Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 46
Filter
1.
Article in Chinese | WPRIM | ID: wpr-921790

ABSTRACT

Cambodia is rich in medicinal plant resources. One hundred and thirty-three medicinal material samples, including the hole herb, root, stem/branch, leaf, flower, fruit, seed, and resin, were collected from the Orussey Herbal Market in Phnom Penh, Cambodia, and then authenticated by ITS and psbA-trnH. A total of 46 samples were identified based on ITS sequences, belonging to 24 families, 40 genera, and 42 species. A total of 100 samples were identified by psbA-trnH sequences to belong to 42 families, 77 genera, and 84 species. A total of 103 samples were identified by two DNA barcodes. According to the morphological characteristics of the medicinal materials, 120 samples classified into 50 species, 86 genera, and 86 families were identified, and the majority of them were from Zingiberaceae, Fabaceae, and Acanthaceae. Such samples have been commonly used in traditional Cambodian medicine, Ayurvedic medicine, Unani medicine, traditional Chinese medicine, and ethnomedicine, but different medical systems focus on different functional aspects of the same medicinal material. The results of this study have demonstrated that DNA barcoding has a significant advantage in identifying herbal products, and this study has provided basic data for understanding the traditional medicinal materials used in Cambodia.


Subject(s)
Cambodia , DNA Barcoding, Taxonomic , DNA, Plant/genetics , Humans , Plant Leaves , Plants, Medicinal/genetics
2.
Article in Chinese | WPRIM | ID: wpr-879175

ABSTRACT

Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". According to the national drug standard, it's derived from the resin extracted from the wood of Dracaena cochinchinensis, a Liliaceae plant. In addition, a variety of Dracaena species all over the world can form red resins, and there is currently no molecular identification method that can efficiently identify the origin of Dracaena medicinal materials. In this study, seven species of Dracaena distributed in China were selected as the research objects. Four commonly used DNA barcodes(ITS2, matK, rbcL and psbA-trnH), and four highly variable regions(trnP-psaJ, psbK-psbI, trnT-trnL, clpP) in chloroplast genome were used to evaluate the identification efficiency of Dracaena species. The results showed that clpP sequence fragment could accurately identify seven species of Dracaena plants. However, due to the long sequence of clpP fragment, there were potential problems in the practical application process. We found that the combined fragment "psbK-psbI+ trnP-psaJ" can also be used for accurate molecular identification of the Resina Draconis origin plants and relative species of Dracaena, which were both relatively short sequences in the combined fragment, showing high success rates of amplification and sequencing. Therefore, the "psbK-psbI+ trnP-psaJ" combined fragment can be used as the DNA barcode fragments for molecular identification of Resina Dracon's origin plants and relative species of Dracaena. Research on the identification of Dracaena species, the results of this study can be used to accurately identify the original material of Resina Draconis, and providing effective means for identification, rational development and application of Resina Draconis base source.


Subject(s)
China , DNA Barcoding, Taxonomic , DNA, Plant/genetics , Dracaena/genetics , Plants , Resins, Plant , Sequence Analysis, DNA
3.
Biol. Res ; 53: 21, 2020. tab, graf
Article in English | LILACS | ID: biblio-1124206

ABSTRACT

BACKGROUND: Liriodendron chinense ranges widely in subtropical China and northern Vietnam; however, it inhabits several small, isolated populations and is now an endangered species due to its limited seed production. The objective of this study was to develop a set of nuclear SSR (simple sequence repeats) and multiple chloroplast genome markers for genetic studies in L. chinense and their characterization in diverse germplasm. RESULTS: We performed low-coverage whole genome sequencing of the L. chinense from four genotypes, assembled the chloroplast genome and identified nuclear SSR loci by searching in contigs for SSR motifs. Comparative analysis of the four chloroplast genomes of L. chinense revealed 45 SNPs, 17 indels, 49 polymorphic SSR loci, and five small inversions. Most chloroplast intraspecific polymorphisms were located in the interspaces of single-copy regions. In total, 6147 SSR markers were isolated from low-coverage whole genome sequences. The most common SSR motifs were dinucleotide (70.09%), followed by trinucleotide motifs (23.10%). The motif AG/TC (33.51%) was the most abundant, followed by TC/AG (25.53%). A set of 13 SSR primer combinations were tested for amplification and their ability to detect polymorphisms in a set of 109 L. chinense individuals, representing distinct varieties or germplasm. The number of alleles per locus ranged from 8 to 28 with an average of 21 alleles. The expected heterozygosity (He) varied from 0.19 to 0.93 and the observed heterozygosity (Ho) ranged from 0.11 to 0.79. CONCLUSIONS: The genetic resources characterized and tested in this study provide a valuable tool to detect polymorphisms in L. chinense for future genetic studies and breeding programs.


Subject(s)
Polymorphism, Genetic/genetics , Genome, Plant/genetics , Liriodendron/genetics , Genome, Chloroplast/genetics , DNA Primers/genetics , DNA, Plant/genetics , Microsatellite Repeats , Alleles , Whole Genome Sequencing , Genotype
4.
Biol. Res ; 51: 42, 2018. tab
Article in English | LILACS | ID: biblio-1038782

ABSTRACT

BACKGROUND: Polymorphic microsatellite markers were developed for Gaultheria pumila (Ericaceae) to evaluate genetic diversity and population structure within its native range in Chile. This is a very important Ericaceae endemic to Chile with a large commercial potential. Its resistance to different abiotic conditions makes it a valuable target for genetic improvement. RESULTS: Ten polymorphic simple sequence repeat (SSR) loci were isolated from Gaultheria pumila using new-generation 454 FLX Titanium pyrosequencing technology. The mean number of alleles per locus ranged from 2 to 4. Observed and expected heterozygosity ranged from 0.00 to 1.0 and 0.00 to 0.64, respectively. CONCLUSIONS: From 10 SSR markers developed for G. pumila, 9 markers are promising candidates for analyzing genetic variation within or between natural populations of G. pumila and other species from the same genus.


Subject(s)
DNA, Plant/genetics , Microsatellite Repeats/genetics , Gaultheria/genetics , Polymorphism, Genetic , Genetic Variation , Alleles
5.
Electron. j. biotechnol ; 30: 6-11, nov. 2017. ilus, tab
Article in English | LILACS | ID: biblio-1021043

ABSTRACT

Background: Penthorum chinense Pursh (P. chinense) is a well-known traditional Chinese medicine (TCM) plant, which has long been used for the prevention and treatment of hepatic diseases. This study aimed to genetically characterize the varieties of P. chinense from different geographic localities of China by random amplification of polymorphic DNA (RAPD)-PCR technique and verified with inter-simple sequence repeat (ISSR) markers. Results: The P. chinense samples were collected from nine different geographic localities. Previously improved RAPD and ISSR markers were utilized for genetic analysis using DNA amplification. The genetic relationship dendrogram was obtained by conducting cluster analysis to the similarity coefficient of improved RAPD and ISSR markers. Improved RAPD yielded 185 scorable amplified products, of which 68.6% of the bands were polymorphic, with an average amplification of 9.25 bands per primer. The ISSR markers revealed 156 alleles with 7.8 bands per primers, where 59.7% bands were polymorphic. Furthermore, the similarity coefficient ranges of RAPD and ISSR markers were 0.71­0.91 and 0.66­0.89, respectively. Conclusions: This study indicated that improved RAPD and ISSR methods are useful tools for evaluating the genetic diversity and characterizing P. chinense. Our findings can provide the theoretical basis for cultivar identification, standardization, and molecular-assisted breeding of P. chinense for medicinal use.


Subject(s)
Plants, Medicinal/genetics , Magnoliopsida/genetics , Polymorphism, Genetic , Genetic Variation , Genetic Markers , China , DNA, Plant/genetics , Random Amplified Polymorphic DNA Technique , Microsatellite Repeats , Medicine, Chinese Traditional
6.
Electron. j. biotechnol ; 27: 1-7, May. 2017. tab, ilus, graf
Article in English | LILACS | ID: biblio-1009846

ABSTRACT

Background: Genetic diversity studies are important for the selection of parents with a greater combination capacity that, when crossed, increase the chances of obtaining superior genotypes. Thus, 26 polymorphic simple sequence repeat (SSR) primers were used to assess the genetic diversity of 140 individual samples from 12 diploid sugar beet pollinators (pollen parents) and two cytoplasmic male sterile (cms) lines (seed parents). Eight pollinators originated from three research centers in the United States Department of Agriculture, while four pollinators and cms lines were from the Institute of Field and Vegetable Crops, Novi Sad, Serbia. Results: In total, 129 alleles were obtained, with a mean of 3.2 alleles per SSR marker. The observed heterozygosity ranged from 0.00 to 0.87 (mean = 0.30). Expected heterozygosity and Shannon's information index were the lowest for marker BQ590934 and the highest for markers SB15s and FDSB502s; the same markers were the most informative, with PIC values of 0.70 and 0.69, respectively. Three private alleles were found in pollinator EL0204; two in pollinator C51; and one in pollinators NS1, FC221, and C93035. Molecular variance showed that 77.34% of the total genetic variation was attributed to intrapopulation variability. Cluster and correspondence analysis grouped sugar beet pollinators according to the breeding centers, with few exceptions, which indicate that certain amount of germplasm was shared, although centers had their own breeding programs. Conclusions: The results indicate that this approach can improve the selection of pollinators as suitable parental components and could further be applied in sugar beet breeding programs.


Subject(s)
Pollen/genetics , Genetic Variation , Beta vulgaris/genetics , Polymorphism, Genetic , Seeds/genetics , Selection, Genetic , Breeding , Polymerase Chain Reaction , DNA, Plant/genetics , Microsatellite Repeats , Pollination , Genotype
7.
Braz. j. microbiol ; 45(3): 977-983, July-Sept. 2014. ilus, tab
Article in English | LILACS | ID: lil-727029

ABSTRACT

Extraneous DNA interferes with PCR studies of endophytic fungi. A procedure was developed with which to evaluate the removal of extraneous DNA. Wheat (Triticum aestivum) leaves were sprayed with Saccharomyces cerevisiae and then subjected to physical and chemical surface treatments. The fungal ITS1 products were amplified from whole tissue DNA extractions. ANOVA was performed on the DNA bands representing S. cerevisiae on the agarose gel. Band profile comparisons using permutational multivariate ANOVA (PERMANOVA) and non-metric multidimensional scaling (NMDS) were performed on DGGE gel data, and band numbers were compared between treatments. Leaf surfaces were viewed under variable pressure scanning electron microscopy (VPSEM). Yeast band analysis of the agarose gel showed that there was no significant difference in the mean band DNA quantity after physical and chemical treatments, but they both differed significantly (p < 0.05) from the untreated control. PERMANOVA revealed a significant difference between all treatments (p < 0.05). The mean similarity matrix showed that the physical treatment results were more reproducible than those from the chemical treatment results. The NMDS showed that the physical treatment was the most consistent. VPSEM indicated that the physical treatment was the most effective treatment to remove surface microbes and debris. The use of molecular and microscopy methods for the post-treatment detection of yeast inoculated onto wheat leaf surfaces demonstrated the effectiveness of the surface treatment employed, and this can assist researchers in optimizing their surface sterilization techniques in DNA-based fungal endophyte studies.


Subject(s)
Endophytes/isolation & purification , Microbiological Techniques/methods , Sterilization/methods , Triticum/microbiology , Denaturing Gradient Gel Electrophoresis , DNA, Fungal/genetics , DNA, Fungal/isolation & purification , DNA, Plant/chemistry , DNA, Plant/genetics , DNA, Ribosomal Spacer/genetics , DNA, Ribosomal Spacer/isolation & purification , Microscopy, Electron, Scanning , Polymerase Chain Reaction , Plant Leaves/microbiology , Plant Leaves/ultrastructure , Surface Properties , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/isolation & purification , Saccharomyces cerevisiae/ultrastructure , Triticum/ultrastructure
8.
Rev. biol. trop ; 61(1): 311-320, Mar. 2013. ilus
Article in English | LILACS | ID: lil-674083

ABSTRACT

The Thar Desert, a very inhospitable place, accommodates only plant species that survive acute drought, unpredictable precipitation, and those can grow in the limited moisture of sandy soils. Capparis decidua is among one of the few plants able to grow well under these conditions. This species is highly exploited and has been naturally taken, as local people use it for various purposes like food, timber and fuel, although, no management or conservation efforts have been established. The present study was conducted in this arid area of Western Rajasthan (India) with the aim to obtain preliminary molecular information about this group of plants. We evaluated diversity among 46 samples of C. decidua using chemical parameters and random amplified polymorphic DNA (RAPD) markers. Fourteen chemical parameters and eight minerals (total 22 variables) of this species fruits were estimated. A total of 14 RAPD primers produced 235 band positions, of which 81.27% were polymorphic. Jaccard s similarity coefficients for RAPD primers ranged from 0.34 to 0.86 with a mean genetic similarity of 0.50. As per observed coefficient of variation, NDF (Neutral Detergent Fiber) content was found to be the most variable trait followed by starch and soluble carbohydrate. The Manhattan dissimilarity coefficient values for chemical parameters ranged between 0.02-0.31 with an average of 0.092. The present study revealed a very low correlation (0.01) between chemical parameters and RAPD-based matrices. The low correlation between chemical- and RAPD-based matrices indicated that the two methods were different and highly variable. The chemical-based diversity will assist in selection of nutritionally rich samples for medicinal purpose, while genetic diversity to face natural challenges and find sustainable ways to promote conservation for future use.


El desierto de Thar, un lugar muy inhóspito, alberga sólo a las especies de plantas capaces de resistir a condiciones de sequía extrema, a las precipitaciones impredecibles, y a las plantas que pueden crecer en la humedad limitada de los suelos arenosos. Capparis decidua se encuentra entre una las pocas plantas capaces de crecer bien en estas condiciones. Esta especie es altamente explotada y se ha tomado de forma natural, así los habitantes locales las han usado para varios propósitos, como alimento, madera y combustible, aunque sin ningún programa de manejo o esfuerzo por conservación. El presente estudio se llevó a cabo en esta zona árida del oeste de Rajastán (India) con el objetivo de obtener información molecular preliminar sobre este grupo de plantas. Se evaluó la diversidad entre 46 muestras de C. decidua usando parámetros químicos y marcadores de ADN polimórfico amplificado al azar (RAPD por sus siglas en inglés). Catorce parámetros químicos y ocho minerales (22 variables en total) de los frutos de esta especie fueron estimados. Un total de 14 cebadores para RAPD produjeron 235 posiciones de bandas, de las cuales 81.27% fueron polimórficas. El coeficiente de similitud de Jaccard para los cebadores del RAPD varió entre 0.34 y 0.86 con un promedio de similitud genética de 0.50. De acuerdo con el coeficiente de variación observado, se encontró que el contenido de NDF fue el rasgo más variable, seguido por el almidón y los carbohidratos solubles. Los valores del coeficiente de disimilitud de Manhattan para los parámetros químicos osciló entre 0.02-0.31 con un promedio de 0.092. El presente estudio reveló una correlación muy baja (0.01) entre los parámetros químicos y las matrices basadas en RAPD. La baja correlación entre las matrices químicas y la basada en RAPD indicó que los dos métodos fueron diferentes y altamente variables. El estudio de la diversidad basada en su química ayudará en la selección de muestras nutricionalmente ricas para fines medicinales, mientras que la diversidad genética ayudará a enfrentar los desafíos naturales y encontrar formas sostenibles para promover la conservación de esta plana para uso futuro.


Subject(s)
Biodiversity , Capparis/chemistry , Capparis/genetics , DNA, Plant/genetics , Capparis/classification , Desert Climate , India , Random Amplified Polymorphic DNA Technique
9.
Braz. j. microbiol ; 44(2): 657-665, 2013. ilus, tab
Article in English | LILACS | ID: lil-688573

ABSTRACT

Stone pine (Pinus pinea L.), like other conifers, forms ectomycorrhizas (ECM), which have beneficial impact on plant growth in natural environments and forest ecosystems. An in vitro co-culture of stone pine microshoots with pure mycelia of isolated ECM sporocarps was used to overcome the root growth cessation not only in vitro but also to improve root development during acclimation phase. Pisolithus arhizus (Scop.) Rauschert and Lactarius deliciosus (L. ex Fr.) S.F. Gray fungi, were collected, pure cultured and used in in vitro co-culture with stone pine microshoots. Samples of P. arhizus and L. deliciosus for the in vitro co-cultures were collected from the pine stands southwest Portugal. The in situ characterization was based on their morphotypes. To confirm the identity of the collected material, ITS amplification was applied using the pure cultures derived from the sporocarps. Additionally, a molecular profile using PCR based genomic fingerprinting comparison was executed with other genera of Basidiomycetes and Ascomycetes. Our results showed the effectiveness of the techniques used to amplify DNA polymorphic sequences, which enhances the ­characte­rization of the genetic profile of ECM fungi and also provides an option to verify the fungus identity at any stage of plant mycorrhization.


Subject(s)
Mycorrhizae/classification , Mycorrhizae/isolation & purification , Pinus/microbiology , Cluster Analysis , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Plant/chemistry , DNA, Plant/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Molecular Sequence Data , Mycorrhizae/genetics , Mycorrhizae/growth & development , Phylogeny , Portugal , Sequence Analysis, DNA
10.
Article in English | IMSEAR | ID: sea-144080

ABSTRACT

WRKY transcription factor proteins play important roles in diverse stress responses. In this study, we first cloned a novel WRKY from our constructed bacteriophage full-length cDNA library for cotton (Gossypium barbadense). The plants were stressed by exposure to a defoliating strain of Verticillium dahliae. The capacity of primary cDNA library was 1.28 × 106 PFU and the titer of the amplified cDNA library was >1010 PFU mL–1. The recombination rate of the library was 94% and average insert size was about 1.1 kb. This novel gene, named GbWRKY1 was 1971 bp long and encodes a protein of 489 amino acids. It contains two characteristic WRKY domains and two zinc finger motifs. The sub-cellular assay indicated that GbWRKY1–GFP fusion protein was localized in the nucleus. Furthermore, Northern blot analysis showed that expression pattern of GbWRKY1 was similar among tissue types (roots, stems and leaves), but differed between pathogen-infiltrated and Czapek medium-infiltrated (untreated control) plants. Quantitative real-time PCR showed that GbWRKY1 could also be induced by salicylic acid (SA), methyl jasmonate (MeJA) and 1-aminocyclopropane-1-carboxylic acid (ACC). These findings clearly suggest that as a pathogen-inducible transcription factor GbWRKY1 plays an important role in plant defense responses.


Subject(s)
DNA/chemistry , Genes/analysis , Gossypium/genetics , Plant Proteins/genetics , Plant Proteins/isolation & purification , Verticillium/isolation & purification , Genes, Plant , DNA, Plant/genetics
11.
Rev. biol. trop ; 60(4): 1463-1478, Dec. 2012. graf, mapas, tab
Article in English | LILACS | ID: lil-662221

ABSTRACT

The study of the genetic structure of wild plant populations is essential for their management and conservation. Several DNA markers have been used in such studies, as well as isozyme markers. In order to provide a better comprehension of the results obtained and a comparison between markers which will help choose tools for future studies in natural populations of Oryza glumaepatula, a predominantly autogamous species, this study used both isozymes and microsatellites to assess the genetic diversity and genetic structure of 13 populations, pointing to similarities and divergences of each marker, and evaluating the relative importance of the results for studies of population genetics and conservation. A bulk sample for each population was obtained, by sampling two to three seeds of each plant, up to a set of 50 seeds. Amplified products of eight SSR loci were electrophoresed on non-denaturing polyacrylamide gels, and the fragments were visualized using silver staining procedure. Isozyme analyses were conducted in polyacrylamide gels, under a discontinuous system, using six enzymatic loci. SSR loci showed higher mean levels of genetic diversity (A=2.83, p=0.71, A P=3.17, Ho=0.081, He=0.351) than isozyme loci (A=1.20, p=0.20, A P=1.38, Ho=0.006, He=0.056). Interpopulation genetic differentiation detected by SSR loci (R ST=0.631, equivalent to F ST=0.533) was lower than that obtained with isozymes (F ST=0.772). However, both markers showed high deviation from Hardy-Weinberg expectations (F IS=0.744 and 0.899, respectively for SSR and isozymes). The mean apparent outcrossing rate for SSR ( =0.14) was higher than that obtained using isozymes ( =0.043), although both markers detected lower levels of outcrossing in Amazonia compared to the Pantanal. The migrant number estimation was also higher for SSR (Nm=0.219) than isozymes (Nm=0.074), although a small number for both markers was expected due to the mode of reproduction of this species, defined ...


El estudio de la estructura genética de poblaciones de plantas silvestres es esencial para su manejo y conservación. Varios marcadores de ADN e isoenzimas se han utilizado en este tipo de análisis. Con el fin de proporcionar una mejor comprensión de los resultados obtenidos y saber que marcador codominante elegir para futuros estudios en poblaciones naturales de Oryza glumaepatula, este trabajo busco evaluar y comparar dos marcadores de ADN, isoenzimas y microsatélites, en la diversidad y estructura genética de 13 poblaciones, destacando las similitudes y divergencias de cada marcador, así como la importancia relativa de los resultados en genética de poblaciones y conservación. Para los SSR, ocho loci SSR fueron evaluados, y los fragmentos se visualizaron utilizando el procedimiento de coloración con plata. Los análisis de isoenzimas se realizaron en geles de poliacrilamida, en los seis loci enzimáticos. Los loci SSR mostraron mayores niveles de diversidad genética que los loci isoenzimáticos, en promedio. La diferenciación genética entre los loci SSR (R ST=0.631, equivalente a F ST=0.533) fue inferior a la obtenida con las isoenzimas (F ST=0.772). Ambos marcadores mostraron alta desviación del equilibrio de Hardy-Weinberg (F IS=0.744 y 0.899, respectivamente, para SSR e isoenzimas). La tasa media aparente de cruzamiento para SSR ( =0.14) fue mayor que la obtenida con isoenzimas ( =0.043), aunque ambos marcadores detectaron niveles más bajos en la tasa de fecundación cruzada para la Amazonia, en comparación con la región del Pantanal. La estimación de número de migrantes también fue mayor para los SSR (Nm=0.219) que en isoenzimas (Nm=0.074). No se obtuvo ninguna correlación entre las distancias genéticas y geográficas para los SSR, y para las isoenzimas se obtuvo una correlación positiva entre las distancias genéticas y geográficas. Llegamos a la conclusión de que estos marcadores son divergentes en la detección de los parámetros de la diversidad genética en O. glumaepatula y que los microsatélites son más eficientes para detectar la información a nivel intra-poblacional, mientras que las isoenzimas son más potentes para detectar la diversidad entre poblaciones.


Subject(s)
Genetic Variation/genetics , Isoenzymes/analysis , Microsatellite Repeats/genetics , Oryza/enzymology , Oryza/genetics , Brazil , DNA, Plant/genetics , Genetic Markers , Polymorphism, Genetic
12.
Electron. j. biotechnol ; 15(4): 3-3, July 2012. ilus, tab
Article in English | LILACS | ID: lil-646953

ABSTRACT

Background: Cassava (Manihot esculenta Crantz) is a crop that is high in carbohydrates in the roots and in protein in the leaves, important for both human consumption and animal feed, and also has a significant industrial use for its starches. In this study we evaluated the genetic variability with molecular markers in different stages in micropropagated plants from somatic embryos of Venezuelan native clone 56. Results: Three markers were used: ISTR, AFLP and SSR, finding that ISTR showed the highest polymorphism among individuals tested. With AFLP a high similarity between the evaluated individuals was observed and with SSR total monomorphism was seen. Using cluster analysis it was found that individuals from an embryo labeled as fasciated at the beginning of the somatic embryogenesis process were grouped as independent of the other plants when analyzed at the acclimatization stage. The differences found with the different markers used are discussed. In field trials, micropropagated plants had a yield between 4 and 5 times the average yield of cassava in Venezuela. Conclusion: Despite variability in terms of DNA markers, somatic embryogenesis is suitable for mass propagation of highly performing cassava clones.


Subject(s)
Manihot/embryology , Manihot/genetics , DNA, Plant/genetics , Amplified Fragment Length Polymorphism Analysis , Biomarkers , Embryonic Development , Microsatellite Repeats
13.
An. acad. bras. ciênc ; 83(3): 993-1006, Sept. 2011. ilus, graf, tab
Article in English | LILACS | ID: lil-595526

ABSTRACT

Cytogenetic analyses, of pollen viability, nuclear DNA content and RAPD markers were employed to study three chemotypes of Lippia alba (Mill.) (Verbenaceae) in order to understand the genetic variation among them. Different ploidy levels and mixoploid individuals were observed. This work comprises the first report of different chromosome numbers (cytotypes) in L. alba. The chromosome numbers of La2-carvone and La3-linalool chemotypes suggested that they are polyploids. Flow cytometric analysis showed an increase of nuclear DNA content that was not directly proportional to ploidy level variation. A cluster analysis based on RAPD markers revealed that La3-linalool shares genetic markers with La1-citral and La2-carvone. The analysis showed that the majority of genetic variation of La3-linalool could be a consequence of ixoploidy. ur data indicates that sexual reproduction aong those three chemotypes is unlikely and suggests the beginning of reproductive isolation. The results demonstrated that chromosome analysis, nuclear DNA content estimation and RAPD markers constitute excellent tools for detecting genetic variation among L. alba chemotypes.


Análises citogenéticas, de viabilidade do pólen, do conteúdo de DNA nuclear e marcadores RAPD foram empregadas no estudo de três quimiotipos de Lippia alba (Mill.) (Verbenaceae) visando contribuir para o entendimento da variação genética entre os mesmos. Diferentes níveis de ploidia e indivíduos mixoploides foram observados. Este trabalho compreende o primeiro relato de diferentes números cromossômicos (citótipos) em L. alba. Os números cromossômicos dos quimiotipos La2-carvona e La3-linalol sugere que eles seja poliploides. A análise da citometria de fluxo mostrou um aumento do conteúdo de DNA nuclear que não foi diretamente proporcional à variação no nível de ploidia. A análise de agrupamento baseada nos marcadores RAPD demonstrou que La3-linalol compartilha marcadores genéticos com La1-citral e La2-carvona. A análise mostrou que a maior parte da variação genética de La3-linalol pode ser consequência da mixoploidia. Nossos dados indicam que a reprodução sexual entre os três quimiotipos parece improvável, sugerindo o início de isolamento reprodutivo. Os resultados demonstraram que a análise cromossômica, a quantificação do DNA nuclear estimado e os marcadores RAPD constituem excelentes ferramentas para detecção de variação genética entre quimiotipos de L. alba.


Subject(s)
DNA, Plant/genetics , Genetic Markers/genetics , Genetic Variation/genetics , Karyotype , Lippia/genetics , Lippia/classification , Random Amplified Polymorphic DNA Technique
14.
Rev. biol. trop ; 59(2): 607-617, jun. 2011. ilus, tab
Article in English | LILACS | ID: lil-638107

ABSTRACT

In Coffea arabica (arabica coffee), the phenotypic as well as genetic variability has been found low because of the narrow genetic basis and self fertile nature of the species. Because of high similarity in phenotypic appearance among the majority of arabica collections, selection of parental lines for inter-varietals hybridization and identification of resultant hybrids at an early stage of plant growth is difficult. DNA markers are known to be reliable in identifying closely related cultivars and hybrids. Sequence Related Amplified Polymorphism (SRAP) is a new molecular marker technology developed based on PCR. In this paper, sixty arabica-hybrid progenies belonging to six crosses were analyzed using 31 highly polymorphic SRAP markers. The analysis revealed seven types of SRAP marker profiles which are useful in discriminating the parents and hybrids. The number of bands amplified per primer pair ranges from 6.13 to 8.58 with average number of seven bands. Among six hybrid combinations, percentage of bands shared between hybrids and their parents ranged from 66.29% to 85.71% with polymorphic bands varied from 27.64% to 60.0%. Percentage of hybrid specific fragments obtained in various hybrid combinations ranged from 0.71% to 10.86% and ascribed to the consequence of meiotic recombination. Based on the similarity index calculation, it was observed that F1 hybrids share maximum number of bands with the female parent compared to male parent. The results obtained in the present study revealed the effectiveness of SRAP technique in cultivar identification and hybrid analysis in this coffee species. Rev. Biol. Trop. 59 (2): 607-617. Epub 2011 June 01.


En Coffea arabica (café arabica), el fenotipo y la variabilidad genética son bajos debido a la estrecha base genética y la autofecundación de la especie. Por su alta similitud fenotípica entre la mayoría de las colecciones de arábica, la selección de líneas parentales para hibridación entre variedades, y la identificación de los híbridos resultantes en una fase inicial de crecimiento, es difícil. Para la identificación de variedades estrechamente relacionadas y sus híbridos, los marcadores de ADN son confiables, pero los polimorfismos de amplificación de secuencias relacionadas (SRAP, por sus siglas en inglés) constituyen una nueva tecnología de marcadores moleculares basada en PCR. En este trabajo, sesenta progenies arábica híbridas, pertenecientes a seis cruces, fueron analizadas utilizando 31 marcadores altamente polimórficos. El análisis reveló siete tipos de perfiles de marcadores que son útiles en la discriminación de los progenitores y los híbridos. El número de bandas amplificadas por pares de cebadores estuvo entre 6.13 a 8.58 con un promedio de siete bandas. Entre las seis combinaciones de híbridos, el porcentaje de bandas compartidas entre híbridos y sus progenitores estuvo entre 66.29% y 85.71% con bandas polimórficas que variaron entre 27.64% y 60.0%. El porcentaje de fragmentos híbridos específicos obtenidos en diversas combinaciones híbridas varió entre 0.71% y 10.86% lo que se atribuye a la recombinación meiótica. Con base en el cálculo del índice de similitud, se observó que los híbridos F1 compartieron un número máximo de bandas con el progenitor femenino que con el masculino. Los resultados obtenidos en este estudio muestran la eficacia de la técnica de SRAP en la identificación de cultivos e híbridos de esta especie de café.


Subject(s)
Coffea/genetics , DNA, Plant/genetics , Hybridization, Genetic/genetics , Polymorphism, Genetic/genetics , Genetic Markers/genetics , Phenotype , Polymerase Chain Reaction , Random Amplified Polymorphic DNA Technique
15.
Electron. j. biotechnol ; 14(3): 3-3, May 2011. ilus, tab
Article in English | LILACS | ID: lil-602980

ABSTRACT

Increasing scarcity of irrigation water is a major threat to sustainable production of cotton (Gossypium hirsutum L.). Identifying genomic regions contributing to abiotic stress tolerance will help develop cotton cultivars suitable for water-limited regions through molecular marker-assisted breeding. A molecular mapping F2 population was derived from an intraspecific cross of the drought sensitive G. hirsutum cv. FH-901 and drought tolerant G. hirsutum cv. RH-510. Field data were recorded on physiological traits (osmotic potential and osmotic adjustment); yield and its component traits (seedcotton yield, number of bolls/plant and boll weight); and plant architecture traits (plant height and number of nodes per plant) for F2, F2:3 and F2:4 generations under well-watered versus water-limited growth conditions. The two parents were surveyed for polymorphism using 6500 SSR primer pairs. Joinmap3.0 software was used to construct linkage map with 64 polymorphic markers and it resulted into 35 markers mapped on 12 linkage groups. QTL analysis was performed by composite interval mapping (CIM) using QTL Cartographer2.5 software. In total, 7 QTLs (osmotic potential 2, osmotic adjustment 1, seedcotton yield 1, number of bolls/plant 1, boll weight 1 and plant height 1) were identified. There were three QTLs (qtlOP-2, qtlOA-1, and qtlPH-1) detected only in water-limited conditions. Two QTLs (qtlSC-1 and qtlBW-1) were detected for relative values. Two QTLs (qtlOP-1 and qtlBN-1) were detected for well-watered treatment. Significant QTLs detected in this study can be employed in MAS for molecular breeding programs aiming at developing drought tolerant cotton cultivars.


Subject(s)
Droughts , Gossypium/physiology , Gossypium/genetics , Quantitative Trait Loci , Adaptation, Physiological , DNA, Plant/genetics , Genetic Variation , Gossypium/growth & development , Chromosome Mapping/methods , Osmosis , Polymorphism, Genetic , Software
16.
Electron. j. biotechnol ; 13(6): 4-5, Nov. 2010. ilus, tab
Article in English | LILACS | ID: lil-591908

ABSTRACT

A preliminary characterization was undertaken to describe genetic structure of mango ginger (Curcuma amada) acquired from farmers and ex situ genebank in Myanmar using neutral (rice SSR based RAPDs) and functional genomic (P450 based analog) markers. The high polymorphism (> 91 percent) depicted has displayed existence of genetic variability in the germplasm investigated. Large number of source-specific alleles (neutral-markers = 78, functional-markers = 63) was amplified which revealed that neutral regions of the mango ginger were more variable compared with the functional regions. The major fraction of the molecular variance (neutral-markers = 85 percent, functional-markers = 93 percent) was explained within germplasm acquisition sources and this tendency was also supported by the estimate of gene diversity. The genebank accessions have shown comparatively more genetic variability than farmers' accessions. The variability observed in mango ginger may possibly be associated with the long history of its cultivation under diverse ecological conditions. The two marker systems elucidated their high resolving power which detected variability even in fewer genotypes assayed. As the target sites of these markers are different, therefore, the variability detected is believed to cover diverse part of the genome together with neutral and functional regions. We found the concurrent use of the different types of molecular markers valuable to comprehend a dependable variability pattern in the germplasm assayed.


Subject(s)
Curcuma/genetics , Ginger/genetics , DNA, Plant/genetics , Genetic Markers , Genetic Variation , Myanmar , Polymorphism, Genetic , Random Amplified Polymorphic DNA Technique
17.
Electron. j. biotechnol ; 13(6): 5-6, Nov. 2010. ilus, tab
Article in English | LILACS | ID: lil-591909

ABSTRACT

Cultivated peanut (Arachis hypogaea L.) is an oilseed crop of economic importance. It is native to South America, and it is grown extensively in the semi-arid tropics of Asia, Africa, and Latin America. Given an extremely narrow genetic base, efforts are being made to develop simple sequence repeat (SSR) markers to provide useful genetic and genomic tools for the peanut research community. A SSR-enriched library to isolate trinucleotide (GGC)n SSRs in peanut was constructed. A total of 143 unique sequences containing (GGC)n repeats were identified. One hundred thirty eight primer pairs were successfully designed at the flanking regions of SSRs. A suitable polymerase was chosen to amplify these GC-rich sequences. Although a low level of polymorphism was observed in cultivated peanut by these new developed SSRs, a high level of transferability to wild species would be beneficial to increasing the number of SSRs in wild species.


Subject(s)
Arachis/genetics , Microsatellite Repeats , DNA, Plant/genetics , Agricultural Cultivation , Genetic Markers , Polymorphism, Genetic , Trinucleotide Repeats
18.
Electron. j. biotechnol ; 13(6): 7-8, Nov. 2010. ilus, tab
Article in English | LILACS | ID: lil-591911

ABSTRACT

In order to saturate a sunflower genetic map and facilitate marker-assisted selection (MAS) breeding for stress response, it is necessary to enhance map saturation with molecular markers localized in linkage groups associated to genomic regions involved in these traits. This work describes the identification and characterization of 1,134 simple sequence repeat (SSR) containing expressed sequence tags (EST) from unigenes available databases. Twelve of these functional markers as well as 41 public SSR markers were successfully localized in linkage groups, thus contributing to the saturation of specific regions on a reference genetic-linkage-map derived from recombinant inbred lines (RIL) mapping population from the cross between PAC2 x RHA266 lines. The enriched map includes 547 markers (231 SSR, 9 EST-SSR, 3 insertions/deletions (InDel) and 304 amplified fragment length polymorphisms (AFLP) distributed in 17 linkage groups (LG), spanning genetic size to 1,942.3 cM and improving its mean density to 3.6 cM per locus. As consequence, no gaps longer than 13.2 cM remain uncovered throughout the entire map, which increases the feasibility of detecting genes or traits of agronomic importance in sunflower.


Subject(s)
Chromosome Mapping , Helianthus/genetics , DNA, Plant/genetics , Agronomy , Amplified Fragment Length Polymorphism Analysis , Breeding , Genetic Linkage , Genetic Markers , INDEL Mutation , Microsatellite Repeats , Polymerase Chain Reaction , Polymorphism, Genetic
19.
An. acad. bras. ciênc ; 82(3): 779-789, Sept. 2010. ilus, mapas, tab
Article in English | LILACS | ID: lil-556812

ABSTRACT

Caryocar brasiliense (pequi), is one of the main species at the biome of the Brazilian savannah due to its use in culinary, popular medicine, industry in general, and iron and steel industry. At São José do Xingu (MT), a tree of C. brasiliense without thorn at the endocarp was found, which enables the improvement of C. brasiliense not only for consumption but also to the high appreciation it already has. To detect the existing differences between the pequi with and without the thorn at the endocarp, RADP markers were used. The generated polymorphisms were cloned and sequenced in order to identify the sequences that are responsible for the fenotypical alteration. It was observed that the pequi without thorn is genetically isolated from the other populations of pequi with thorn at the endocarp, proving that this characteristic is related to the genetic divergence of the species. Analysis in BLASTn evidenced the similarity of the Dof1 genes of Zea mays to its gene of phosphinotricin acetyl transferase. In the analysis of BLASTx, the similarity was verified to the proteins responsible for the deficiency in ferric reductase 4, and catalase.


Pequi, Caryocar brasiliense, é uma das espécies de destaqueno bioma do cerrado brasileiro, devido a sua utilização na medicina, na culinária popular, indústria em geral, e na do ferro e do aço. Na região de São José do Xingu (MT), uma árvore de pequi sem espinho no endocarpo foi encontrado e isso permite melhorar pequi não só para o consumo, aproveitando a alta apreciação que já possui. Para detectar as diferenças existentes entre o genoma de pequi com e sem espinho no endocarpo, marcadores moleculares RAPD foram utilizados. Os polimorfismos gerados foram clonados e sequenciados, a fim de identificar as sequências responsáveis pela alteração fenotípica. Observou-se que o pequi sem espinho é geneticamente isolado de outras populações de pequi com espinho no endocarpo, provando que essa característica está relacionada com a divergência genética da espécie. Análise em Blastn evidenciou a similaridade dos genes Dof1 e com o gene da fosfinotricina-acetiltransferase de Z. mays. Na análise da BLASTx, a similaridade foi verificada com as proteínas responsáveis pela deficiência de ferro 4 redutase e catalase.


Subject(s)
DNA, Plant/genetics , Genome, Plant/genetics , Polymorphism, Genetic/genetics , Ericales/genetics , Random Amplified Polymorphic DNA Technique , Ericales/anatomy & histology , Ericales/classification
20.
Electron. j. biotechnol ; 13(5): 17-18, Sept. 2010. ilus, tab
Article in English | LILACS | ID: lil-591899

ABSTRACT

Genetically modified (GMO) rapeseed (Brassica napus) is not grown commercially in European Union, but several lines have been approved for production and use as food and feed. A case-specific monitoring of herbicide-tolerant rapeseed, events RT73, RF3 and T45 was established by Ministry of Agriculture of Republic of Serbia. The objectives of the present study were to introduce methods for detection of herbicide-tolerant GM oilseed rape, investigate occurrence and monitor the presence of GM rapeseed in seed and the feed products, as well as to develop a protocol for quantification. The study was based on 48 samples, rapeseed (33) and feed (15) products, imported from EU countries (Germany, Belgium, France, Czech Republic, Austria) and from domestic market. Seven positive feed samples and no positive seed samples have found. The percent of GMO in feed samples, estimated on semi-quantitative way, was below labelling threshold. Adventitious presence of GM materials in non-GM grain, derived food and feedstuffs is a concern to international grain trade and needs continuous monitoring.


Subject(s)
DNA, Plant/analysis , Brassica rapa/genetics , Monitoring , Organisms, Genetically Modified , DNA, Plant/genetics , Polymerase Chain Reaction , Yugoslavia
SELECTION OF CITATIONS
SEARCH DETAIL