A simple protocol for transient gene expression in ripe fleshy fruit mediated by Agrobacterium.

Fleshy fruits represent a very important economic resource and, therefore, they are an ideal target for biotechnological ameliorations. However, because of their physiological and anatomical characteristics, ripe fleshy fruits represent an extremely difficult material for transient gene expression assays aimed at the study of gene promoters in a short time. To this purpose, a fast and efficient Agrobacterium-mediated transient gene expression system was developed for ripe fleshy fruits. A beta-glucuronidase reporter gene interrupted by an intron was used in order to prevent the possible expression of GUS activity by the Agrobacterium cells. The contemporary use of another reporter gene was used to check the transformation efficiency. This method is based on the injection of an Agrobacterium suspension into the fruits, and allows both qualitative and quantitative assays in a wide range of fruits to be carried out.     

The involvement of auxin in the ripening of climacteric fruits comes of age: the hormone plays a role of its own and has an intense interplay with ethylene in ripening peaches

Ethylene has long been regarded as the main regulator of ripening in climacteric fruits. The characterization of a few tomato mutants, unable to produce climacteric ethylene and to ripen their fruits even following treatments with exogenous ethylene, has shown that other factors also play an important role in the control of climacteric fruit ripening. In climacteric peach and tomato fruits it has been shown that, concomitant with ethylene production, increases in the amount of auxin can also be measured. In this work a genomic approach has been used in order to understand if such an auxin increase is functional to an independent role played by the hormone during ripening of the climacteric peach fruits. Besides the already known indirect activity on ripening due to its up-regulation of climacteric ethylene synthesis, it has been possible to show that auxin plays a role of its own during ripening of peaches. In fact, the hormone has shown the ability to regulate the expression of a number of different genes. Moreover, many genes involved in biosynthesis and transport and, in particular, the signalling (receptors, Auxin Response Factors and Aux/IAA) of auxin had increased expression in the mesocarp during ripening, thus strengthening the idea that this hormone is actively involved in the ripening of peaches. This study has also demonstrated the existence of an important cross-talk between auxin and ethylene, with genes in the auxin domain regulated by ethylene and genes in the ethylene domain regulated by auxin.     

Cellulase involvement in the abscission of peach and pepper leaves is affected by salicylic acid

Phenolic compounds appear to be involved in a number of regulatoryactivities in plants. During the last decade an increasing amountof evidence has brought to light the role played by salicylicacid in a number of physiological processes. Particularly interestingis the inhibitory effect of salicylic acid on ethylene biosynthesis,which might make this natural plant compound a useful tool forcontrolling some of the responses usually promoted by ethylene.Our data show that salicylic acid is actually able to reduceleaf abscission in both peach and pepper plants. Biochemicalanalyses have revealed that the enzyme usually involved in thisphenomenon (cellulase, EC 3.2.1.4 [EC] .) does not increase followingactivation of leaf abscission in plants treated with salicylicacid. In contrast, control plants show a marked increase inthe levels of both enzyme activity and cellulase protein. Flushingplants with exogenous ethylene in the presence of salicylicacid induces an increase in cellulase expression which, however,does not equal the level induced in plants without salicylicacid. Key words: Cellulase, leaf abscission, peach, pepper, salicylic acid     

Cell-specific expression of tryptophan decarboxylase and 10-hydroxygeraniol oxidoreductase,key genes involved in camptothecin biosynthesis in <Emphasis Type="Italic">Camptotheca acuminata</Emphasis> Decne (Nyssaceae)

Background  

Camptotheca acuminata is a major natural source of the terpenoid indole alkaloid camptothecin (CPT). At present, little is known about the cellular distribution of the biosynthesis of CPT, which would be useful knowledge for developing new strategies and technologies for improving alkaloid production.