Concentration and Distribution of Nuclei and Plastids in Xylem Cells in Cunninghamia lanceolata and Aquilaria sinensis

Zhang, Rong; Xu, Kun; Ke-lin, YE

doi:10.15376/biores.10.1.1304-1317

Cited by 2 publications

(2 citation statements)

References 19 publications

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“…The limiting factor for the overall development and adoption of DNA methods for wood identification is the recalcitrance of wood as a DNA source. In the living tree, most cells comprising wood are the product of programmed cell death, and thus lack nuclei and other organelles at functional maturity [ 9 – 11 ]. Additionally, in many species, the presence of extractives, secondary metabolites associated with heartwood formation, can interfere with the extraction and amplification of DNA.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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Background 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.

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Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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“…In these steps, DNA is quickly degraded and the molecules can be attacked by bacteria, fungi and insects resulting in contaminations (MAGEL, 2001). Zhang et al (2015) observed and quantified nuclei and plastids adhered to the walls of parenchyma cells in heartwood and sapwood. They also verified these structures continue to be degraded after the tree is cut and while the logs are stored or dried.…”

Section: Introductionmentioning

confidence: 99%

The use of ITS region as a tool for wood identification in Brazil: A case study of a tree from the genus Hovenia sp.

Schardosin¹,

Cademartori²,

Morrone³

et al. 2019

RCM

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The knowledge of botanical identity of wood allows its adequate application as raw material in the industry and has importance related to deterring the illegal trade in protected species. This paper reports the species identification of seized logs, based on anatomical characteristics and ITS (Internal Transcribed Spacer) region. Three wood samples came from an apprehension in deforestation in Paraná State, Brazil, by the Federal Police, resulting from actions to combat illegal logging. First anatomical identification was done based on the macroscopic and microscopic characteristics of material. After, DNA extraction was done with the DNeasy Mericon Food Kit® with some adaptation. Wood presented growth rings distinct, semiring porous, vessels solitary predominant, in radial multiples of 2-3, occasionally clustered, without deposits in vessels, axial parenchyma unilateral and vasicentric, tending to lozenge-aliform, confluent and in marginal bands, heterogeneous rays, uniseriate and multiseriate, not storied, fibers thin to thick-walled, non-septate and was identified as from genus Hovenia. ITS1 region resulted in the same genus and allowed the identification of the seized sample, which is an exotic and invader species that can be explored.

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Concentration and Distribution of Nuclei and Plastids in Xylem Cells in Cunninghamia lanceolata and Aquilaria sinensis

Cited by 2 publications

References 19 publications

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

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

The use of ITS region as a tool for wood identification in Brazil: A case study of a tree from the genus Hovenia sp.

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