Development and Validation of Sterility Systems for Trees

Strauss, Steve ; Shevchenko, Olga ; Ma, Caiping (Cathleen)

关键词: ABLATION;    ARABIDOPSIS;    CONTAINMENT;    GENES;    MONITORING;    MUTANTS;    POPLARS;    PROMOTERS;    PROTEINS;    RNA;    SEASONS;    STERILITY;    TRANSFORMATIONS;    TREES;    VALIDATION;    WOOD containment;    biotechnolo;   

DOI  :  10.2172/901554
RP-ID  :  DOE/ID/13552
PID  :  OSTI ID: 901554
Others  :  Other: FC07-97ID13552
Others  :  TRN: US200714%%253

美国|英语

来源: SciTech Connect

【 摘 要 】

The overall goal of this project was to develop and validate sterility systems in poplar with the ultimate goal of fulfilling the basic requirements for commercial use. For this, sterility must be complete and stable over multiple growing seasons, cause no detrimental effects on vegetative growth, and successful transformation events must be identifiable via molecular tests when trees are still juvenile. Because of the inherent difficulties in achieving and demonstrating complete sterility in trees, our approach was to study alternate sterility systems in Arabidopsis and/or early-flowering tree systems. The public benefit from this work is the capacity for containment of genes or exotic forms of trees so they can be of benefit for industry for production of wood, energy, and renewable products, while having minimal impact on wild populations of trees. We tested three methods for engineering sterility: dominant negative mutant (DNM) proteins, floral tissue ablation, and RNA interference (RNAi) to suppress the expression of several floral regulatory genes. The ultimate goal of this work was to produce a number of transgenic poplars that could be outplanted to enable future assessments of the effectiveness of these transgenic sterility methods. Our attempts to produce ablation constructs that did not interfere with tree health were partially successful. Using the poplar LEAFY gene promoter and the barnase/barstar system, we were able to regenerate plants that grew well in the greenhouse, but they showed poor health in the field. Four of seven DNM genes tested were considered promising enough, based on results in Arabidopsis, to produce transgenic poplars. Single, double, and triple RNAi genes were produced and transformed into poplar. Over all, we produced 1,964 PCR-confirmed transgenic events with 19 different kinds of sterility genes and several kinds of control genes. We propagated 5,640, 6,820, and 7,055 trees for each of three test poplar genotypes, and field plantings were begun in Spring of 2003 and will be finished in Spring 2007. Continued field studies and monitoring will be required to establish if any of the approaches we have taken will prove to be safe for tree health, stable, and provide reliable containment.