RESUMO
The overall aims of the research are to develop genetically engineered alfalfa producing high levels of industrially important enzymes and to develop rapid methods for extracting and purifying these enzymes from alfalfa juice. Using a reporter gene beta-glucuronidase (GUS) as a model system, we were able to demonstrate production of a foreign protein in alfalfa and gain valuable insight into the molecular approaches required for the expression and accumulation of foreign proteins in leaf tissue. GUS activity varied among individual transformants, and GUS was expressed in all plant tissues. GUS activity was shown to segregate in sexual progeny. There was no correlation between copy number of the GUS gene and activity. We have recently demonstrated the production of Mn-dependent lignin peroxidase and alpha-amylase in transgenic alfalfa. Concurrent research in the agricultural engineering aspects of this feasibility study focused on extraction strategies for the recovery of alfalfa juice, and on an evaluation of methods for processing and concentrating the juice. Thus, we are in a position to use plants expressing enzymes that have current or potential industrial importance to complete a feasibility study, and determine whether we can indeed economically recover target enzymes from field-grown transgenic alfalfa plants. The technology developed for these enzymes can be used to extract other value-added products from plants in the future.
