In order to investigate the genetic basis of morphological variation of tetraploid plantlets of Atractylodes macrocephala, diploid plantlets were taken as experimental material, sterile filtration colchicine was used to soak 0.5-1.0 cm long buds. The difference between morphology and stomatal of diploid and tetraploid of A. macrocephala was compared, and genome polymorphism was explored by AFLP. The results showed that the buds dipped in 0.1% colchicine solution for 36 h was optimal conditions to induce tetraploid of A. macrocephala with induction rate of 32.0%. Morphological indexes such as leaf area index, leaf length and width, the density of stomas and the number of chloroplast of tetraploid were distinctly different from diploid. Four hundred and fifty-one bands ranging with 80-500 bp were amplified with 24 pairs of primers, the rate of polymorphism was 32.59%. These amplification sites of diploid were different from tetraploid of A. macrocephala, and the differences in morphology of them were reflected in the DNA polymorphism.
Amplified Fragment Length Polymorphism Analysis ; methods ; Atractylodes ; genetics ; Sequence Analysis, DNA ; Tetraploidy

Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.
Amplified Fragment Length Polymorphism Analysis ; methods ; Cytoplasm ; metabolism ; DNA, Mitochondrial ; genetics ; DNA, Plant ; genetics ; Gene Expression Profiling ; Genotype ; Plant Infertility ; genetics ; Triticum ; genetics

The Direct Amplification of Length Polymorphisms was applied to assess genetic diversity and structure of 7 populations of Dendrobium nobile, comparing one population of D. lituflorum. The five primer combinations were amplified to produce 140 clear bands, and 102 polymorphic bands had been detected with each pair of primer producing 20.4 polymorphic bands on average. At species level, the percentage of polymorphic bands (PPB) was 72.86%, the Nei's gene diversity index (H) was 0.288 9, and the Shannon's information index (I) was 0.424 2. At population level, the average PPB was 47.96%, H was 0.1861, and I was 0.273 9 in 7 populations. The coefficient of gene differentiation (Gst) was 0.338 6 among populations of D. nobile. It showed that 33.86% of the total genetic diversity was attributable to genetic differentiation among populations, while the rest 66.14% was resided between individuals within population.
Amplified Fragment Length Polymorphism Analysis ; methods ; China ; Dendrobium ; classification ; genetics ; Genetic Variation ; Phylogeny ; Polymorphism, Genetic

OBJECTIVETo study the application of the multiplex ligation-dependent probe amplification (MLPA) method in genetic and prenatal diagnosis for spinal muscular atrophy (SMA).

METHODSFour patients, 16 parents and 4 fetuses from 8 SMA pedigrees were included. MLPA was performed for molecular analysis, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for the mutation detection of the 4 patients.

RESULTSFor all the four patients, the same homozygous deletion of the exons 7 and 8 of the survival motor neuron 1 (SMN1) gene, was detected by PCR-RFLP and MLPA. All fourteen parents from the 8 pedigrees were carriers of the SMN1 gene heterozygous deletion, except the mothers in pedigrees 1 and 4 in whom the mutations were different.

CONCLUSIONMLPA is a simple and efficient quantitative method for copy number analysis of the SMN genes. It can be used for the genetic diagnosis and prenatal diagnosis of the SMA patients and carriers.

Adult ; Amplified Fragment Length Polymorphism Analysis ; Exons ; Female ; Genetic Carrier Screening ; Heterozygote ; Humans ; Ligase Chain Reaction ; methods ; Male ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; Pedigree ; Polymorphism, Restriction Fragment Length ; Pregnancy ; Prenatal Diagnosis ; methods ; Sequence Deletion ; Survival of Motor Neuron 1 Protein ; genetics ; Young Adult

OBJECTIVE: To detect DNA polymorphism of Papaver somniferum L using fluorescent Amplified Fragment Length Polymorphism. METHODS: Genomic DNA was isolated using the AxyPrep DNA Kit, double-digested by two restrictional endonucleases (Eco RI and Mse I) and ligated to oligonucleotide adapters. After Pre-amplification and selective amplification, the DNA fragments were separated by capillary electrophoresis using the CEQ8000 DNA Fragment Analyzer. RESULTS: More than 20 fragments of highly polymorphic products were obtained by 8 pairs of primer from 64 selective amplifying primer pairs. CONCLUSION The fluorescent AFLP technique can be used to detect the DNA polymorphism of Papaver somniferum.
Amplified Fragment Length Polymorphism Analysis/methods* ; DNA, Plant/genetics* ; Fluorescent Dyes ; Forensic Genetics ; Papaver/genetics* ; Polymorphism, Genetic

OBJECTIVE: To screen the AFLP primers with good diversity to distinguish various species of Cannabis. METHODS: The AFLP was used to analyze the genetic diversity of 12 species of Cannabis using 55 primer combinations. RESULTS: A total of 285 AFLP bands were obtained using five primer combinations with better diversity, among which 99 bands were polymorphic and 10 bands were special, with 47-76 bands amplified in each pair of primers. CONCLUSION AFLP may has good resolution in the diversity study of Cannabis. It may provide an essential basis for further study of the genetic diversity of Cannabis.
Amplified Fragment Length Polymorphism Analysis/methods* ; Cannabis/genetics* ; DNA, Plant/genetics* ; Forensic Genetics ; Genetic Variation ; Polymorphism, Genetic

Amplified fragment length polymorphism (AFLP) is a new molecular marker to detect genomic polymorphism. This new technology has advantages of high resolution, good stability, and reproducibility. Great achievements have been derived in recent years in AFLP related technologies with several AFLP expanded methodologies available. AFLP technology has been widely used in the fields of plant, animal, and microbes. It has become one of the hotspots in Forensic Botany. This review focuses on the recent advances of AFLP and its applications in forensic biology.
Amplified Fragment Length Polymorphism Analysis/trends* ; Botany/methods* ; DNA, Plant/genetics* ; Forensic Genetics ; Plants/genetics*

Forensic botany is a study of judicial plant evidence. Recently, researches on DNA labeling technology have been a mainstream of forensic botany. The article systematically reviews various types of DNA labeling techniques in forensic botany with enumerated practical cases, as well as the potential forensic application of each individual technique. The advantages of the DNA labeling technology over traditional morphological taxonomic methods are also summarized.
Amplified Fragment Length Polymorphism Analysis ; Botany/methods* ; DNA Fingerprinting/methods* ; DNA, Plant/analysis* ; Forensic Genetics ; Minisatellite Repeats ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo develop fluorescent amplified fragment length polymorphism (AFLP) method and to evaluate the its typing capability with pulsed-field gel electrophoresis (PFGE) in molecular typing of Vibrio cholerae.

METHODSForty-seven strains of V. cholerae, with different PFGE patterns, were selected as the reference group to optimize the selective primers of AFLP analysis. Eighty-three strains including 20 strains from one epidemic episode, isolated from different provinces during 1961 and 2005, were used to compare the typing abilities of AFLP and PFGE. LI-COR4300 DNA sequencing system was used for AFLP electrophoresis. The images were recorded by Saga(MX) software and transferred to BioNumerics for clustering analysis. A standard protocol for V. cholerae from PulseNet was used in PFGE.

RESULTSWhen comparison was made with different selective primers on AFLP based on the 47 strains, results showed that the optimized selective primer pair was EcoR I-G/Mse I-T, and the reproducibility of the tests was 99.2%. Eighty-three isolates showed 52 AFLP patterns and 44 PFGE patterns, with D values as 0.9545 (AFLP) and 0.9251 (PFGE) respectively.

CONCLUSIONThe protocol of fluorescent AFLP on V. cholerae typing was established. AFLP was higher than PFGE in discrimination of V. cholerae which could be used for molecular typing. When combined with PFGE, AFLP became a more insightful tool to identify genome difference of different isolates.

Amplified Fragment Length Polymorphism Analysis ; methods ; Electrophoresis, Gel, Pulsed-Field ; Genotype ; Phylogeny ; Vibrio cholerae ; classification ; genetics

Sequence related amplified polymorphism (SRAP) technique was used to identify SRAP fragment linked to Carthamus tinctorius L. spines of outer involucral bract (OIB) , experimental evidence for molecular marker assistant breeding of Carthamus tinctorius L. has been provided. Based on the strategy of bulk segregate analysis (BSA), two gene pools were separately constructed according to the extreme trait of OIB with many long spines and no spines from Carthamus tinctorius L. Forty-five pairs of SRAP primers were selected and screened from two parents and two gene pools, and one SRAP marker M3E3 was found to be linked to the spines in segregating F2 population confirmation. M3E3 SRAP band was excised, cloned and sequenced. In 20 spininess individuals, this marker was present in 16 spininess individuals and absent in 4 individuals. This band was absent in the 15 spineless F, segregating individuals, which accounted for 11.4% recombination. The M3E3 extract length was 349bp, of which the base components of A + T accounted for 41. 08%. One SRAP marker M3E3 linked to the spines in Carthamus tinctorius L. will be of good use for breeding spineless cultivars at the molecular level in the future.
Amplified Fragment Length Polymorphism Analysis ; methods ; Base Sequence ; Carthamus tinctorius ; anatomy & histology ; genetics ; DNA Primers ; DNA, Plant ; genetics ; Genetic Markers ; Molecular Sequence Data ; Plant Leaves ; anatomy & histology ; genetics ; Polymorphism, Genetic ; Sequence Analysis, DNA