Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues

-This study was carried out to assess the genetic variability of ten "cagaita" tree (Eugenia dysenterica) populations in Southeastern Goiás. Fifty-four randomly amplified polymorphic DNA (RAPD) loci were used to characterize the population genetic variability, using the analysis of molecular variance (AMOVA). A φ ST value of 0.2703 was obtained, showing that 27.03% and 72.97% of the genetic variability is present among and within populations, respectively. The Pearson correlation coefficient (r) among the genetic distances matrix (1 -Jaccard similarity index) and the geographic distances were estimated, and a strong positive correlation was detected. Results suggest that these populations are differentiating through a stochastic process, with restricted and geographic distribution dependent gene flow.Index terms: Myrtaceae, Cerrado, tropical species, genetic diversity. Estrutura genética e fluxo gênico em populações naturais de cagaitaResumo -Este trabalho teve por objetivo o estudo da variabilidade genética em 10 populações de cagaiteiras (Eugenia dysenterica), da região Sudeste do Estado de Goiás. Foram identificados 54 locos marcadores RAPD, para a caracterização da variabilidade genética, avaliada por meio da análise da variância molecular (AMOVA). Foi verificado que 27,03% da variabilidade genética está entre populações, e 72,97% dentro de populações, índices obtidos a partir do valor de φ ST igual a 0,2703. Foi estimado o coeficiente de correlação de Pearson (r) entre a matriz de distâncias genéticas (1 -índice de similaridade de Jaccard) e de distâncias geográficas, tendo sido encontrada forte correlação positiva. Os resultados sugerem que essas populações estão se diferenciando por um processo estocástico havendo fluxo restrito dependente da distribuição geográfica.Termos para indexação: Myrtaceae, Cerrado, espécies tropicais, diversidade genética, Eugenia dysenterica.

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“…The DNA was extracted by the CTAB (cetyltrime thylammonium bromide) method described by Murray & Thompson (1980) and Rogers & Bendich (1985).…”

Section: Methodsmentioning

confidence: 99%

Genetic structure and gene flow of Eugenia dysenterica natural populations

Zucchi

Pinheiro

Chaves

et al. 2005

Pesq. agropec. bras.

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“…Genomic DNA from transformed and non-transformed soilgrown plants were extracted using the CTAB extraction method according to Rogers [18]. Approximately 10 mg of DNA from each sample were digested with HindIII, BamHI, EcoRI, respectively, non-transformed plant was digested with EcoRI, then separated by electrophoresis 0.8% (w/v) agarose gel, transferred from the agarose gel to Hybond + nylon membrane and cross-linked to the membrane by UV light for 3 min.…”

Section: Southern Blot Hybridizationmentioning

confidence: 99%

Regeneration of plants from callus cultures of roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi

Wang

Luo

et al. 2001

Cell Res

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The legume forage Alhagi pseudoalhagi was transformed by the Agrobacterium rhizogenes strain A4 using cotyledon and hypocotyl segments as infection materials. Plant regeneration was achieved from sterile calluses derived from hairy roots, which occurred at or near the infection sites. The hairy root regenerants were characterized by normal leaf morphology and stem growth but a shallow and more extensive root system than normal plants. Opine synthesis, PCR and Southern blot confirmed that T-DNA had been intergrated into the A. pseudoalhagi genome. Acetosyringone (AS) was found to be vital for successful transformation of A. pseudoalhagi.

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“…DNA was isolated from callus and root tissues according to the method of Rogers and Bendich [20]. Ten micrograms of DNA digested with EcoRI/HindIII was separated by agarose gel electrophoresis.…”

Section: Dna Analysismentioning

confidence: 99%