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Cloning, characterization and impact of up- and down-regulating subabul cinnamyl alcohol dehydrogenase (CAD) gene on plant growth and lignin profiles in transgenic tobacco
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2011 (English)In: Plant growth regulation (Print), ISSN 0167-6903, E-ISSN 1573-5087, Vol. 66, no 3, p. 239-253Article in journal (Refereed) Published
Abstract [en]

Both cDNA including 5′UTR and 3′UTR and genomic clones of cinnamyl alcohol dehydrogenase (CAD) were isolated and characterized from a pulp-yielding leguminous tree Leucaena leucocephala (LlCAD1). The deduced amino acid sequence shared high identity with orthologous sequences of Acacia mangium × Acacia auriculiformis (83%), Medicago sativa (83%), Nicotiana tabaccum (83%) and Aralia cordata (81%). Full length cDNA contained 78 bases of 5′UTR and 283 bases of 3′UTR, while the genomic clone contained 5 exons and 4 introns. Western blot analysis revealed elevated expression of LlCAD1 in seedling roots and shoots compared to leaves. Sense and antisense CAD tobacco transgenics showed increased and reduced CAD activity accompanied by a change in monomeric lignin composition. Histochemical staining of lignin in down-regulated plants suggested an increase in aldehyde units and a decrease in S/G ratio. Down-regulation of CAD resulted in accumulation of syringic, ferulic, p-coumaric and sinapic acids compared to untransformed controls. These observations were validated by anatomical studies of down-regulated transgenic stems which showed thin walled, elongated phloem and xylem fibres, accompanied by a reduction in the density of vessel elements and amount of secondary xylem when compared to untransformed plants. Furthermore, Klason lignin analysis of CAD antisense transgenics showed 7–32% reduced lignin and normal phenotype as compared to untransformed plants. Such a reduction was not noticed in up-regulated transgenics. These results demonstrate a unique opportunity to explore the significant role that down-regulation of CAD gene plays in reducing lignin content thereby offering potential benefits to the pulp and paper industry.

Place, publisher, year, edition, pages
Springer Nature , 2011. Vol. 66, no 3, p. 239-253
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Natural Sciences
Research subject
Biotechnology
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URN: urn:nbn:se:kth:diva-335453DOI: 10.1007/s10725-011-9647-1ISI: 000302573400003Scopus ID: 2-s2.0-84858450927OAI: oai:DiVA.org:kth-335453DiVA, id: diva2:1794894
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QC 20230908

Available from: 2023-09-06 Created: 2023-09-06 Last updated: 2023-09-08Bibliographically approved

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Sivan, Pramod

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