BackgroundPhytophthora nicotianae Breda de Haan (Phytophthora parasitica Dastur) causes severe damage to citrus crops worldwide. A population of citrandarins was created from the cross between the susceptible parent Citrus sunki Hort. Ex Tan. and the resistant parent Poncirus trifoliata (L.) Raf. cv. Rubidoux, both parents and two reference rootstocks (Rangpur lime and Swingle citrumelo) were grafted in a greenhouse on Rangpur lime. Inoculations were performed at 10 cm and 15 cm above the grafting region and the resulting lesions were evaluated by measuring the lesion length 60 days after inoculation. As control, non-inoculated plants of each genotype were used. In addition, we evaluated the expression of 19 candidate genes involved in citrus defense response 48 h after pathogen infection by quantitative Real-Time PCR (qPCR). We mapped genomic regions of Quantitative Trait Loci (QTLs) and Expression Quantitative Trait Loci (eQTLs) associated with resistance to P. parasitica in the linkage groups (LGs) of the previously constructed maps of C. sunki and P. trifoliata.ResultsWe found disease severity differences among the generated hybrids, with lesion lengths varying from 1.15 to 11.13 mm. The heritability of the character was 65%. These results indicate that there is a great possibility of success in the selection of resistant hybrids within this experiment. The analysis of gene expression profile demonstrated a great variation of responses regarding the activation of plant defense pathways, indicating that citrandarins have several defense strategies to control oomycete infection. The information of the phenotypic and gene expression data made possible to detect genomic regions associated with resistance. Three QTLs and 84 eQTLs were detected in the linkage map of P. trifoliata, while one QTL and 110 eQTLs were detected in C. sunki.ConclusionsThis is the first study to use eQTLs mapping in the Phytophthora-citrus interaction. Our results from the QTLs and eQTLs mapping allow us to conclude that the resistance of some citrandarins to the infection by P. parasitica is due to a favorable combination of QTLs and eQTLs transmitted by both parents.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4888-2) contains supplementary material, which is available to authorized users.
Mitochondrial Uncoupling Proteins (UCPs) are mitochondrial inner membrane proteins that dissipate the proton electrochemical gradient generated by the respiratory chain complexes. In plants, these proteins are crucial for maintaining mitochondrial reactive oxygen species (ROS) homeostasis. In this study, single T-DNA insertion mutants for two (AtUCP1and AtUCP2) out of the three UCP genes present in Arabidopsis thaliana were employed to elucidate their potential roles in planta. Our data revealed a significant increase in the ATP/ADP ratios of both mutants, indicating clear alterations in energy metabolism, and a reduced respiratory rate in atucp2. Phenotypic characterization revealed that atucp1 and atucp2 plants displayed reduced primary root growth under normal and stressed conditions. Moreover, a reduced fertility phenotype was observed in both mutants, which exhibited increased number of sterile siliques and lower seed yield compared with wild-type plants. Reciprocal crosses demonstrated that both male and female fertility were compromised in atucp1, while such effect was exclusively observed in the male counterpart in atucp2. Most strikingly, a pronounced accumulation of hydrogen peroxide in the reproductive organs was observed in all mutant lines, indicating a disturbance in ROS homeostasis of mutant flowers. In line, the atucp1 and atucp2 mutants exhibited higher levels of ROS in pollen grains. Also in support, alternative oxidase 1a was highly induced in mutant flowers, while the expression profiles of transcription factors implicated in gene regulation during female and male reproductive organ/tissue development were perturbed. Overall, these data give support for an important role for AtUCP1 and AtUCP2 in flower oxidative homeostasis and overall plant fertility.
Huanglongbing (HLB) is a severe disease of citrus caused by the bacterium Candidatus Liberibacter. In America, the most common species is Candidatus Liberibacter asiaticus (CLas). In a previous study of the Citrus-HLB pathosystem, our group found differences in CLas multiplication in a population of hybrids obtained by hybridization between Citrus sunki Hort. Ex Tan. and Poncirus trifoliata (L.) Raf. cv. Rubidoux. The bacterial concentration was higher in C. sunki than in P. trifoliata. Thus, this study aims to map phenotypic (QTL) and gene expression (eQTL) data associated with host response to CLas in the linkage groups (LGs) of the previously constructed parental maps of C. sunki and P. trifoliata cv. Rubidoux. For the phenotypic analysis, we used a population of 79 F1 hybrids (termed citrandarins) between C. sunki and P. trifoliata. CLas and starch were quantified in the leaves of the plants two years after pathogen inoculation, allowing the classification of hybrids as resistant, tolerant, and susceptible. The expression of 14 candidate genes was measured in 72 hybrids of the population and used as expression data for the eQTL mapping. We located nine QTL and 52 eQTL in the C. sunki map and 17 QTL and 40 eQTL in the P. trifoliata map. The overlap of eQTL of the majority of genes with QTL from the phenotypic data indicates that the genes are related to the phenotype and are probably related to pathogen infection.
Phytophthora nicotianae Breda de Haan (syn. Phytophthora parasitica Dastur), causal agent of citrus gummosis disease, has caused great damage to citrus orchards throughout the world. While chemical and horticultural measures do not guarantee the preventive or curative control of citrus gummosis, the use of resistant rootstocks is the most reliable management strategy against the disease. Aiming at the development of citrus rootstocks resistant to gummosis and to better elucidate the Phytophthora-citrus pathosystem, citrus breeding programs have been ongoing worldwide, mostly employing directed crosses. These studies have succeeded in identifying differences in symptom development between resistant and susceptible rootstocks, as well as in the progeny of their crosses. In addition, differentially expressed genes were assessed, which ultimately should lead to the identification of markers involved in resistance to P. nicotianae. In this review we summarize the current knowledge of the molecular basis of citrus gummosis and the main strategies employed to obtain genetically resistant rootstocks.
Plant dicarboxylate carriers (DICs) transport a wide range of dicarboxylates across the mitochondrial inner membrane. The Arabidopsis thalianaDIC family is composed of three genes (AtDIC1, 2 and 3), whereas two genes (EgDIC1 and EgDIC2) have been retrieved in Eucalyptus grandis. Here, by combining in silico and in planta analyses, we provide evidence that DICs are partially redundant, important in plant adaptation to environmental stresses and part of a low-oxygen response in both species. AtDIC1 and AtDIC2 are present in most plant species and have very similar gene structure, developmental expression patterns and absolute expression across natural Arabidopsis accessions. In contrast, AtDIC3 seems to be an early genome acquisition found in Brassicaceae and shows relatively low (or no) expression across these accessions. In silico analysis revealed that both AtDICs and EgDICs are highly responsive to stresses, especially to cold and submergence, while their promoters are enriched for stress-responsive transcription factors binding sites. The expression of AtDIC1 and AtDIC2 is highly correlated across natural accessions and in response to stresses, while no correlation was found for AtDIC3. Gene ontology enrichment analysis suggests a role for AtDIC1 and AtDIC2 in response to hypoxia, and for AtDIC3 in phosphate starvation. Accordingly, the investigated genes are induced by submergence stress in A. thaliana and E. grandis while AtDIC2 overexpression improved seedling survival to submergence. Interestingly, the induction of AtDIC1 and AtDIC2 is abrogated in the erfVII mutant that is devoid of plant oxygen sensing, suggesting that these genes are part of a conserved hypoxia response in Arabidopsis.
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