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

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.

Fast and novel botanical exploration of a 320-km transect in eastern Amazonia using DNA barcoding

“…Compared to wood, it was found that the cambium had a higher concentration of DNA than the heartwood or sapwood, although it also had larger amounts of PCR reaction inhibitors (Tang et al 2011). The cambium samples that we collected were rapidly dried with silica gel, corresponding to the best long-term approach for DNA preservation (Mangaravite et al 2020), and we indeed found that DNA Table 2. List of tree families identified in seven 25x25-m plots along the Brazil-French Guiana border.…”

Análise quali-quantitativa do DNA genômico extraído de folha e estipe de x Butyagrus nabonnandii (Prosch.) Vorster

“…A extração desse tipo de amostra foi realizada aproximadamente 15 dias após a coleta, quando o material já estava bem desidratado. Amostras de tecidos do estipe: os métodos para a coleta e armazenagem de tecido interno do estipe se basearam nos trabalhos de Mello et al (2015) e Mangaravite et al (2020), respectivamente. Amostras foram coletadas sempre que possível a 1,30 m do solo, por meio de um trado de incremento (Sonda Pressler) (Figura 2c).…”

“…Tradicionalmente, as folhas são usadas como fonte principal de DNA genômico (Colpaert et al, 2005;Novaes et al, 2009;Aydin et al, 2020). Materiais alternativos, entretanto, como a casca e tecidos do caule, vêm sendo utilizados por serem mais fáceis de serem coletados, especialmente em espécies arbóreas onde a altura das folhas dificulta sua amostragem (Mangaravite et al, 2020).…”