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.
