Zhikong scallop(Chlamys farreri) is an important maricultured species in China.Many researches on this species,such as population genetics and QTL fine-mapping,need a large number of molecular markers.In this study,ba...Zhikong scallop(Chlamys farreri) is an important maricultured species in China.Many researches on this species,such as population genetics and QTL fine-mapping,need a large number of molecular markers.In this study,based on the expressed sequence tags(EST),a total of 300 putative single nucleotide polymorphisms(SNPs) were selected and validated using high resolution melting(HRM) technology with unlabeled probe.Of them,101(33.7%) were found to be polymorphic in 48 individuals from 4 populations.Further evaluation with 48 individuals from Qingdao population showed that all the polymorphic loci had two alleles with the minor allele frequency ranged from 0.046 to 0.500.The observed and expected heterozygosities ranged from 0.000 to 0.925 and from 0.089 to 0.505,respectively.Fifteen loci deviated significantly from Hardy-Weinberg equilibrium and significant linkage disequilibrate was detected in one pair of markers.BLASTx gave significant hits for 72 of the 101 polymorphic SNP-containing ESTs.Thirty four polymorphic SNP loci were predicted to be non-synonymous substitutions as they caused either the change of codons(33 SNPs) or pretermination of translation(1 SNP).The markers developed can be used for the population studies and genetic improvement on Zhikong scallop.展开更多
In the monocot rice species Oryza sativa L., one of the most striking morphological processes during reproductive development is the concurrence of panicle development with the sequential elongation of upper internod...In the monocot rice species Oryza sativa L., one of the most striking morphological processes during reproductive development is the concurrence of panicle development with the sequential elongation of upper internodes (UPIs). To elucidate the underlying molecular mechanisms, we cloned the rice gene NECK LEAF 1 (NL1), which when mutated results in delays in flowering time, smaller panicles with overgrown bracts and abnormal UPI elongation patterns. The NL1 gene encodes a GATA-type transcription factor with a single zinc finger domain, and its transcripts are de- tected predominantly in the bract primordia, which normally degenerate in the wild-type plants. Overexpression of NL1 in transgenic plants often gives rise to severe growth retardation, less vegetative phytomers and smaller leaves, suggesting that NL1 plays an important role in organ differentiation. A novel mutant allele of PLASTOCHRON1 (PLAD, a gene known to play a key role in regulating leaf initiation, was identified in this study. Genetic analysis demonstrated an interaction between nil and plal, with NL1 acting upstream of PLA1. The expression level and spatial pattern of PLA1 were found to be altered in the nil mutant. Furthermore, the expression of two regulators of flowering, Hd3a and OsMADS1, was also affected in the nil mutant. On the basis of these findings, we propose that NL1 is an intrinsic factor that modulates and coordinates organogenesis through regulating the expression of PLA1 and other regulatory genes during reproductive development in rice.展开更多
Buffaloes (Bubalus bubalis) are widely distributed and were introduced to Brazil in 1895. Most of the molecular genetic characterization of buffaloes has been done with cross-specific (cattle) markers, but few of ...Buffaloes (Bubalus bubalis) are widely distributed and were introduced to Brazil in 1895. Most of the molecular genetic characterization of buffaloes has been done with cross-specific (cattle) markers, but few of them include Brazilian populations. Nineteen commonly used cattle microsatellites were tested to develop a multiplexed set of microsatellites and characterize Brazilian buffalo. Three PCR mixes were finally developed with the 11 markers that succeed in amplify and were polymorphic (58%). The average number of alleles was 5.42, with an average observed and expected heterocigozity of 0.441 and 0.695, respectively. As it was expected, Brazilian buffalo variability was lower than the previously reported from the domestication centres (China and India), but higher than the seriously selected European populations. The exclusion power calculated for the eleven markers in Brazilian buffalo was 0.9999999996, this allows its use in DNA based traceability.展开更多
Abstract Objectives To investigate the relationship between the β-fibrinogen gene -455A/G polymorphism and plasma fibrinogen level and to determine the influence of the mutation on ischemic stroke. Methods Ninety-o...Abstract Objectives To investigate the relationship between the β-fibrinogen gene -455A/G polymorphism and plasma fibrinogen level and to determine the influence of the mutation on ischemic stroke. Methods Ninety-one patients (63.5±10.1 years) with ischemic stroke and 74 elderly control subjects (60.6±10.8 years) without any thromboembolic events and 98 healthy blood donators as young control (37.5±13.3 years) were enrolled in this trial. The β-fibrinogen gene -455A/G polymorphism was analyzed for all subjects by PCR-RFLP with the restrictive enzyme Hae Ⅲ, while plasma fibrinogen levels were obtained from the prothrombin time (PT) assay. For statistical analysis, the parameters were compared between any two different groups by the unpaired Student’s t test and the Chi-square test. Before analysis, log transformations for concentrations of fibrinogen were carried out.Results H2 allele frequency was higher in male ischemic stroke patients than in the elderly control (22.7% vs 7.1%, χ 2=5.56, P【0.02). There was no significant difference between the female groups. In those patients without any thromboembolic events (both elderly and young control groups), the frequency of H2 decreased with age (≤40, 21.3%; 41-59, 15.4%; and ≥60, 10.2%). In the male elderly and young control groups, the level of plasma fibrinogen was lower in the H1H1 genotype (287±96*!mg/dl and 234±58*!mg/dl) than in H1H2 and H2H2 (331±44*!mg/dl and 307±55*!mg/dl; t=2.53 and 9.67, P【0.05). In the female elderly groups, this tendency was not found.Conclusion Plasma fibrinogen expression is affected by the β-fibrinogen gene -455A/G polymorphism, and the H2 allele may be a risk factor for ischemic stroke in Chinese males.展开更多
Brassinosteroids (BRs) are important hormones that regulate plant development and physiology. Substantial progresses have been made in BR-related studies, and especially an increasing number of new genes involved in...Brassinosteroids (BRs) are important hormones that regulate plant development and physiology. Substantial progresses have been made in BR-related studies, and especially an increasing number of new genes involved in BR biosynthesis have been identified. Here, we characterize a BR-related rice mutant, small grain 4 (sg4), obtained from callus culture of japonica cultivar Nipponbare. This mutant showed multiple phenotypes such as dark green, rugose erect leaves and small round grains. It was higher than the wild type, different from the majority of BR- and gibberellin-related mutants. Genetic analysis showed that the mutant phenotypes are controlled by a single recessive locus. The gene was fine-mapped to a 90.7-kb interval with 1,100 F2 recessive individuals by means of map-basedcloning. Totally 11 open reading frames were found in this interval, only one of which was detected with an 8-bp in- sertion in the 5rUTR region by sequencing. Functional complementation test revealed that a DWARFll allele, LOC_OsO4g39430, is answer for the mutant phenotype of sg4. Tissue-specific response to BR and decreased expression levels of BR biosynthetic genes suggest that sg4 is a weak BR-deficient mutant. These results are beneficial to understanding the physiological action of sg4 in a more comprehensive way.展开更多
Plant height and tillering are crucial factors determining rice plant architecture and influencing rice grain production. In this study, rnulti-tillering dwarf1 (mtdl), a stable multi-tiller and dwarf mutant, was sc...Plant height and tillering are crucial factors determining rice plant architecture and influencing rice grain production. In this study, rnulti-tillering dwarf1 (mtdl), a stable multi-tiller and dwarf mutant, was screened from the ethylmethane sulfonate-treated japonica rice variety Wuyunging7. Compared with the wild type, mtdl mutant exhibited pleiotropic phenotypes, including dwarf- ism, more tillers, brittle culms and delayed heading date. By employing map-based cloning strategy, the gene MTD1 was finally mapped to an approximately 66-kb region on the short arm of chromosome 9. Sequencing results showed that the gene LOCOsO9g02650 (BC12) in mtdl mutant had a single nucleotide substitution (G to A), which gen- erated a premature translation stop. Over-expressing MTD1/BC12 coding sequ(nce rescued all the phenotypes of mtdl mutants including plant height and tillers, which confirms that BC12 is the mutated gene in mtdl mutant. Quantitative reverse tran,-eription-PCR analysis showed that MTDI/BCI2 could negatively regulate the expression of MONOCULM 1, IDEAL PLANT ARCHITECTURE1 and Tillering and Dwarf 1, and control rice tillering. Remark- ably, a-amylase activity analysis and gibberellic acid (GA) treatment showed that the dwarf phenotype of mtdl mutant was dependent on GA biosynthesis pathway. These results facilitated to further uncover the molecular mechanism of the growth and development in rice.展开更多
基金the National Natural Science Foundation of China(No.31130054)the National Basic Research Program of China(973 Program,2010CB126406 and 2010CB126402)+2 种基金the National High-Tech R&D Program(863 Program,2012AA10A402 and 2012AA10A405)the National Key Technology R&D Program(2011BAD13B06)the Earmarked Fund for Modern Agro-Industry Technology Research
文摘Zhikong scallop(Chlamys farreri) is an important maricultured species in China.Many researches on this species,such as population genetics and QTL fine-mapping,need a large number of molecular markers.In this study,based on the expressed sequence tags(EST),a total of 300 putative single nucleotide polymorphisms(SNPs) were selected and validated using high resolution melting(HRM) technology with unlabeled probe.Of them,101(33.7%) were found to be polymorphic in 48 individuals from 4 populations.Further evaluation with 48 individuals from Qingdao population showed that all the polymorphic loci had two alleles with the minor allele frequency ranged from 0.046 to 0.500.The observed and expected heterozygosities ranged from 0.000 to 0.925 and from 0.089 to 0.505,respectively.Fifteen loci deviated significantly from Hardy-Weinberg equilibrium and significant linkage disequilibrate was detected in one pair of markers.BLASTx gave significant hits for 72 of the 101 polymorphic SNP-containing ESTs.Thirty four polymorphic SNP loci were predicted to be non-synonymous substitutions as they caused either the change of codons(33 SNPs) or pretermination of translation(1 SNP).The markers developed can be used for the population studies and genetic improvement on Zhikong scallop.
文摘In the monocot rice species Oryza sativa L., one of the most striking morphological processes during reproductive development is the concurrence of panicle development with the sequential elongation of upper internodes (UPIs). To elucidate the underlying molecular mechanisms, we cloned the rice gene NECK LEAF 1 (NL1), which when mutated results in delays in flowering time, smaller panicles with overgrown bracts and abnormal UPI elongation patterns. The NL1 gene encodes a GATA-type transcription factor with a single zinc finger domain, and its transcripts are de- tected predominantly in the bract primordia, which normally degenerate in the wild-type plants. Overexpression of NL1 in transgenic plants often gives rise to severe growth retardation, less vegetative phytomers and smaller leaves, suggesting that NL1 plays an important role in organ differentiation. A novel mutant allele of PLASTOCHRON1 (PLAD, a gene known to play a key role in regulating leaf initiation, was identified in this study. Genetic analysis demonstrated an interaction between nil and plal, with NL1 acting upstream of PLA1. The expression level and spatial pattern of PLA1 were found to be altered in the nil mutant. Furthermore, the expression of two regulators of flowering, Hd3a and OsMADS1, was also affected in the nil mutant. On the basis of these findings, we propose that NL1 is an intrinsic factor that modulates and coordinates organogenesis through regulating the expression of PLA1 and other regulatory genes during reproductive development in rice.
文摘Buffaloes (Bubalus bubalis) are widely distributed and were introduced to Brazil in 1895. Most of the molecular genetic characterization of buffaloes has been done with cross-specific (cattle) markers, but few of them include Brazilian populations. Nineteen commonly used cattle microsatellites were tested to develop a multiplexed set of microsatellites and characterize Brazilian buffalo. Three PCR mixes were finally developed with the 11 markers that succeed in amplify and were polymorphic (58%). The average number of alleles was 5.42, with an average observed and expected heterocigozity of 0.441 and 0.695, respectively. As it was expected, Brazilian buffalo variability was lower than the previously reported from the domestication centres (China and India), but higher than the seriously selected European populations. The exclusion power calculated for the eleven markers in Brazilian buffalo was 0.9999999996, this allows its use in DNA based traceability.
文摘Abstract Objectives To investigate the relationship between the β-fibrinogen gene -455A/G polymorphism and plasma fibrinogen level and to determine the influence of the mutation on ischemic stroke. Methods Ninety-one patients (63.5±10.1 years) with ischemic stroke and 74 elderly control subjects (60.6±10.8 years) without any thromboembolic events and 98 healthy blood donators as young control (37.5±13.3 years) were enrolled in this trial. The β-fibrinogen gene -455A/G polymorphism was analyzed for all subjects by PCR-RFLP with the restrictive enzyme Hae Ⅲ, while plasma fibrinogen levels were obtained from the prothrombin time (PT) assay. For statistical analysis, the parameters were compared between any two different groups by the unpaired Student’s t test and the Chi-square test. Before analysis, log transformations for concentrations of fibrinogen were carried out.Results H2 allele frequency was higher in male ischemic stroke patients than in the elderly control (22.7% vs 7.1%, χ 2=5.56, P【0.02). There was no significant difference between the female groups. In those patients without any thromboembolic events (both elderly and young control groups), the frequency of H2 decreased with age (≤40, 21.3%; 41-59, 15.4%; and ≥60, 10.2%). In the male elderly and young control groups, the level of plasma fibrinogen was lower in the H1H1 genotype (287±96*!mg/dl and 234±58*!mg/dl) than in H1H2 and H2H2 (331±44*!mg/dl and 307±55*!mg/dl; t=2.53 and 9.67, P【0.05). In the female elderly groups, this tendency was not found.Conclusion Plasma fibrinogen expression is affected by the β-fibrinogen gene -455A/G polymorphism, and the H2 allele may be a risk factor for ischemic stroke in Chinese males.
基金supported by the National Natural Science Foundation of China(31201194,31171532,91335105and 31201183)
文摘Brassinosteroids (BRs) are important hormones that regulate plant development and physiology. Substantial progresses have been made in BR-related studies, and especially an increasing number of new genes involved in BR biosynthesis have been identified. Here, we characterize a BR-related rice mutant, small grain 4 (sg4), obtained from callus culture of japonica cultivar Nipponbare. This mutant showed multiple phenotypes such as dark green, rugose erect leaves and small round grains. It was higher than the wild type, different from the majority of BR- and gibberellin-related mutants. Genetic analysis showed that the mutant phenotypes are controlled by a single recessive locus. The gene was fine-mapped to a 90.7-kb interval with 1,100 F2 recessive individuals by means of map-basedcloning. Totally 11 open reading frames were found in this interval, only one of which was detected with an 8-bp in- sertion in the 5rUTR region by sequencing. Functional complementation test revealed that a DWARFll allele, LOC_OsO4g39430, is answer for the mutant phenotype of sg4. Tissue-specific response to BR and decreased expression levels of BR biosynthetic genes suggest that sg4 is a weak BR-deficient mutant. These results are beneficial to understanding the physiological action of sg4 in a more comprehensive way.
基金supported by the National Natural Science Foundation of China (31401464, 31201183)Zhejiang Provincial Natural Science Foundation of China (Y3110194, LY16C130001)+1 种基金China Postdoctoral Science Foundation (2014M561108)the Open Foundation from Zhejiang Provincial Top Key Discipline of Biology (KFJJ2014006)
文摘Plant height and tillering are crucial factors determining rice plant architecture and influencing rice grain production. In this study, rnulti-tillering dwarf1 (mtdl), a stable multi-tiller and dwarf mutant, was screened from the ethylmethane sulfonate-treated japonica rice variety Wuyunging7. Compared with the wild type, mtdl mutant exhibited pleiotropic phenotypes, including dwarf- ism, more tillers, brittle culms and delayed heading date. By employing map-based cloning strategy, the gene MTD1 was finally mapped to an approximately 66-kb region on the short arm of chromosome 9. Sequencing results showed that the gene LOCOsO9g02650 (BC12) in mtdl mutant had a single nucleotide substitution (G to A), which gen- erated a premature translation stop. Over-expressing MTD1/BC12 coding sequ(nce rescued all the phenotypes of mtdl mutants including plant height and tillers, which confirms that BC12 is the mutated gene in mtdl mutant. Quantitative reverse tran,-eription-PCR analysis showed that MTDI/BCI2 could negatively regulate the expression of MONOCULM 1, IDEAL PLANT ARCHITECTURE1 and Tillering and Dwarf 1, and control rice tillering. Remark- ably, a-amylase activity analysis and gibberellic acid (GA) treatment showed that the dwarf phenotype of mtdl mutant was dependent on GA biosynthesis pathway. These results facilitated to further uncover the molecular mechanism of the growth and development in rice.