RESUMEN
ERFs (ethylene-responsive factors) are known to play a key role in orchestrating cold stress signal transduction. However, the regulatory mechanisms and target genes of most ERFs are far from being well deciphered. In this study, we identified a cold-induced ERF, designated as PtrERF110, from trifoliate orange (Poncirus trifoliata L. Raf., also known as Citrus trifoliata L.), an elite cold-hardy plant. PtrERF110 is a nuclear protein with transcriptional activation activity. Overexpression of PtrERF110 remarkably enhanced cold tolerance in lemon (Citrus limon) and tobacco (Nicotiana tabacum), whereas VIGS (virus-induced gene silencing)-mediated knockdown of PtrERF110 drastically impaired the cold tolerance. RNA sequence analysis revealed that PtrERF110 overexpression resulted in global transcriptional reprogramming of a range of stress-responsive genes. Three of the genes, including PtrERD6L16 (early responsive dehydration 6-like transporters), PtrSPS4 (sucrose phosphate synthase 4), and PtrUGT80B1 (UDP-glucose: sterol glycosyltransferases 80B1), were confirmed as direct targets of PtrERF110. Consistently, PtrERF110-overexpressing plants exhibited higher levels of sugars and sterols compared to their wild type counterparts, whereas the VIGS plants had an opposite trend. Exogenous supply of sucrose restored the cold tolerance of PtrERF110-silencing plants. In addition, knockdown of PtrSPS4, PtrERD6L16, and PtrUGT80B1 substantially impaired the cold tolerance of P. trifoliata. Taken together, our findings indicate that PtrERF110 positively modulates cold tolerance by directly regulating sugar and sterol synthesis through transcriptionally activating PtrERD6L16, PtrSPS4, and PtrUGT80B1. The regulatory modules (ERF110-ERD6L16/SPS4/UGT80B1) unraveled in this study advance our understanding of the molecular mechanisms underlying sugar and sterol accumulation in plants subjected to cold stress.
Asunto(s)
Citrus , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Factores de Transcripción , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Citrus/genética , Citrus/fisiología , Citrus/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Plantas Modificadas Genéticamente , Nicotiana/genética , Nicotiana/fisiología , Nicotiana/metabolismo , Frío , Azúcares/metabolismo , Esteroles/metabolismo , Respuesta al Choque por Frío/genéticaRESUMEN
As self-incompatibility is a major issue in pummelo breeding and production, its mechanism in citrus was analyzed to improve breeding efficiency and reduce production costs. Rutaceae belongs to S-RNase type of gametophytic self-incompatibility. While the function of S-RNase/SLF and the mechanism of self-incompatibility have been studied extensively, the transcriptional regulation of S-RNase has been less studied. We performed transcriptome sequencing with the styles of 'Shatian' pummelo on the day of anthesis and 1-5 days before anthesis, and found that the transcript level of S-RNase gradually decreased with flower development. By analyzing differentially expressed genes and correlation with the expression trend of S-RNase, we identified a candidate gene, CgHSFB1, and utilized biochemical experiments such as yeast one-hybrid assay, electrophoretic mobility shift assay and dual-luciferase assay, as well as transient transformation of citrus calli and Citrus microcarpa and demonstrated that CgHSFB1 could directly bind to the S1-RNase promoter and repress the expression of S1-RNase, which is involved in the pummelo self-incompatibility response. In contrast, CgHSFB1 did not bind to the promoter of S2-RNase, and there was specificity in the regulation of S-RNase.
Asunto(s)
Citrus , Flores , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Ribonucleasas , Autoincompatibilidad en las Plantas con Flores , Citrus/genética , Citrus/fisiología , Citrus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Flores/genética , Flores/fisiología , Flores/crecimiento & desarrollo , Autoincompatibilidad en las Plantas con Flores/genética , Ribonucleasas/genética , Ribonucleasas/metabolismo , Regiones Promotoras Genéticas/genética , Transcriptoma , Perfilación de la Expresión GénicaRESUMEN
Citrus plants are grown in diverse regions of the world, from subtropical to semi-arid and humid tropical areas. Through mechanisms essential for their survival, they adapt to the environmental conditions to which they are subjected. Although there is vast literature on adaptation of citrus plants to individual stresses, plant responses to interaction among different types of stresses have not been clearly examined. Abiotic or biotic stresses, or a combination of these stresses, result in reorganization of plant energy resources for defense, whether it be for resistance, tolerance, or prevention of stress. Plants generally respond to these stress factors through production of secondary metabolites, such as volatile compounds, derived from different biosynthesis and degradation pathways, which are released through distinct routes. Volatile compounds vary among plant species, meeting the specific needs of the plant. Simultaneous exposure to the stress factors of water deficit and herbivory leads to responses such as qualitative and quantitative changes in the emission of secondary metabolites, and compounds may accumulate within the leaves or predispose the plant to more quickly respond to the stress brought about by the herbivore. The genetic makeup of citrus plants can contribute to a better response to stress factors; however, studies on the emission of volatile compounds in different citrus genotypes under simultaneous stresses are limited. This review examines the effects of abiotic stress due to water deficit and biotic stress due to herbivory by Diaphorina citri in citrus plants and examines their connection with volatile compounds. A summary is made of advances in knowledge regarding the performance of volatile compounds in plant defense against both stress factors, as well as the interaction between them and possible findings in citrus plants. In addition, throughout this review, we focus on how genetic variation of the citrus species is correlated with production of volatile compounds to improve stress tolerance.
Asunto(s)
Citrus , Herbivoria , Metabolismo Secundario , Compuestos Orgánicos Volátiles , Citrus/metabolismo , Citrus/fisiología , Citrus/genética , Compuestos Orgánicos Volátiles/metabolismo , Animales , Adaptación Fisiológica , Estrés Fisiológico , Escarabajos/fisiología , Deshidratación , Sequías , HemípterosRESUMEN
BACKGROUND: Ichang papeda (Citrus ichangensis), a wild perennial plant of the Rutaceae family, is a cold-hardy plant. WRKY transcription factors are crucial regulators of plant growth and development as well as abiotic stress responses. However, the WRKY genes in C. ichangensis (CiWRKY) and their expression patterns under cold stress have not been thoroughly investigated, hindering our understanding of their role in cold tolerance. RESULTS: In this study, a total of 52 CiWRKY genes identified in the genome of C. ichangensis were classified into three main groups and five subgroups based on phylogenetic analysis. Comprehensive analyses of motif features, conserved domains, and gene structures were performed. Segmental duplication plays a significant role in the CiWRKY gene family expansion. Cis-acting element analysis revealed the presence of various stress-responsive elements in the promoters of the majority of CiWRKYs. Gene ontology (GO) analysis and protein-protein interaction predictions indicate that the CiWRKYs exhibit crucial roles in regulation of both development and stress response. Expression profiling analysis demonstrates that 14 CiWRKYs were substantially induced under cold stress. Virus-induced gene silencing (VIGS) assay confirmed that CiWRKY31, one of the cold-induced WRKYs, functions positively in regulation of cold tolerance. CONCLUSION: Sequence and protein properties of CiWRKYs were systematically analyzed. Among the 52 CiWRKY genes 14 members exhibited cold-responsive expression patterns, and CiWRKY31 was verified to be a positive regulator of cold tolerance. These findings pave way for future investigations to understand the molecular functions of CiWRKYs in cold tolerance and contribute to unravelling WRKYs that may be used for engineering cold tolerance in citrus.
Asunto(s)
Citrus , Respuesta al Choque por Frío , Regulación de la Expresión Génica de las Plantas , Filogenia , Proteínas de Plantas , Factores de Transcripción , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Citrus/genética , Citrus/fisiología , Respuesta al Choque por Frío/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Genoma de Planta , Perfilación de la Expresión Génica , Genes de Plantas , FríoRESUMEN
Citrus is a model plant for studying adventitious embryos, a form of asexual reproduction controlled by a single dominant gene, RWP. This gene has been identified as the causal gene for nucellar embryogenesis, but its function has not yet been fully understood. In this study, we used the fast-growing Fortunella hindsii as a system to explore chromatin accessibility during the nucellar embryony initiation, emphasizing elevated chromatin accessibility in polyembryonic (PO) genotypes compared to monoembryonic ones (MO). Notably, a higher level of accessible chromatin was observed in one allele of the promoter region of FhRWP, consistent with increased expression of the allele carrying the causal structural variant. By independently performing RNAi and gene editing experiments on PO genotypes, we found the downregulation of FhRWP expression could reduce the number of nucellar embryos, while its knockout resulted in abnormal axillary bud development. In overexpression experiments, FhRWP was identified as having the unique capability of inducing the embryogenic callus formation in MO stem segments, possibly through the regulation of the WUS-CLV signaling network and the ABA and cytokinin pathway, marking the inaugural demonstration of FhRWP's potential to reignite somatic cells' embryogenic fate. This study reveals the pleiotropic function of RWP in citrus and constructs a regulatory network during adventitious embryo formation, providing a new tool for bioengineering applications in plant regeneration.
Asunto(s)
Citrus , Regulación de la Expresión Génica de las Plantas , Fenotipo , Proteínas de Plantas , Citrus/genética , Citrus/fisiología , Citrus/crecimiento & desarrollo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Semillas/genética , Semillas/crecimiento & desarrollo , Edición Génica , Genes de Plantas/genética , GenotipoRESUMEN
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
Embolism resistance of xylem tissue varies among species and is an important trait related to drought resistance, with anatomical attributes like pit membrane thickness playing an important role in avoiding embolism spread. Grafted Citrus trees are commonly grown in orchards, with the rootstock being able to affect the drought resistance of the whole plant. Here, we evaluated how rootstocks affect the vulnerability to embolism resistance of the scion using several rootstock/scion combinations. Scions of 'Tahiti' acid lime, 'Hamlin', 'Pera' and 'Valencia' oranges grafted on a 'Rangpur' lime rootstock exhibit similar vulnerability to embolism. In field-grown trees, measurements of leaf water potential did not suggest significant embolism formation during the dry season, while stomata of Citrus trees presented an isohydric response to declining water availability. When 'Valencia' orange scions were grafted on 'Rangpur' lime, 'IAC 1710' citrandarin, 'Sunki Tropical' mandarin or 'Swingle' citrumelo rootstocks, variation in intervessel pit membrane thickness of the scion was found. The 'Rangpur' lime rootstock, which is known for its drought resistance, induced thicker pit membranes in the scion, resulting in higher embolism resistance than the other rootstocks. Similarly, the rootstock 'IAC 1710' citrandarin generated increased embolism resistance of the scion, which is highly relevant for citriculture.
Asunto(s)
Citrus , Raíces de Plantas , Xilema , Citrus/fisiología , Xilema/fisiología , Raíces de Plantas/fisiología , Agua/metabolismo , Sequías , Hojas de la Planta/fisiología , Hojas de la Planta/anatomía & histología , Estomas de Plantas/fisiologíaRESUMEN
Plants face a wide range of biotic and abiotic stress conditions, which are further intensified by climate change. Among these stressors, increased irradiation in terms of intensity and wavelength range can lead to detrimental effects, such as chlorophyll degradation, destruction of the PSII reaction center, generation of ROS, alterations to plant metabolism, and even plant death. Here, we investigated the responses of two citrus genotypes, Citrus macrophylla (CM), and Troyer citrange (TC) to UV-B light-induced stress, by growing plants of both genotypes under control and UV-B stress conditions for 5 days to evaluate their tolerance mechanisms. TC seedlings had higher sensitivity to UV-B light than CM seedlings, as they showed more damage and increased levels of oxidative harm (indicated by the accumulation of MDA). In contrast, CM seedlings exhibited specific adaptive mechanisms, including accumulation of higher levels of proline under stressful conditions, and enhanced antioxidant capacity, as evidenced by increased ascorbate peroxidase activity and upregulation of the CsAPX2 gene. Phytohormone accumulation patterns were similar in both genotypes, with a decrease in ABA content in response to UV-B light. Furthermore, expression of genes involved in light perception and response was specifically affected in the tolerant CM seedlings, which exhibited higher expression of CsHYH/CsHY5 and CsRUP1-2 genes. These findings underscore the importance of the antioxidant system in citrus plants subjected to UV-B light-induced stress and suggest that CsHYH/CsHY5 and CsRUP1-2 could be considered genes associated with tolerance to such challenging conditions.
Asunto(s)
Antioxidantes , Citrus , Prolina , Plantones , Rayos Ultravioleta , Citrus/efectos de la radiación , Citrus/genética , Citrus/fisiología , Citrus/metabolismo , Prolina/metabolismo , Antioxidantes/metabolismo , Plantones/efectos de la radiación , Plantones/fisiología , Plantones/genética , Plantones/metabolismo , Estrés Fisiológico , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Genotipo , Reguladores del Crecimiento de las Plantas/metabolismo , Estrés Oxidativo/efectos de la radiación , Adaptación Fisiológica/efectos de la radiación , Adaptación Fisiológica/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genéticaRESUMEN
Self-incompatibility (SI) is a crucial mechanism that prevents self-fertilization and inbreeding in flowering plants. Citrus exhibits SI regulated by a polymorphic S-locus containing an S-RNase gene and multiple S-locus F-box (SLF) genes. It has been documented that S-RNase functions as the pistil S determinant, but there is no direct evidence that the SLF genes closely linked with S-RNase function as pollen S determinants in Citrus. This study assembled the genomes of two pummelo (Citrus grandis) plants, obtained three novel complete and well-annotated S-haplotypes, and isolated 36 SLF or SLF-like alleles on the S-loci. Phylogenetic analysis of 138 SLFs revealed that the SLF genes were classified into 12 types, including six types with divergent or missing alleles. Furthermore, transformation experiments verified that the conserved S6-SLF7a protein can lead to the transition of SI to self-compatibility by recognizing non-self S8-RNase in 'Mini-Citrus' plants (S7S8 and S8S29, Fortunella hindsii), a model plant for citrus gene function studies. In vitro assays demonstrated interactions between SLFs of different S haplotypes and the Skp1-Cullin1-F-box subunit CgSSK1 protein. This study provides direct evidence that SLF controls the pollen function in Citrus, demonstrating its role in the 'non-self recognition' SI system.
Asunto(s)
Citrus , Proteínas F-Box , Filogenia , Proteínas de Plantas , Polen , Ribonucleasas , Autoincompatibilidad en las Plantas con Flores , Citrus/genética , Citrus/fisiología , Citrus/metabolismo , Autoincompatibilidad en las Plantas con Flores/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Polen/genética , Polen/fisiología , Proteínas F-Box/genética , Proteínas F-Box/metabolismo , Ribonucleasas/metabolismo , Ribonucleasas/genética , Secuencia de AminoácidosRESUMEN
Flowering is an essential process in fruit trees. Flower number and timing have a substantial impact on the yield and maturity of fruit. Ethylene and gibberellin (GA) play vital roles in flowering, but the mechanism of coordinated regulation of flowering in woody plants by GA and ethylene is still unclear. In this study, a lemon (Citrus limon L. Burm) 1-aminocyclopropane-1-carboxylic acid synthase gene (CiACS4) was overexpressed in Nicotiana tabacum and resulted in late flowering and increased flower number. Further transformation of citrus revealed that ethylene and starch content increased, and soluble sugar content decreased in 35S:CiACS4 lemon. Inhibition of CiACS4 in lemon resulted in effects opposite to that of 35S:CiACS4 in transgenic plants. Overexpression of the CiACS4-interacting protein ETHYLENE RESPONSE FACTOR3 (CiERF3) in N. tabacum resulted in delayed flowering and more flowers. Further experiments revealed that the CiACS4-CiERF3 complex can bind the promoters of FLOWERING LOCUS T (CiFT) and GOLDEN2-LIKE (CiFE) and suppress their expression. Moreover, overexpression of CiFE in N. tabacum led to early flowering and decreased flowers, and ethylene, starch, and soluble sugar contents were opposite to those in 35S:CiACS4 transgenic plants. Interestingly, CiFE also bound the promoter of CiFT. Additionally, GA3 and 1-aminocyclopropanecarboxylic acid (ACC) treatments delayed flowering in adult citrus, and treatment with GA and ethylene inhibitors increased flower number. ACC treatment also inhibited the expression of CiFT and CiFE. This study provides a theoretical basis for the application of ethylene to regulate flower number and mitigate the impacts of extreme weather on citrus yield due to delayed flowering.
Asunto(s)
Citrus , Etilenos , Flores , Regulación de la Expresión Génica de las Plantas , Giberelinas , Proteínas de Plantas , Plantas Modificadas Genéticamente , Giberelinas/metabolismo , Citrus/genética , Citrus/fisiología , Citrus/crecimiento & desarrollo , Flores/genética , Flores/fisiología , Flores/crecimiento & desarrollo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Etilenos/metabolismo , Nicotiana/genética , Nicotiana/fisiología , Nicotiana/crecimiento & desarrollo , Liasas/metabolismo , Liasas/genéticaRESUMEN
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
Citrus production is severely threatened by the devastating Huanglongbing (HLB) disease globally. By studying and analyzing the defensive behaviors of an HLB-tolerant citrus cultivar 'Shatangju', we discovered that citrus can sense Candidatus Liberibacter asiaticus (CLas) infection and induce immune responses against HLB, which can be further strengthened by both endogenously produced and exogenously applied methyl salicylate (MeSA). This immune circuit is turned on by an miR2977-SAMT (encoding a citrus Salicylate [SA] O-methyltransferase) cascade, by which CLas infection leads to more in planta MeSA production and aerial emission. We provided both transgenic and multi-year trail evidences that MeSA is an effective community immune signal. Ambient MeSA accumulation and foliage application can effectively induce defense gene expression and significantly boost citrus performance. We also found that miRNAs are battle fields between citrus and CLas, and about 30% of the differential gene expression upon CLas infection are regulated by miRNAs. Furthermore, CLas hijacks host key processes by manipulating key citrus miRNAs, and citrus employs miRNAs that coordinately regulate defense-related genes. Based on our results, we proposed that miRNAs and associated components are key targets for engineering or breeding resistant citrus varieties. We anticipate that MeSA-based management, either induced expression or external application, would be a promising tool for HLB control.
Asunto(s)
Citrus , MicroARNs , Rhizobiaceae , Citrus/fisiología , Enfermedades de las Plantas , Fitomejoramiento , Salicilatos/metabolismo , Liberibacter/genética , MicroARNs/genética , MicroARNs/metabolismoRESUMEN
Bordeaux mixture (Bm) is a copper (Cu)-based pesticide that has been widely used for controlling citrus scab and citrus canker. However, frequent spraying of Bm is toxic to citrus. To our knowledge, few studies are available that discuss how the photosynthetic characteristics and chloroplast ultrastructure of citrus leaves are affected by Cu toxicity induced by excessive Bm. In the study, two-year-old seedlings of Citrus grandis (C. grandis) and Citrus sinensis (C. sinensis), which were precultured in pots, were foliar-sprayed with deionized water (as control) or Bm diluted 500-fold at intervals of 7 days for 6 times (4 times as recommended by the manufacturer) to investigate the leaf Cu absorption, photosynthesis, chloroplast ultrastructure and antioxidant enzymatic activities. Bm foliar-sprayed 6 times on citrus seedlings increased the leaf Cu content, decreased the photosynthetic pigments content and destroyed the chloroplast ultrastructure, which induced leaf chlorosis and photosynthetic inhibition. A lower Cu absorption, a higher light photon-electron transfer efficiency, a relative integrity of chloroplast ultrastructure and a promoted antioxidant protection contributed to a higher photosynthetic activity of C. grandis than C. sinensis under excessive spraying of Bm. The present study provides crucial references for screening and selecting citrus species with a higher tolerance to Cu toxicity induced by excessive Bm.
Asunto(s)
Citrus , Antioxidantes , Cloroplastos , Citrus/fisiología , Cobre/toxicidad , Fotosíntesis , Hojas de la Planta/fisiología , Plantones/fisiologíaRESUMEN
Morpho-physiological strategies to deal with water deficit vary among citrus species and the chemical signaling through ABA and anatomical, hydraulic, and physiological traits were evaluated in saplings of Rangpur lime, Swingle citrumelo and Valencia sweet orange. Trunk and roots of Swingle citrumelo presented lower vessel diameter and higher vessel frequency as compared to the other species. However, relative water content at the turgor loss point (RWCTLP), the osmotic potential at full turgor (Ψ0), the osmotic potential at the turgor loss point (ΨTLP), bulk modulus of elasticity (ε) and the xylem water potential when hydraulic conductivity is reduced by 50% (Ψ50) and 88% (Ψ88) indicated similar hydraulic traits among citrus species, with Rangpur lime showing the highest hydraulic safety margin. Roots of Rangpur lime and Swingle citrumelo were more water conductive than ones of Valencia sweet orange, which was linked to higher stomatal conductance. Chemical signaling through ABA prevented shoot dehydration in Rangpur lime under water deficit, with this species showing a more conservative stomatal behavior, sensing, and responding rapidly to low soil moisture. Taken together, our results suggest that Rangpur lime - the drought tolerant species - has an improved control of leaf water status due to chemical signaling and effective stomatal regulation for reducing water loss as well as decreased root hydraulic conductivity for saving water resources under limiting conditions.
Asunto(s)
Citrus , Deshidratación , Citrus/fisiología , Sequías , Hojas de la Planta/fisiología , Xilema/fisiologíaRESUMEN
Heat stress is a major abiotic stress for plants, which can generate a range of biochemical and genetic responses. In 'Ponkan' mandarin fruit, hot air treatment (HAT) accelerates the degradation of citric acid. However, the transcriptional regulatory mechanisms of citrate degradation in response to HAT remain to be elucidated. Here, 17 heat shock transcription factor sequences were isolated, and dual-luciferase assays were employed to investigate whether the encoded proteins that could trans-activate the promoters of key genes in the GABA shunt, involved in citrate metabolism. We identified four heat shock transcription factors (CitHsfA7, CitHsfA3, CitHsfA4b and CitHsfA8) that showed trans-activation effects on CitAco3, CitIDH3 and CitGAD4, respectively. Transient expression of the CitHsfs in citrus fruits indicated that CitHsfA7 was the only factor that resulted in a significant lowering of the citric acid content, and these results were confirmed by a virus-induced gene silencing system (VIGS). Sub-cellar localization showed that CitHsfA7 is located in the nucleus and is capable of binding directly to a putative HSE in the CitAco3 promoter and enhance its expression. We proposed that the induction of CitHsfA7 transcript level contributes to citric acid degradation in citrus fruit, via modulation of CitAco3 in response to HAT.
Asunto(s)
Ácido Cítrico/metabolismo , Citrus/metabolismo , Factores de Transcripción del Choque Térmico/metabolismo , Respuesta al Choque Térmico/fisiología , Aire , Citrus/fisiología , Regulación de la Expresión Génica de las Plantas , Silenciador del Gen , Factores de Transcripción del Choque Térmico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiones Promotoras Genéticas , Ácido gamma-Aminobutírico/genética , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Consequences of warming and postwarming events on photosynthetic thermotolerance (PT ) and photoprotective responses in tropical evergreen species remain elusive. We chose Citrus to answer some of the emerging questions related to tropical evergreen species' PT behaviour including (i) how wide is the genotypic variation in PT ? (ii) how does PT respond to short-term warming and (iii) how do photosynthesis and photoprotective functions respond over short-term warming and postwarming events? A study on 21 genotypes revealed significant genotypic differences in PT , though these were not large. We selected five genotypes with divergent PT and simulated warming events: Tmax 26/20°C (day-time highest maximum/night-time lowest maximum) (Week 1) < Tmax 33/30°C (Week 2) < Tmax 36/32°C (Week 3) followed by Tmax 26/16°C (Week 4, recovery). The PT of all genotypes remained unaltered despite strong leaf megathermy (leaf temperature > air temperature) during warming events. Though moderate warming showed genotype-specific stimulation in photosynthesis, higher warming unequivocally led to severe loss in net photosynthesis and induced higher nonphotochemical quenching. Even after a week of postwarming, photoprotective mechanisms strongly persisted. Our study points towards a conservative PT in evergreen citrus genotypes and their need for sustaining higher photoprotection during warming as well as postwarming recovery conditions.
Asunto(s)
Citrus/fisiología , Termotolerancia/fisiología , Citrus/genética , Genotipo , Calor , Fotosíntesis/fisiología , Hojas de la Planta/fisiología , Clima TropicalRESUMEN
Citrus are economically important fruit crops to which infectious diseases like citrus canker caused by Xanthomonas citri subs. citri, citrus variegated chlorosis caused by Xylella fastidiosa, "huanglongbing" associated with the presence of Candidatus liberibacter species, anthracnose caused by Colletotrichum gloeosporioides and citrus black spot caused by Phyllosticta citricarpa, impose significant losses. Control measures involve chemical treatment of orchards but often, eradication of infected plants is unavoidable. To circumvent the environmental impacts of pesticides and the socio-economic impacts of eradication, innovative antimicrobial approaches like photodynamic inactivation are being tested. There is evidence of the susceptibility of Xanthomonas citri subs. citri and C. gloeosporioides to photodynamic damage. However, the realistic assessment of perspectives for widespread application of photodynamic inactivation in the control of citrus diseases, necessarily implies that other microorganisms are also considered. This review intends to provide a critical summary of the current state of research on photodynamic inactivation of citrus pathogens and to identify some of the current limitations to the widespread use of photodynamic treatments in citrus crops.
Asunto(s)
Citrus/microbiología , Productos Agrícolas/microbiología , Fármacos Fotosensibilizantes , Enfermedades de las Plantas/microbiología , Antiinfecciosos , Citrus/fisiología , Colletotrichum/efectos de la radiación , Xanthomonas/efectos de la radiación , XylellaRESUMEN
Senescence is a gradual physiological process involving the integration of numerous internal and environmental signals. Abscisic acid (ABA) is a well-known inducer of senescence. However, the regulatory mechanisms underlying ABA-mediated senescence remain largely unknown. Here, we report that the citrus homeodomain leucine zipper I (HD-ZIP I) transcription factor CsHB5 functions as a regulator of ABA-triggered senescence. CsHB5 acts as a nucleus-localized transcriptional activator, the expression of which appeared to be closely associated with citrus senescence. Overexpression of CsHB5 in citrus calli upregulated the expression of ABA- and reactive oxygen species (ROS)-related genes, and significantly increased the content of ABA and hydrogen peroxide (H2 O2 ), whereas silencing CsHB5 in citrus calli downregulated the expression of ABA-related genes. Additionally, heterogenous overexpression of CsHB5 in Solanum lycopersicum (tomato) and Arabidopsis thaliana (Arabidopsis) leads to early leaf yellowing under dark-induced senescence conditions. Meanwhile, the levels of ABA and H2 O2 in transgenic tomatoes increased significantly and the lycopene content decreased. Transcriptome analysis of CsHB5-overexpressing citrus calli and tomato showed that CsHB5 was involved in multiple senescence-associated processes, including chlorophyll degradation, nutrient compound biosynthesis and transport, as well as ABA and ROS signal transduction. The results of yeast one-hybrid assays, electrophoretic mobility shift assays and dual luciferase assays indicated that CsHB5 directly binds to the promoters of ABA biosynthetic genes, including ß-carotene hydroxylase 1 (BCH1) and 9-cis-epoxycarotenoid dioxygenase 2 (NCED2), thereby activating their transcription. Our findings revealed that CsHB5 participates in senescence, at least partly, by directly controlling ABA accumulation. Our work provides insight into the regulatory mechanisms underlying ABA-mediated senescence.
Asunto(s)
Ácido Abscísico/metabolismo , Citrus/genética , Regulación de la Expresión Génica de las Plantas/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Factores de Transcripción/metabolismo , Arabidopsis/genética , Arabidopsis/fisiología , Clorofila/metabolismo , Citrus/fisiología , Expresión Génica , Proteínas de Homeodominio , Leucina Zippers , Solanum lycopersicum/genética , Solanum lycopersicum/fisiología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Senescencia de la Planta , Regiones Promotoras Genéticas/genética , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Factores de Transcripción/genética , Regulación hacia ArribaRESUMEN
Ethylene-responsive factors (ERFs) are plant-specific transcription factors involved in cold stress response, and raffinose is known to accumulate in plants exposed to cold. However, it remains elusive whether ERFs function in cold tolerance by modulating raffinose synthesis. Here, we identified a cold-responsive PtrERF108 from trifoliate orange (Poncirus trifoliata (L.) Raf.), a cold-tolerant plant closely related to citrus. PtrERF108 is localized in the nucleus and has transcriptional activation activity. Overexpression of PtrERF108 conferred enhanced cold tolerance of transgenic lemon, whereas virus-induced gene silencing (VIGS)-mediated knockdown of PtrERF108 in trifoliate orange greatly elevated cold sensitivity. Transcriptome profiling showed that PtrERF108 overexpression caused extensive reprogramming of genes associated with signaling transduction, physiological processes and metabolic pathways. Among them, a raffinose synthase (RafS)-encoding gene, PtrRafS, was confirmed as a direct target of PtrERF108. RafS activity and raffinose content were significantly increased in PtrERF108-overexpressing transgenic plants, but prominently decreased in the VIGS plants under cold conditions. Meanwhile, exogenous replenishment of raffinose could recover the cold tolerance of PtrERF108-silenced plants, whereas VIGS-mediated knockdown of PtrRafS resulted in cold-sensitive phenotype. Taken together, the current results demonstrate that PtrERF108 plays a positive role in cold tolerance by modulation of raffinose synthesis via regulating PtrRafS. Our findings reveal a new transcriptional module composed of ERF108-RafS underlying cold-induced raffinose accumulation in plants.
Asunto(s)
Respuesta al Choque por Frío/fisiología , Galactosiltransferasas/genética , Proteínas de Plantas/genética , Poncirus/fisiología , Rafinosa/biosíntesis , Núcleo Celular/genética , Núcleo Celular/metabolismo , Citrus/genética , Citrus/fisiología , Galactosiltransferasas/metabolismo , Regulación de la Expresión Génica de las Plantas , Silenciador del Gen , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Poncirus/efectos de los fármacos , Regiones Promotoras Genéticas , Rafinosa/genética , Rafinosa/farmacología , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Physiologically dropped immature Citrus reticulata Blanco fruits are regarded as waste and discarded in the citrus orchard but are a good source of bioactive compounds including flavonoids, antioxidants and total phenols. A study was undertaken to identify and quantify these bioactive compounds and to investigate the influence of different drying techniques, namely freeze drying and hot air oven drying, on flavonoids namely flavanone glycosides, antioxidant potential and total phenol content in immature dropped fruits of Citrus reticulata Blanco. Flavonoids were quantified in high-performance liquid chromatography (HPLC). The antioxidant activity were investigated with three assays azino-bis [3-ethylbenzthiazoline-6-sulfonic acid]) (ABTS), 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), Ferric Reducing Ability of Plasma (FRAP) and total phenol content was determined. Freeze dried samples of 12 and 14 mm size retained maximum hesperidin flavonoid content (27.03% and 27.20%) as compared to the hot air dried samples (17.99%) and retained higher phenolic content ranged from 50.54-54.19 mg GAEL-1. The antioxidant activity in freeze dried fruits was from 12.21-13.55 mM L-1 Trolox and 15.27-16.72 mM L-1 Trolox with ABTS, DPPH assay and FRAP values ranging from 7.31-9.07 mM L-1 Trolox. Significant positive correlation was found between the flavonoid hesperidin with antioxidant assays and total phenolic content (TPC). The results showed that waste citrus fruits can act as potential source of bioflavonoids, especially hesperidin, and antioxidants for pharmaceutical as well as nutraceutical industry.