Down-Regulation of Lignin Biosynthesis in Transgenic Leucaena leucocephala Harboring O-Methyltransferase Gene

摘要

In the present study,a 0.47 kb OMT gene construct from aspen,encoding for an enzyme O-methyltransferase (OMT,EC 2.1.1.6),in antisense orientation was used to down-regulate lignin biosynthesis in Leucaena leucocephala.The plants were transformed with Agrobacterium tumefaciens strain harboring the antisense gene,and the transformation was confirmed by PCR amplification of the npt II gene.The integration of a heterologous antisense OMT gene construct in transformed plants led to a maximum of 60% reduction in OMT activity relative to control.The evaluation of total lignin content by the Klason method revealed a maximum of 28% reduction.Histochemical analyses of stem sections depicted a reduction in lignin content and normal xylem development.The results also suggested a probable increase in aldehyde levels and a decrease in syringyl units.Lignin down-regulation was accompanied by an increase in methanol soluble phenolics to an extent that had no impact on wood discoloration,and the plants displayed a normal phenotype.Concomitantly,an increase of up to 9% in cellulose content was also observed.Upon alkali extraction,modified lignin was more extractable as evident from reduced Klason lignin in saponified residue and increased alkali soluble phenolics.The results together suggested that the extent of down-regulation of OMT activity achieved may lead to quality amelioration of Leucaena with respect to its applicability in pulp and paper manufacture as well as nutritive and easily digestible forage production.