ANATOMICAL, PHYSICAL AND MECHANICAL PROPERTIES OF TRANSGENIC LOBLOLLY PINE (PINUS TAEDA L.) MODIFIED FOR INCREASED DENSITY

Traditional breeding methods are often constrained by the reproductive cycles of tree species and the difficulty in achieving significant improvements to complex traits; therefore, genetic manipulation of complex traits such as wood properties has the potential to resolve those issues. The objectives of this study were to analyze MOE, MOR, and the physical and anatomical properties of 2- to 3-yr-old field-grown transgenic Pinus taeda trees modified for increased density. This investigation consisted of a total of 55 sample trees in two separate experiments. Transgenic trees from sets OX41 and OX55, modified for increased density using two variants of the same HAP5 gene, exhibited higher mechanical properties with smaller stem diameter and tracheid lumen diameter than their set of control trees. In addition, set OX55 exhibited increased cell wall thickness. In the second experiment, the transgenic group WVK249, modified for higher density using an unrelated MYB gene, exhibited similar diameter growth and increased cell wall thickness and lower lumen/cell wall ratios but no change in mechanical properties compared with its control

Ptr-miR397a is a negative regulator of laccase genes affecting lignin content in Populus trichocarpa

Laccases, as early as 1959, were proposed to catalyze the oxidative polymerization of monolignols. Genetic evidence in support of this hypothesis has been elusive due to functional redundancy of laccase genes. An Arabidopsis double mutant demonstrated the involvement of laccases in lignin biosynthesis. We previously identified a subset of laccase genes to be targets of a microRNA (miRNA) ptr-miR397a in Populus trichocarpa. To elucidate the roles of ptr-miR397a and its targets, we characterized the laccase gene family and identified 49 laccase gene models, of which 29 were predicted to be targets of ptr-miR397a. We overexpressed Ptr-MIR397a in transgenic P. trichocarpa. In each of all nine transgenic lines tested, 17 PtrLACs were down-regulated as analyzed by RNA-seq. Transgenic lines with severe reduction in the expression of these laccase genes resulted in an ∼40% decrease in the total laccase activity. Overexpression of Ptr-MIR397a in these transgenic lines also reduced lignin content, whereas levels of all monolignol biosynthetic gene transcripts remained unchanged. A hierarchical genetic regulatory network (GRN) built by a bottom-up graphic Gaussian model algorithm provides additional support for a role of ptr-miR397a as a negative regulator of laccases for lignin biosynthesis. Full transcriptome–based differential gene expression in the overexpressed transgenics and protein domain analyses implicate previously unidentified transcription factors and their targets in an extended hierarchical GRN including ptr-miR397a and laccases that coregulate lignin biosynthesis in wood formation. Ptr-miR397a, laccases, and other regulatory components of this network may provide additional strategies for genetic manipulation of lignin content