RESUMEN
MAIN CONCLUSION: Overexpression of VvmybA1 transcription factor in 'Hamlin' citrus enhances cold tolerance by increasing anthocyanin accumulation. This results in improved ROS scavenging, altered gene expression, and stomatal regulation, highlighting anthocyanins' essential role in citrus cold acclimation. Cold stress is a significant threat to citrus cultivation, impacting tree health and productivity. Anthocyanins are known for their role as pigments and have emerged as key mediators of plant defense mechanisms against environmental stressors. This study investigated the potential of anthocyanin overexpression regulated by grape (Vitis vinifera) VvmybA1 transcription factor to enhance cold stress tolerance in citrus trees. Transgenic 'Hamlin' citrus trees overexpressing VvmybA1 were exposed to a 30-day cold stress period at 4 °C along with the control wild-type trees. Our findings reveal that anthocyanin accumulation significantly influences chlorophyll content and their fluorescence parameters, affecting leaf responses to cold stress. Additionally, we recorded enhanced ROS scavenging capacity and distinct expression patterns of key transcription factors and antioxidant-related genes in the transgenic leaves. Furthermore, VvmybA1 overexpression affected stomatal aperture regulation by moderating ABA biosynthesis, resulting in differential responses in a stomatal opening between transgenic and wild-type trees under cold stress. Transgenic trees exhibited reduced hydrogen peroxide levels, enhanced flavonoids, radical scavenging activity, and altered phytohormonal profiles. These findings highlighted the role of VvmybA1-mediated anthocyanin accumulation in enhancing cold tolerance. The current study also underlines the potential of anthocyanin overexpression as a critical regulator of the cold acclimation process by scavenging ROS in plant tissues.
Asunto(s)
Antocianinas , Citrus sinensis , Respuesta al Choque por Frío , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Plantas Modificadas Genéticamente , Antocianinas/metabolismo , Citrus sinensis/genética , Citrus sinensis/metabolismo , Citrus sinensis/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Respuesta al Choque por Frío/genética , Especies Reactivas de Oxígeno/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vitis/genética , Vitis/fisiología , Vitis/metabolismo , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Hojas de la Planta/fisiología , Clorofila/metabolismo , Frío , Estomas de Plantas/fisiología , Estomas de Plantas/genética , Ácido Abscísico/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismoRESUMEN
BACKGROUND: Abiotic stress, such as salinity, affects the photosynthetic apparatus of plants. It is reported that the use of selenium nanoparticles (Se NPs), and biochemical compounds such as chitosan (CS) increase the tolerance of plants to stress conditions. Therefore, this study aimed to elucidate the potential of Se NPs, CS, and their composite (CS + Se NPs) in improving the photosynthetic apparatus of C. sinensis under salt stress in greenhouse conditions. The grafted seedlings of C. sinensis cv. Valencia after adapting to the greenhouse condition, were imposed with 0, 50, and 100 mM NaCl. After two weeks, the plants were foliar sprayed with distilled water (control), CS (0.1% w/v), Se NPs (20 mg L- 1), and CS + Se NPs (10 and 20 mg L- 1). Three months after treatment, the levels of photosynthetic pigments, leaf gas exchange, and chlorophyll fluorescence in the treated plants were evaluated. RESULTS: Under salinity stress, total chlorophyll, carotenoid, and SPAD values decreased by 31%, 48%, and 28% respectively, and Fv/Fm also decreased compared to the control, while the ratio of absorption flux (ABS), dissipated energy flux (DI0) and maximal trapping rate of PSII (TR0) to RC (a measure of PSII apparent antenna size) were increased. Under moderate (50 mM NaCl) and intense (100 mM NaCl) salinity stress, the application of CS + Se NPs significantly increased the levels of photosynthetic pigments and the Fv/Fm value compared to plants treated with distilled water. CONCLUSIONS: It may be inferred that foliar treatment with CS + Se NPs can sustain the photosynthetic ability of C. sinensis under salinity stress and minimize its deleterious effects on photosynthesis.
Asunto(s)
Quitosano , Citrus sinensis , Nanocompuestos , Fotosíntesis , Estrés Salino , Selenio , Fotosíntesis/efectos de los fármacos , Estrés Salino/efectos de los fármacos , Citrus sinensis/efectos de los fármacos , Citrus sinensis/fisiología , Clorofila/metabolismo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/fisiologíaRESUMEN
Source-sink balance in plants determines carbon distribution, and altering it can impact carbon fixation, transport, and allocation. We aimed to investigate the effect of altered source-sink ratios on carbon fixation, transport, and distribution in 'Valencia' sweet orange (Citrus x sinensis) by various defoliation treatments (0%, 33%, 66%, and 83% leaf removal). Gas exchange parameters were measured on 0 and 10 days after defoliation using A/Ci response curves, and leaf export was measured two days after defoliation using radioisotope tracer techniques. Greater defoliation increased the maximum rate of carboxylation (Vcmax), electron transport rate (J1200), and triose-phosphate utilization rate (TPU). Leaf export was unaffected by defoliation but increased in leaves closer to the shoot apex. Basipetal translocation velocity in the trunk remained unaltered, indicating that more photosynthates remained in the shoot rather than being transported directly to the root sink. Defoliated plants initiated more new flush shoots but accumulated less shoot biomass per plant after 8 weeks. Carbon allocation to fine roots was smaller in defoliated plants, suggesting defoliation led to retention of carbohydrates in aboveground organs such as the trunk and other shoots from previous growing cycles. In conclusion, the low source-sink ratio increased carbon fixation without impacting individual leaf export in citrus. The results suggest that intermediate sinks such as the aboveground perennial organs play a role in mediating the translocation velocity. Further research is necessary to better understand the dynamics of source-sink regulation in citrus trees.
Asunto(s)
Carbono , Citrus , Fotosíntesis , Hojas de la Planta , Hojas de la Planta/metabolismo , Carbono/metabolismo , Fotosíntesis/fisiología , Citrus/metabolismo , Citrus/fisiología , Citrus/crecimiento & desarrollo , Ciclo del Carbono , Raíces de Plantas/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Brotes de la Planta/metabolismo , Brotes de la Planta/crecimiento & desarrollo , Biomasa , Árboles/metabolismo , Árboles/fisiología , Citrus sinensis/metabolismo , Citrus sinensis/crecimiento & desarrollo , Citrus sinensis/fisiologíaRESUMEN
Magnesium (Mg) and calcium (Ca) are two essential macronutrients in plants; however, the characteristics of Mg and Ca concentrations in organ, subcellular and chemical forms and their relationships in citrus plants, especially under varying Mg supply, are not well understood. In this study, Citrus sinensis seedlings (cv. Xuegan) were cultivated in conditions of Mg deficiency (0 mmol Mg2+ L-1) and Mg sufficiency (2 mmol Mg2+ L-1) to investigate the responses of Mg and Ca homeostasis in different organs and fractions. Compared with Mg sufficiency, Mg deficiency significantly decreased root and shoot growth, with the shoot biomass reduction of branch organs was greater than that of parent organs. In addition to increasing the Ca concentration in the parent stem and lateral root organs, Mg deficiency significantly decreased the concentrations and accumulations of Mg and Ca in citrus seedlings, further altering their distribution in different organs. More than 50% of Ca and Mg were sequestrated in the cell wall and soluble fractions, respectively, with Mg concentration decreasing by 15.4% in roots and 46.9% in leaves under Mg deficiency, while Ca concentration decreased by 27.6% in roots and increased by 23.6% in parent leaves. Approximately 90% of Mg exists in inorganic, water-soluble, and pectate and protein-bound forms, and nearly 90% of Ca exists in water-soluble, pectate and protein-bound, phosphate and oxalate acid forms. Except for the decreased inorganic Mg in roots and water-soluble Mg and Ca in leaves, Mg deficiency increased the proportions of Mg and Ca in all chemical forms. However, Mg deficiency generally increased the Ca/Mg ratio in various organs, subcellular and chemical forms, with negative relationships between Mg concentration and Ca/Mg ratio, and the variations of Mg and Ca were highly separated between Mg supply and organs. In conclusion, our results provide insights into the effects of Mg supply on Mg and Ca homeostasis in citrus plants.
Asunto(s)
Citrus sinensis , Citrus , Citrus/fisiología , Magnesio/farmacología , Plantones/fisiología , Calcio/farmacología , Raíces de Plantas/fisiología , Citrus sinensis/fisiología , Hojas de la Planta/fisiología , Agua/farmacología , HomeostasisRESUMEN
BACKGROUND: Carotenoids and flavonoids are important secondary metabolites in plants, which exert multiple bioactivities and benefits to human health. Although the genes that encode carotenogenesis and flavonoid biosynthetic enzymes are well characterized, the transcriptional regulatory mechanisms that are related to the pathway genes remain to be investigated. In this study, 'Cara cara' navel orange (CNO) fruit at four development stages were used to identify the key genes and TFs for carotenoids and flavonoids accumulation. RESULTS: In this study, CNO was used to investigate the profiles of carotenoids and flavonoids by a combination of metabolomic and transcriptomic analyses. The important stage for the accumulation of the major carotenoid, lycopene was found to be at 120 days after florescence (DAF). The transcripts of five carotenogenesis genes were highly correlated with lycopene contents, and 16, 40, 48, 24 and 18 transcription factors (TFs) were predicted to potentially bind 1-deoxy-D-xylulose-5-phosphate synthase (DXS1), deoxyxylulose 5-phosphate reductoisomerase (DXR), geranylgeranyl diphosphate synthase (GGPPS2), phytoene synthase (PSY1) and lycopene ß-cyclase (LCYB) promoters, respectively. Narirutin was the most abundant flavonoid in the flesh at the early stages, 60 DAF was the most important stage for the accumulation of flavonoids, and 17, 22, 14, 25, 24 and 16 TFs could potentially bind phenylalanine ammonia-lyase (PAL-1 and PAL-4), 4-Coumarate-CoA ligase (4CL-2 and 4CL-5), chalcone synthase (CHS-1) and chalcone isomerase (CHI) promoters, respectively. Furthermore, both sets of 15 candidate TFs might regulate at least three key genes and contribute to carotenoids/flavonoids accumulation in CNO fruit. Finally, a hierarchical model for the regulatory network among the pathway genes and TFs was proposed. CONCLUSIONS: Collectively, our results suggest that DXS1, DXR, GGPPS2, PSY1 and LCYB genes were the most important genes for carotenoids accumulation, while PAL-1, PAL-4, 4CL-2, 4CL-5, CHS-1 and CHI for flavonoids biosynthesis. A total of 24 TFs were postulated as co-regulators in both pathways directly, which might play important roles in carotenoids and flavonoids accumulation in CNO fruit.
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Carotenoides/metabolismo , Citrus sinensis/genética , Citrus sinensis/fisiología , Flavonoides/biosíntesis , Flavonoides/genética , Frutas/genética , Frutas/fisiología , China , Productos Agrícolas/genética , Productos Agrícolas/fisiología , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Metaboloma , Factores de Transcripción , TranscriptomaRESUMEN
Pectinase and cellulase participate in cell wall degradation during secretory cavity formation in Citrus fruits. However, it remains unknown how secretory cavity formation is regulated by pectinase and cellulase genes in a schizolysigenous model. Our Results showed that PCD was involved in the schizolysigenous formation of the secretory cavities, and pectinase was involved in the degradation of the middle lamella while pectinase combined with cellulase were responsible for the degradation of the primary cell wall. Furthermore, the expression levels of CisPG21 and CisCEL16 at the intercellular space-forming and lumen-expanding stages with the continuous degradation of the cell wall were significantly higher than those at the initial cell stage and mature stage. The in situ hybridization (ISH) results also showed that CisPG21 and CisCEL16 were mainly located in the degrading cells of secretory cavities, and signals were very strong at the intercellular space-forming and lumen-expanding stages. In conclusion, pectinase and cellulase are directly involved in the degradation of PCD cell walls during schizolysigenous formation in the secretary cavity of Citrus sinensis (L.) Osbeck fruit, while CisPG21 and CisCEL16 are important regulatory genes of pectinase and cellulose during cell wall degradation.
Asunto(s)
Apoptosis , Pared Celular/metabolismo , Citrus sinensis/genética , Frutas/metabolismo , Genes de Plantas/fisiología , Pared Celular/fisiología , Celulasa/metabolismo , Celulosa/metabolismo , Citrus sinensis/metabolismo , Citrus sinensis/fisiología , Frutas/fisiología , Genes de Plantas/genética , Hibridación in Situ , Etiquetado Corte-Fin in Situ , Pectinas/metabolismo , Poligalacturonasa/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
The blood red color of pigmented orange fruit varieties [Citrus sinensis L. (Osbeck)] is due to the presence of anthocyanin pigments that largely contribute to determine the high organoleptic qualities and the nutritional properties of the fruits. The content of pigments in sweet orange depends primarily on genetic factors and on environmental conditions. In particular, it has been extensively shown that cold temperature induces an increase of anthocyanin content that is achieved by the induction of the related gene expression. The purpose of our work is to understand the mechanism underlying the color variegation occurring inside the blood oranges during the cold induction of anthocyanin biosynthesis, despite the fact that the entire fruit is genotypically programmed to produce pigments. Therefore, the amount of anthocyanin and the expression of both structural and regulatory genes have been monitored in either high-pigmented (HP) or not/low pigmented (NP) segments of the same fruit during the storage at 4 °C for a total experimental period of 25 days. Our results clearly indicate that the anthocyanin content is directly correlated with the levels of gene transcription, with higher pigmented areas showing higher enhancement of gene expression. Furthermore, we analyzed the reshaping of the DNA methylation status at the promoter regions of genes related to anthocyanin biosynthetic pathway, such as DFR and Ruby. Our results unequivocally demonstrate that in the promoter regions of both DFR and Ruby, the amount of cytosine methylation strongly decreases along the cold storage in the HP areas, whereas it increases in the NP areas of the same fruit, probably causing a partial block of the gene transcription. Finally, by measuring the changes in the expression levels of the Citrus DNA demethylases, we found that DML1 might play a crucial role in determining the observed demethylation of DFR and Ruby promoters, with its expression induced by cold in the HP areas of the fruits. This is the first report in which different levels of gene expression implicated in anthocyanin production in blood orange fruit is correlated with an epigenetic control mechanism such as promoter methylation.
Asunto(s)
Antocianinas/biosíntesis , Citrus sinensis/fisiología , Metilación de ADN , Citrus sinensis/genética , Citrus sinensis/crecimiento & desarrollo , Frío , Respuesta al Choque por Frío , Frutas/genética , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Candidatus Liberibacter asiaticus Jagoueix, Bové, and Garnier (Rhizobiales: Rhizobiaceae) is transmitted by the psyllid Diaphorina citri Kuwayama and putatively causes Huanglongbing disease in citrus. Huanglongbing has reduced yields by 68% relative to pre-disease yields in Florida. Disease management is partly through vector control. Understanding vector biology is essential in this endeavor. Our goal was to document differences in probing behavior linked to sex. Based on both a literature review and our results, we conclude that there is either no effect of sex or that identifying such an effect requires a sample size at least four times larger than standard methodologies. Including both color and sex in statistical models did not improve model performance. Both sex and color are correlated with body size, and body size has not been considered in previous studies on sex in D. citri in terms of probing behavior. An effect of body size was found wherein larger psyllids took longer to reach ingestion behaviors and larger individuals spent more time-ingesting phloem, but these relationships explained little of the variability in these data. We suggest that the effects of sex can be ignored when running EPG experiments on healthy psyllids.
Asunto(s)
Citrus sinensis/fisiología , Hemípteros/fisiología , Herbivoria , Pigmentación , Animales , Tamaño Corporal , Color , Conducta Alimentaria , Florida , Frutas/fisiología , Hemípteros/microbiología , Enfermedades de las Plantas/microbiología , Rhizobiaceae/fisiología , Factores SexualesRESUMEN
Xylem vulnerability to embolism represents an important trait to determine species distribution patterns and drought resistance. However, estimating embolism resistance frequently requires time-consuming and ambiguous hydraulic lab measurements. Based on a recently developed pneumatic method, we present and test the "Pneumatron", a device that generates high time-resolution and fully automated vulnerability curves. Embolism resistance is estimated by applying a partial vacuum to extract air from an excised xylem sample, while monitoring the pressure change over time. Although the amount of gas extracted is strongly correlated with the percentage loss of xylem conductivity, validation of the Pneumatron was performed by comparison with the optical method for Eucalyptus camaldulensis leaves. The Pneumatron improved the precision of the pneumatic method considerably, facilitating the detection of small differences in the (percentage of air discharged [PAD] < 0.47%). Hence, the Pneumatron can directly measure the 50% PAD without any fitting of vulnerability curves. PAD and embolism frequency based on the optical method were strongly correlated (r2 = 0.93) for E. camaldulensis. By providing an open source platform, the Pneumatron represents an easy, low-cost, and powerful tool for field measurements, which can significantly improve our understanding of plant-water relations and the mechanisms behind embolism.
Asunto(s)
Diseño de Equipo , Xilema/química , Citrus sinensis/fisiología , Bases de Datos Factuales , Sequías , Eucalyptus , Hojas de la Planta/fisiología , Raíces de Plantas/fisiología , Tallos de la Planta/fisiología , Transpiración de Plantas/fisiología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Árboles/fisiología , Agua/fisiologíaRESUMEN
BACKGROUND: Fruit coloration is one of the main quality parameters of Citrus fruit primarily determined by genetic factors. The fruit of ordinary sweet orange (Citrus sinensis) displays a pleasant orange tint due to accumulation of carotenoids, representing ß,ß-xanthophylls more than 80% of the total content. 'Pinalate' is a spontaneous bud mutant, or somatic mutation, derived from sweet orange 'Navelate', characterized by yellow fruits due to elevated proportions of upstream carotenes and reduced ß,ß-xanthophylls, which suggests a biosynthetic blockage at early steps of the carotenoid pathway. RESULTS: To identify the molecular basis of 'Pinalate' yellow fruit, a complete characterization of carotenoids profile together with transcriptional changes in carotenoid biosynthetic genes were performed in mutant and parental fruits during development and ripening. 'Pinalate' fruit showed a distinctive carotenoid profile at all ripening stages, accumulating phytoene, phytofluene and unusual proportions of 9,15,9'-tri-cis- and 9,9'-di-cis-ζ-carotene, while content of downstream carotenoids was significantly decreased. Transcript levels for most of the carotenoid biosynthetic genes showed no alterations in 'Pinalate'; however, the steady-state level mRNA of ζ-carotene isomerase (Z-ISO), which catalyses the conversion of 9,15,9'-tri-cis- to 9,9'-di-cis-ζ-carotene, was significantly reduced both in 'Pinalate' fruit and leaf tissues. Isolation of the 'Pinalate' Z-ISO genomic sequence identified a new allele with a single nucleotide insertion at the second exon, which generates an alternative splicing site that alters Z-ISO transcripts encoding non-functional enzyme. Moreover, functional assays of citrus Z-ISO in E.coli showed that light is able to enhance a non-enzymatic isomerization of tri-cis to di-cis-ζ-carotene, which is in agreement with the partial rescue of mutant phenotype when 'Pinalate' fruits are highly exposed to light during ripening. CONCLUSION: A single nucleotide insertion has been identified in 'Pinalate' Z-ISO gene that results in truncated proteins. This causes a bottleneck in the carotenoid pathway with an unbalanced content of carotenes upstream to ß,ß-xanthophylls in fruit tissues. In chloroplastic tissues, the effects of Z-ISO alteration are mainly manifested as a reduction in total carotenoid content. Taken together, our results indicate that the spontaneous single nucleotide insertion in Z-ISO is the molecular basis of the yellow pigmentation in 'Pinalate' sweet orange and points this isomerase as an essential activity for carotenogenesis in citrus fruits.
Asunto(s)
Citrus sinensis/fisiología , Frutas/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Isomerasas/genética , Proteínas de Plantas/genética , Alelos , Secuencia de Aminoácidos , Citrus sinensis/genética , Color , Frutas/genética , Isomerasas/química , Isomerasas/metabolismo , Pigmentación/genética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alineación de SecuenciaRESUMEN
Citrus tatter leaf virus (CTLV) threatens citrus production worldwide because it induces bud-union crease on the commercially important Citrange (Poncirus trifoliata × Citrus sinensis) rootstocks. However, little is known about its genomic diversity and how such diversity may influence virus detection. In this study, full-length genome sequences of 12 CTLV isolates from different geographical areas, intercepted and maintained for the past 60 years at the Citrus Clonal Protection Program (CCPP), University of California, Riverside, were characterized using next generation sequencing. Genome structure and sequence for all CTLV isolates were similar to Apple stem grooving virus (ASGV), the type species of Capillovirus genus of the Betaflexiviridae family. Phylogenetic analysis highlighted CTLV's point of origin in Asia, the virus spillover to different plant species and the bottleneck event of its introduction in the United States of America (USA). A reverse transcription quantitative polymerase chain reaction assay was designed at the most conserved genome area between the coat protein and the 3'-untranslated region (UTR), as identified by the full genome analysis. The assay was validated with different parameters (e.g. specificity, sensitivity, transferability and robustness) using multiple CTLV isolates from various citrus growing regions and it was compared with other published assays. This study proposes that in the era of powerful affordable sequencing platforms the presented approach of systematic full-genome sequence analysis of multiple virus isolates, and not only a small genome area of a small number of isolates, becomes a guideline for the design and validation of molecular virus detection assays, especially for use in high value germplasm programs.
Asunto(s)
Citrus sinensis/virología , Flexiviridae/clasificación , Poncirus/virología , Secuenciación Completa del Genoma/métodos , Citrus sinensis/fisiología , Secuencia Conservada , Evolución Molecular , Flexiviridae/genética , Flexiviridae/aislamiento & purificación , Tamaño del Genoma , Genoma Humano , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Filogenia , Fitomejoramiento , Poncirus/fisiologíaRESUMEN
BACKGROUND: Small RNAs regulate a wide variety of processes in plants, from organ development to both biotic and abiotic stress response. Being master regulators in genetic networks, their biogenesis and action is a fundamental aspect to characterize in order to understand plant growth and development. Three main gene families are critical components of RNA silencing: DICER-LIKE (DCL), ARGONAUTE (AGO) and RNA-DEPENDENT RNA POLYMERASE (RDR). Even though they have been characterized in other plant species, there is no information about these gene families in Citrus sinensis, one of the most important fruit species from both economical and nutritional reasons. While small RNAs have been implicated in the regulation of multiple aspects of plant growth and development, their role in the abscission process has not been characterized yet. RESULTS: Using genome-wide analysis and a phylogenetic approach, we identified a total of 13 AGO, 5 DCL and 7 RDR genes. We characterized their expression patterns in root, leaf, flesh, peel and embryo samples using RNA-seq data. Moreover, we studied their role in fruit abscission through gene expression analysis in fruit rind compared to abscission zone from samples obtained by laser capture microdissection. Interestingly, we determined that the expression of several RNA silencing factors are down-regulated in fruit abscission zone, being particularly represented gene components of the RNA-dependent DNA Methylation pathway, indicating that repression of this process is necessary for fruit abscission to take place in Citrus sinensis. CONCLUSIONS: The members of these 3 families present characteristic conserved domains and distinct expression patterns. We provide a detailed analysis of the members of these families and improved the annotation of some of these genes based on RNA-seq data. Our data suggests that the RNA-dependent DNA Methylation pathway is involved in the important fruit abscission process in C. sinensis.
Asunto(s)
Citrus sinensis/fisiología , Metilación de ADN/fisiología , Frutas/crecimiento & desarrollo , Genes de Plantas/fisiología , Genoma de Planta/fisiología , Citrus sinensis/genética , Citrus sinensis/crecimiento & desarrollo , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Familia de Multigenes , FilogeniaRESUMEN
The HVA22 gene has been isolated for the first time from the aleurone layer of barley (Hordeum vulgare). Here, we characterized the HVA22 family from citrus (C. clementina and C. sinensis). Twelve genes, 6 in each species, were identified as well as duplication events for some of them. The ORF size ranged from 235 to 804 bp and the protein molecular weight from 94 to 267â¯kDa. All the citrus HVA22 protein presented transmembrane location and conserved TB2/DP1/HVA22 region. Phylogenetic and gene expression analyses suggested that some citrus HVA22 play a role in flower and fruit development, and that gene expression may be regulated by hormone or environmental conditions. Other regulation levels were also predicted, such as alternative splicing and post-translational modifications. The overall data indicated that citrus HVA22 may be involved in vesicular traffic in stressed cells, and that CcHVA22d could be involved in dehydration tolerance.
Asunto(s)
Citrus/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Citrus/fisiología , Citrus sinensis/genética , Citrus sinensis/fisiología , Deshidratación , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Genes de Plantas/fisiología , Peróxido de Hidrógeno/metabolismo , Filogenia , Proteínas de Plantas/fisiología , Plantas Modificadas Genéticamente , Regiones Promotoras Genéticas/genética , Estrés Fisiológico , Nicotiana/genética , TranscriptomaRESUMEN
Citrus leprosis virus C (CiLV-C) is an economically important pathogen and the main causative agent of leprosis disease in citrus orchards. The main vector of this disease, the mite Brevipalpus yothersi, is widely distributed in Mexican orchards on a wide range of citrus species. Despite the importance of both the virus and the mite, field studies recording their occurrence and co-occurrence are practically non-existent. We systematically sampled orange orchards for both CiLV-C and B. yothersi throughout the year. The distribution of the CiLV-C and B. yothersi was evaluated on each sampling occasion and their spatiotemporal associations were determined. Specifically, 100-112 orange trees, distributed in 18 rows (five or six trees per row), were sampled monthly between March 2017 and February 2018 (11 sampling dates). Twenty leaves per tree were sampled on each occasion. The number of mites per tree and the percentage of leaves per tree with disease symptoms were recorded. On each sampling occasion, spatiotemporal associations between mites and disease were determined using the Spatial Analysis by Distance Indices (SADIE) method. CiLV-C and B. yothersi were identified using molecular methods. Throughout the study, the distribution of CiLV-C was aggregated and the distribution of B. yothersi was random. No association was found between the virus and the mite on any of the sampling dates. In total, 173 mites were collected, but only 43 mites were found to be carrying CiLV-C. The reason for this lack of association between the virus and the mite, as well as the impact of our findings on the epidemiology of the disease in orange orchards, are discussed.
Asunto(s)
Distribución Animal , Ácaros/fisiología , Virus de Plantas/fisiología , Animales , Citrus sinensis/crecimiento & desarrollo , Citrus sinensis/fisiología , Citrus sinensis/virología , México , Enfermedades de las Plantas/virología , Hojas de la Planta/fisiología , Hojas de la Planta/virología , Dinámica Poblacional , Análisis Espacio-TemporalRESUMEN
Citrus fruit ripening is a complex process involving biochemical, physiological and molecular events that differ between the flesh and the peel of the fruit. We characterized sweet orange peel maturation by means of a comparative transcriptomic analysis between Navelate orange (Citrus sinensis L. Osbeck) and its mutant fruit Pinalate, which presents a severe blockage at early steps of the carotenoid biosynthetic pathway and consequently reduced ABA levels. Peel ripening involved the decrease of the photosynthetic activity and the transmembrane transport processes, as well as the buildup of starch and cuticular waxes and the cell wall modification. In addition, a number of biotic and abiotic stress responses, including the defense response, and the response to blue light, water deprivation and abscisic acid stimulus were modulated in a ripening-stage specific manner. The regulation of energy-related processes and secondary metabolism pathways was attenuated in Pinalate, while the molecular mechanisms underlying stress responses displayed dependency on ABA levels. These results indicate that ABA is a key signal inducing stress responses along orange peel ripening, which might determine the fruit postharvest performance.
Asunto(s)
Citrus sinensis/fisiología , Mutación , Proteínas de Plantas/genética , Ácido Abscísico/metabolismo , Carotenoides/metabolismo , Citrus sinensis/genética , Citrus sinensis/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Fotosíntesis , Metabolismo SecundarioRESUMEN
BACKGROUND: Magnesium (Mg)-deficiency is one of the most prevalent physiological disorders causing a reduction in Citrus yield and quality. 'Xuegan' (Citrus sinensis) seedlings were irrigated for 16 weeks with nutrient solution containing 2 mM (Mg-sufficiency) or 0 mM (Mg-deficiency) Mg(NO3)2. Thereafter, we investigated the Mg-deficient effects on gas exchange and chlorophyll a fluorescence in the upper and lower leaves, and Mg, reactive oxygen species (ROS) and methylglyoxal (MG) metabolisms in the roots, lower and upper leaves. The specific objectives were to corroborate the hypothesis that the responses of ROS and MG metabolisms to Mg-deficiency were greater in the lower leaves than those in the upper leaves, and different between the leaves and roots. RESULTS: Mg level was higher in the Mg-deficient upper leaves than that in the Mg-deficient lower leaves. This might be responsible for the Mg-deficiency-induced larger alterations of all the measured parameters in the lower leaves than those in the upper leaves, but they showed similar change patterns between the Mg-deficient lower and upper leaves. Accordingly, Mg-deficiency increased greatly their differences between the lower and upper leaves. Most of parameters involved in ROS and MG metabolisms had similar variation trends and degrees between the Mg-deficient lower leaves and roots, but several parameters (namely glutathione S-transferase, sulfite reductase, ascorbate and dehydroascorbate) displayed the opposite variation trends. Obviously, differences existed in the Mg-deficiency-induced alterations of ROS and MG metabolisms between the lower leaves and roots. Although the activities of most antioxidant and sulfur metabolism-related enzymes and glyoxalase I and the level of reduced glutathione in the Mg-deficient leaves and roots and the level of ascorbate in the leaves were kept in higher levels, the levels of malonaldehyde and MG and/or electrolyte leakage were increased in the Mg-deficient lower and upper leaves and roots, especially in the Mg-deficient lower leaves and roots. CONCLUSIONS: The ROS and MG detoxification systems as a whole did not provide sufficient detoxification capacity to prevent the Mg-deficiency-induced production and accumulation of ROS and MG, thus leading to lipid peroxidation and the loss of plasma membrane integrity, especially in the lower leaves and roots.
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Citrus sinensis/fisiología , Magnesio/metabolismo , Piruvaldehído/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Antioxidantes/metabolismo , Clorofila A/metabolismo , Fluorescencia , Hojas de la Planta/fisiología , Raíces de Plantas/fisiología , Plantones/fisiología , Azufre/metabolismoRESUMEN
BACKGROUND: Citrus plants are commercially propagated by grafting, with the rootstock variety influencing a number of horticultural traits, including drought tolerance. Among the different rootstock varieties available for citrus propagation, 'Rangpur' lime is known to confer enhanced tolerance to drought as compared to other citrus rootstocks. The objective of this study was to investigate the poorly understood molecular responses underlying the rootstock-induced drought tolerance in sweet orange. RESULTS: RNA-Seq transcriptome analysis was carried out in leaves of sweet orange grafted on 'Rangpur' lime subjected to control and drought-stress treatments, under greenhouse conditions, using the Illumina HiSeq platform. A total of 41,827 unique transcripts were identified, among which 1764 transcripts showed significant variation (P ≤ 0.001) between the treatments, with 1081 genes induced and 683 repressed by drought-stress treatment. The transcripts were distributed in 44 different categories of cellular component, molecular function and biological process. Several genes related to cell metabolism, including those involved in the metabolisms of cell wall, carbohydrates and antioxidants, light reactions, biotic and abiotic stress responses, as well as genes coding for transcription factors (TFs), protein kinases (PKs) and proteins involved in the abscisic acid (ABA) and ethylene signaling pathways, were differentially regulated by drought stress. RNA-Seq data were validated by quantitative real-time PCR (qPCR) analysis and comparative analysis of expression of the selected genes between sweet orange grafted on drought-tolerant and -sensitive rootstocks revealed new candidate genes for drought tolerance in citrus. CONCLUSIONS: In conclusion, our results showed that only a relatively small but functionally diverse fraction of the sweet orange transcriptome, with functions in metabolism, cellular responses and regulation, was differentially regulated by drought stress. The data suggest that the rootstock-induced drought tolerance in sweet orange includes the transcriptional activation of genes related to the cell wall, soluble carbohydrate and antioxidant metabolisms, biotic and abiotic stress responses, TFs, PKs and ABA signaling pathway, and the downregulation of genes involved in the starch metabolism, light reactions and ethylene signaling. Future efforts to elucidate their functional roles and explore their potential in the citrus genetic improvement should benefit from this data.
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Citrus sinensis/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Estrés Fisiológico , Ácido Abscísico/metabolismo , Citrus sinensis/metabolismo , Citrus sinensis/fisiología , Sequías , Análisis de Secuencia de ARN , Transducción de Señal , Factores de TranscripciónRESUMEN
This study investigated the effects of cross-pollination by 'Murcott' tangor on the fruit quality of 'Qicheng52' navel orange, including the physicochemical properties, bioactive compounds and antioxidant capacities. There were no significant differences on the fruit weight, juice yield and pH value of juice between self- and cross-pollinated fruits. However, cross-pollination could significantly improve the fruit quality of 'Qicheng52' fruits by increasing the total soluble solid content from 11.12⯱â¯1.02⯰Brix to 13.86⯱â¯1.17⯰Brix. The results of high performance liquid chromatography analysis of three sugar components indicated that the increase of total sugar was mainly contributed by the increase of fructose and sucrose. Cross-pollination exhibited no effect on the flavonoids content, while the total phenolics content was increased from 210.09⯱â¯18.55â¯mg/L to 298.25⯱â¯29.10â¯mg/L, which contributed to the higher antioxidant capacity in the cross-pollination fruit juice.
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Antioxidantes/análisis , Citrus sinensis/fisiología , Polinización , Citrus , FrutasRESUMEN
MAIN CONCLUSION: In the current study, we showed that exogenous GABA supplementation increases the endogenous GABA level, several amino acids, and phytohormones in citrus plants, suggesting that GABA works in harmony with phytohormones. Gamma-aminobutyric acid (GABA) plays a key role in cytosolic regulation of pH, controlling of carbon and nitrogen metabolism, and protection against biotic and abiotic stresses. Although it is well-known that GABA is implicated in plant defense and it could act as a signaling molecule, its effect on phytohormones is not completely understood. In this study, we investigated the effect of exogenous GABA on citrus phytohormones using gas chromatography-mass spectrometry. A significant increase in endogenous GABA was observed in GABA-treated plants. The highest increase in GABA was recorded in plants treated with 10 mM 7 days post-treatment. In addition, we observed a moderate increase in several amino acids including glycine, L-alanine, L-proline, L-asparagine, and L-glutamine. The levels of benzoic acid, cinnamic acid, salicylic acid, trans-jasmonic acid, indole acetic acid, indole propionic acid, and abscisic acid were significantly increased in GABA-treated plants compared to the control. The gene expression showed that GABA transaminase (GABA-T) and succinic semialdehyde dehydrogenase (SSADH) were induced in GABA-treated plants, indicating a conversion of GABA to succinate. In addition, the gene expression of the regulatory enzymes of the TCA cycle (malate dehydrogenase and succinic dehydrogenase) was upregulated in GABA-treated plants, indicating an induction of respiration. In agreement with the chemical analysis, the gene expression results showed that most of the genes implicated in the biosynthesis of phytohormones were also upregulated in GABA-treated plants. Our results indicated that GABA works in harmony with phytohormones and suggested that regulation of phytohormones by exogenous GABA could play a key role in reducing plant stress.
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Aminoácidos/metabolismo , Citrus sinensis/efectos de los fármacos , Reguladores del Crecimiento de las Plantas/metabolismo , Ácido gamma-Aminobutírico/farmacología , Ácido Abscísico/metabolismo , Citrus sinensis/fisiología , Ciclopentanos , Ácidos Indolacéticos/metabolismo , Oxilipinas , Ácido Salicílico/metabolismo , Succionato-Semialdehído Deshidrogenasa/metabolismoRESUMEN
The basic helix-loop-helix (bHLH) transcription factors (TFs) comprise one of the largest gene families in plants, and participate in various physiological processes, but the physiological role and regulatory function of the majority of bHLHs remain poorly understood. Here, a total of 56 putative CsbHLH genes were identified in sweet orange (Citrus sinensis) based on a genome-wide analysis. The CsbHLH genes, except four members, were distributed throughout nine chromosomes and divided into 19 subgroups. Most of the CsbHLH genes were responsive to cold stress, with the greatest up-regulation being observed in CsbHLH18. CsbHLH18 is localized in the nuclei and has transcriptional activation activity. Overexpression of CsbHLH18 conferred enhanced cold tolerance in transgenic tobacco. The transgenic plants accumulated significantly less reactive oxygen species (ROS), concurrent with increased activities and transcript levels of antioxidant enzymes. In contrast, knockdown of bHLH18 by RNAi in trifoliate orange promoted cold susceptibility, accompanied by down-regulation of antioxidant genes and accumulation of more ROS. Protein-DNA interaction assays demonstrate that CsbHLH18 directly and specifically binds to and activates the promoter of CsPOD. Taken together, these findings indicate that CsbHLH18 plays a positive role in cold tolerance through, at least partly, modulation of ROS homeostasis by directly regulating the antioxidant gene.