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Among the important pulse-yielding plants Vigna radiata is important. In the Indian subcontinent, it is an essential source of dietary protein particularly in the vegetarian population. It contains abundant nutrients with biological activities. Soaked seeds are eaten as a good source of protein and mature seeds are cooked or can be sprouted for human consumption. Mung beans and its sprouts contain chemical constituents like flavonoids, phenolic acids, organic acids, amino acids, carbohydrates, and lipids. It is related to different biological activities like antioxidant, antimicrobial, antitumor, anti-inflammatory, anti-diabetic, and antihypertensive effects, etc. The seeds are a traditional source of cures for paralysis, rheumatism, coughs, fevers, and liver ailments. As a green manure or cover crop, the plant can be grown, enriching the soil with the nitrogen formed on its roots. The powerful Amplified Fragment Length Polymorphism (AFLP) is a molecular marker used for a variety of applications like assessing genetic diversity within species or among closely related species, inferring population-level phylogenies, generating genetic maps, and determining the relatedness among cultivars. AFLP has become extremely beneficial in the study of taxa including bacteria, fungi, and plants, where much is still unknown about the genomic makeup of various organisms. A study was made to assess the genetic diversity and phylogenetic relationships of a set of five varieties of Vigna radiata (L) Wilczek using the AFLP technique. Five different varieties of Vigna radiata viz. B1, TARM 2, PDM 84, TM 99, and TM 98 were subjected to AFLP analysis. A total of 471 fragments were scored across all the 12 AFLP primer sets used and the results were used to plot a dendrogram. It was observed that these five varieties formed three distinct groups among which the aromatic B1 variety formed a distinct group.

Bu calismanin amaci hiyar genotiplerinin morfolojik ve molekuler cesitliligini degerlendirmektir. Bu amacla, alti adet morfolojik ozellik (bitki morfolojisi, bitki boyu, yaprak ayasi buyuklugu, meyve uzunlugu, meyve capi ve meyve sapi uzunlugu) ve sekiz AFLP markeri kullanilarak 18 hiyar hatti genetik cesitlilik acisindan degerlendirilmistir. Kullanilan AFLP primer kombinasyonlari polimorfizm gostermistir. Calisma sonucunda 1975 AFLP fragmani elde edilmis ve 1468 fragmanin polimorfik oldugu gorulmustur (%75.34). Dendrogramlar, aritmetik ortalamalar kullanilarak UPGMA (Unweighted Pair Group Method) yontemiyle cizilmis ve UPGMA dendrogramina gore hiyar genotipleri iki ana gruba ayrilmistir. Dendrogramin genetik mesafeleri 0.92 ile 0.96 arasinda degismistir. Morfolojik verilere dayanan kume analizinde ise UPGMA dendrogrami tum hatlari uc ana kumeye ayirmistir. Calismada kullanilan hiyar hatlarinin benzerlik katsayisinin 0.888 ile 0.982 arasinda degisen dusuk bir genetik varyasyon seviyesine sahip oldugu belirlenmistir. 8 cift AFLP primeri ile olusturulan filogenetik dendogramlarla, morfolojik taksonomi ile yapilan UPGMA kumeleme analizleriyle olusturulan dendogramlar ortusmektedir.

The genetic diversity of 123 apple accessions from the collection of the research and production base Pushkin and Pavlovsk Laboratories of VIR, including landraces, Soviet cultivars, and commercial Russian and foreign cultivars, was for the first time estimated using the AFLP technique. A total of 355 fragments were obtained, 319 of which were polymorphic (89.86%), and each of the accessions had a unique set of AFLP fragments. The level of diversity of modern commercial cultivars was lower (74.11%, Не = 0.202, I = 0.302) as compared with landraces (89.20%, Не = 0.238, I = 0.366). A wide range of genetic differences between the studied accessions (0.99–0.63) was demonstrated. The statistical analysis of the data obtained made it possible to divide the studied samples into two groups. The first group included most of the analyzed landraces; with no clear differentiation of these landraces by origin. The second group included modern commercial Russian and foreign apple cultivars, Soviet cultivars, and some landraces.

The efficient use of genetic resources- stored in germplasm collections can be maximized if morphoagronomic and molecular information on the accessions is made available. To achieve this, a collection that is well-structured, well-curated and easily accessible (the core collection) is required. Consequently, the objective of the current study was to characterize 80 landrace accessions from the maize core collection of the Federal University of Viçosa (UFV), and assay thenngenetic diversity of the various landraces, considering grain type and ecogeographic origin. For this, AFLP analysis was performed using 12 primer combinations. Genetic diversity of the collection was quantified with the UPGMA method, using the Jaccard Index to quantify dissimilarity. The core collection was divided into four sub-populations by grain type, and into six sub-populations based on ecogeographic origin. Genetic diversity analysis was performed both within and between sub-populations. A high level of genetic variability was found among the landrace accessions of UFV Core Collection, principally among those accessions with dentate type grains.Classification by grain type andecogeographic origin allowed genetically divergent groups to be distinguished., The efficient use of genetic resources- stored in germplasm collections can be maximized if morphoagronomic and molecular information on the accessions is made available. To achieve this, a collection that is well-structured, well-curated and easily accessible (the core collection) is required. Consequently, the objective of the current study was to characterize 80 landrace accessions from the maize core collection of the Federal University of Viçosa (UFV), and assay thenngenetic diversity of the various landraces, considering grain type and ecogeographic origin. For this, AFLP analysis was performed using 12 primer combinations. Genetic diversity of the collection was quantified with the UPGMA method, using the Jaccard Index to quantify dissimilarity. The core collection was divided into four sub-populations by grain type, and into six sub-populations based on ecogeographic origin. Genetic diversity analysis was performed both within and between sub-populations. A high level of genetic variability was found among the landrace accessions of UFV Core Collection, principally among those accessions with dentate type grains.Classification by grain type and ecogeographic origin allowed genetically divergent groups to be distinguished.

The first molecular genetic analysis of the apple species and varieties from Russian collections with the AFLP marker system was performed in order to study the genetic diversity of the genus Malus, as well as to clarify the phylogeny and solve some systematic issues of the genus. Nienty-one apple accessions, including species from five sections of the genus Malus and hybrid species, were examined. The level of polymorphism constituted 90.2%. It was demonstrated that the classical taxonomy of the genus Malus, which identifies five sections based on differences in their morphological characters, is valid and may be used to classify apple species. The species assignment of the Antonovka landraces was established. All of them belonged to the species M. domestica. It was demonstrated that the Yakutskaya apple variety was a domesticated species of the section Gymnomeles, presumably, M. baccata. AFLP analysis confirmed the hybrid nature of many species. The relationships between apple varieties of the Golden group with American wild species were demonstrated. The data suggest that the species M. sieversii was the ancestor of not only the domestic apple but also of other species of the Malus sections.

Strawberry (Fragaria ananassa Duch.) is one of the most important berries in the world. Amplified fragment length polymorphism (AFLP) was used to evaluate the genetic diversity of strawberry in Motuo County of the Tibet Plateau. Seventy strawberry samples grown in Motuo County were collected for AFLP analysis using 256 primer combinations. A set of 10 primer combinations produced 374 clearly identifiable AFLP loci. The polymorphism ranged from 50% to 64% with an average of 58.8%. Nei's genetic differentiation index for AFLP was 0.75, and the estimate of gene flow was less than 1 (Nm, 0.41). The unweighted pair group method with arithmetic mean dendrogram and Bayesian analysis clustered 70 samples into six groups. These findings clearly indicated high genetic diversity in wild strawberry from the Motuo County. These results could lay the foundation for further breeding and conservation of strawberry.

Sasa borealis (Hack.) Makino & Shibata is widely distributed in South Korea. With amplified fragment lengthpolymorphism (AFLP) markers, we analyzed the genetic diversity of S. borealis to predict and measure the phytogeograph-ical factors of these populations. Relatively high levels of genetic diversity (PPL = 37.2%, h= 0.143, I= 0.205) and geneticdifferentiation (G st = 0.324, θ B = 0.395) were confirmed in populations of S. borealis. Moreover, an analysis of molecular vari-ance (AMOVA) showed that the rate of differentiation among the populations was 47.7%. The results showed that geneticdiversity is inversely proportional to the latitude of the S. borealis populations, indicating that the distribution of S. borealismay have extended from lower to higher latitudes. This method of investigating the correlation between genetic diversity andlatitude presents critical information for estimating changes in distributions and plant conservation due to climate change. Keywords

Biodiversity of Rottboellia cochinchinensis from different areas were studied by morphological traits and amplified fragment length polymorphism (AFLP) analysis for classify of an allelopathic ability. The correlation of the similarity/distance between AFLP markers (Jaccard coefficient) and morphological traits (Euclidean distance) was significant with r = − 0.84**. Itchgrass could be divided into two groups from both UPGMA and STRUCTURE analyses: the group A consisted of itchgrass from Chaehom - Lampang, Si Thep - Phetchabun, Phrom Phiram - Phitsanulok, Amphur Muang - Nakhon Sawan, Kamalasai - Kalasin, Amphur Muang - Chachoengsao and Bang Yai - Nonthaburi, whereas itchgrass from Amphur Muang - Chiang Mai, Pak Chong - Nakhon Ratchasima and Kamphaeng Saen - Nakhon Pathom constituted the group B. Allelopathic properties of itchgrass as representative from different group were determined in bioassay test, the result showed that the aqueous extract of itchgrass from Chaehom - Lampang area has a strong allelopathic ability on growth of Echinochloa crus-galli L., Bidens pilosa L. and Lactuca sativa L. than the other group. The molecular analysis was strongly supported in morphological analysis clustering with bioassay test for allelopathic ability, specific morphological traits were soft trichomes, and the dark purple stems and roots which can be used for the preliminary classification of allelopathic ability. Our findings suggest that classification of itchgrass by morphological traits is related to the analysis of the genetic relationship of itchgrass with AFLP analysis, that allowing the assessment of the bio - diversity of itchgrass and their allelopathic potentials.

In this work, a set of pea accessions obtained from the Vavilov Research Institute (VIR) collection comprising 83 P. sativum samples, including representatives of three subspecies, was studied using the AFLP method. Local cultivars for different uses with maximum ecological and geographical diversity (including those from the centers of origin of the species) were predominantly chosen for the study; a number of their morphological and biological characteristics were evaluated. We obtained 382 polymorphic AFLP fragments; each sample was characterized by a unique set of fragments. The genetic diversity of the studied material was evaluated, and a wide range of genetic differences in the investigated samples (0.07–0.27) was demonstrated. The affiliation of the samples to the certain subspecies was not confirmed by the obtained data; the ecogeographical differentiation of the samples was not reflected by the data. Factor analysis allowed us to identify the sample groups of European and Asian origin and the intermediate nature of most of the samples from the studied set of pea accessions.

Biligetu, B., Schellenberg, M. P. and Fu, Y.B. 2013. Genetic diversity of side-oats grama [Bouteloua curtipendula (Michx.) Torr.] populations as revealed by amplified fragment length polymorphism markers. Can. J. Plant Sci. 93: 1105–1114. Side-oats grama [Bouteloua curtipendula (Michx.) Torr.] is a warm-season grass widely distributed in North America and is considered as an important grass for reclamation and summer forage production in drier regions. This study assessed genetic diversity of nine wild populations of side-oats grama grass, their corresponding balanced multiple-site composite (BMSC), and a population selected for forage type (FT), using the amplified fragment length polymorphism (AFLP) technique. Five AFLP primer pairs were employed to genotype 157 plants, and 312 polymorphic AFLP bands were detected. The frequencies of AFLP bands ranged from 0.01 to 0.99, and averaged 0.39. The AFLP analysis revealed 6% of the total AFLP variation present among the nine wild populations and 94% of variation within populations. The Minto population had the largest within-population diversity, while the FT population the lowest based on AFLP band richness and polymorphic loci. The BMSC population displayed significant genetic differentiation from the wild populations, but still captured substantial genetic diversity. Bayesian cluster analysis using BAPS and STRUCTURE programs revealed three and four optimal clusters of populations that explained 9.8 and 9.5% of the total AFLP variation, respectively. A genetic clustering of individual plants displayed no clear genetic separations among wild, FT, and BMSC populations, but the FT population showed some level of genetic shift, indicating the initial impact of artificial selection. These findings are significant for our understanding of the genetic diversity of side-oats grama. Large genetic variation present within populations provides a potential for further genetic enhancement.

Fil: Velez, Maria Laura. Centro de Investigacion y Extension Forestal Andino Patagonico; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas; Argentina

Qin, Y., Kabir, M. A., Wang, H. W., Lee, Y. H., Hong, S. H., Kim, J. Y., Yook, M. J., Kim, D. S., Kim, C. S., Kwon, H. and Kim, W. 2013. Assessment of genetic diversity and relationships based on RAPD and AFLP analyses in Miscanthus genera landraces. Can. J. Plant Sci. 93: 171–182. In this study, molecular markers, random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) as well as combined RAPD and AFLP analysis were used to assess genetic diversity in a reference set of 38 Miscanthus accessions of which 32 were collected from South Korea and 6 from foreign countries. Using 30 selected RAPD primers, 197 amplified products were generated with an average of 6.6 bands, of which 135 bands were polymorphic (68.6%). A total of 1150 bands were detected by four-primer AFLP combinations with an average of 287.5 bands, out of which 923 bands were polymorphic (80.3%) across all the accessions. Analysis of molecular variance (AMOVA) indicated that a high proportion of the genetic variation (56% for RAPD and 58% for AFLP) was found among the Miscanthus species in South Korea. Genetic relationship was estimated using the Jaccard's coefficient values between different accessions, ranging from 0.23 to 0.93 in RAPD and 0.34 to 0.94 in AFLP. The un-weighted pair group method with arithmetic mean (UPGMA) analysis demonstrated less difference between RAPD and AFLP when alternative similarity coefficient was applied. The principal coordinates (PCO) analysis also revealed the significant geographic structure in the tested accessions. Among the accessions, SNU-M-040, 074 and 157 were highly divergent. Pattern of isolation by distance was observed in Miscanthus accessions, indicating that significant genetic differentiation among accessions might be due to the geographic distance.

Genetic variation among 45 Rhizopogon roseolus isolates from 21 different regions of Japan were inferred using amplified fragment length polymorphism (AFLP) markers. Using three primer pair combinations, AFLP analysis reproducibly produced a total of 223 DNA fragments, 74.4% of which were polymorphic. Pairwise dissimilarity of AFLP patterns between isolates ranged from 0.043 to 0.228. Cluster analysis and principal coordinate analysis of AFLP data generally showed four major clusters from geographically distinct areas. The findings suggested that the Japanese populations of R. roseolus from different geographical regions can be distinguished based on AFLP characters.

Determination of the minimum population size is an important component for the ex situ conservation of an endangered species. Here, we present the identification of collection strategies that most efficiently captured the genetic diversity of Brasenia schreberi J.F. Gmelin (water-shield) in natural populations from the mainland (MGC) and Jeju Island (JNS) of South Korea, using amplified fragment length polymorphism (AFLP) markers. A total of 313 and 383 polymorphic bands were detected in the MGC and JNS populations, respectively. All of the 140 sampled ramets were distinguishable by the presence of distinct AFLP phenotypes. According to the simulation of the individual sampling by maximization sampling, 25 and 28 individuals captured all of the genetic diversity in the MGC population (mainland of South Korea) and the JNS population (Jeju Island), respectively. The level of genetic diversity of the core collections was similar to the entire collection, indicating that the core collections very well represent the diversity of the entire collection. We therefore suggest a management unit of B. schreberi based on the genetic information for assessing the minimum population size for its ex situ conservation.

AFLP fingerprinting of the 98 main sweetpotato varieties planted in China has been constructed. Using 17 AFLP primer combinations which were selected from 1 208 primer combinations and generated the most amounts of polymorphic bands, AFLP analysis of the 98 main sweetpotato varieties gave a total of 410 clear polymorphic bands with an average of 24.12 polymorphic bands per primer combination. Each one of the 98 sweetpotato varieties could be clearly distinguished by Eco R I-cta /Mse I-ggc primer combination which generated the most polymorphic bands. AFLP-based genetic distance ranged from 0.0546 to 0.5709 with an average of 0.3799. The dendrogram based on AFLP markers indicated that sweetpotato varieties coming from the same regions or having same parents were clustered in the same groups. Analysis of molecular variance (AMOVA) revealed greater variations within regions (94.08%) than among regions (5.92%). Thus, the genetic variations mainly existed within regions, while the variations among regions were very low in the tested sweetpotato varieties. Significant genetic variations existed between “Northern” and “Southern” sweetpotato varieties when Yangtze River was used as the dividing line.

Echinacea is an allogamous genus, thus its cultivars or populations are genetically heterogeneous. Using amplified fragment length polymorphism (AFLP) to estimate the genetic diversity of Echinacea is generally limited by the large number of individual plants and the higher cost that need to be processed. In the present study, effectiveness of several sizes of DNA bulking (10, 15, 20, 25 and 30 individuals) with 20, 36 and 55 primer pairs was compared using AFLP in determining the genetic diversity of Echinacea species. The results indicated that the use of bulked DNA-based AFLP analysis by using the selected eight primer pairs was capable of detecting genetic diversity between the tested Echinacea species, provided that the potential presence of low frequency variants was ignored and a possible bias in the estimates of genetic similarity was accepted. The assessments showed that a bulk of 15 individuals could detect sufficient AFLP variations at most genomic sites. Additionally, 20 primer pairs could generate sufficient polymorphic fragments to achieve high resolving power of AFLP for the tested Echinacea species.

AFLP fingerprinting analysis of Fusarium circinatum, Nirenberg and O’Donnell and relative species was carried out. Ten primer-pairs that could generate abundant polymorphism fragments were screened. A total of 298 nucleotide acid fragments were amplified with the primers from the template of the 17 strains of Fusarium spp., among which 283 fragments were polymorphic. Percentage of polymorphic loci produced by each pair of AFLP primer-pair was 94.97% in average and varied from 89.29% to 100%. All these data indicated that considerable genetic variation existed among F. circinatum and relative species at DNA level. Molecular genetic distances among Fusarium spp. were calculated, and the relationship among them was described quantitatively. Compared with biological species, the result of cluster analysis was basically similar to the phenotypic classification of species. Genetic diversity of E-AT/M-CAA AFLP fingerprinting of Fusarium spp. was analyzed, and specific and difference bands for each species and all Fusarium section Liseola tested were identified based on the E-AT/M-CAA AFLP fingerprinting.

We analyzed with an amplified fragment length polymorphism (AFLP) marker system the genetic diversity and relationships among 67 Monilinia laxa isolates obtained from different host plants. From a total of 1,089 amplified bands scored using 20 primer combinations with two selective nucleotides, 354 were polymorphic and further used in genetic diversity analysis. Genetic relationships among isolates were assessed with different phenetic approaches, including unweighted pair group method with arithmetic mean clustering and principal coordinate analysis; the population's differentiation estimate was analyzed by molecular variance; and model-based clustering was employed to infer population structure. All four analyses clearly showed significant differences between isolates from apple trees and isolates from other host plants. No further grouping according to any other host plant was observed. The results indicate host specialization of apple isolates and support the taxonomic grouping of apple isolates.

Amplified fragment length polymorphism (AFLP) is a powerful DNA fingerprinting technique for studying genetic relationships and genetic diversity in insects. However, the crucial prerequisite for AFLP analysis is to extract DNA of high quality. In this study, we evaluate four different protocols (SDS method, improved SDS method, CTAB method and a complex method with SDS and CTAB) for isolating DNA from the seabuckthorn carpenter moth (Holcocerrus hippophaecolus (Lepidoptera: Cossidae)). The results indicate that the CTAB method does not produce DNA suitable for AFLP analysis. The SDS method and the complex method with SDS and CTAB are comparatively time-consuming and resulted in low yields of DNA and were therefore not used for AFLP assay. The improved SDS method is recommended for preparing DNA templates from H. hippophaecolus for AFLP analysis.

In Korean waters, there are two color types (blue and purple) of the blue crab Portunus trituberculatus. The blue type is common, but the ratio of the purple type has increased in landings. To determine whether there were significant morphometric or genetic differences between the blue and purple types, crabs caught from the West Sea of Korea were examined. Based on covariance analysis, there were significant differences in 1 of 10 morphometric characteristics of males between the two types, in none of the ten characteristics for females. Using amplified fragment length polymorphism (AFLP) DNA fingerprinting, no specific AFLP marker was detected for each type. The heterozygosity and genetic diversity were very low. Analyses of pairwise distance, the Fst index, and genetic similarity revealed similar results, with very low genetic differentiation. Therefore, there is no significant difference between blue and purple types of the crab from the West Sea of Korea, and the two types in the West Sea can be managed as one stock.

Two hexaploid species of Solanum sect. Solanum are present in Africa: Solanum scabrum and S. nigrum. Solanum scabrum is a widely cultivated species and is used as a leafy vegetable, as a source of medicine and as a source of ink dye. In previous studies a wide range of morphological diversity has been reported in this species and in some studies subspecies have been proposed. Subspecies are also recognized in S. nigrum. However, it has not been established whether or not the morphological differences are reflected at the genomic level. The present study applies AFLPs to study the genetic diversity in S. scabrum and its relationship to geographical provenance, morphological differences and the possible existence of subspecies within S. scabrum and S. nigrum. The data obtained were analyzed with cluster analysis (using UPGMA and NJ). The results indicate that the genetic variation within S. scabrum was higher within accessions than between accessions. Accessions did not cluster according to their geographical provenance, indicating that accessions from different geographical areas were not significantly different genetically. The clustering reflected neither morphological differences nor domestication status (cultivated or wild). The morphological differences exhibited by S. scabrum could be due to selection by farmers for different plant types. The AFLP derived clustering pattern did not segregate the subspecies recognized in S. scabrum and S. nigrum into separate subclusters.

Genetic diversity among some important Syrian wheat cultivars was estimated using Amplified Fragment Length Polymorphism (AFLP) markers. Five Triticum aestivum L. and 10 Triticum turgidum ssp. durum were analyzed with 11 EcoRI–MseI primer pair combinations. Of the approximately 525 detected AFLP markers, only 46.67% were polymorphic. Cluster analysis with the entire AFLP data divided all cultivars into two major groups reflecting their origins. The first one contained T. aestivum L. cultivars, and the T. turgidum ssp. durum cultivars and landraces were grouped in the second. Narrow genetic diversity among all cultivars was detected with an average genetic similarity of 0.884. The lowest similarity index (0.9) was found between Cham5 and Hamary (durum wheat), whereas this value was 0.93 between Salamony and Bouhouth 4 (T. aestivum L.). The narrow genetic diversity level indicates that these genotypes could be originated from the same source. AFLP analysis provides crucial information for studying genetic variation among wheat cultivars and provides important information for plant improvement.

Fil: Espinoza, Francisco. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Nordeste. Instituto de Botanica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botanica del Nordeste; Argentina

Several varieties of Agave tequilana Weber have been described, but only A. tequilana Weber var. “azul” is accepted for the production of tequila. Discrimination between the different varieties is difficult and the relationships between them at the genotype level are unclear. In addition genetic diversity within and between populations of A. tequilana Weber var. “azul” has been shown to be low. AFLP analysis of nine A. tequilana varieties indicated that certain varieties were genotypically close to A. tequilana Weber var. “azul”. A significant level of diversity was also observed among A. tequilana Weber var. “azul” samples obtained from the same plantation and from different plantations in Guanajuato State, Mexico.

To evaluate the genetic diversity and to clarify the genetic relationships of Japanese peach cultivars, we analyzed the amplified fragment length polymorphism (AFLP) and traced the pedigree of 17 Japanese commercial peach cultivars and six traditional accessions. Sixteen AFLP primer combinations produced a total of 837 fragments and 146 polymorphic bands with a polymorphism percentage of 17.5%. All of the peach accessions could be identified from differences in at least 10 polymorphic bands. A cluster analysis showed that all the Japanese commercial peach cultivars, except ‘Kiyomi’ and ‘Jichigetsuto’, formed a major group consisting of three sub-groups. Of the six traditional accessions, four were genetically distant from the Japanese commercial peach cultivars while two accessions from China were classified into the Japanese commercial peach cultivars group. Both the AFLP analysis and pedigree tracing suggested that Japanese commercial peach cultivars are mainly derived from ‘Shanhai Suimitsuto’, one of the traditional accessions from China. Although the genetic relationships revealed by AFLP were generally in agreement with those shown by the pedigree information, some contradictions were found. Combining the AFLP results and pedigree information can provide a better understanding of the genetic relationships of Japanese peach cultivars.

SummaryHigh-throughput amplified fragment length polymorphism (AFLP) analysis, employing a two-dye automated DNA sequencer, was used to genotype accessions of four important Prunus spp. fruit crops in China, fruiting-mei (P. mume), apricot (P. armeniaca), plum (P. salicina, and P. ussuriensis) and peach (P. persica). Nine primer pairs generated a total of 2,089 AFLP bands, of which 94.4% were polymorphic. Levels of polymorphism and the genetic relationships among the crops were studied.The fixation index (Fst) of the four different fruit trees ranged from 0.53 – 0.82, and the average genetic diversity (AGD) in these species ranged from 3.2% to 12.8%. These four stone fruit species could be clustered into four clear groups on a phylogenetic tree based on bootstrap analysis. Phylogenetically, fruiting-mei was shown to be closest to apricot and most distant from peach.

Analyse AFLP de la diversité génétique de Meloidogyne chitwoodi et M. fallax, ravageurs importants des cultures. Les populations de M. chitwoodi et M. fallax se regroupent séparément des autres espèces étudiées. La diversité génétique observée chez M. incognita, M. arenaria, M. javanica, M. hapla et M. mayaguensis est corrélée avec les espèces établies. Deux groupes principaux sont identifiés à l'intérieur de l'ensemble M. chitwoodi / M. fallax, le premier comprenant uniquement des populations de M. chitwoodi, et le second des populations de M. chitwoodi et M. fallax. De plus, M. chitwoodi montre une diversité génétique supérieure à celle de M. fallax et se structure en plusieurs sous-ensembles.

Kenaf (Hibiscus cannabinus L.) is one of the world's most economically important fiber crops. In order to identify different varieties, and investigate its diversity and genetic relationships, twenty-three kenaf accessions and two accessions of its relative, roselle (H. sabdariffa var. altissima), were analyzed by morphological characterization and AFLP fingerprinting. It is very difficult to identify kenaf accessions based merely on morphological characters, due to their limited variation. For the AFLP study, a total of 505 polymorphic markers (out of 560) were produced by six selected AFLP primer combinations. The AFLP fingerprinting was effective in identifying all kenaf accessions included in the study. Kenaf and roselle are independent species with close relationships, and great genetic diversity was also detected among the kenaf accessions with different origins, based on the analysis of the AFLP markers. The AFLP analysis strongly supports the opinion that kenaf originated in Africa. It also demonstrated that the dissemination of kenaf was from Africa through Asia to Central and North America.

Two species, Trifolium glomeratum and T. nigrescens, from Sardinia, Italy, were analysed for genetic diversity using amplified fragment length polymorphism (AFLP). Variation between and within populations was compared between the inbreeder, T. glomeratum, and the outbreeder, T. nigrescens. Four AFLP primer combinations resulted in a total of 292 loci, of which 75% were polymorphic in T. glomeratum and 85% in Trifolium nigrescens. Variation was highest between populations in both species, but the difference between populations was greater in T. glomeratum (Fst = 0.17), compared with T. nigrescens (Fst = 0.02). Cluster analysis and principal coordinates analysis were used to verify the relationships found. The high level of genetic variation within populations in both species is attributed to the movement of sheep between paddocks, the existence of both species in Sardinia for thousands of years and the persistence of a long-lived seedbank due to the production of large numbers of small seeds with high levels of hard seededness.

We used amplified fragment length polymorphisms (AFLPs) to conduct a study of the genetic diversity of Flemish populations of Iris pseudacorus in two hydrologically isolated river basins. AFLP analysis using four primer combinations was carried out on 88 different ramet samples, collected at nine different locations. A total of 148 polymorphic markers were scored. Samples from the basin of the Ijzer river were different from those of the basin of the Scheldt river, require can be explained by the geographical and hydrological isolation of both the basins. A high degree of genetic diversity was detected in the populations, although the AFLP technique allowed us to identify three ramets belonging to one clone. The different genetic background in the basins of the rivers Ijzer and Scheldt enforce the use of local material in re-plantation projects for the conservation of natural genetic resources of yellow flag in both the basins.

The experiment was conducted to assess the genetic diversity of genetically improved farmed tilapia (GIFT) in order to evaluate and preserve this strain as a germplasm resource. The experiment study focused on the genetic diversity of the newly bred F16 population of GIFT tilapia obtained from Guanghui Farm Limited Company, Huazhou (Guangdong, China) was analyzed using AFLP. 187 bands were detected with 5 pairs of AFLP primers (E-AGG/M-CTT, E-AGG/M-CTG, E-AAC/M-CAG, E-ACA/M-CTG, E-ACA/M-CAG) based on 27 individuals, and among them 163 (87.17%) were polymorphic with an average of 32.6 bands for per AFLP primer. Average Nei’s gene diversity was 0.288,6, average Shannon’s information index was 0.433,9. These results showed that the experiment group deposit had rich genetic diversity and maintained a high genetic heterozygosity, had further breeding and development value.

Amplified fragment length polymorphism (AFLP) analysis was used to assess the biodiversity of one of the most important dipteran pests of cereals, the Asian rice gall midge (Orseolia oryzae Wood Mason). Larvae and pupae were collected at 15 locations in five Asian countries and preserved in 95% ethanol for storage, shipment, and DNA extraction using cetyltrimethylammonium bromide (CTAB). Although only ~1 µg of DNA was extracted from a single pupa or larva, the use of several AFLP primers in various combinations meant that this amount of DNA was sufficient to allow many DNA fingerprints to be made per individual. Fingerprints were sufficiently reproducible, especially during selective amplification, to allow the genetic diversity within a field population to be characterized. Extraction of DNA from a pool of 20 insects yielded AFLP fingerprints in which variation among individuals was sacrificed in favor of detecting differences among populations. For each location, pooled DNA was amplified with three primer pairs. A total of 261 distinct AFLP bands were identified for the 45 fingerprints. Cluster analysis, performed by the unweighted pair-group method (UPGMA), separated the populations into two distinct groups. Group I included two populations from Guangdong province of southern China and one each from Laos and Imphal in northeastern India, while group II was comprised of eleven populations from elsewhere in India (Assam, Orissa, Madhya Pradesh, Andhra Pradesh, and Kerala) and from Nepal and Sri Lanka. AFLP analysis provided insight into the origins of gall midge biotypes. In 1992, the prevailing biotype in Imphal changed from Indian biotype 3 to a new biotype 3M. Our data show that biotype 3M belongs to group I and did not arise by a recent mutation from biotype 3, which belongs to group II. By contrast, Indian biotypes 2 and 4 are likely to have diverged through recent mutation and selection, as are Chinese biotypes 1 and 4. The almost simultaneous emergence of new biotypes in Kerala and Sri Lanka during 1985-1988 was most probably coincidental, because these biotypes are not closely related. AFLP fingerprints were also able to detect sexual dimorphism in the DNA of adult gall midges and to distinguish gall midge from its major parasite Platygaster oryzae. Key words: biotypes, Cecidomyiidae, insect, Oryza sativa, Platygaster oryzae, population, sexual dimorphism.

To assess diversification of indica rice, fingerprinting of 42 Indian rice varieties (Oryza sativa L) and one accession of O. brachyantha Chev. et Roehr. was taken up using amplified fragment length polymorphism (AFLP). Six primer combinations of PstI and MseI were used for this purpose. On denaturing polyacrylamide sequencing gel, on average 41 scorable AFLP bands were obtained of which 81% were polymorphic. Each variety revealed a distinct fingerprint. Data were scored on the basis of presence and absence of bands and cluster analysis was performed using Dice similarity coefficients. The resultant dendrogram showed four major clusters and traits. Some varieties especially landraces revealed more polymorphism and unique loci. Such study will be a valuable tool for proper choice of parents in mapping populations or breeding programs to produce heterotic several inexplicable minor clusters. Some varieties were grouped according to parentages of the cross while some were grouped according to physiological combinations apart from providing a characteristic fingerprint.

Analysis of amplified fragment length polymorphism (AFLP) has the potential to become a powerful new DNA fingerprinting technique for studying genetic relationships and genetic diversity in arthropods. Since DNA of high quality is a crucial prerequisite for AFLP analysis we evaluated the applicability of six protocols (one fast and four complex methods with phenol-chloroform treatments as well as one CTAB-based method) for extracting DNA from insect material and three additional DNA purification steps. The most rapid DNA isolation method did not produce DNA suitable for AFLP analysis. Among four complex methods tested, two protocols resulted in comparatively low yields of DNA that was therefore not used as template for AFLP analysis. The other two complex methods with phenol treatments and a CTAB-based DNA extraction protocol provided DNA suitable for AFLP assay. An additional purification of the DNA using spermine precipitation revealed a few extra bands in an AFLP gel that were masked in unpurified DNA. Therefore spermine precipitation is recommended for AFLP templates.

An assessment of genetic diversity within and between Saccharum, Old World Erianthus sect. Ripidium, and North American E.giganteus (S.giganteum) was conducted using Amplified Fragment Length Polymorphism (AFLP(TM)) markers. An automated gel scoring system (GelCompar(TM)) was successfully used to analyse the complex AFLP patterns obtained in sugarcane and its relatives. Similarity coefficient calculations and clustering revealed a genetic structure for Saccharum and Erianthus sect. Ripidium that was identical to the one previously obtained using other molecular marker types, showing the appropriateness of AFLP markers and the associated automated analysis in assessing genetic diversity in sugarcane. A genetic structure that correlated with cytotype (2n=30, 60, 90) was revealed within the North American species, E. giganteus (S.giganteum). Complex relationships among Saccharum, Erianthus sect. Ripidium, and North American E.giganteus were revealed and are discussed in the light of a similar study which involved RAPD markers.

Chlorophytum borivilianum is a traditional medicinal plant distributed throughout the tropics and subtropics. In the present investigation, AFLP analysis was used to assess the genetic similarity among 34 accessions. Nine primer sets of AFLP amplified 612 fragments, of which 246 fragments were found to be polymorphic. The average number of polymorphic bands per AFLP primer pair was 27.33. The amplified fragments ranged from 50 base pairs to 600 base pairs. Significant correlation was observed between total number of amplified fragments and polymorphic bands (p > 0.05) per primers. Cluster analysis based on AFLP data revealed limited genetic variation within the thirty four accessions collected from various parts of Central Indian forests.

AFLP-system of multipoint marking was used for estimation of genetic diversity in 45 varieties of Capsicum annuum L. As a result, it was selected 8 primer combinations, which were the most informative for marking of paprika genome. Due to these primers the authors obtained the spe-cific DNA-spectrums for each variety. In total, 956 polymorphous AFLP-fragments were detected, of which 182 fragments characterize the individual varietal genotypes. The calculated coefficients of intervarietal genetic diversity are varying from 0.005 to 0.064. On dendrogram all varieties form general cluster with weak intrinsic differentiation that indicates the low degree of genetic polymorphism. So the forming of distance basal branches, revealed in closely related cultural species C. frutescens, C. chinense and C. baccatum, reflects the potential of their genetic diversity which may be used in paprika breeding. Keywords: Capsicum annuum, genetic diversity, genetic polymorphism, AFLP-markers.

OBJECTIVE To study the genetic background of Evodia rutaecarpa by AFLP, and analyze the genetic diversity of E. rutaecarpa from different areas. METHOD E. rutaecarpa genomic DNA was extracted. The AFLP reaction system was established and AFLP primer pairs were chosen for the analysis. Forty-six individuals of E. rutaecarpa which from five provinces were analyzed by AFLP. The NTSYS-pc 2.1 software was used for cluster analysis. RESULT Six out of the original 72 pairs of primers were optimized for the study; AFLP analysis revealed the similarity coefficient of 0.53, the samples of E. rutaecarpa var. officinalis from Zhejiang province was separated from other accessions; E. rutaecarpa var. officinalis also showed more pronounced genetic variation than the E. rutaecarpa, and strong geo-related relevance. CONCLUSION Variance of genetic background of E. rutaecarpa are large, AFLP analysis method can obviously identify different varieties of E. rutaecarpa, and can detect the genetic characteristics of inter-regional differences.

AFLP technique was used to elucidate systematic relationships among 26 cultivars of Tree Peony. Six AFLP primer combinations (M+3 and P+3) were used to produce recognizable 757 bands, among which 623 bands are polymorphic. The percentage of polymorphic bands was 82%, suggesting abundant genetic diversity among sect. Moutan. Clustering analysis showed that some cultivars of dwarf and tall peony types clustered separately, the cultivars of the same follower former only clustered partially. The clustering result based on DNA bands among the cultivars was not consistent with that based on the plant highness and flower former.