In cultivated rice ( Oryza sativa L.), F-1 pollen sterility is controlled by at least 6 loci of the F, pollen sterility genes. To map S-b, one of loci, rice variety Taichung 65 (T65) carrying S-b(j)/S-b(j) and its nea...In cultivated rice ( Oryza sativa L.), F-1 pollen sterility is controlled by at least 6 loci of the F, pollen sterility genes. To map S-b, one of loci, rice variety Taichung 65 (T65) carrying S-b(j)/S-b(j) and its near-isogenic line TIST2 carrying S-b(i)/S-b(i) were used to develop the mapping population. One hundred and fifty-eight microsatellite markers were selected to survey T65 and TISL2. RM13 on chromosome 5 was found to be polymorphic between them. Cosegregation indicated that RM13 was closely linked with locus S-b. Eleven RFLP markers were selected on the corresponding region from the genetic map of Rice Genome Research Program (RGP) of Japan to convert into sequence-tagged site (STS) markers. Amplicon length polymorphism (ALP) was carried out, but none of them was found to be polymorphic between T65 and TISL2. Then PCR-based RFLP (PBR) was done using six 4-nucleotide recognizing restriction endonucleases. Polymorphism was detected when PCR products of R830STS and R2213SSTS were digested with Taq I. Genetic analysis indicated that the distance between locus S-b and markers, R830STS, RM13 and R2213SSTS were 3.3 cM (centi-Morgan), 5.2 cM and 5.5 cM, respectively. These PCR-based markers could be directly used in marker-assisted selection. The technical system combining genetic mapping and PCR-based marker-assisted selection will facilitate the development of molecular breeding.展开更多
Powdery mildew is a serious disease of wheat in China. As part of ITEC (International Triteace EST cooperation), EST (expressed sequence tags) technique was used to explore the gene expression in leaf induced by Ery...Powdery mildew is a serious disease of wheat in China. As part of ITEC (International Triteace EST cooperation), EST (expressed sequence tags) technique was used to explore the gene expression in leaf induced by Erysiphe graminis DC. A conventional cDNA library was constructed, and a total of 1 500 clones picked randomly from the library were sequenced, three hundred and eighty_seven ESTs of them were unique, which got the Accession Number in GenBank. About 49.4% ESTs showed significant similarity to functions of known sequences in GenBank. There are 196 ESTs' with functions not able to be determined, and eighty_four ESTs were demonstrated to be novel sequences. High_density dot membranes from unique clones were produced, and several disease resistance related genes were screened by differential hybridization.展开更多
Rhododendron simsii（Ericaeae：Rhododendron） has high ornamental value and ecological value.In this study,7 pairs of novel EST-SSR markers were developed from the genomic sequence of R.simsii,and they were used to in...Rhododendron simsii（Ericaeae：Rhododendron） has high ornamental value and ecological value.In this study,7 pairs of novel EST-SSR markers were developed from the genomic sequence of R.simsii,and they were used to investigate the genetic diversity of 32 natural R.simsii samples from Guifeng Mountain,Hubei Province.Results showed that a total of 31 polymorphic bands were amplified with allele number per locus of 4.43.Mean values of heterozygosity（Ho） and expected heterozygosity（He） were 0.679 58 and 0.723 14,respectively.This research will not only enrich the existing SSR database,but also lay a foundation for subsequent studies about molecular marker-assisted breeding,genetic diversity analysis,genetic structure analysis and phylogenetic analysis.展开更多
Segmental duplications on rice (Oryza sativa L.) chromosomes 8, 9, 11, and 12 were studied by examining the distributions of sequences resolved by 13 probes detecting multiple copies of DNA sequences. Four of the hyb...Segmental duplications on rice (Oryza sativa L.) chromosomes 8, 9, 11, and 12 were studied by examining the distributions of sequences resolved by 13 probes detecting multiple copies of DNA sequences. Four of the hybridization bands detected by a repetitive sequence probe, rTRS, were mapped to the ends of all the four chromosomes. Two or three of the bands detected by each of the other 12 probes were also mapped to different chromosomes. The bands detected by the same probe usually occurred in similar locations of different chromosomes. Loci detected by different DNA probes were often similarly arranged on different chromosomes. Chromosomes 8 and 9 showed colinearity of marker loci arrangement indicating a possible common origin. A segment on chromosome 9 was also very similar to the previously reported duplicated fragments on the ends of chromosomes 11 and 12 which were also detected in this study, indicating a likely common origin. Moreover, the various degrees of distributional similarity of the segments suggest a complex relationship among the chromosomes in the evolution of the rice genome. These results support the proposition that chromosome duplication and diversification may be a mechanism for the origin and evolution of the chromosomes in the rice genome.展开更多
Objctive The aim was to make better use of the EST-SSR resources of kiwifruit for further molecular biological studies and new EST-SSR marker development by screening and mining the SSR repeats in the EST database of ...Objctive The aim was to make better use of the EST-SSR resources of kiwifruit for further molecular biological studies and new EST-SSR marker development by screening and mining the SSR repeats in the EST database of kiwifruit （Actinidia spp. ） [ Method] 56 400 of EST sequences were randomly selected from EST（ Expressed Sequence Tag）sequences of kiwifruit in the database of NCBI. EST sequences were analyzed and the SSR（Microsatellite） was screened by using the SSRHunter software. [ Result] 7 939 SSRs were identified from the ran- domly selected kiwifruit EST resources, among which there were 5 131 （64.63%） dinucleotide, 1 237 （ 15.58% ） trinucleotide, 284 （ 3.58% ） tetra nucleotide, 397 （5.00%） pentanucleotide and 890 （ 11.21% ） hexanucleotide SSRs. Among the dinucleotide sequences, AG/CT repeat motif was accounted for 4 654（90.70% ）. The frequency of SSRs was approximately 1/2.48 kb, which could exist to 1 SSR in 7 unigenes. [ Conclusion] The dinucleotide repeats appeared to be the most abundant SSRs, followed by the trinucleotide and hexanucleotide repeats. Among them the repeat motif such as AG/CT was predominant in each type of SSRs.展开更多
The objectives of this study were to isolate and characterize microsatellites from a heat tolerant variety of snap bean (Phaseolus vulgaris L.) in order to generate polymorphic genetic markers linked to quantitative t...The objectives of this study were to isolate and characterize microsatellites from a heat tolerant variety of snap bean (Phaseolus vulgaris L.) in order to generate polymorphic genetic markers linked to quantitative trait loci for heat tolerance. A genomic library contained 400-800 bp inserts was constructed and screened for the presence of (GA/CT) n and (CA/GT) n repeats. The proportion of positive clones yielded estimated of 3.72×10 4 such dinucleotide repeats per genome, roughly comparable to the abundance reported in other eukaryotic genomes. Twenty_six positive clones were sequenced. In contrast to mammalian genomes, the (GA/CT) n motif was much more abundant than the (CA/GT) n motif in these clones. The (GA/CT) n repeats also showed longer average repeat length (mean n =10.4 versus 6.5), suggesting that they are better candidates for yielding polymorphic genetic markers in the snap bean genome.展开更多
LK783 was found to be a good fertility restorer for K-type male sterility of wheat (Triticum aestivum L.). RAPD and ISSR (inter-simple sequence repeat polymorphism) markers were employed to map the major restoring gen...LK783 was found to be a good fertility restorer for K-type male sterility of wheat (Triticum aestivum L.). RAPD and ISSR (inter-simple sequence repeat polymorphism) markers were employed to map the major restoring gene in LK783. Maintainer and restorer DNA pools were established using the extreme sterile and fertile plants among KJ5418A//911289/LK783 F 1 population, respectively. Four hundred and eighteen RAPD primers and 33 ISSR primers were used for screening polymorphisms between the two pools, and amplification bands using a RAPD primer of OPK18 and an ISSR primer of UBC-845 were found polymorphic between the two pools. Linkage analysis showed that OPK18 450 and UBC-845 800 were linked to the restoring gene in LK783. The distance between the restoring gene and OPK18 450 was (15.07±6.28) cM (centiMorgan), with the distance between the restoring gene and UBC-845 800 being (8.20±4.85) cM. The marker of UBC-845 800 was located on chromosome 1BS by amplifying nulli-tetrasomics and 1B ditelosomics of Chinese Spring with the primer of UBC-845, indicating that the restoring gene in LK783 was located on 1BS. The breeding for new fertility restorer lines of K-type cytoplasmic male sterility of wheat would be facilitated by using the two markers.展开更多
A joint direction of arrival (DOA) estimation and phase calibration for synchronous CDMA system with decorrelator are presented. Through decorrelating processing DOAs of the desired users can be estimated independentl...A joint direction of arrival (DOA) estimation and phase calibration for synchronous CDMA system with decorrelator are presented. Through decorrelating processing DOAs of the desired users can be estimated independently and all other resolved signal interferences are eliminated. Emphasis is directed to applications in which sensor phases may be in error. It is shown that accurate phase calibration in conjunction with their use in high resolution DOA estimation can be achieved for the decoupled signals.展开更多
An improved estimation of motion vectors of feature points is proposed for tracking moving objects of dynamic image sequence. Feature points are firstly extracted by the improved minimum intensity change （MIC） algor...An improved estimation of motion vectors of feature points is proposed for tracking moving objects of dynamic image sequence. Feature points are firstly extracted by the improved minimum intensity change （MIC） algorithm. The matching points of these feature points are then determined by adaptive rood pattern searching. Based on the random sample consensus （RANSAC） method, the background motion is finally compensated by the parameters of an affine transform of the background motion. With reasonable morphological filtering, the moving objects are completely extracted from the background, and then tracked accurately. Experimental results show that the improved method is successful on the motion background compensation and offers great promise in tracking moving objects of the dynamic image sequence.展开更多
From the NCBI database, 81,518 Lactuca sativa EST sequences were downloaded, from which 61,757 non-redundant sequences were obtained. In total, 2040 SSR loci were identified, with the frequency of 3.3%. Trinucleotide ...From the NCBI database, 81,518 Lactuca sativa EST sequences were downloaded, from which 61,757 non-redundant sequences were obtained. In total, 2040 SSR loci were identified, with the frequency of 3.3%. Trinucleotide repeat was dominant, followed by dinucieotide and hexanucleotide repeats. Among 181 types of repeat motifs, （AGA）n and （ATG）n were the dominant repeat motif types, taking up 12.01% and 3.53%, respectively. Totally, four material-special EST-SSR markers were characterized. This study enriches molecular markers of L. sativa, and also lays a theoretical foundation for following germplasm identification, molecular marker assisted breeding and genetic map construction.展开更多
Seventy-five previously known plant microRNAs (miRNAs) were classified into 14 families according to their gene sequence identity. A total of 18,694 plant expressed sequence tags (EST) were found in the GenBank EST da...Seventy-five previously known plant microRNAs (miRNAs) were classified into 14 families according to their gene sequence identity. A total of 18,694 plant expressed sequence tags (EST) were found in the GenBank EST databases by comparing all previously known Arabidopsis miRNAs to GenBank’s plant EST databases with BLAST algorithms. After removing the EST sequences with high numbers (more than 2) of mismatched nucleotides, a total of 812 EST contigs were identified. After predicting and scoring the RNA secondary structure of the 812 EST sequences using mFold software, 338 new potential miRNAs were identified in 60 plant species. miRNAs are widespread. Some microRNAs may highly conserve in the plant kingdom, and they may have the same ancestor in very early evolution. There is no nucleotide substitution in most miRNAs among many plant species. Some of the new identified potential miRNAs may be induced and regulated by environmental biotic and abiotic stresses. Some may be preferentially expressed in specific tissues, and are regulated by developmental switching. These findings suggest that EST analysis is a good alternative strategy for identifying new miRNA candidates, their targets, and other genes. A large number of miRNAs exist in different plant species and play important roles in plant developmental switching and plant responses to environmental abiotic and biotic stresses as well as signal transduction. Environmental stresses and developmental switching may be the signals for synthesis and regulation of miRNAs in plants. A model for miRNA induction and expression, and gene regulation by miRNA is hypothesized.展开更多
Common carp expressed sequence tags （ESTs） were analyzed for the existence of microsatellites, or simple sequence repeats （SSRs）. In the NCBI dbEST database, a total of 10612 sequences were registered before Decem...Common carp expressed sequence tags （ESTs） were analyzed for the existence of microsatellites, or simple sequence repeats （SSRs）. In the NCBI dbEST database, a total of 10612 sequences were registered before December 31, 2004. A complete search of 2-6 nucleotide microsatellites resulted in the identification of 513 SSR-containing ESTs, accounting for 4.8% of the total. Cluster analysis indicated that 73 sequences of SSR-containing ESTs fell into 27 groups and the remaining 440 ESTs were independent. A total of 467 unique SSR-containing ESTs were identified. These EST-SSRs contained a vari- ety of simple sequence types, and di- and tri-nucleotide repeats were the most abundant, accounting for 42.1% and 27.9% of the whole, respectively. Of the dinucleotide repeats, CA/TG was the most abundant, followed by GA/TC. BLASTx search showed that 38.1% of the SSR loci could be associated with genes or proteins of known or unknown function. BLASTx searches of SSR-containing ESTs also showed high frequencies （98/179） of hits on zebrafish sequences.展开更多
文摘In cultivated rice ( Oryza sativa L.), F-1 pollen sterility is controlled by at least 6 loci of the F, pollen sterility genes. To map S-b, one of loci, rice variety Taichung 65 (T65) carrying S-b(j)/S-b(j) and its near-isogenic line TIST2 carrying S-b(i)/S-b(i) were used to develop the mapping population. One hundred and fifty-eight microsatellite markers were selected to survey T65 and TISL2. RM13 on chromosome 5 was found to be polymorphic between them. Cosegregation indicated that RM13 was closely linked with locus S-b. Eleven RFLP markers were selected on the corresponding region from the genetic map of Rice Genome Research Program (RGP) of Japan to convert into sequence-tagged site (STS) markers. Amplicon length polymorphism (ALP) was carried out, but none of them was found to be polymorphic between T65 and TISL2. Then PCR-based RFLP (PBR) was done using six 4-nucleotide recognizing restriction endonucleases. Polymorphism was detected when PCR products of R830STS and R2213SSTS were digested with Taq I. Genetic analysis indicated that the distance between locus S-b and markers, R830STS, RM13 and R2213SSTS were 3.3 cM (centi-Morgan), 5.2 cM and 5.5 cM, respectively. These PCR-based markers could be directly used in marker-assisted selection. The technical system combining genetic mapping and PCR-based marker-assisted selection will facilitate the development of molecular breeding.
文摘Powdery mildew is a serious disease of wheat in China. As part of ITEC (International Triteace EST cooperation), EST (expressed sequence tags) technique was used to explore the gene expression in leaf induced by Erysiphe graminis DC. A conventional cDNA library was constructed, and a total of 1 500 clones picked randomly from the library were sequenced, three hundred and eighty_seven ESTs of them were unique, which got the Accession Number in GenBank. About 49.4% ESTs showed significant similarity to functions of known sequences in GenBank. There are 196 ESTs' with functions not able to be determined, and eighty_four ESTs were demonstrated to be novel sequences. High_density dot membranes from unique clones were produced, and several disease resistance related genes were screened by differential hybridization.
文摘Rhododendron simsii（Ericaeae：Rhododendron） has high ornamental value and ecological value.In this study,7 pairs of novel EST-SSR markers were developed from the genomic sequence of R.simsii,and they were used to investigate the genetic diversity of 32 natural R.simsii samples from Guifeng Mountain,Hubei Province.Results showed that a total of 31 polymorphic bands were amplified with allele number per locus of 4.43.Mean values of heterozygosity（Ho） and expected heterozygosity（He） were 0.679 58 and 0.723 14,respectively.This research will not only enrich the existing SSR database,but also lay a foundation for subsequent studies about molecular marker-assisted breeding,genetic diversity analysis,genetic structure analysis and phylogenetic analysis.
文摘Segmental duplications on rice (Oryza sativa L.) chromosomes 8, 9, 11, and 12 were studied by examining the distributions of sequences resolved by 13 probes detecting multiple copies of DNA sequences. Four of the hybridization bands detected by a repetitive sequence probe, rTRS, were mapped to the ends of all the four chromosomes. Two or three of the bands detected by each of the other 12 probes were also mapped to different chromosomes. The bands detected by the same probe usually occurred in similar locations of different chromosomes. Loci detected by different DNA probes were often similarly arranged on different chromosomes. Chromosomes 8 and 9 showed colinearity of marker loci arrangement indicating a possible common origin. A segment on chromosome 9 was also very similar to the previously reported duplicated fragments on the ends of chromosomes 11 and 12 which were also detected in this study, indicating a likely common origin. Moreover, the various degrees of distributional similarity of the segments suggest a complex relationship among the chromosomes in the evolution of the rice genome. These results support the proposition that chromosome duplication and diversification may be a mechanism for the origin and evolution of the chromosomes in the rice genome.
基金Supported by the National Natural Science Foundation of China(30660113,30860167)~~
文摘Objctive The aim was to make better use of the EST-SSR resources of kiwifruit for further molecular biological studies and new EST-SSR marker development by screening and mining the SSR repeats in the EST database of kiwifruit （Actinidia spp. ） [ Method] 56 400 of EST sequences were randomly selected from EST（ Expressed Sequence Tag）sequences of kiwifruit in the database of NCBI. EST sequences were analyzed and the SSR（Microsatellite） was screened by using the SSRHunter software. [ Result] 7 939 SSRs were identified from the ran- domly selected kiwifruit EST resources, among which there were 5 131 （64.63%） dinucleotide, 1 237 （ 15.58% ） trinucleotide, 284 （ 3.58% ） tetra nucleotide, 397 （5.00%） pentanucleotide and 890 （ 11.21% ） hexanucleotide SSRs. Among the dinucleotide sequences, AG/CT repeat motif was accounted for 4 654（90.70% ）. The frequency of SSRs was approximately 1/2.48 kb, which could exist to 1 SSR in 7 unigenes. [ Conclusion] The dinucleotide repeats appeared to be the most abundant SSRs, followed by the trinucleotide and hexanucleotide repeats. Among them the repeat motif such as AG/CT was predominant in each type of SSRs.
文摘The objectives of this study were to isolate and characterize microsatellites from a heat tolerant variety of snap bean (Phaseolus vulgaris L.) in order to generate polymorphic genetic markers linked to quantitative trait loci for heat tolerance. A genomic library contained 400-800 bp inserts was constructed and screened for the presence of (GA/CT) n and (CA/GT) n repeats. The proportion of positive clones yielded estimated of 3.72×10 4 such dinucleotide repeats per genome, roughly comparable to the abundance reported in other eukaryotic genomes. Twenty_six positive clones were sequenced. In contrast to mammalian genomes, the (GA/CT) n motif was much more abundant than the (CA/GT) n motif in these clones. The (GA/CT) n repeats also showed longer average repeat length (mean n =10.4 versus 6.5), suggesting that they are better candidates for yielding polymorphic genetic markers in the snap bean genome.
文摘LK783 was found to be a good fertility restorer for K-type male sterility of wheat (Triticum aestivum L.). RAPD and ISSR (inter-simple sequence repeat polymorphism) markers were employed to map the major restoring gene in LK783. Maintainer and restorer DNA pools were established using the extreme sterile and fertile plants among KJ5418A//911289/LK783 F 1 population, respectively. Four hundred and eighteen RAPD primers and 33 ISSR primers were used for screening polymorphisms between the two pools, and amplification bands using a RAPD primer of OPK18 and an ISSR primer of UBC-845 were found polymorphic between the two pools. Linkage analysis showed that OPK18 450 and UBC-845 800 were linked to the restoring gene in LK783. The distance between the restoring gene and OPK18 450 was (15.07±6.28) cM (centiMorgan), with the distance between the restoring gene and UBC-845 800 being (8.20±4.85) cM. The marker of UBC-845 800 was located on chromosome 1BS by amplifying nulli-tetrasomics and 1B ditelosomics of Chinese Spring with the primer of UBC-845, indicating that the restoring gene in LK783 was located on 1BS. The breeding for new fertility restorer lines of K-type cytoplasmic male sterility of wheat would be facilitated by using the two markers.
文摘A joint direction of arrival (DOA) estimation and phase calibration for synchronous CDMA system with decorrelator are presented. Through decorrelating processing DOAs of the desired users can be estimated independently and all other resolved signal interferences are eliminated. Emphasis is directed to applications in which sensor phases may be in error. It is shown that accurate phase calibration in conjunction with their use in high resolution DOA estimation can be achieved for the decoupled signals.
文摘An improved estimation of motion vectors of feature points is proposed for tracking moving objects of dynamic image sequence. Feature points are firstly extracted by the improved minimum intensity change （MIC） algorithm. The matching points of these feature points are then determined by adaptive rood pattern searching. Based on the random sample consensus （RANSAC） method, the background motion is finally compensated by the parameters of an affine transform of the background motion. With reasonable morphological filtering, the moving objects are completely extracted from the background, and then tracked accurately. Experimental results show that the improved method is successful on the motion background compensation and offers great promise in tracking moving objects of the dynamic image sequence.
基金Supported by General Project of Natural Science Foundation of Hubei Province(2015CFC816)~~
文摘From the NCBI database, 81,518 Lactuca sativa EST sequences were downloaded, from which 61,757 non-redundant sequences were obtained. In total, 2040 SSR loci were identified, with the frequency of 3.3%. Trinucleotide repeat was dominant, followed by dinucieotide and hexanucleotide repeats. Among 181 types of repeat motifs, （AGA）n and （ATG）n were the dominant repeat motif types, taking up 12.01% and 3.53%, respectively. Totally, four material-special EST-SSR markers were characterized. This study enriches molecular markers of L. sativa, and also lays a theoretical foundation for following germplasm identification, molecular marker assisted breeding and genetic map construction.
文摘Seventy-five previously known plant microRNAs (miRNAs) were classified into 14 families according to their gene sequence identity. A total of 18,694 plant expressed sequence tags (EST) were found in the GenBank EST databases by comparing all previously known Arabidopsis miRNAs to GenBank’s plant EST databases with BLAST algorithms. After removing the EST sequences with high numbers (more than 2) of mismatched nucleotides, a total of 812 EST contigs were identified. After predicting and scoring the RNA secondary structure of the 812 EST sequences using mFold software, 338 new potential miRNAs were identified in 60 plant species. miRNAs are widespread. Some microRNAs may highly conserve in the plant kingdom, and they may have the same ancestor in very early evolution. There is no nucleotide substitution in most miRNAs among many plant species. Some of the new identified potential miRNAs may be induced and regulated by environmental biotic and abiotic stresses. Some may be preferentially expressed in specific tissues, and are regulated by developmental switching. These findings suggest that EST analysis is a good alternative strategy for identifying new miRNA candidates, their targets, and other genes. A large number of miRNAs exist in different plant species and play important roles in plant developmental switching and plant responses to environmental abiotic and biotic stresses as well as signal transduction. Environmental stresses and developmental switching may be the signals for synthesis and regulation of miRNAs in plants. A model for miRNA induction and expression, and gene regulation by miRNA is hypothesized.
文摘Common carp expressed sequence tags （ESTs） were analyzed for the existence of microsatellites, or simple sequence repeats （SSRs）. In the NCBI dbEST database, a total of 10612 sequences were registered before December 31, 2004. A complete search of 2-6 nucleotide microsatellites resulted in the identification of 513 SSR-containing ESTs, accounting for 4.8% of the total. Cluster analysis indicated that 73 sequences of SSR-containing ESTs fell into 27 groups and the remaining 440 ESTs were independent. A total of 467 unique SSR-containing ESTs were identified. These EST-SSRs contained a vari- ety of simple sequence types, and di- and tri-nucleotide repeats were the most abundant, accounting for 42.1% and 27.9% of the whole, respectively. Of the dinucleotide repeats, CA/TG was the most abundant, followed by GA/TC. BLASTx search showed that 38.1% of the SSR loci could be associated with genes or proteins of known or unknown function. BLASTx searches of SSR-containing ESTs also showed high frequencies （98/179） of hits on zebrafish sequences.
