Processing and subcellular localization of the leader papain-like proteinase of Beet yellows closterovirus.
ORF 1a of Beet yellows closterovirus (BYV) encodes the domains of the papain-like proteinase (PCP), methyltransferase (MT) and RNA helicase. BYV cDNA inserts encoding the PCP-MT region were cloned in pGEX vectors next to the glutathione S-transferase gene (GST). In a 'double tag' construct, the GST-PCP-MT cDNA was flanked by the 3'-terminal six histidine triplets. Following expression in E. coli, the fusion proteins were specifically self-cleaved into the GST-PCP and MT fragments. MT-His(6) was purified on Ni-NTA agarose and its N-terminal sequence determined by Edman degradation as GVEEEA, thus providing direct evidence for the Gly(588)/Gly(589) bond cleavage. The GST-PCP fragment purified on glutathione S-agarose was used as an immunogen to produce anti-PCP monoclonal antibodies (mAbs). On Western blots of proteins from virus-infected Tetragonia expansa, the mAbs recognized the 66 kDa protein. Immunogold labelling of BYV-infected tissue clearly indicated association of the PCP with the BYV-induced membranous vesicle aggregates, structures related to closterovirus replication. (+info)
Invasion dynamics of two alien Carpobrotus (Aizoaceae) taxa on a Mediterranean island: I. Genetic diversity and introgression.
This study, based on morphological and isozyme analysis, clearly discriminates two invasive Carpobrotus taxa, C. edulis and C. acinaciformis, in the Hyeres archipelago off the southeastern coast of France. However, three different allelic combinations demonstrate the presence of intermediate individuals resulting from an introgression of part of the C. edulis genome into that of C. acinaciformis. Both taxa have higher than average genetic (C. edulis: P(0.95)=62.5%, A=2.25+/-0.70, H(o)=0.329+/-0.324; C. acinaciformis: P(0.95)=75%, A=2.38+/-0.42, H(o)=0.645+/-0.109) and clonal diversities (C. edulis: IP=0.37; C. acinaciformis: IP=0.48). Furthermore, C. acinaciformis has an excess of heterozygotes (F=-0.585+/-0.217), probably due to introgression. The relationship between the probability of clonal identity for two individuals and distance indicates that C. acinaciformis relies more on clonal reproduction than on sexual recruitment (seed recruitment/vegetative propagation=u/v=0.027), in contrast to C. edulis, whose probability of clonal identity did not vary with distance. The overwhelming clonal growth and high genetic diversities of C. acinaciformis and the previously recorded invasion potential for C. edulis raises concern for intensified invasion via hybridisation. (+info)
Invasion dynamics of two alien Carpobrotus (Aizoaceae) taxa on a Mediterranean island: II. Reproductive strategies.
This study compares sexually and asexually produced fruit set, seed production, biomass, germination, and seedling size in Carpobrotus acinaciformis and C. edulis following controlled pollination experiments in order to evaluate the potential role of reproductive traits with respect to the invasive potential of these taxa. C. edulis is slightly agamospermic, completely self-fertile, slightly preferentially self-compatible, experiences no inbreeding depression, and has low hybrid vigour. In contrast, C. acinaciformis does not have reliable agamospermy, is only slightly self-fertile and self-compatible, experiences a slight inbreeding depression, and has a strong hybrid vigour. Both taxa have relatively low, although significantly different germination frequencies, and insignificantly different seedling sizes. Owing to the high performance in hybridisation as compared to all other controlled pollinations in C. acinaciformis, as well as a large amount of previously demonstrated introgression, we refer to the population studied on the island of Bagaud (France) as C. affine acinaciformis. We conclude that both C. edulis and C. affine acinaciformis should be considered as harmful invasive plants in the Mediterranean Basin, the former because of the flexibility of its mating system and high seed production, and the latter because of its strong clonality, high hybrid vigour, and potential for continued introgression from C. edulis genes. These differences require different control strategies, while the avoidance of sympatry is a distinct priority. (+info)
High invasive pollen transfer, yet low deposition on native stigmas in a Carpobrotus-invaded community.