48 research outputs found
Two highly divergent alcohol dehydrogenases of melon exhibit fruit ripening-specific expression and distinct biochemical characteristics
Alcohol dehydrogenases (ADH) participate in
the biosynthetic pathway of aroma volatiles in fruit by
interconverting aldehydes to alcohols and providing substrates
for the formation of esters. Two highly divergent
ADH genes (15% identity at the amino acid level) of
Cantaloupe Charentais melon (Cucumis melo var. Cantalupensis)
have been isolated. Cm-ADH1 belongs to the
medium-chain zinc-binding type of ADHs and is highly
similar to all ADH genes expressed in fruit isolated so far.
Cm-ADH2 belongs to the short-chain type of ADHs. The
two encoded proteins are enzymatically active upon
expression in yeast. Cm-ADH1 has strong preference for
NAPDH as a co-factor, whereas Cm-ADH2 preferentially
uses NADH. Both Cm-ADH proteins are much more active
as reductases with Kms 10–20 times lower for the conversion
of aldehydes to alcohols than for the dehydrogenation
of alcohols to aldehydes. They both show strong preference
for aliphatic aldehydes but Cm-ADH1 is capable of
reducing branched aldehydes such as 3-methylbutyraldehyde,
whereas Cm-ADH2 cannot. Both Cm-ADH genes are
expressed specifically in fruit and up-regulated during
ripening. Gene expression as well as total ADH activity are
strongly inhibited in antisense ACC oxidase melons and in
melon fruit treated with the ethylene antagonist 1-methylcyclopropene
(1-MCP), indicating a positive regulation by
ethylene. These data suggest that each of the Cm-ADH
protein plays a specific role in the regulation of aroma
biosynthesis in melon fruit
Functional characterization of a melon alcohol acyl-transferase gene family involved in the biosynthesis of ester volatiles. Identification of the crucial role of a threonine residue for enzyme activity
Volatile esters, a major class of compounds contributing to the aroma of many fruit, are synthesized by
alcohol acyl-transferases (AAT). We demonstrate here that, in Charentais melon (Cucumis melo var.
cantalupensis), AAT are encoded by a gene family of at least four members with amino acid identity ranging
from 84% (Cm-AAT1/Cm-AAT2) and 58% (Cm-AAT1/Cm-AAT3) to only 22% (Cm-AAT1/Cm-AAT4).
All encoded proteins, except Cm-AAT2, were enzymatically active upon expression in yeast and show
differential substrate preferences. Cm-AAT1 protein produces a wide range of short and long-chain acyl
esters but has strong preference for the formation of E-2-hexenyl acetate and hexyl hexanoate. Cm-AAT3
also accepts a wide range of substrates but with very strong preference for producing benzyl acetate.
Cm-AAT4 is almost exclusively devoted to the formation of acetates, with strong preference for cinnamoyl
acetate. Site directed mutagenesis demonstrated that the failure of Cm-AAT2 to produce volatile esters is
related to the presence of a 268-alanine residue instead of threonine as in all active AAT proteins. Mutating
268-A into 268-T of Cm-AAT2 restored enzyme activity, while mutating 268-T into 268-A abolished
activity of Cm-AAT1. Activities of all three proteins measured with the prefered substrates sharply increase
during fruit ripening. The expression of all Cm-AAT genes is up-regulated during ripening and inhibited in
antisense ACC oxidase melons and in fruit treated with the ethylene antagonist 1-methylcyclopropene
(1-MCP), indicating a positive regulation by ethylene. The data presented in this work suggest that the
multiplicity of AAT genes accounts for the great diversity of esters formed in melon
Integrated health messaging for multiple neglected zoonoses : approaches, challenges and opportunities in Morocco
Integrating the control of multiple neglected zoonoses at the community-level holds great potential, but critical data is missing to inform the design and implementation of different interventions. In this paper we present an evaluation of an integrated health messaging intervention, using powerpoint presentations, for five bacterial (brucellosis and bovine tuberculosis) and dog-associated (rabies, cystic echinococcosis and leishmaniasis) zoonotic diseases in Sidi Kacem Province, northwest Morocco. Conducted by veterinary and epidemiology students between 2013 and 2014, this followed a process-based approach that encouraged sequential adaptation of images, key messages, and delivery strategies using auto-evaluation and end-user feedback. We describe the challenges and opportunities of this approach, reflecting on who was targeted, how education was conducted, and what tools and approaches were used. Our results showed that: (1) replacing words with local pictures and using "hands-on" activities improved receptivity; (2) information "overload" easily occurred when disease transmission pathways did not overlap; (3) access and receptivity at schools was greater than at the community-level; and (4) piggy-backing on high-priority diseases like rabies offered an important avenue to increase knowledge of other zoonoses. We conclude by discussing the merits of incorporating our validated education approach into the school curriculum in order to influence long-term behaviour change