To confirm a hybrid swarm population of Pinus densiflora × P. sylvestris in Jilin, China, we used needles and seeds from P. densiflora, P. sylvestris, and P. densiflora × P. sylvestris collected from natural...To confirm a hybrid swarm population of Pinus densiflora × P. sylvestris in Jilin, China, we used needles and seeds from P. densiflora, P. sylvestris, and P. densiflora × P. sylvestris collected from natural stands or experimental stations to study whether shoot apex morphology of 4-year old seedlings can be correlated with the sequence of a chloroplast DNA simple sequence repeat marker （cpDNA SSRs）. Total genomic DNA was extracted and subjected to sequence analysis of the pine cpDNA SSR marker Pt15169. Results show that morphological characters from 4-year old seedlings did not correlate with sequence variants of this marker. Marker haplotypes from all P. sylvestris trees had a CTAT element that was absent from all sampled P. densiflora trees. However, both haplotype classes involving this insertion/deletion element were found in a P. densiflora × P. sylvestris population and its seedling progeny. It was concluded that the P. densiflora × P. sylvestris accessions sampled from Jilin, China resulted from bi-directional crosses, as evidenced by both species’ cpDNA haplotypes within the hybrid swarm population.展开更多
The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6V...The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.展开更多
Recently, many researchers have used nature inspired metaheuristicalgorithms due to their ability to perform optimally on complex problems. Tosolve problems in a simple way, in the recent era bat algorithm has becomef...Recently, many researchers have used nature inspired metaheuristicalgorithms due to their ability to perform optimally on complex problems. Tosolve problems in a simple way, in the recent era bat algorithm has becomefamous due to its high tendency towards convergence to the global optimummost of the time. But, still the standard bat with random walk has a problemof getting stuck in local minima. In order to solve this problem, this researchproposed bat algorithm with levy flight random walk. Then, the proposedBat with Levy flight algorithm is further hybridized with three differentvariants of ANN. The proposed BatLFBP is applied to the problem ofinsulin DNA sequence classification of healthy homosapien. For classificationperformance, the proposed models such as Bat levy flight Artificial NeuralNetwork (BatLFANN) and Bat levy Flight Back Propagation (BatLFBP) arecompared with the other state-of-the-art algorithms like Bat Artificial NeuralNetwork (BatANN), Bat back propagation (BatBP), Bat Gaussian distribution Artificial Neural Network (BatGDANN). And Bat Gaussian distributionback propagation (BatGDBP), in-terms of means squared error (MSE) andaccuracy. From the perspective of simulations results, it is show that theproposed BatLFANN achieved 99.88153% accuracy with MSE of 0.001185,and BatLFBP achieved 99.834185 accuracy with MSE of 0.001658 on WL5.While on WL10 the proposed BatLFANN achieved 99.89899% accuracy withMSE of 0.00101, and BatLFBP achieved 99.84473% accuracy with MSE of0.004553. Similarly, on WL15 the proposed BatLFANN achieved 99.82853%accuracy with MSE of 0.001715, and BatLFBP achieved 99.3262% accuracywith MSE of 0.006738 which achieve better accuracy as compared to the otherhybrid models.展开更多
The relationship between embB mutation of Mycobacterium tuberculosis and ethambutol （EMB） resistance of the clinical isolates of tuberculous patients in China was investigated by reversedot blot hybridization （RDBH...The relationship between embB mutation of Mycobacterium tuberculosis and ethambutol （EMB） resistance of the clinical isolates of tuberculous patients in China was investigated by reversedot blot hybridization （RDBH） in addition to evaluating the clinical value with application of PCR-RDBH technique to detect EMB resistance. In the present study, the genotypes of the 258 bp fragments of embB genes from 196 clinical isolates of M. tuberculosis were analysed with RDBH and DNA sequencing. It was demonstrated that 60 out of 91 phenotypically EMB-resistant isolates （65.9%） showed 5 types of missense mutations at codon 306 of embB gene, resulting in the replacement of the Met residue of the wild type strain with Val, Ile or Leu residues. In these mutations, the GTP mutation （38/91, 41.8% ） and the ATA mutation （16/91, 17.6% ） were the most encountered genotypes. The embB mutation at codon 306 could also be found in 69 isolates of phenotypically EMB-sensitive but resistant to other anti-tuberculous drugs, but no such gene mutation could be found in 36 strains of drug-sensitive isolates. Meanwhile, the concordance with the results of DNA sequencing fcr one wide-type probe and 5 probes for specific mutations was 100%. It was concluded that the EMB-resistance occurring in most M. tuberculosis is due to appearance of embB mutation at codon 306, and the PCR-RDBH assay was proved to be a rapid, simple and reliable method for the detection of gene mutations, which might be a good alternative for the drug-resistance screening.展开更多
Interspecific hybridization can result in significant shifts in allele frequencies. The objective of the present study was to assess the level of genetic variation in populations of P. mariana × P. rubens hybrids...Interspecific hybridization can result in significant shifts in allele frequencies. The objective of the present study was to assess the level of genetic variation in populations of P. mariana × P. rubens hybrids derived from artificial crosses. Progenies from backcross populations created through a series of controlled pollinations among P. mariana and P. rubens trees across the hybridization index were analyzed. Several Inter Simple Sequence Repeat (ISSR) and Random Amplified Polymorphic DNA (RAPD) primers were used to amplify genomic DNA samples from each population. ISSR primers produced from 30% to 52% polymorphic loci. The level of polymorphism was higher with RAPD markers, ranging from 57% to 76%. Overall, the two marker systems generated similar levels of polymorphic loci for P. mariana and P. rubens populations. No significant differences were found among the P. mariana × P. rubens populations analyzed and between the hybrids and the parental populations regardless of the molecular marker used. This confirms the genetic closeness of P. mariana and P. rubens species.展开更多
基金supported by a grant from the Next-Generation BioGreen 21 Program, Rural Development Administration, Republic of Korea (PJ009052)
文摘To confirm a hybrid swarm population of Pinus densiflora × P. sylvestris in Jilin, China, we used needles and seeds from P. densiflora, P. sylvestris, and P. densiflora × P. sylvestris collected from natural stands or experimental stations to study whether shoot apex morphology of 4-year old seedlings can be correlated with the sequence of a chloroplast DNA simple sequence repeat marker （cpDNA SSRs）. Total genomic DNA was extracted and subjected to sequence analysis of the pine cpDNA SSR marker Pt15169. Results show that morphological characters from 4-year old seedlings did not correlate with sequence variants of this marker. Marker haplotypes from all P. sylvestris trees had a CTAT element that was absent from all sampled P. densiflora trees. However, both haplotype classes involving this insertion/deletion element were found in a P. densiflora × P. sylvestris population and its seedling progeny. It was concluded that the P. densiflora × P. sylvestris accessions sampled from Jilin, China resulted from bi-directional crosses, as evidenced by both species’ cpDNA haplotypes within the hybrid swarm population.
基金国家"8 6 3"计划资助项目 (Z 17 0 4 0 1) 国家转基因植物研究与产业化资助项目 (J0 0 A 0 0 2 )~~
文摘The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.
基金This research is supported by Tier-1 Research Grant, vote no. H938 by ResearchManagement Office (RMC), Universiti Tun Hussein Onn Malaysia and Ministry of Higher Education,Malaysia.
文摘Recently, many researchers have used nature inspired metaheuristicalgorithms due to their ability to perform optimally on complex problems. Tosolve problems in a simple way, in the recent era bat algorithm has becomefamous due to its high tendency towards convergence to the global optimummost of the time. But, still the standard bat with random walk has a problemof getting stuck in local minima. In order to solve this problem, this researchproposed bat algorithm with levy flight random walk. Then, the proposedBat with Levy flight algorithm is further hybridized with three differentvariants of ANN. The proposed BatLFBP is applied to the problem ofinsulin DNA sequence classification of healthy homosapien. For classificationperformance, the proposed models such as Bat levy flight Artificial NeuralNetwork (BatLFANN) and Bat levy Flight Back Propagation (BatLFBP) arecompared with the other state-of-the-art algorithms like Bat Artificial NeuralNetwork (BatANN), Bat back propagation (BatBP), Bat Gaussian distribution Artificial Neural Network (BatGDANN). And Bat Gaussian distributionback propagation (BatGDBP), in-terms of means squared error (MSE) andaccuracy. From the perspective of simulations results, it is show that theproposed BatLFANN achieved 99.88153% accuracy with MSE of 0.001185,and BatLFBP achieved 99.834185 accuracy with MSE of 0.001658 on WL5.While on WL10 the proposed BatLFANN achieved 99.89899% accuracy withMSE of 0.00101, and BatLFBP achieved 99.84473% accuracy with MSE of0.004553. Similarly, on WL15 the proposed BatLFANN achieved 99.82853%accuracy with MSE of 0.001715, and BatLFBP achieved 99.3262% accuracywith MSE of 0.006738 which achieve better accuracy as compared to the otherhybrid models.
文摘The relationship between embB mutation of Mycobacterium tuberculosis and ethambutol （EMB） resistance of the clinical isolates of tuberculous patients in China was investigated by reversedot blot hybridization （RDBH） in addition to evaluating the clinical value with application of PCR-RDBH technique to detect EMB resistance. In the present study, the genotypes of the 258 bp fragments of embB genes from 196 clinical isolates of M. tuberculosis were analysed with RDBH and DNA sequencing. It was demonstrated that 60 out of 91 phenotypically EMB-resistant isolates （65.9%） showed 5 types of missense mutations at codon 306 of embB gene, resulting in the replacement of the Met residue of the wild type strain with Val, Ile or Leu residues. In these mutations, the GTP mutation （38/91, 41.8% ） and the ATA mutation （16/91, 17.6% ） were the most encountered genotypes. The embB mutation at codon 306 could also be found in 69 isolates of phenotypically EMB-sensitive but resistant to other anti-tuberculous drugs, but no such gene mutation could be found in 36 strains of drug-sensitive isolates. Meanwhile, the concordance with the results of DNA sequencing fcr one wide-type probe and 5 probes for specific mutations was 100%. It was concluded that the EMB-resistance occurring in most M. tuberculosis is due to appearance of embB mutation at codon 306, and the PCR-RDBH assay was proved to be a rapid, simple and reliable method for the detection of gene mutations, which might be a good alternative for the drug-resistance screening.
基金国家自然科学基金重点面上资助项目(the National Natural Science Foundation of China No.60533010+1 种基金No.30670540)国家高技术研究发展计划(863)(the National High-Tech Research and Development Plan of China under Grant No.2006AA01Z104)
基金National Natural Science Foundation of China(No.30170733,No.30330480)State Key Basic Research(973)Project of China(No.2001CB109006)Training Project of Excellent Young Research of State Education Ministry of China(No.200248)
文摘Interspecific hybridization can result in significant shifts in allele frequencies. The objective of the present study was to assess the level of genetic variation in populations of P. mariana × P. rubens hybrids derived from artificial crosses. Progenies from backcross populations created through a series of controlled pollinations among P. mariana and P. rubens trees across the hybridization index were analyzed. Several Inter Simple Sequence Repeat (ISSR) and Random Amplified Polymorphic DNA (RAPD) primers were used to amplify genomic DNA samples from each population. ISSR primers produced from 30% to 52% polymorphic loci. The level of polymorphism was higher with RAPD markers, ranging from 57% to 76%. Overall, the two marker systems generated similar levels of polymorphic loci for P. mariana and P. rubens populations. No significant differences were found among the P. mariana × P. rubens populations analyzed and between the hybrids and the parental populations regardless of the molecular marker used. This confirms the genetic closeness of P. mariana and P. rubens species.
