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Gene silencing of cinnamyl alcohol dehydrogenase in Pinus radiata callus cultures. | LitMetric

AI Article Synopsis

  • Researchers transformed Pinus radiata cell cultures with an RNAi construct to silence the cinnamyl alcohol dehydrogenase (CAD) enzyme, which is crucial for lignin biosynthesis.
  • Most transformed clones showed decreased CAD activity, with those retaining about 20% CAD still unable to release high levels of vanillin after treatment.
  • Activating the phenylpropanoid pathway in these modified clones led to observable changes and accumulation of certain metabolites, indicating that significantly lowering CAD levels is essential for it to become a bottleneck in lignin production in conifers.

Article Abstract

Xylem-derived Pinus radiata cell cultures, which can be induced to differentiate tracheary elements (TEs), were transformed with an RNAi construct designed to silence cinnamyl alcohol dehydrogenase (CAD), an enzyme involved in the biosynthesis of monolignols. Quantitative enzymatic CAD measurements revealed reduced CAD activity levels in most transclones generated. TEs from transclones with approximately 20% residual CAD activity did not release elevated levels of vanillin, which was derived from coniferyl-aldehyde through a mild alkali treatment. However, the activation of the phenylpropanoid pathway in transclones with approximately 20% residual CAD activity through the application of non-physiological concentrations of sucrose and l-phenylalanine produced phenotypic changes. The accumulation of metabolites such as dihydroconiferyl-alcohol (DHCA), which also accumulates in the P. taeda CAD mutant cad-n1, was observed. These results indicate that a substantial reduction in CAD activity is necessary for this enzyme to become a rate-limiting step in lignin biosynthesis in conifers such as P. radiata and confirm that transformable P. radiata callus cultures can be useful to investigate the function of xylogenesis-related genes in conifers.

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Source
http://dx.doi.org/10.1016/j.plaphy.2005.11.001DOI Listing

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