Purified proteins, recombinant DNA sequences and processes for controlling the ripening of coffee plants

摘要

The invention establishes that coffee fruit ripening is climacteric. The invention further provides techniques to isolate substantially pure RNA from coffee fruit even though the fruit contains high levels of phenolic compounds and carbohydrate which would otherwise interfere with obtaining clean RNA preparations from this tissue. The invention provides purified proteins, nucleic acid sequences that code on expression therefore and recombinant DNA molecules, including hosts transformed therewith, and methods for transforming coffee plants to suppress the expression of coffee fruit-expressed ACC synthase and/or coffee fruit-expressed ACC oxidase necessary for ethylene biosynthesis and the ripening of coffee fruit. Coffee plants are transformed with vectors containing coffee fruit-expressed ACC synthase and/or with ACC oxidase DNA sequences that code on expression for the respective RNA that is antisense or sense to the mRNA for the respective ACC synthase and/or ACC oxidase. The result is that the expression of the respective enzyme is eliminated and the transformed plants are incapable of synthesizing ethylene during coffee fruit ripening, although other aspects of their metabolism is not affected. The invention further provides methods for controlling the ripening of coffee fruit from the transformed coffee plants by applying exogenous ethylene.