Preliminary functional characterization, cloning and primary sequence of Fastuosain, a cysteine peptidase isolated from fruits of Bromelia fastuosa

The present work reports the characterization of Fastuosain, a novel cysteine protease of 25kDa, purified from the unripe fruits of Bromelia fastuosa, a wild South American Bromeliaceae. Proteolytic activity, measured using casein and synthetic substrates, was dependent on the presence of thiol reagents, having maximum activity at pH 7.0. The present work reports cDNA cloning of Fastuosain; cDNA was amplified by PCR using specific primers. The product was 1096pb long. Mature fastuosain has 217 residues, and with the proregion has a total length of 324 residues. Its primary sequence showed high homology with ananain(74%), stem bromelain (66%) and papain (44%).

Floral biology and late-acting self-incompatibility in Jacaranda racemosa (Bignoniaceae)

Breeding-system studies have been conducted with 38 of the approximately 800 species of Bignoniaceae, and self-incompatibility was found in 31 of these. In species for which the site of self-incompatibility barrier was studied, self-pollinated flowers consistently failed to develop into fruits, even though pollen tubes grew down to the ovary and penetrated most of the ovules. In this study, we have investigated the. oral biology and the breeding system in Jacaranda racemosa Chamisso, with hand-pollination experiments and the histology of post-pollination events. Flower anthesis lasted 1-3 days, and although the frequency of flower visitation was extremely low, natural pollination seemed to be effected mainly by medium-sized bees. Because the conspicuous staminodium favours eventual pollination by small bees, a possible role of the staminodium in the increase of potential pollinators is suggested. Hand-pollinations indicated that J. racemosa is a self-sterile species. Histological analysis of post-pollination events indicated the occurrence of a kind of late-acting self-incompatibility in which the processes of ovule penetration, fertilisation and endosperm initiation were slower in selfed than in crossed pistils. Until the time of self-pollinated pistil abscission, no signs of endosperm malfunction or proembryo development were observed in selfed pistils. Therefore, inbreeding depression is an unlikely explanation for self-sterility in J. racemosa.