A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment

Um método viável para extrair DNA do câmbio de árvores de dossel alto: da coleta à aplicação

Érica MANGARAVITE Vanessa TERRA Eric Koiti Okiyama HATTORI Thaís Carolina da Silva DAL’SASSO Leonardo Lopes BHERING Luiz Orlando de OLIVEIRA About the authors

ABSTRACT

Many tropical trees have high canopies and their leaves are not accessible. Thus, the use of tissue from a more accessible organ (cambium) for DNA extraction may be an alternative for molecular studies. We adapted a feasible methodology for extracting genomic DNA from cambium tissue harvested in the field for the assessment with PCR. We tested three storage conditions (two buffers and a silica gel) and four periods of time after harvest. We used previously described protocols and tested them on three species that occur in Amazonian forests and other biomes: Anadenanthera peregrina var. peregrina, Cedrela fissilis, and Ceiba speciosa. Our protocol obtained suitable PCR-grade genomic DNA for DNA sequencing and microsatellite genotyping. We recommend the use of silica for long-term storage and the buffer with ascorbic acid for short-term storage.

KEYWORDS:
ascorbic acid; dithiothreitol; DNA isolation

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