Citrus Junos Research Articles

Identification of CjSPL gene family in Ziyang Xiangcheng rootstock and expression pattern analysis

Squamosa promoter binding protein-like (SPL) family is a group of important transcription factors involved in the regulation of plant growth and development and the response to environmental stress, but there are few studies in perennial fruit trees such as citrus. In this study, Ziyang Xiangcheng (Citrus junos Sib.ex Tanaka), an important rootstock of Citrus, was used as the material for analysis. Based on plantTFDB transcription factor database and sweet orange genome database, 15 SPL family members were genome-widely identified and cloned from Ziyang Xiangcheng, and named CjSPL1-CjSPL15. Sequence analysis showed that the open reading frame (ORF) length of CjSPLs ranged from 393 bp to 2 865 bp, encoding 130-954 amino acids. Phylogenetic tree divided 15 CjSPLs into 9 subfamilies. Gene structure and conserved domain analysis predicted 20 different conserved motifs and SBP basic domains. Analysis of cis-acting promoter elements predicted 20 different promoter elements, including those related to plant growth and development, abiotic stress and secondary metabolites. The expression patterns of CjSPLs under drought, salt and low temperature stresses were analyzed by real-time fluorescence quantitative PCR (qRT-PCR), and many CjSPLs were significantly up-regulated after stress treatment. This study provides a reference for further study on the function of SPL family transcription factors in citrus and other fruit trees.

Molecular and biological characterization of a novel citrus tristeza virus isolate that causes severe symptoms in Citrus junos cv. Ziyangxiangcheng.

The complete genomic sequence of a novel citrus tristeza virus (CTV) isolate, CT91-A1, from Orah tangor grafted on Citrus junos cv. Ziyangxiangcheng rootstock in China was determined by transcriptome sequencing. Sequence alignments showed that isolate CT91-A1 shared 83.3 to 95.5% nucleotide sequence identity with extant CTV genotypes at the whole-genome level, with the highest similarity to the S1 genotype. Phylogenetic analysis revealed that CT91-A1 clustered in a unique subclade with the S1 genotype. Isolate CT91-A1 induced severe stem pitting in Mexican lime and C. junos cv. Ziyangxiangcheng and moderate stem pitting in Guanximiyou pummelo and Duncan grapefruit. It was successfully transmitted by Aphis citricidus, and it can potentially cause significant damage to the citrus industry in China.

Effects of enzymatic treatments on yuzu (<i>Citrus junos</i>) juice residue under low temperature by adding small amounts of enzyme solutions on paste yields, and their volatile compositions

Effects of enzymatic treatments on yuzu (<i>Citrus junos</i>) juice residue under low temperature by adding small amounts of enzyme solutions on paste yields, and their volatile compositions

Skin Improvement with Antioxidant Effect of Yuja (Citrus junos) Peel Fractions: Wrinkles, Moisturizing, and Whitening.

Yuja (Citrus junos) has been cultivated and used for food and medicinal purposes in China and Korea. Its antioxidant, anti-wrinkle, moisturizing, and whitening effects were evaluated in HaCaT, HDF, and B16F10 cells. UVB has been known to cause cellular stress and the production of reactive oxygen species (ROS). Ambivalence of oxidative stress has been reported; however, excessive levels of ROS contribute to skin aging through the loss of elasticity and collagen fibers of connective tissue in the dermis. Skin aging is one of the biological processes that is affected by various factors, including UVB. Pro-Collagen I and hyaluronic acid contents were measured in UVB-irradiated HaCaT and HDF cells to evaluate the anti-wrinkle and moisturizing effects of Yuja-peel (YJP) fractions in -EA (ethyl acetate), -Hex (hexane), and -BuOH (butanol). The expression of matrix metalloproteinases (MMPs) involved in collagen degradation was confirmed to be inhibited by YJP fractions at both the protein and mRNA levels. Filaggrin and serine palmitoyltransferase (SPT), which are moisturizing factors, were induced by YJP fractions. B16F10 cells were treated with α-MSH to induce hyperpigmentation, and then the whitening efficacy of YJP fractions was verified by observing a decrease in melanin content. Overall, our results contribute to the development of various novel skin-improving cosmetics and pharmaceuticals with YJP fractions as active ingredients.

Citrus junos Tanaka Peel Extract Ameliorates HDM-Induced Lung Inflammation and Immune Responses In Vivo

In the wake of the COVID-19 pandemic, lung disorders have become a major health concern for humans. Allergic asthma is the most prevalent form of asthma, and its treatments target the inflammation process. Despite significant developments in the diagnosis and management of allergic asthma, side effects are a major concern. Additionally, its extreme heterogeneity impedes the efficacy of the majority of treatments. Thus, newer, safer therapeutic substances, such as natural products, are desired. Citrus junos Tanaka has traditionally been utilized as an anti-inflammatory, sedative, antipyretic, and antitoxic substance. In this study, the protective effects of Citrus junos Tanaka peel extract (B215) against lung inflammation were examined, and efforts were made to understand the underlying protective mechanism using an HDM-induced lung inflammation murine model. The administration of B215 reduced immune cell infiltration in the lungs, plasma IgE levels, airway resistance, mucus hypersecretions, and cytokine production. These favorable effects alleviated HDM-induced lung inflammation by modulating the NF-κB signaling pathway. Hence, B215 might be a promising functional food to treat lung inflammation without adverse effects.

Comparative Transcriptomic Analysis of Inarching Invigorating Rootstock onto Incompatible Grafts in Citrus

Grafting is a technique that is widely used in citrus production. Graft incompatibility often occurs in the orchard. Inarching can effectively improve the vigor of incompatible grafts, but its mechanism remains poorly understood. Our previous studies investigated the scion-rootstock interaction of citrus and highlighted the role of hormonal balance and genes in abscisic acid biosynthesis regulation. To further elucidate the mechanism of inarched grafts rejuvenation, Hm/Pt combination (Citrus maxima (Burm.) Merrill cv. 'Hongmian miyou' grafted onto Poncirus trifoliata) were inarched with 'Pujiang Xiangcheng' (a novel citrus rootstock cultivar recently selected from wild Citrus junos populations), and comprehensive analysis was performed to compare the inarched grafts and controls. Compared with incompatible grafts, the results revealed that inarching could recover the leaf metabolism balance, including reducing starch content, increasing chlorophyll content and restoring the cell structure. Additionally, our results corroborated that hormonal balance and hormone-related genes played a central role in inarching compatibility. Furthermore, the roles of Hsf4, ERF1, NCED3 and PYL were highlighted, and a model for explaining inarched grafts recovery invigoration was proposed. This study shed light on the mechanism of inarching regulation tree vigor and offered deep insights into the scion-rootstock interaction in citrus.

A Study of Scion Phenotypes in Pummelo Grafted onto a New Citrus Rootstock Citrus junos ‘Pujiang Xiangcheng’

Citrus is one of the most important fruit crops in the world. Citrus junos Siebold ex Tanaka cv. Pujiang Xiangcheng is a new rootstock with a good grafting affinity and multiple abiotic stress tolerance. In this study, the effects of ‘Pujiang Xiangcheng’ and two commonly used rootstocks for Citrus maxima (Burm.) Merrill cv. Guanxi Miyou, considering their growth, photosynthetic performance, phytohormones and fruit quality, were evaluated. There was little difference between three graft combinations in the height and diameter of the scions. However, the rootstock ‘Pujiang Xiangcheng’ can cause an up-regulated photosynthetic capacity. In addition, ‘Guanxi Miyou’ on ‘Pujiang Xiangcheng’ had the highest total sugar (TS) content (129.28 mg·g−1) and the lowest titratable acid (TA) content (0.78 g·100 mL−1) in 2020 and the highest TSS (11.84%) in 2021. There was no significant difference in the TSS-to-acid ratio between the fruit of ‘Guanxi Miyou’ on trifoliate orange (13.41) and ‘Pujiang Xiangcheng’ (15.68), but it was significantly higher than that of ‘Guanxi Miyou’ on sour pummelo (11.61). Moreover, the comprehensive fruit quality of ‘Guanxi Miyou’ on ‘Pujiang Xiangcheng’ was better than on sour pummelo and trifoliate orange based on two-year statistics. We suggest that ‘Pujiang Xiangcheng’ might be the appropriate rootstock of ‘Guanxi Miyou’ in production compared with sour pummelo and trifoliate orange.

Identification of Putative Parental Species of Yuzu (Citrus junos Sieb. Ex Tanaka) by Comparative Analyses of Variations in Chloroplast Genomes and Nuclear Single-copy Genes of Citrus

Identification of Putative Parental Species of Yuzu (Citrus junos Sieb. Ex Tanaka) by Comparative Analyses of Variations in Chloroplast Genomes and Nuclear Single-copy Genes of Citrus

Nomilin from Yuzu Seed Has In Vitro Antioxidant Activity and Downregulates Melanogenesis in B16F10 Melanoma Cells through the PKA/CREB Signaling Pathway.

Yuzu (Citrus junos) is a citrus plant native to Asian countries, including Korea, Japan, and China. Yuzu peel and seed contain abundant vitamin C, citric acid, and polyphenols. Although the antioxidative and antimelanogenic activities of other citrus fruits and yuzu extract have been reported, the tyrosinase inhibitory activity of the limonoid aglycone contained in yuzu seed extract is unknown. We separated yuzu seeds into the husk, shell, and meal and evaluated antioxidant activity of each. The limonoid glucoside fraction of the husk identified nomilin, a novel tyrosinase inhibitor. We performed tyrosinase inhibitory activity and noncompetitive inhibition assays and docking studies to determine nomilin binding sites. Furthermore, we evaluated the antioxidative mechanism and antimelanogenic activity of nomilin in B16F10 melanoma cells. The concentration of nomilin that did not show toxicity was <100 µg/mL. Nomilin suppressed protein expression of TYR, TRP-1, TRP-2, and microphthalmia-associated transcription factor (MITF) in a concentration-dependent manner. Nomilin significantly reduced the levels of p-CREB and p-PKA at the protein level and decreased the levels of skin-whitening-related factors MITF, tyrosinase, TRP-1, and TRP-2 at the mRNA level in a concentration-dependent manner. Thus, nomilin from yuzu seed husk can be used as a skin-whitening agent in cosmetics.

Metabolomic Analysis Revealed Distinct Physiological Responses of Leaves and Roots to Huanglongbing in a Citrus Rootstock.

Huanglongbing (HLB) is an obstinate disease in the citrus industry. No resistant citrus resources were currently available, but various degrees of Huanglongbing tolerance exist in different germplasm. Citrus junos is emerging as one of the popular rootstocks widely used in the citrus production. However, its responses to the HLB causal agent, Candidatus Liberibacter asiaticus (CLas), were still elusive. In the current study, we investigated the physiological, anatomical, and metabolomic responses of a C. junos rootstock ‘Pujiang Xiangcheng’ by a controlled CLas grafting inoculation. The summer flushes and roots were impaired at 15 weeks after inoculation, although typical leaf symptomatic phenotypes were not obvious. The chlorophyll pigments and the photosynthetic rate were compromised. The phloem sieve tubes were still working, despite the fact that the callose was deposited and the starch granules were accumulated in the phloem cells. A wide, targeted metabolomic analysis was carried out to explore the systematic alterations of the metabolites at this early stage of infection in the leaves and root system. The differentially accumulated metabolites in the CLas-affected leaves and roots compared with the mock-inoculation control tissues revealed that distinct responses were obvious. Besides the commonly observed alteration of sugar and amino acids, the active break down of starch in the roots was discovered. The different types of fatty acids were altered in the two tissues, with a more pronounced content decline in the roots. Our results not only provided fundamental knowledge about the response of the C. junos rootstock to the HLB disease, but also presented new insights into the host–pathogen interaction in the early stages.

Complete Genome Sequence of Citrus Vein Enation Virus Identified from a Korean Yuja Tree.

ABSTRACTWe determined the complete genome sequence of the citrus vein enation virus (CVEV) collected from a Korean yuja tree (Citrus junos). The CVEV genome has 5,983 nucleotides, showing 97.3 to 98.6% identity with complete genomic sequences of other CVEV isolates, with the highest similarity being to the isolate PCJ.

Ameliorative Effect of Citrus junos Tanaka Waste (By-Product) Water Extract on Particulate Matter 10-Induced Lung Damage.

Citrus junos Tanaka (CJ)-related products are well-accepted by consumers worldwide; thus, they generate huge amounts of waste (peel, pulp, and seed) through CJ processing. Although some CJ by-products (CJBs) are recycled, their use is limited owing to the limited understanding of their nutritional and economic value. The exposure to particulate matter (PM) increases the risk of respiratory diseases. In this study, we investigated the ameliorative effects of CJB extracts (100, 200 mg/kg/day, 7 days) on PM10-induced (10 mg/kg, intranasal, 6 h) lung damage in BALB/c mice. Cell type-specific signaling pathways are examined using the A549 (PM10, 200 μg/mL, 6 h) and RAW264.7 (LPS, 100 ng/mL, 6 h) cell lines. The CJB extracts significantly attenuated PM10-induced pulmonary damage and inflammatory cell infiltration in a mouse model. The essential protein markers in inflammatory signaling pathways, such as AKT, ERK, JNK, and NF-κB for PM10-induced phosphorylation, were dramatically reduced by CJB extract treatment in both the mouse and cell models. Furthermore, the CJB extracts reduced the production of reactive oxygen species and nitric oxide in a dose-dependent manner in the cells. Comprehensively, the CJB extracts were effective in reducing PM10-induced lung injuries by suppressing pulmonary inflammation, potentially due to their anti-inflammatory and antioxidant properties.

Citrus junos Tanaka Peel Extract and Its Bioactive Naringin Reduce Fine Dust-Induced Respiratory Injury Markers in BALB/c Male Mice.

Particulate matter (PM) 10 refers to fine dust with a diameter of less than 10 µm and induces apoptosis and inflammatory responses through oxidative stress. Citrus junos Tanaka is a citrus fruit and contains bioactive flavonoids including naringin. In the present study, we aimed to identify the preventive effect of Citrus junos Tanaka peel extract (CPE) against PM10-induced lung injury. As a proof of concept, NCI-H460 cells were treated with CPE (800 μg/mL, 12 h) in conjunction with PM10 to examine intracellular antioxidative capacity in the pulmonary system. In an in vivo model, male BALB/c mice (n = 8/group) were randomly assigned into five groups: NEG (saline-treated), POS (PM10 only), NAR (PM10 + naringin, 100 mg/kg), CPL (PM10 + CPE low, 100 mg/kg), and CPH (PM10 + CPE high, 400 mg/kg). Intervention groups received dietary supplementations for 7 days followed by PM10 exposure (100 mg/kg, intranasal instillation). Compared to the NEG, the CPE decreased to 22% of the ROS generation and significantly increased cell viability in vitro. The histological assessments confirmed that pulmonary damages were alleviated in the PM10 + CPL group compared to the POS. Pro-inflammatory cytokines and NF-κB/apoptosis signaling-related markers were decreased in the PM10 + CPL group compared to the POS. These results indicated that CPE showed promising efficacy in preventing pulmonary injuries in vivo. Such protection can be explained by the anti-oxidative capacity of CPE, likely due to its bioactives, including naringin (7.74 mg/g CPE). Follow-up human intervention, as well as population-level studies, will further shed light on the preventive efficacy of CPE against pulmonary damage in humans.

Effect of essential oils of yuzu (Citrus junos Sieb. ex Tanaka) and its aroma components on postharvest quality in strawberry fruit

Effect of essential oils of yuzu (<i>Citrus junos</i> Sieb. ex Tanaka) and its aroma components on postharvest quality in strawberry fruit

Beneficial Effects of a Combination of Curcuma longa L. and Citrus junos Against Beta-Amyloid Peptide-Induced Neurodegeneration in Mice.

Both Curcuma longa (CL) and Citrus junos Tanaka (CJ) have been used to treat various diseases due to their anti-inflammatory and antioxidative stress activities. In this study, we investigated the ameliorative effect of the combination of CL extract and CJ extract (CCC) against beta-amyloid (Aβ) peptide-induced neurological damage. CCC prevented neurocytotoxicity in vitro. In addition, it was confirmed that abnormal alternation behavior and memory impairment caused by Aβ peptide were reversed by treatment with CCC. Furthermore, CCC treatment led to recovery of the cholinergic system and reactive oxygen species (ROS) oxidative damage defense system. CCC induced expressions of cyclic-adenosine monophosphate response element-binding protein (CREB)-responsive element-binding protein and brain-derived neurotrophic factor were confirmed as was the significantly improved processing in the hippocampus of the mouse Aβ peptides. Accordingly, these results suggest that CCC can prevent and/or reverse neurocytotoxicity and cognitive deficits in a mouse model of Alzheimer's disease.

Enantioselective synthesis of 6-methyloctanal and 8-methyldecanal, the characteristic aroma components in yuzu Citrus junos, and the analysis of their enantiomeric compositions in yuzu essential oil.

6-Methyloctanal and 8-methyldecanal are the characteristic aroma components of yuzu Citrus junos. However, their absolute configurations and enantiomeric compositions in yuzu essential oil have not been analyzed. A concise enantioselective synthesis of both aldehydes was successfully carried out to determine their absolute configurations and enantiomeric compositions. Both aldehydes in yuzu essential oil were found to be (S)-form with high enantiomeric excess.

Application of integrative physiological approach to evaluate human physiological responses to the inhalation of essential oils of Japanese citrus fruits iyokan (Citrus iyo) and yuzu (Citrus junos).

This study investigated the effects of essential oil odors from Japanese citrus fruits, iyokan (Citrus iyo) and yuzu (Citrus junos), on human psychology and both the autonomic and central nervous systems. The inhalation of both essential oils significantly increased miosis rate and fingertip temperature and could induce parasympathetic dominance by suppressing sympathetic nerve activity. Oxyhemoglobin concentration in the prefrontal cortex increased after the inhalation of yuzu essential oil and decreased after the inhalation of iyokan essential oil. Subjectively, the inhalation of both essential oils reduced the feelings of fatigue and improved the feelings of refreshment, suggesting that the effect of autonomic nervous activity might involve in these psychological changes directly. Moreover, we observed that task performance improved after inhaling yuzu essential oil, which may be due to the increase in oxyhemoglobin concentration in the prefrontal cortex.

A Narrative Review of Human Clinical Trials on the Impact of Phenolic-Rich Plant Extracts on Prediabetes and Its Subgroups.

Phenolic-rich plant extracts have been demonstrated to improve glycemic control in individuals with prediabetes. However, there is increasing evidence that people with prediabetes are not a homogeneous group but exhibit different glycemic profiles leading to the existence of prediabetes subgroups. Prediabetes subgroups have been identified as: isolated impaired fasting glucose (IFG), isolated impaired glucose tolerance (IGT), and combined impaired fasting glucose and glucose intolerance (IFG/IGT). The present review investigates human clinical trials examining the hypoglycemic potential of phenolic-rich plant extracts in prediabetes and prediabetes subgroups. Artemisia princeps Pampanini, soy (Glycine max (L.) Merrill) leaf and Citrus junos Tanaka peel have been demonstrated to improve fasting glycemia and thus may be more useful for individuals with IFG with increasing hepatic insulin resistance. In contrast, white mulberry (Morus alba Linn.) leaf, persimmon (Diospyros kaki) leaf and Acacia. Mearnsii bark were shown to improve postprandial glycemia and hence may be preferably beneficial for individuals with IGT with increasing muscle insulin resistance. Elaeis guineensis leaf was observed to improve both fasting and postprandial glycemic measures depending on the dose. Current evidence remains scarce regarding the impact of the plant extracts on glycemic control in prediabetes subgroups and therefore warrants further study.

Different Host Plants Distinctly Influence the Feeding Ability of the Brown Citrus Aphid Toxoptera citricida.

Simple SummaryThe brown citrus aphid, Toxoptera citricida, is an important pest of citrus and causes serious damage in the main production areas. Host plant resistance is an environmentally friendly method to manage aphid infestations and is becoming increasingly important as aphids develop greater resistance to insecticides. The aim of this study was to assess T. citricida resistance on seven widespread and common hosts using electropenetrography combined with a population development test. We showed that the feeding parameters of the brown citrus aphid differed significantly depending on the host plants. Toxoptera citricida spent more time in the pathway stage and less time in the phloem stage on Eureka, while the opposite was observed on Yuzu and Rough Lemon. Measurements of population development of aphids showed that on the Eureka, aphids developed more slowly. Our data suggest that different host plants distinctly influence the ability of T. citricida to feed. The brown citrus aphid did not prefer to feed on Eureka Lemon compared to the other six host plants.Piercing–sucking insects are important crop pests, and an understanding of their feeding behavior and population development plays a crucial role in studying insect population dynamics and crop resistance. In our study, we examined the probing behavior of the brown citrus aphid, Toxoptera citricida, using electropenetrography and assessed its population development after 8 days on seven host plants: Yuzu, Citrus junos Sieb. ex Tanaka; Rough Lemon, C. jambhiri Lush.; ‘Luofu’ kumquat, Fortunella margarita Swingle; ‘Olinda’ valencia orange, C. sinensis (L.) Osbeck; ‘Yanxiwanlu’ Ponkan, C. reticulata Blanco; ‘Rohde Red’ valencia orange, C. sinensis; and ‘Eureka’ lemon, C. limon (L.) Osbeck. The results demonstrated that probing by the brown citrus aphid differed significantly according to the target hosts. Toxoptera citricida produced significantly more pathway activities on Eureka than on Rough Lemon and Yuzu. Toxoptera citricida spent more time from the first probe to first salivation into phloem sieve elements on Eureka compared to Yuzu. In addition, the total duration of ingestion from sieve cells of each aphid in the phloem-feeding phase was shortest on Eureka, and this was significantly shorter than that on Yuzu, Rough Lemon, Luofu, and Olinda. The population number of T. citricida on Eureka after 8 days was significantly lower than that on the other hosts. Overall, Eureka was found to have obvious resistance to T. citricida, whereas Yuzu and Rough Lemon were susceptible host plants. These results provide a theoretical basis for exploring aphid-resistant fruit tree resources using resistant varieties.

Suppressive Effect of Yuzu (&lt;i&gt;Citrus junos&lt;/i&gt;) Peel Extract on Fatty Liver Steatosis Induced by a High-sucrose Diet in Rats

Suppressive Effect of Yuzu (&lt;i&gt;Citrus junos&lt;/i&gt;) Peel Extract on Fatty Liver Steatosis Induced by a High-sucrose Diet in Rats