High-throughput DNA extraction from forest trees

Abstract

Trees are typically more demanding than other plants to extract pure, high quality DNA for molecular genetics and may not necessarily be amenable to advances in extraction methodology suitable for other plants. A recently available, commercial, high throughput DNA extraction system utilising a silica binding matrix for purification and a multi-sample mixer mill for tissue disruption was evaluated for its suitability with Eucalyptus spp., Pinus spp. and Araucaria cunninghamii (hoop pine). DNA suitable for a range of molecular biology applications was successfully extracted from all genera. The method was highly reliable when tested in over 500 preparations and could be adapted to different tree species with relatively minor modifications to the standard protocols.3

A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis

“…For characterization of germplasm of Terminalia arjuna, T. bellerica and T. chebula, isolation of purified DNA proved to be a major bottleneck, especially in case of T. arjuna, as has also been experienced with other species like Theobroma cacao (Haymes et al 2004), Vitis vinifera (Hanania et al 2004), Pinus radiata (Crowley et al 2003), Tagetes minuta (Hills and van Staden, 2002) Eucalyptus spp., Pinus spp. and Araucaria cunninghamii (Shepherd et al 2002), Davidia involuctata (Li et al 2002) Anthurium andreanum (Buldewo and Jaufeerally-Fakim, 2002), Drosera rotundifolia, Artemisia dracunculus (Pirttila et al 2001). …”

DNA isolation protocol for the medicinal plant lemon balm (Melissa officinalis, Lamiaceae)

“…(Michiels et al, 2003). Although various protocols for simple isolation of high-quality DNA from plant tissues are available (Saghai Maroof et al, 1984;Doyle and Doyle, 1990;Scott and Playford, 1996;Sharma et al, 2000;Pirttilä et al, 2001;Drábková et al, 2002;Shepherd et al, 2002;Mogg and Bond, 2003;Haymes et al, 2004), none of them is fully applicable for a large range of plants. In the present study, we focused on the analysis of PCR and PCR temperature program, and their requirements for efficient PCR quality DNA extraction.…”

“…To isolate pure and intact DNA from plant tissues, numerous protocols have been established (Saghai Maroof et al, 1984;Doyle and Doyle, 1990;Scott and Playford, 1996;Sharma et al, 2000;Pirttilä et al, 2001;Drábková et al, 2002;Shepherd et al, 2002;Mogg and Bond, 2003;Haymes et al, 2004). However, these DNA extraction protocols are not suitable for all medicinal plants, since each medicinal plant species contains specific secondary DNA isolation protocol for Melissa officinalis metabolites (Ribeiro and Lovato, 2007).…”

An efficient and rapid DNA minipreparation procedure suitable for PCR/SSR and RAPD analyses in tropical forest tree species

“…Several different methods and technologies are available for the isolation of plant genomic DNA, however, the better results can be observed with those based in use of CTAB (Doyle and Doyle, 1990;Kidwell and Osborn, 1992;Ferreira and Grattapaglia, 1995;Dilworth and Frey, 2000;Shepherd et al, 2002;Kang and Yang, 2004;Narayanan et al, 2006). In general, all methods consists of the following major steps: (1) grinding of samples, (2) phenol:chloroform:isoamyl alcohol extraction, and (3) DNA precipitation.…”

“…However, in tree leaf samples, cross contamination of DNA due to secondary metabolites such as terpenes, polyphenols, tannins and polysaccharides, which are often abundant in the foliage of perennials species, were related (Scott and Playford, 1996). As a consequence, many tree species require more complex extraction methods than annual plants (Shepherd et al, 2002). In this study, an efficient mini-scale DNA extraction method, modified from CTAB procedure (Doyle and Doyle, 1990) for rapid isolation of DNA genomic from five tropical forest tree species, Copaifera langsdorffii, Hymenaea courbaril, Eugenia uniflora, Tabebuia roseo alba and Cariniana estrellensis was developed.…”