Abstract
一、The conservation and application of the genomics and production of secondary products in forestry (2/4)
（一）We have obtained hairy root lines of Taxus sumatrana, Nothopodytes nimmoniana, Camptotheca acuminata , and Cephalotaxus wilsoniana which transformed with rol gene. These root lines grew rapidly and produced stably secondary metabolites greater and more easily to preserve than that from cell cultures. The limitation is that they are difficult to scale up in batch cultures. This study is trying to improve culture techniques including to enhance transformation rates of hairy roots, to increase biomass coupling with high concentration of secondary metabolites, to design suitable bioreactors to grow root cultures, and to establish a cooperative platform among research, application, and biotech companies.（二）For developing the forestry genomic research, the nucleotide sequencing plays the important role. In this project, the specific tree species' genome and transcriptome will be sequencing by using next generation sequencing. The sequencing data will provide useful information for developing molecular marker and improve the studies of the population genetics, molecular breeding and species identification. In addition, this project will manage and preserve the valuable forestry genomic germplasm resources which we had (or the species with highly application potential) in Germplasm Bank. According to characters of superior tree varieties, we will develop preservation methods in vitro to prevent loss of specific genes. We also will provide the strategy of value added regulations for conservation, storage and application of these genomic germplasm.二、Cloning of xylem cellulose synthase gene and breeding of genetically modified trees(4/5)
（一） Genetics, breeding and biosafety assessment of genetically modified Eucalyptus trees:
This 4-year research project is proposed to develop gene-stacking technology for the breeding of new Eucalyptus varieties in which multiple genetic characteristics are improved to meet the demand in industry. Preservation technology is also proposed to develop for the establishment of mutant bank containing varied transgenic lines of Eucalyptus that will bring the molecular breeding into the new research area of functional genomics. In this fourth years project, gene stacking by sequential transformation will be developed to raise transgenic E. camaldulensis plants with the antisense Pt4CL1 and the sense CAld5H transgenes. The transgenic E. camaldulensis plants previously transformed with the sense CAld5H transgenes will be sequentially transferred with recombinant SiRNA of the CAD gene to produced transgenic callus or shoots with double transgenes. In co-transformation, transgenic Eucalyptus plants with the antisense Euc4CL1 and/or the sense CALd5H transgene will be produced. The transgenic E. camaldulensis plants previously transformed with the sense CAld5H transgenes will be selected and transferred with recombinant SiRNA of the CAD gene to produce transgenic callus or shoots with multiple transgenes. In addition to these, another 2 cellulose synthase (CesA) genes will be cloned from E. camaldulensis readily for the further co-transformation work in the following year. In controlled pollination of gene stacking, transgenic seedlings with C4H and GUS transgenes derived from pollination between transgenic E. grandis x E. urophylla and E. camaldulensis will be raised. Transgenic Eucalyptus previously transformed with either the antisense Euc4CL1or the sense CAld5H transgene will be induced to flower for making more controlled pollination. For rapidly measuring monomeric lignol of transgenic wood, an analytical technique by using pyrolyzer coupled with GC-MS will be developed. The low-temperature preservation technique developed in the last year's project will be used to preserve varied transgenic shoots produced from this or previous projects.
（二） Lignin biosynthesis-related genes isolated strains of Eucalyptus field trials: according the regulation of "Investigation on genetic characteristics and biosafety evaluation of transgentic plants", the trangenic eucalypytus will be conducted in isolated field.三、Research of Breeding and Silviculture and Utility for Short Rotation Tree Species:
The amount of logs for culturing mushroom in Taiwan is estimated about 350 thousands tones annually. Most logs felled from stands in Taiwan are gradually depleted. Mushroom industry urged Taiwan government to face the problems. The project was drawn studies on silvicultural techniques of fast growing tree species with nitrogen fixation, key techniques related to mushroom industrial chain gap, soil management to sustain tree growth, and using sawdust of Leucaena leucocephala to culture mushroom. Based on the mushroom chain gap, there are 4 topics in this project: 1. Study on tree breeding and sivilcultural techniques of Acacia confusa, Alnus formomsana , and Trema orientalis to increase biomass production in forestation areas. 2. Study on sawdust of L. leucocephala as a part of ones to mix A. confusa sawdust to grow mushroom possible. 3. Study on reforestation in invasive L. leucocehala areas when they are removed and replaced with A. confusa trees for expanding the supplement of mushroom sawdust. 4. Study on the changes of lignocellulose contents of sawdust during the period of mushroom cultures at initial, medium, and end stages. The potential of spent sawdust to be transformed into biofuel will be evaluated. From the understanding the production chain from seedling nursing, forestation, logging, sawdust production and mediation, mushroom culture and harvest, and finally into spent sawdust composition, the well consideration of sustainable management can be established.