of terminal restriction fragment (TRF) showed significant variation in telomere length among cotton species. The telomere length of cultivated cotton was close to 20 kb and was larger than those of wild cotton species whose telomere length rahged from 6 to 20 kb.
文摘Objective To identify the deletions in Duchenne/Becker muscular dystrophy (DMD/ BMD) by using fluorescence in situ hybridization (FISH)Methods The exon-specific cosmid DNA probes (representing 18 exons) were used to perform one-color FISH on metaphase and interphase preparations. The peripheral blood samples from 9 normal people (4 males and 5 females) and 5 females from independent deletion DMD/BMD families, as well as 2 amniotic fluid specimens and 2 chorionic villus samples (CVS) from normal pregnant females were analyzed. Results 72%-100% of peripheral blood lymphocyte metaphases or interphases, 60% -70% of amniocyte interphases, and 95 - 99% of chorionic villus cell interphases showed expected signals. One suspected female was identified as deletion carriers and two were excluded.Conclusion FISH in combination with other available techniques allows efficient screening of DMD/BMD deletion carriers, which also lay the ground work for prenatal diagnosis for potential fetal carriers.
基金Supported by the National Natural Science Foundation of China(Nos.41376136,31201992)the Double First-Class Discipline of Fisheries Science in China。
文摘It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of primers were designed for amplification to verify this linkage of two genes in a kelp cultivar of Saccharina japonica,one of species in Ochrophyta.This result supplemented the previous report that 5S rDNA was unlinked to 25S rDNA in this kelp.In order to simultaneously visualize this unlinkage of two genes,dual-color fluorescence in situ hybridization(FISH)technique was applied to the cytogenetics of S.japonica.Dual-color FISH images showed that two and four hybridization signals were present in the kelp gametophyte and sporophyte,respectively,metaphase nuclei hybridized simultaneously with the labeled probes of 18S rDNA and 5S rDNA.Both haploid and diploid karyotypes in decreasing length of chromosomes showed that 18S-5.8S-25S rDNA was localized at the interstitial region of Chromosome 23,whereas 5S rDNA resided at the sub-telomeric region of Chromosome 27.These karyotypes suggested that the kelp nuclear genome had only one locus of each rRNA gene,and their loci on different chromosomes indicated the physical unlinkage of 5S rDNA to 18S-5.8S-25S rDNA in this kelp.Therefore,dual-color FISH seems to be a powerful technique for the discrimination and pairing of chromosomes featured in both small size and nearly identical shape in S.japonica.
基金Supported by Project of National Natural Science Foundation(31160292)Crop Discipline Construction Project of Tibet Agricultural and Animal Husbandry College(2015ZWXKJS&2016ZWXKJS)
文摘In order to explore researches about the chromosome karyotype analysis and fluorescence in situ hybridization(FISH) technology in China,using the bibliometric method,taking " fluorescence in situ hybridization(FISH) " and " chromosome" as key words,this paper made a statistical analysis on the literature published in China National Knowledge Infrastructure(CNKI) during 2002-2016.The results indicated that the number of papers published in 2002 was the smallest(37),while the number of papers published in 2012 was the largest(125).In terms of the distribution of organizations of authors,in 1201 papers,11 organizations published papers ≥15,accounting for 21.65%.In terms of distribution of papers published by different periodicals,11 periodicals published papers ≥10,accounting for 17.65%.In terms of the papers supported by foundation projects,in all papers searched,377 papers were supported by foundation projects,accounting for 31.39%.In terms of the distribution of doctoral and master's dissertations,259 papers were master's dissertations,accounting for 21.57%;92 papers were doctoral dissertations,accounting for 7.66%.
文摘The karyotype of the primary wheat wheatgrass alien addition line TAI 27 was 2 n=44 in which all of the chromosomes were metacentric and submetacentric.However,in the progeny of TAI 27 a pair of chromosomes had become small chromosomes in the two morphologically different plants.Fluorescence in situ hybridization(FISH)technique was used to analyze the two different plants.The observations indicate that a pair of small chromosomes in one variation line are from wheatgrass.In another variation line,a pair of small chromosomes are also from wheatgrass,while another pair of wheatgrass chromosomes have substituted the wheat chromosomes.TAI 27 and its variant lines showed a high level of resistance to barley yellow dwarf virus(BYDV).The possible explanation for such a variation and the potential use of the variant lines were discussed briefly.
文摘It has been reported that endosperm undergoes programmed cell death (PCD) during maize kernel development.Both bz1 (bronze ) and bz2 are anthocyanin biosynthetic genes,and related to development of aleuronic layer of maize seeds.Tyramide signal amplification fluorescence in situ hybridization (TSA FISH) is a novel and high sensitive FISH technique,which is suitable for routine application in plant cytogenetic research.Using this technique,we physically mapped the bz1 gene onto the short arm of chromosome 9 and the long arm of chromosome 1;the percentage distances from centromere to hybridization site were 40.2,75.4 respectively,and the bz2 onto the long arm of chromosome 1 and the short arm of chromosome 5;the percentage distances from centromere to hybridization site were 21.6,15.3 separately.The TSA FISH techniques of small low copy DNA sequences for plants are discussed.
基金supported by the National Basic Research Program of China (Grant No. 2008CB418000)the National Water Pollution Control and Management Technology Major Project (Grant No. 2009ZX07527- 005)+1 种基金the Natural Science Foundation of China-Yunnan Project (Grant No. U0833604)the Knowledge Innovative Program of the Chinese Academy of Sciences (Grant No. KZCX1-YW-14-1)
文摘Toxic cyanobacterial blooms constitute a threat to human safety because Microcystis sp. releases microcystins during growth, and particularly during cell death. Therefore, analysis of toxic and nontoxic Microcystis in natural communities is required in order to assess and predict bloom dynamics and toxin production by these organisms. In this study, an analysis combining fluorescence in situ hybridization (FISH) with flow cytometry (FCM) was used to discriminate between toxic and nontoxic Microcystis and also to quantify the percentage of toxic Microcystis present in blooms. The results demonstrate that the combination of FISH and flow cytometry is a useful approach for studying the ecology of Microcystis toxin production and for providing an early warning for toxic Microcystis blooms.
基金Supported by the National Natural Science Foundation of China (39928003).
文摘The localization of 18S ribosomal RNA genes (rDNA) by fluorescence in situ hybridization (FISH) had been performed for some species of Paeonla. However, the pattern of 18S rDNA loci among populations Is Indistinct. In the present study, we localized 18S rDNA loci on meiotic or mitotic chromosomes of six populations of Paeonla obovata Maxim. (Paeonlaceae). Different numbers of rDNA loci were found with different diploid (2n=10) populations, namely eight (Lushl and Mt. JIuhua populations), 10 (Mt. Talbal population), and seven (Mt. Guandl population), whereas tetraplold (2n=20) populations were all found with 16 loci. Aii rDNA loci were mapped near teiomeres of mitotic chromosomes and there was no chromosome with two loci. The present results show that molecular cytological polymorphlsm exists among P. obovata diploid populations, Indicating that structural variations occurred frequently during the evolutionary history of this species, accompanied with differentiation among populations.
文摘Objective To compare the roles of fluorescence in situ hybridization (FISH) and karyotype analysis on chorionic villus in spontaneous abortion. Methods A total of 201 cases were included in this study and were randomly divided into 2 groups by the same gestational age. The villi from 100 cases (group A) were cultured and used for karyotype analysis, while the uncultured villi from 101 women (group B) were used for FISH. A case was randomly selected from pregnant patients of 6-11 weeks at each gestational week in each group, 6 cases from each group and a total of 12 cases were analyzed by FISH and karyotype at the same time. Results The successful karyotype analysis rate was 66. 0%, and the abnormality karyotype rate was 30.3%; FISH success analysis rate was 100%, and the abnormality rate was 46. 5%; there were significant differences between FISH and karyotype analysis (P=0.036). Conclusion There were obvious differences between the two techniques. FISH was more successfully analyzed, and was used to more precisely determine fetal chromo- some number. It is an effective way to determine abnormal chromosome by integrating FISH and karyotype analysis in spontaneous abortion.
基金Project at the Alfred Wegener Institute, Helmholtz Centre for Polar and Marine Research (Germany)the funding from the China Scholarship Council+1 种基金Jutta Niggemann and Thorsten Dittmar (ICBM COU Oldenburg) who supported the preparation of the DOM free seawater strongly with material and technical supportthe crew of the AADE research vessel for providing samples as well as the entire team of the AWI Food Web Project
文摘Jellyfish blooms have increased worldwide, and the outbreaks of jellyfish population not only affect the food web structures via voracious predation but also play an important role in the dynamics of nutrients and oxygen in planktonic food webs. However, it remains unclear whether specific carbon compounds released through jellyfish metabolic processes have the potential to shape bacterial community composition. Therefore, in this study, we aimed to investigate the compositional succession of the bacterioplankton community in response to the dissolved organic matter (DOM) released by the live Scyphomedusae Cyanea lamarckii and Chrysaora hysoscella collected from Helgoland Roads of the North Sea. The bacterial community was significantly stimulated by the DOM released form live jellyfish and different dominant phylotypes were observed for these two Scyphomedusae species. Furthermore, the bacterial community structures in the different DOM sources, jellyfish-incubated media, Kabeltonne seawater, and artificial seawater (DOM-free) were significantly different, as revealed by automated ribosomal intergenic spacer analysis fingerprints. Catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) revealed a rapid species-specific shift in bacterial community composition. Gammaproteobacteria dominated the community instead of the Bacteroidetes community for C. lamarckii, whereas Gammaproteobacteria and Bacteroidetes dominated the community for C. hysoscella. The significant differences in the bacterial community composition and succession indicate that the components of the DOM released by jellyfish might differ with jellyfish species.
基金Project (No. 2006BAC19B04) supported by the National Key Technology R&D Program of China
文摘Performance of biological phosphorus removal in the oxic-settling-anaerobic(OSA) process was investigated. Cell staining and fluorescent in situ hybridization(FISH) were used to analyze characteristics and microbial community of sludge. Experimental results showed that phosphorus removal efficiency was near 60% and the amount of biological phosphorus accumulation in aerobic sludge of the OSA system was up to 26.9 mg/g. Biological phosphorus removal efficiency was partially inhibited by carbon sources in the continuous OSA system. Contrasted to the OSA system,biological phosphorus removal efficiency was enhanced by 14% and the average total phosphorus(TP) contents of aerobic sludge were increased by 0.36 mg/g when sufficient carbon sources were supplied in batch experiments. Staining methods indicated that about 35% of microorganisms had typical characteristics of phosphorus accumulating organisms(PAOs) . FISH analysis demonstrated that PAOMIX-binding bacteria were predominant microbial communities in the OSA system,which accounted for around 28% of total bacteria.