AFLP technology has been widely used in molecular biology due to its integration of several advantages of high throughput, high efficiency and requiring no sequence information, etc. Great changes have been achieved in recent years in AFLP-related technologies and platforms. There are several AFLP-expanded technologies available. These improved technologies are capable of distinguishing the heterozygote from the homozygote and of converting any AFLP band of interest, without much effort, into locus-specific markers, which can be deployed for massive locus detection and for gene isolation. This review focuses on these favorable changes from conventional AFLP technology into more effective and more practicable AFLP-related ones. Understanding these advancements and AFLP-expanded technologies will facilitate the achievement of our research goals.
Amplified Fragment Length Polymorphism Analysis ; methods ; Microsatellite Repeats ; Polymorphism, Single Nucleotide ; Quantitative Trait Loci

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

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

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*

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

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

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 explore the reasonable sample size and the number of molecular marker loci in the study of amplified fragment length polymorphism (AFLP) being used to analyze the genetic diversity of Ocomelania hupensis.

METHODSThe ribbed-shelled snails coming from Yueyang, Hunan province, were selected to analyze the relationship of the number of AFLP molecular marker loci and sample size with the reliability of information on genetic variation for Ocomelania hupensis by AFLP method.

RESULTSCorrelations found among the numbers of AFLP molecular marker loci and the sample size with reliable information on genetic variation for Ocomelania hupensis. When sample size was less than 7 individuals, the total number of AFLP loci, the number of polymorphic loci, Nei's gene diversity and Shannon's information index appeared great changes. However, when sample size was bigger than 30 individuals, the values of these indices tended to be stabilized. When the number of AFLP loci was less than 128, the frequency of polymorphic loci, Nei's gene diversity, Shannon's information index and the standard deviation of these two indices changed greatly. Again, when the number of loci was bigger than 338, the values of these indices tended to be stabilized.

CONCLUSIONWhen the genetic diversity of Ocomelania hupensis were analyzed by AFLP method, the sample size coming from each snail population should not be less than 30 individuals and the number of molecular loci analyzed not less than 338.

Amplified Fragment Length Polymorphism Analysis ; methods ; Animals ; Gene Frequency ; Genetic Loci ; Genetic Markers ; Genetic Variation ; Polymorphism, Genetic ; Reproducibility of Results ; Sample Size ; Snails ; genetics

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