Organellar microcapture to extract nuclear and plastid DNA from recalcitrant wood specimens and trace evidence

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Autor(es): dc.contributorMississippi State University-
Autor(es): dc.contributorUniversity of Wisconsin-
Autor(es): dc.contributorForest Products Laboratory-
Autor(es): dc.contributorChinese Academy of Forestry-
Autor(es): dc.contributorPurdue University-
Autor(es): dc.contributorUniversidade Estadual Paulista (UNESP)-
Autor(es): dc.creatorCosta, Adriana-
Autor(es): dc.creatorGiraldo, Giovanny-
Autor(es): dc.creatorBishell, Amy-
Autor(es): dc.creatorHe, Tuo-
Autor(es): dc.creatorKirker, Grant-
Autor(es): dc.creatorWiedenhoeft, Alex C.-
Data de aceite: dc.date.accessioned2025-08-21T19:44:10Z-
Data de disponibilização: dc.date.available2025-08-21T19:44:10Z-
Data de envio: dc.date.issued2023-03-01-
Data de envio: dc.date.issued2023-03-01-
Data de envio: dc.date.issued2022-11-30-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.1186/s13007-022-00885-z-
Fonte completa do material: dc.identifierhttp://hdl.handle.net/11449/240858-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/11449/240858-
Descrição: dc.descriptionBackground: Illegal logging is a global crisis with significant environmental, economic, and social consequences. Efforts to combat it call for forensic methods to determine species identity, provenance, and individual identification of wood specimens throughout the forest products supply chain. DNA-based methodologies are the only tools with the potential to answer all three questions and the only ones that can be calibrated “non-destructively” by using leaves or other plant tissue and take advantage of publicly available DNA sequence databases. Despite the potential that DNA-based methods represent for wood forensics, low DNA yield from wood remains a limiting factor because, when compared to other plant tissues, wood has few living DNA-containing cells at functional maturity, it often has PCR-inhibiting extractives, and industrial processing of wood degrades DNA. To overcome these limitations, we developed a technique—organellar microcapture—to mechanically isolate intact nuclei and plastids from wood for subsequent DNA extraction, amplification, and sequencing. Results: Here we demonstrate organellar microcapture wherein we remove individual nuclei from parenchyma cells in wood (fresh and aged) and leaves of Carya ovata and Tilia americana, amyloplasts from Carya wood, and chloroplasts from kale (Brassica sp.) leaf midribs. ITS (773 bp), ITS1 (350 bp), ITS2 (450 bp), and rbcL (620 bp) were amplified via polymerase chain reaction, sequenced, and heuristic searches against the NCBI database were used to confirm that recovered DNA corresponded to each taxon. Conclusion: Organellar microcapture, while too labor-intensive for routine extraction of many specimens, successfully recovered intact nuclei from wood samples collected more than sixty-five years ago, plastids from fresh sapwood and leaves, and presents great potential for DNA extraction from recalcitrant plant samples such as tissues rich in secondary metabolites, old specimens (archaeological, herbarium, and xylarium specimens), or trace evidence previously considered too small for analysis.-
Descrição: dc.descriptionU.S. Department of State-
Descrição: dc.descriptionDepartment of Sustainable Bioproducts Mississippi State University-
Descrição: dc.descriptionDepartment of Botany University of Wisconsin-
Descrição: dc.descriptionForest Products Laboratory-
Descrição: dc.descriptionDepartment of Wood Anatomy and Utilization Chinese Research Institute of Wood Industry Chinese Academy of Forestry-
Descrição: dc.descriptionWood Collections (WOODPEDIA) Chinese Academy of Forestry-
Descrição: dc.descriptionDepartment of Forestry and Natural Resources Purdue University-
Descrição: dc.descriptionDepartamento de Ciências Biológicas (Botânica) Universidade Estadual Paulista–Botucatu-
Descrição: dc.descriptionDepartamento de Ciências Biológicas (Botânica) Universidade Estadual Paulista–Botucatu-
Descrição: dc.descriptionU.S. Department of State: 19318814Y0010-
Idioma: dc.languageen-
Relação: dc.relationPlant Methods-
???dc.source???: dc.sourceScopus-
Palavras-chave: dc.subjectNucleus isolation-
Palavras-chave: dc.subjectOrganellar microcapture-
Palavras-chave: dc.subjectPlastid isolation-
Palavras-chave: dc.subjectSingle-cell sequencing-
Palavras-chave: dc.subjectWood identification-
Título: dc.titleOrganellar microcapture to extract nuclear and plastid DNA from recalcitrant wood specimens and trace evidence-
Tipo de arquivo: dc.typelivro digital-
Aparece nas coleções:Repositório Institucional - Unesp

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