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

   
   

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.

A invenção estabelece que ripening de fruta do café é climacteric. A invenção mais adicional fornece técnicas para isolar o RNA substancialmente puro da fruta do café mesmo que a fruta contenha níveis elevados dos compostos phenolic e do hidrato de carbono que interfeririam de outra maneira com obter preparações limpas do RNA deste tecido. A invenção fornece proteínas purified, as seqüências do ácido nucleic que codificam na expressão conseqüentemente e em moléculas recombinant do DNA, including os anfitriões transformados therewith, e em métodos transformando plantas do café para suprimir a expressão do synthase fruta-expressado café do ACUMULADOR e/ou do oxidase fruta-expressado café do ACUMULADOR necessários para o biosynthesis do etileno e ripening da fruta do café. As plantas do café são transformadas com os vetores que contêm o synthase fruta-expressado café do ACUMULADOR e/ou com seqüências do DNA do oxidase do ACUMULADOR que codificam na expressão para o RNA respectivo que é antisense ou sentido ao mRNA para o synthase do ACUMULADOR e/ou o oxidase respectivos do ACUMULADOR. O resultado é que a expressão do enzyme respectivo está eliminada e as plantas transformadas são incapable de synthesizing o etileno durante ripening de fruta do café, embora outros aspectos de seu metabolism não sejam afetados. A invenção mais adicional fornece métodos controlando ripening da fruta do café das plantas transformadas do café aplicando o etileno exogenous.

 
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