RESUMO
Root-knot nematode (RKN) Meloidogyne species are major polyphagous pests of most crops worldwide, and cultivars with durable resistance are urgently needed because of nematicide bans. The Ma gene from the Myrobalan plum (Prunus cerasifera) confers complete-spectrum, heat-stable, and high-level resistance to RKN, which is remarkable in comparison with the Mi-1 gene from tomato (Solanum lycopersicum), the sole RKN resistance gene cloned. We report here the positional cloning and the functional validation of the Ma locus present at the heterozygous state in the P.2175 accession. High-resolution mapping totaling over 3,000 segregants reduced the Ma locus interval to a 32-kb cluster of three Toll/Interleukin1 Receptor-Nucleotide Binding Site-Leucine-Rich Repeat (LRR) genes (TNL1-TNL3), including a pseudogene (TNL2) and a truncated gene (TNL3). The sole complete gene in this interval (TNL1) was validated as Ma, as it conferred the same complete-spectrum and high-level resistance (as in P.2175) using its genomic sequence and native promoter region in Agrobacterium rhizogenes-transformed hairy roots and composite plants. The full-length cDNA (2,048 amino acids) of Ma is the longest of all Resistance genes cloned to date. Its TNL structure is completed by a huge post-LRR (PL) sequence (1,088 amino acids) comprising five repeated carboxyl-terminal PL exons with two conserved motifs. The amino-terminal region (213 amino acids) of the LRR exon is conserved between alleles and contrasts with the high interallelic polymorphisms of its distal region (111 amino acids) and of PL domains. The Ma gene highlights the importance of these uncharacterized PL domains, which may be involved in pathogen recognition through the decoy hypothesis or in nuclear signaling.
Assuntos
Genes de Plantas/genética , Imunidade Inata/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/química , Prunus/genética , Prunus/parasitologia , Tylenchoidea/fisiologia , Alelos , Sequência de Aminoácidos , Animais , Cromossomos Artificiais Bacterianos/genética , Éxons/genética , Estudos de Associação Genética , Teste de Complementação Genética , Loci Gênicos/genética , Íntrons/genética , Proteínas de Repetições Ricas em Leucina , Dados de Sequência Molecular , Família Multigênica/genética , Mapeamento Físico do Cromossomo , Doenças das Plantas/genética , Doenças das Plantas/parasitologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/imunologia , Raízes de Plantas/parasitologia , Regiões Promotoras Genéticas/genética , Estrutura Terciária de Proteína , Proteínas/química , Prunus/imunologia , Sequências Repetitivas de Aminoácidos/genética , Reprodutibilidade dos Testes , Especificidade da EspécieRESUMO
Fruit trees are likely to suffer from the effects of severe drought in the future; however, sound criteria for evaluating the species' ability to survive these extreme conditions are largely missing. Here, we evaluated the feasibility of using xylem cavitation resistance as a tool for screening Prunus species for drought resistance. Ten different Prunus species were selected to cover a large range of water requirements, from hydrophilic to xerophilic types. Shoot cavitation resistance was evaluated with the new Cavitron technique. At this inter-specific level, cavitation resistance was related to species drought resistance, with xerophilic species being less vulnerable to cavitation. The Cavitron technique enabled species characterization that required a short time and small amounts of plant material. This technique could be used to evaluate the drought resistance of a limited number of fruit tree genotypes. Genotype screening on a larger scale, however, would likely require another approach. Out of a number of anatomical traits tested, a significant correlation was found between cavitation resistance and inter-vessel wall thickness across species. This anatomical trait is, therefore, suggested as a possible alternative to direct cavitation estimates and could be included as a screening criterion in breeding programs for drought resistance of Prunus genotypes.