Protein changes in the albedo of citrus fruits on postharvesting storage.

In this work, major protein changes in the albedo of the fruit peel of Murcott tangor (tangerine x sweet orange) during postharvest ageing were studied through 2D PAGE. Protein content in matured on-tree fruits and in fruits stored in nonstressing [99% relative humidity (RH) and 25 degrees C], cold (99% RH and 4 degrees C), and drought (60% RH and 25 degrees C) conditions was initially determined. Protein identification through MS/MS determinations revealed in all samples analyzed the occurrence of manganese superoxide dismutase (Mn SOD), actin, ATP synthase beta subunit (ATPase), citrus salt-stress associated protein (CitSap), ascorbate peroxidase (APX), translationally controlled tumor protein (TCTP), and a cysteine proteinase (CP) of the papain family. The latter protein was identified in two different gel spots, with different molecular mass, suggesting the simultaneous presence of the proteinase precursor and its active form. While Mn SOD, actin, ATPase, and CitSap were unchanged in the assayed conditions, TCTP and APX were downregulated during the postharvest ageing process. Ageing-induced APX repression was also reversed by drought. CP contents in albedo, which were similar in on- and off-tree fruits, were strongly dependent upon cold storage. The active/total CP protein ratio significantly increased after cold exposure. This proteomic survey indicates that major changes in protein content in the albedo of the peel of postharvest stored citrus fruits are apparently related to the activation of programmed cell death (PCD).

Engineering of gibberellin levels in citrus by sense and antisense overexpression of a GA 20-oxidase gene modifies plant architecture.

Carrizo citrange (Citrus sinensisxPoncirus trifoliata) is a citrus hybrid widely used as a rootstock, whose genetic manipulation to improve different growth characteristics is of high agronomic interest. In this work, transgenic Carrizo citrange plants have been produced overexpressing sense and antisense CcGA20ox1 (a key enzyme of GA biosynthesis) under control of the 35S promoter to modify plant architecture. As expected, taller (sense) and shorter (antisense) phenotypes correlated with higher and lower levels, respectively, of active GA1 in growing shoots. In contrast, other phenotypic characteristics seemed to be specific to citrus, or different from those described for similar transgenics in other species. For instance, thorns, typical organs of citrus at juvenile stages, were much longer in sense and shorter in antisense plants, and xylem tissue was reduced in leaf and internode of sense plants. Antisense plants presented a bushy phenotype, suggesting a possible effect of GAs on auxin biosynthesis and/or transport. The main foliole of sense plants was longer, although total leaf area was reduced. Leaf thickness was smaller in sense and larger in antisense plants due to changes in the spongy parenchyma. Internode cell length was not altered in transgenic plants, indicating that, in citrus, GAs regulate cell division rather than cell elongation. Interestingly, the phenotypes described were not apparent when transgenic plants were grafted on non-transgenic rootstock. This suggests that roots contribute to the GA economy of aerial parts in citrus and opens the possibility of using the antisense plants as dwarfing rootstocks.

The dwarfing mechanism of citrus rootstocks F&A 418 and #23 is related to competition between vegetative and reproductive growth.

The annual development of Navelina (Citrus sinensis (L.) Osbeck) trees budded on three hybrid citrus rootstocks was studied. Two rootstocks, named #23 and #24, were obtained from the cross of Troyer citrange (C. sinensis x Poncirus trifoliata (L.) Raf.) x Cleopatra mandarin (C. reshni Hort. ex Tan.). The third rootstock, named F&A 418, came from a cross of Troyer citrange x common mandarin (C. deliciosa Ten.). Rootstocks #23 and F&A 418 are dwarfing rootstocks and reduce the size of the scion by about 75%. Rootstock #24 yields a standard size scion. Major growth differences that influenced tree size were apparent during the first summer after grafting and appeared to be related to fruit productivity, because defruiting the dwarfed scions caused a significant increase in vegetative shoot development, including summer sprouting. The reduced growth of the dwarfed scions was not restored by hormone application, indicating that a hormonal deficiency is unlikely to be the primary reason for scion dwarfing, although differences in gibberellin concentrations were found in actively growing shoots. Leaf photosynthesis was similar in scions on all three rootstocks, but the carbohydrate accumulation in fruits and fibrous roots during the summer sprouting period was significantly greater in the dwarfed trees than in the standard trees. Our results suggest that the dwarfing mechanism induced by the F&A 418 and #23 rootstocks is mediated by enhanced reproductive development and fruit growth, resulting in reduced vegetative development in the summer. Thus, a change in the pattern of assimilate distribution appears to be one of the main components of the dwarfing mechanism.