Poncirus Research Articles

Poncirus trifoliata Aqueous Extract Protects Cardiomyocytes against Doxorubicin-Induced Toxicity through Upregulation of NAD(P)H Dehydrogenase Quinone Acceptor Oxidoreductase 1.

Doxorubicin (DOX), an anthracycline-based chemotherapeutic agent, is widely used to treat various types of cancer; however, prolonged treatment induces cardiomyotoxicity. Although studies have been performed to overcome DOX-induced cardiotoxicity (DICT), no effective method is currently available. This study investigated the effects and potential mechanisms of Poncirus trifoliata aqueous extract (PTA) in DICT. Changes in cell survival were assessed in H9c2 rat cardiomyocytes and MDA-MB-231 human breast cancer cells. The C57BL/6 mice were treated with DOX to induce DICT in vivo, and alterations in electrophysiological characteristics, serum biomarkers, and histological features were examined. The PTA treatment inhibited DOX-induced decrease in H9c2 cell viability but did not affect the MDA-MB-231 cell viability. Additionally, the PTA restored the abnormal heart rate, R-R interval, QT interval, and ST segment and inhibited the decrease in serum cardiac and hepatic toxicity indicators in the DICT model. Moreover, the PTA administration protected against myocardial fibrosis and apoptosis in the heart tissue of mice with DICT. PTA treatment restored DOX-induced decrease in the expression of NAD(P)H dehydrogenase quinone acceptor oxidoreductase 1 in a PTA concentration-dependent manner. In conclusion, the PTA inhibitory effect on DICT is attributable to its antioxidant properties, suggesting the potential of PTA as a phytotherapeutic agent for DICT.

Enhancing soil health and nutrient availability for Carrizo citrange (X Citroncirus sp.) through bokashi and biochar amendments: An exploration into indoor sustainable soil ecosystem management

This study investigated the efficacy of organic soil amendments: bokashi (Bok), biochar (BC), and their combination (Bok_BC) in promoting soil health, nutrient availability, and growth of Carrizo citrange (X Citroncirus sp. Rutaceae, Parentage Citrus sinensis x Poncirus trifoliata) under indoor greenhouse settings. Results indicate significant alterations in soil parameters like total carbon (C), total nitrogen (N), and C:N ratio due to Bok, BC, and Bok_BC treatments. BC treatments boosted total C, while Bok increased total N, compared to controls. A noteworthy 25 % average decrease in C:N ratio was observed with Bok and Bok_BC, nearing the optimal 24:1 C:N for microbial growth. This highlights the potential of waste by-products in balancing nutrient release to benefit soil health and plant development. Analysis of nitrite (NO2-), nitrate (NO3-), and ammonium (NH4-N) levels revealed a dynamic relationship between soil treatments and time. Bok and Bok_BC amendments combined with both fertilizer doses [700 and 1400 Electrical Conductivity, EC] showed an initial NH4-N spike (averaging 1513 and 1288 μg N/g dry, respectively), outperforming control soils (average 503 μg N/g dry). Other key elements like phosphorus, potassium, calcium, and chlorine also experienced initial surges in Bok and Bok_BC soils before declining, suggesting a gradual nutrient release. The concentration of potentially toxic elements remained mostly stable or inconclusive, warranting further exploration. Bok, BC, and Bok_BC treatments considerably influenced germination rate and plant growth. The germination rate averaged 24.2 %, 23 %, and 22.5 % for Bok, BC, and Bok_BC, compared to the 15.9 % control. Plant height increased with Bok, BC, and Bok_BC to 18.4 cm, 18.7 cm, and 16.4 cm, respectively, from the 14.8 cm control. The results remained consistent across fertilizer doses, emphasizing the soil amendments' role in bolstering soil and plant health. In summary, the research underscores the potential of carbon-based amendments like bokashi and biochar in enhancing soil health, reducing reliance on synthetic fertilizers, and fostering sustainable soil ecosystems. The insights are pivotal for advancing sustainable agriculture in indoor greenhouse settings for nursery plant production.

A C2H2-type zinc finger protein ZAT12 of Poncirus trifoliata acts downstream of CBF1 to regulate cold tolerance.

The Cys2/His2 (C2H2)-type zinc finger family has been reported to regulate multiple aspects of plant development and abiotic stress response. However, the role of C2H2-type zinc finger proteins in cold tolerance remains largely unclear. Through RNA-sequence analysis, a cold-responsive zinc finger protein, named as PtrZAT12, was identified and isolated from trifoliate orange (Poncirus trifoliata L. Raf.), a cold-hardy plant closely related to citrus. Furthermore, we found that PtrZAT12 was markedly induced by various abiotic stresses, especially cold stress. PtrZAT12 is a nuclear protein, and physiological analysis suggests that overexpression of PtrZAT12 conferred enhanced cold tolerance in transgenic tobacco (Nicotiana tabacum) plants, while knockdown of PtrZAT12 by virus-induced gene silencing (VIGS) increased the cold sensitivity of trifoliate orange and repressed expression of genes involved in stress tolerance. The promoter of PtrZAT12 harbors a DRE/CRT cis-acting element, which was verified to be specifically bound by PtrCBF1 (Poncirus trifoliata C-repeat BINDING FACTOR1). VIGS-mediated silencing of PtrCBF1 reduced the relative expression levels of PtrZAT12 and decreased the cold resistance of trifoliate orange. Based on these results, we propose that PtrZAT12 is a direct target of CBF1 and plays a positive role in modulation of cold stress tolerance. The knowledge gains new insight into a regulatory module composed of CBF1-ZAT12 in response to cold stress and advances our understanding of cold stress response in plants.

Effect of Natural Variation and Rootstock on Fruit Quality and Volatile Organic Compounds of ‘Kiyomi tangor’ (Citrus reticulata Blanco) Citrus

In this study, we compared the fruit quality and color of ‘Kiyomi’ (WT) and its mutant (MT) grafted on Ziyang xiangcheng (Cj) (WT/Cj, MT/Cj), and the MT grafted on Trifoliate orange (Pt) (MT/Pt). The differences in sugar, organic acid, flavonoids, phenols, and volatile substances of the three materials were also analyzed by high performance liquid chromatography (HPLC) and headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC–MS). The results showed significant differences in the appearance of WT/Cj, MT/Cj, and MT/Pt. MT/Pt, compared to WT/Cj, MT/Cj, had lower sugar, acid, phenol and flavonoid contents in the pulp. However, MT/Pt pulp was higher in vitamin C (VC), and the peel had significantly higher total phenol and flavonoid contents. In terms of pulp, WT/Cj had the greatest diversity of volatile organic compounds (VOCs). 4-methyl-1-pentanol was significantly higher in MT/Cj pulp, while MT/Pt pulp had a unique octanoic acid, methyl ester. VOCs were more diverse in the peels of the three materials. β-Myrcene and valencen were significantly higher in MT/Cj peels. In contrast, 16 unique VOCs were detected in MT/Pt, and D-limonene content was significantly higher than in WT/Cj and MT/Cj. The results suggest Trifoliate orange is a suitable rootstock for MT.

Comparative proteomic analysis identifies proteins associated with arbuscular mycorrhizal symbiosis in Poncirus trifoliata

Comparative proteomic analysis identifies proteins associated with arbuscular mycorrhizal symbiosis in Poncirus trifoliata

Physiological and Biochemical Evaluation of Salt Stress Tolerance in a Citrus Tetraploid Somatic Hybrid

Somatic hybridization has emerged as a valuable tool for developing novel genetic combinations in citrus breeding programs, including the creation of salt-tolerant rootstocks. In this study, the performance of a tetraploid somatic hybrid, obtained by fusing protoplasts derived from salt-tolerant Cleopatra mandarin (Citrus reshni hort. ex Tanaka) and salt-sensitive Carrizo citrange (Citrus sinensis L. Osbeck × Poncirus trifoliata L. Raf), was assessed under in vitro salt stress. Hybrid plants were characterized by leaf morphology, and ploidy level by flow cytometry and molecular markers. In vitro shoots were generated from the micropropagation of mature stem pieces of the somatic hybrid and its parents, and these were challenged by exposure to NaCl (0, 50, or 100 mM) supplemented to the media for three weeks to induce salt stress. The leaves of the somatic hybrid display intermediate morphology compared to the parental Cleopatra mandarin and Carrizo citrange rootstocks. All molecular markers successfully amplified DNA from the three cultivars; however, only 11 of 14 unequivocally confirmed somatic hybridity. The physiological and biochemical parameters, including chlorophyll content, lipid peroxidation, total phenolic compounds, antioxidants activity and proline content, were measured in the leaves. The somatic hybrid exhibited superior salt stress tolerance compared to the parent varieties, as evidenced by the reduced cellular membrane damage indicated by the lower levels of malondialdehyde and electrolyte leakage, particularly under 100 mM NaCl treatment. The somatic tetraploid hybrid also displayed higher total phenolic content than either parent, while Cleopatra mandarin exhibited the highest proline levels under 50 mm NaCl. These results demonstrate the enhanced salinity stress tolerance of the somatic hybrid compared to its parent lines, highlighting its potential as a valuable candidate for developing salt-tolerant citrus rootstocks.

IMPACT OF VARIOUS CONCENTRATIONS OF NACL ON MORPHOLOGICAL ATTRIBUTES OF DIFFERENT CITRUS ROOTSTOCKS UNDER FIELD CONDITIONS

Citrus (Poncirus trifoliata), a widely cultivated fruit crop, is known for its salt sensitivity. Salinity poses a significant challenge to citrus production across various regions worldwide, where it acts as a limiting factor. When citrus trees are irrigated with saline water, their yields are significantly reduced. The accumulation of excessive chloride ions (Cl-) and sodium ions (Na+) can lead to specific ion toxicities, further exacerbating salinity's negative impact on citrus plants. However, this issue can be minimized by selecting proper rootstocks. Six treatments were administered to the plants in a study to assess the effects of different stress levels on citrus plants. These treatments involved varying concentrations of salt stress: 0mM, 10mM, 20mM, 30mM, 60mM, and 80mM. By subjecting the plants to these different stress levels, researchers aimed to understand the response of citrus plants to increasing salinity. Water salinity presents a significant issue for citrus cultivation due to its detrimental influence on crop yields. The high salt content in irrigation water negatively affects the growth of citrus trees and induces physiological disorders. In addition to impeding growth, salinity harms plant height and root development. These combined effects of salinity contribute to the overall decline in citrus productivity. Among the various citrus rootstocks tested in the study, C-35 exhibited the most significant susceptibility to salinity stress. As the salt concentrations increased, C-35 experienced the most severe adverse effects on growth and productivity. On the other hand, Poncirus trifoliata, a commonly used rootstock, demonstrated a higher degree of tolerance to salinity. Poncirus trifoliata remained relatively unaffected even at higher salt concentrations, making it a more suitable choice for citrus cultivation in saline environments.

Serendipita indica mitigates drought-triggered oxidative burst in trifoliate orange by stimulating antioxidant defense systems.

Soil drought is detrimental to plant growth worldwide, particularly by triggering reactive oxygen species (ROS) burst. Serendipita indica (Si), a culturable root-associated endophytic fungus, can assist host plants in dealing with abiotic stresses; however, it is unknown whether and how Si impacts the drought tolerance of citrus plants. To unravel the effects and roles of Si on drought-stressed plants, trifoliate orange (Poncirus trifoliata L. Raf.; a citrus rootstock) seedlings were inoculated with Si and exposed to soil drought, and growth, gas exchange, ROS levels, antioxidant defense systems, and expression of genes encoding antioxidant enzymes and fatty acid desaturases in leaves were measured. Soil drought suppressed plant biomass, whereas Si inoculation significantly increased plant biomass (10.29%-22.47%) and shoot/root ratio (21.78%-24.68%) under ample water and drought conditions, accompanied by improved net photosynthetic rate (105.71%), water use efficiency (115.29%), chlorophyll index (55.34%), and nitrogen balance index (63.84%) by Si inoculation under soil drought. Soil drought triggered an increase in leaf hydrogen peroxide and superoxide anion levels, while Si inoculation significantly reduced these ROS levels under soil drought, resulting in lower membrane lipid peroxidation with respect to malondialdehyde changes. Furthermore, Si-inoculated seedlings under soil drought had distinctly higher levels of ascorbate and glutathione, as well as catalase, peroxidase, and glutathione peroxidase activities, compared with no-Si-inoculated seedlings. Si inoculation increased the expression of leaf PtFAD2, PtFAD6, PtΔ9, PtΔ15, PtFe-SOD, PtCu/Zn-SOD, PtPOD, and PtCAT1 genes under both ample water and soil drought conditions. Overall, Si-inoculated trifoliate orange plants maintained a low oxidative burst in leaves under drought, which was associated with stimulation of antioxidant defense systems. Therefore, Si has great potential as a biostimulant in enhancing drought tolerance in plants, particularly citrus.

The Antioxidant and Anti-Complementary Activities of Crude Polysaccharides from Trifoliate Orange (Poncirus trifoliate) Seeds.

In this study, I extracted the crude polysaccharides from trifoliate orange (Poncirus trifoliate) seeds, known as TSCP, using water extraction and ethanol precipitation. The monosaccharide composition of TSCP was in the following order: arabinose (28.28 mol%)> galactose (16.76 mol%)> galacturonic acid+glucuronic acid (13.6 mol%)> glucose (12.45 mol%)> rhamnose (4.18 mol%)> mannose (0.57 mol%)> fucose (0.32 mol%). Its total polyphenol contents were 28.66 and 70.96 μg/mL at 1 and 10 mg/mL, respectively (P<0.01). Further, the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity of 10 mg/mL TSCP (31.67%) was higher than that of 1 mg/mL TSCP (8.07%; P<0.01) and also higher than its 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (11.97%) at the same concentration (10 mg/mL; P<0.01). The anti-complementary property of TSCP increased in a concentration-dependent manner (P<0.001), and at 1,000 μg/mL, it was comparable (61.36%) to the positive control (60%) consisting of polysaccharide-K. In conclusion, TSCP might be a potential immune modulator.

Introduction pages, NBHA-CN 51(3), 2023

The papers published in Notulae Botanicae Horti Agrobotanici Cluj-Napoca, Issue 3, Volume 51, 2023 represent new exciting research in different topics of life science, respectively in plant science, horticulture, agronomy, crop science, and forestry. Among the interesting articles we invite you to find news about: Comparison of the potential of extracts from the flower, fruit pulp, and seed of Cassia fistula L. on MCF-7 breast cancer cell growth and cell migration; Evaluation of anticancer activity of Plumbago indica root extract on cervical HeLa cancer cells; Metabolome and transcriptome analyses reveal the colouring mechanism of red honeysuckle (Lonicera japonica Thunb.); Ethylene modulates root growth and mineral nutrients levels in trifoliate orange through the auxin-signaling pathway; Green nanotechnology for plant bacterial diseases management in cereal crops: a review on metal-based nanoparticles; Physiological responses of coffee (Coffea arabica L.) plants to biochar application under water deficit conditions; Gene mining and functional analysis related to maize (Zea mays L.) seed size; Phytochemical properties, antioxidant potential and fatty acids profiling of three dragon fruit species grown under sub-tropical climate; Chemical composition and anti-inflammatory effect of Phellodendron amurense Rupr. stem bark extract; Ecotype effects on photosynthesis performance using A/PFFD among Pinus nigra Arn.; Assessment of herbaceous vegetation classification using orthophotos produced from the image acquired with unmanned aerial systems.

Combined metabolomic and transcriptomic analysis reveals variation in phenolic acids and regulatory networks in the peel of sweet orange 'Newhall' (C. sinensis) after grafting onto two different rootstocks

Grafting plays a significant role in the biosynthesis of secondary metabolites in plants. In our study, we examined the impact of grafting on the phenolic acid biosynthesis in sweet orange 'Newhall' peel. Through transcriptome-metabolome analysis, we observed notable differences in total phenolic acid content between the two rootstocks [Poncirus trifoliata (ct) and C. junos Siebold ex Tanaka (cj)]. Total phenolic acid content reached its highest level during the later fruit expansion stage (347.27 µg · g−1 FW) in ct. The metabolome analysis identified 270 phenolic acids, out of which 24 were found to be influenced by rootstock grafting. The transcriptome analysis identified 8,562 differentially expressed genes (DEGs). Further analysis using KEGG and GO enrichment revealed that 36 structural genes enriched in the phenolic acid biosynthesis pathway and showed differential expression in ct and cj. Co-expression analysis indicated that the AP2/ERF-ERF and MYB family members exhibited the highest number of differences in ct and cj, suggesting their potential involvement in phenolic acid biosynthesis. Our findings provide insights into the differential accumulation mechanism of phenolic acids in sweet orange 'Newhall' peel after grafting with different rootstocks. These results enhance our understanding of the regulatory mechanisms underlying phenolic acid biosynthesis and offer valuable information for the effective utilization of citrus peel.

Ameliorating effect of Citrus trifoliata L. fruits extract on motor incoordination, neurodegeneration and oxidative stress in Parkinson’s disease model

ABSTRACT Background Citrus trifoliate fruit (also known as Trifoliate orange) is one of the commercially-cultivated Citrus genus of plants belonging to the Rutaceae family. It has been traditionally-utilized in treatment of neurodegenerative disorders. However, the scientific evidence verifying this utilization needs further elucidation. Aim of the study Characterization of the bioactive constituents of C. trifoliata L. fruits extract and evaluating its effect on Parkinson’s disease (PD) model. Material and methods Rats were classified into 5 groups; control, PD, PD-treated by L-dopa/Carpidopa and PD-treated by oral Citrus trifoliata L. fruits extract (50 and 100 mg/kg). Deterioration in brain functions was evaluated through an in vivo open field, grid and catalepsy tests. The study also assessed the striatal neurotransmitters, oxidative stress markers and histopathological changes. Results Citrus trifoliata L. fruit extract has revealed motor improvement comparable to L-dopa and carbidopa. It has also effectively-improved oxidative stress via reduction of striatal malondialdehyde & nitric oxide along with replenishment of the striatal glutathione and superoxide dismutase. The extract caused significant reduction of the striatal myeloperoxidase activity and restoration of dopamine, γ-amino butyric acid (GABA), and acetylcholinesterase. This effect was further confirmed by amelioration of neuronal apoptosis, microgliosis and peri-neuronal vacuolation. Metabolite profiling revealed 40 constituents, with flavonoids representing the main identified class. Conclusion The neuro-protective effect of Citrus trifoliata extract was achieved through the antioxidant and anti-inflammatory activities of its flavonoids, particularly hesperidin and naringin. This neuro-protective effect was evident at the behavioral, histological and neurotransmitter levels.

Ethylene modulates root growth and mineral nutrients levels in trifoliate orange through the auxin-signaling pathway

Roots are fundamental to plant growth, with their development being profoundly influenced by water, mineral nutrients, and notably, plant hormones. Citrus, a leading horticultural crop cultivated globally, holds immense economic value. This study tried to investigate the roles and mechanisms effect of foliar spray 1.0 μmol/L ethephon (ETH) of ethylene and 5.0 μmol/L AgNO3 (an ethylene inhibitor) its inhibitor with auxin on root growth and mineral nutrients content levels in trifoliate orange under sand culture conditions for 45 days. Our findings indicate that ethephon treatment significantly boosted the number of lateral roots, root fresh weight, and enhanced the mineral nutrient content (N, P, K, B, Ca, Mg, Fe, Mn, and Zn, with the exception of Cu) in both root and shoot tissues. In contrast, the inhibitors notably reduced the length of lateral roots and decreased the content of P, and Fe in both root and shoot, whilst elevating Zn contents. Furthermore, ETH incited the up-regulation of auxin biosynthesis and transportation genes, leading to an increased endogenous auxin content in the lateral root-hair region. whereas, the inhibitors produced an opposite effect. We thus conclude that ethylene modulates auxin transportation and biosynthesis. This activity stimulates the auxin signaling pathway locally-specifically, in the root hair zone in the lateral root, which in turn, regulates root growth and mineral nutrient content.

Insight into Physiological and Biochemical Determinants of Salt Stress Tolerance in Tetraploid Citrus.

Citrus are classified as salt-sensitive crops. However, a large diversity has been observed regarding the trends of tolerance among citrus. In the present article, physiological and biochemical studies of salt stress tolerance were carried out according to the level of polyploidy of different citrus genotypes. We particularly investigated the impact of tetraploidy in trifoliate orange (Poncirus trifoliata (L.) Raf.) (PO4x) and Cleopatra mandarin (Citrus reshni Hort. Ex Tan.) (CL4x) on the tolerance to salt stress compared to their respective diploids (PO2x and CL2x). Physiological parameters such as gas exchange, ions contents in leaves and roots were analyzed. Roots and leaves samples were collected to measure polyphenol, malondialdehyde (MDA), ascorbate and H2O2 contents but also to measure the activities of enzymes involved in the detoxification of active oxygen species (ROS). Under control conditions, the interaction between genotype and ploidy allowed to discriminate different behavior in terms of photosynthetic and antioxidant capacities. These results were significantly altered when salt stress was applied when salt stress was applied. Contrary to the most sensitive genotype, that is to say the diploid trifoliate orange PO2x, PO4x was able to maintain photosynthetic activity under salt stress and had better antioxidant capacities. The same observation was made regarding the CL4x genotype known to be more tolerant to salt stress. Our results showed that tetraploidy may be a factor that could enhance salt stress tolerance in citrus.

Research progress on the anti-tumor effect of Naringin.

Naringin is a kind of natural dihydro flavone, which mainly exists in citrus fruits of the Rutaceae family, as well as traditional Chinese medicines such as trifoliate orange, fingered citron, exocarpium citri grandis, and rhizoma dynamite. Modern pharmacological studies have shown that Naringin has excellent anti-tumor activity. Through reviewing the relevant literature at home and abroad in recent years, we summarized the pharmacological mechanism of Naringin to play an anti-cancer role in blocking tumor cell cycle, inhibiting tumor cell proliferation, inducing tumor cell apoptosis, inhibiting tumor cell invasion and metastasis, inducing tumor cell autophagy, reversing tumor cell drug resistance and enhancing chemotherapeutic drug sensitivity, as well as anti-inflammatory to prevent canceration, alleviate Adverse drug reaction of chemotherapy, activate and strengthen immunity, It provides theoretical basis and reference basis for further exploring the anticancer potential of Naringin and its further development and utilization.

Texas microfungi: a new species of Corynesporopsis (Xylariales) associated with the trifoliate orange

Corynesporopsis ponciri sp. nov. is described from dead twigs, branches, and thorns still attached to trees of Poncirus trifoliata, the trifoliate orange, in Texas, U.S.A. The fungus is characterized by black, punctiform at first, later caespitose and hairy colonies forming large superficial patches covering almost entirely the surface of colonized parts of the host, unbranched conidiophores, solitary or fasciculate and arising from substomatal, compact stromata in loose to dense fascicles and bearing monotretic, mostly determinate, rarely percurrent conidiogenous cells. Conidia are ellipsoidal or fusiform to naviculiform, smooth or verruculose, brown, often with the apical cell light brown, euseptate, with 2, rarely 1 or 3 septa, usually with an associated dark band and forming short, unbranched, acropetal chains of up to four conidia. Multilocus phylogenetic analyses suggested that C. ponciri belongs within the Xylariales and forms a strongly supported monophyletic lineage sister to members of Vamsapriya in Vamsapriyaceae. Corynesporopsis is revealed as polyphyletic based on this and the other two species with available molecular data, C. iberica and C. acaciae. The newly described fungus is compared with morphologically similar species of Corynesporopsis and Heteroconium having 1- or 2-septate conidia and phylogenetic affinities of these polyphyletic anamorphic genera are discussed. The mode and successful colonization of dead parts of the host with conidiophores arising from substomatal stromata suggest that C. ponciri may have a putative endophytic stage and it is capable of switching to a saprobic lifestyle upon senescence or death of the host.

Genome-Wide Identification and Characterization of the Biosynthesis of the Polyamine Gene Family in Citrus unshiu.

Polyamines (PAs) contribute to diverse plant processes, environmental interaction, and stress responses. In citrus, the mechanism underlying the biosynthesis of polyamines is poorly understood. The present study aims to identify the biosynthesis of PA gene family members in satsuma mandarin (Citrus unshiu) and investigate their response against various stresses. The identified biosynthesis of PA genes in C. unshiu showed clustering in six groups, i.e., SPMS, SPDS, ACL5, ADC, ODC, and SAMDC. Syntenic analysis revealed that segmental duplication was prevalent among the biosynthesis of PA genes compared to tandem duplication. Thus, it might be the main reason for diversity in the gene family in C. unshiu. Almost all biosynthesis of PA gene family members in C. unshiu showed syntenic blocks in the genome of Arabidopsis, Citrus sinensis, Poncirus trifoliata, and Citrus reticulata. Analysis of Cis-regulatory elements (CREs) indicated the occurrence of hormones, light, defense, and environmental stress responses as well as the development and other plant mechanisms-related elements in the upstream sequence of the biosynthesis of PA genes. Expression profiling revealed that the biosynthesis of PA gene expression modulates in different organs during various developmental stages and stress in C. unshiu. This information will provide a deep understanding of genomic information and its expression in multiple tissues to better understand its potential application in functional genomics.

Transcriptome and UPLC-MS/MS reveal mechanisms of amino acid biosynthesis in sweet orange 'Newhall' after different rootstocks grafting.

Sweet orange 'Newhall' (C. sinensis) is a popular fruit in high demand all over the world. Its peel and pulp are rich in a variety of nutrients and are widely used in catering, medicine, food and other industries. Grafting is commonly practiced in citrus production. Different rootstock types directly affect the fruit quality and nutritional flavor of citrus. However, the studies on citrus metabolites by grafting with different rootstocks are very limited, especially for amino acids (AAs). The preliminary test showed that there were significant differences in total amino acid content of two rootstocks (Poncirus trifoliata (CT) and C. junos Siebold ex Tanaka (CJ)) after grafting, and total amino acid content in the peel was higher than flesh. However, the molecular mechanism affecting amino acid differential accumulation remains unclear. Therefore, this study selected peel as the experimental material to reveal the amino acid components and differential accumulation mechanism of sweet orange 'Newhall' grafted with different rootstocks through combined transcriptome and metabolome analysis. Metabolome analysis identified 110 amino acids (AAs) and their derivatives in sweet orange 'Newhall' peels, with L-valine being the most abundant. L-asparagine was observed to be affected by both developmental periods and rootstock grafting. Weighted gene co-expression network analysis (WGCNA) combined with Redundancy Analysis (RDA) revealed eight hub structural genes and 41 transcription factors (TFs) that significantly influenced amino acid biosynthesis in sweet orange 'Newhall' peels. Our findings further highlight the significance of rootstock selection in enhancing the nutritional value of citrus fruits and might contribute to the development of functional citrus foods and nutritional amino acid supplements.

Effect of Magnesium on Mineral Nutrition, Chlorophyll, Proline and Carbohydrate Concentrations of Sweet Orange (Citrus sinensis cv. Newhall) Plants

Magnesium is an essential nutrient for the growth and development of plants. Its deficiency is becoming a growing concern in many citrus orchards worldwide, adversely affecting numerous functions in plants and limiting their productivity and quality. Three-year-old orange (Citrus sinensis cv. Newhall) plants grafted on Swingle citrumelo (C. paradisi Macf. × Poncirus trifoliata L.) rootstock were irrigated for 63 days with nutrient solutions containing 0, 12, 24, 48, 96, and 192 mg Mg L−1. Thereafter, Mg deficiency-induced changes were investigated in leaf chlorophyll concentration and fluorescence, and in proline and carbohydrate concentration in leaves and roots, as well as in the nutritional status of leaves, stems, and roots. Magnesium concentration in the nutrient solution was positively correlated with the concentration of Mg (leaves, stems, roots), Ca (rootstock’s stem), K (roots), and Fe (leaves, rootstock’s stem), as well as with the total Mg absorption. However, Mg concentration in the nutrient solution was negatively correlated with the concentration of Mn (rootstock’s stem, roots), Ca (leaves, scion’s stems, roots), and Fe (roots), as well as with the total absorption of Mn, Fe, P, K, and Ca. The lower values of the chlorophyll fluorescence parameters were observed by the effect of the highest concentration of Mg. As far as leaf chlorophyll concentration and carbohydrate and proline content of leaves and roots, they were not affected by the Mg treatments. Concluding, this research highlights the significance of Mg management in citrus farming, offering insights into increasing Mg concentrations, understanding root mechanisms in Mg absorption, and suggesting the benefits of fertilization to address Mg deficiency. It emphasizes the importance of careful Mg fertilizer dosages, considering other nutrient interactions, and provides valuable guidance for optimizing Mg nutrition and overall nutrient management in sweet orange trees.

Phenylalanine-mediated changes in the soil bacterial community promote nitrogen cycling and plant growth

Soil amino acids (AAs) are the most active components of soil N, which can be mineralized or absorbed by bacteria as N and C sources. We hypothesized that exogenous AAs could regulate the bacterial community and affect soil N cycling, and the effect sizes could vary depending on individual AAs. Here, we applied feather (keratin)-based compost rich in AAs to Poncirus trifoliata (L.) to evaluate the regulation of bacterial community by AAs; furthermore, we applied six individual AAs to test their effects. The compost significantly increased soil hydrolysable AA content, ammonia monooxygenase gene abundance, and plant growth and changed bacterial community structure. Redundancy analysis revealed that the effects of AAs on the bacterial community composition were greater than those of soil chemical properties, and phenylalanine (Phe) was the most effective among thirteen individual AAs. When applied individually, Phe caused the greatest increase in N cycling-related enzyme activity and plant growth and most significantly altered the bacterial community structure among the six exogenous AAs. Notably, Phe significantly increased the relative abundances of Burkholderia-Caballeronia-Paraburkholderia, Azospirillum, Cupriavidus, and Achromobacter, whose abundances were significantly positively correlated with plant biomass, and significantly reduced the relative abundances of Arachidicoccus, Pseudopedobacter, Sphingobacterium, and Paenibacillus, whose abundances were significantly negatively correlated with plant biomass. We demonstrate that soil AAs strongly shape the bacterial community. Particularly, Phe enhances N cycling and plant growth by increasing the potentially beneficial bacterial taxa and inhibiting the potentially harmful bacterial taxa, which needs further validation.