RESUMO
Grapevine (Vitis vinifera L.) is a crop of major economic importance. However, grapevine yield is guaranteed by the massive use of pesticides to counteract pathogen infections. Under temperate-humid climate conditions, downy mildew is a primary threat for viticulture. Downy mildew is caused by the biotrophic oomycete Plasmopara viticola Berl. & de Toni, which can attack grapevine green tissues. In lack of treatments and with favourable weather conditions, downy mildew can devastate up to 75% of grape cultivation in one season and weaken newly born shoots, causing serious economic losses. Nevertheless, the repeated and massive use of some fungicides can lead to environmental pollution, negative impact on non-targeted organisms, development of resistance, residual toxicity and can foster human health concerns. In this manuscript, we provide an innovative approach to obtain specific pathogen protection for plants. By using the yeast two-hybrid approach and the P. viticola cellulose synthase 2 (PvCesA2), as target enzyme, we screened a combinatorial 8 amino acid peptide library with the aim to identify interacting peptides, potentially able to inhibit PvCesa2. Here, we demonstrate that the NoPv1 peptide aptamer prevents P. viticola germ tube formation and grapevine leaf infection without affecting the growth of non-target organisms and without being toxic for human cells. Furthermore, NoPv1 is also able to counteract Phytophthora infestans growth, the causal agent of late blight in potato and tomato, possibly as a consequence of the high amino acid sequence similarity between P. viticola and P. infestans cellulose synthase enzymes.
Assuntos
Aptâmeros de Peptídeos/farmacologia , Glucosiltransferases/antagonistas & inibidores , Oomicetos/efeitos dos fármacos , Doenças das Plantas/terapia , Proteínas de Plantas/antagonistas & inibidores , Proteínas Citotóxicas Formadoras de Poros/farmacologia , Sequência de Aminoácidos , Celulose/biossíntese , Glucosiltransferases/química , Oomicetos/enzimologia , Oomicetos/ultraestrutura , Biblioteca de Peptídeos , Fotossíntese , Phytophthora infestans/efeitos dos fármacos , Phytophthora infestans/enzimologia , Phytophthora infestans/ultraestrutura , Doenças das Plantas/parasitologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Proteínas de Plantas/química , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Solanum tuberosum , Técnicas do Sistema de Duplo-Híbrido , VitisRESUMO
Carpels are the female reproductive organs of the flower, organized in a gynoecium, which is arguably the most complex organ of a plant. The gynoecium provides protection for the ovules, helps to discriminate between male gametophytes, and facilitates successful pollination. After fertilization, it develops into a fruit, a specialized organ for seed protection and dispersal. To carry out all these functions, coordinated patterning and tissue specification within the developing gynoecium have to be achieved. In this chapter, we describe different methods to characterize defects in carpel patterning and morphogenesis associated with developmental mutations as well as a list of reporter lines that can be used to facilitate genetic analyses.