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1.
Sci Adv ; 8(17): eabj5586, 2022 04 29.
Article in English | MEDLINE | ID: mdl-35476439

ABSTRACT

The comorbidity of chronic pain and mental dysfunctions such as depression and anxiety disorders has long been recognized, but the underlying mechanisms remain poorly understood. Here, using a mouse model of neuropathic pain, we demonstrated neuronal plasticity in the bed nucleus of the stria terminalis (BNST), which plays a critical role in chronic pain-induced maladaptive anxiety. Electrophysiology demonstrated that chronic pain increased inhibitory inputs to lateral hypothalamus (LH)-projecting BNST neurons. Chemogenetic manipulation revealed that sustained suppression of LH-projecting BNST neurons played a crucial role in chronic pain-induced anxiety. Furthermore, using a molecular genetic approach, we demonstrated that chronic pain elevated the excitability of a specific subpopulation of BNST neurons, which express cocaine- and amphetamine-regulated transcript (CART). The elevated excitability of CART-positive neurons caused the increased inhibitory inputs to LH-projecting BNST neurons, thereby inducing anxiety-like behavior. These findings shed light on how chronic pain induces psychiatric disorders, characterized by maladaptive anxiety.


Subject(s)
Chronic Pain , Septal Nuclei , Anxiety/etiology , Anxiety Disorders , Chronic Pain/etiology , Humans , Neuronal Plasticity , Septal Nuclei/physiology
2.
J Neurosci ; 39(42): 8376-8385, 2019 10 16.
Article in English | MEDLINE | ID: mdl-31451580

ABSTRACT

Although dysfunction of the mesolimbic dopaminergic system has been implicated in chronic pain, the underlying mechanisms remain to be elucidated. We hypothesized that increased inhibitory inputs to the neuronal pathway from the dorsolateral bed nucleus of the stria terminalis (dlBNST) to the ventral tegmental area (VTA) during chronic pain may induce tonic suppression of the mesolimbic dopaminergic system. To test this hypothesis, male Sprague Dawley rats were subjected to spinal nerve ligation to induce neuropathic pain and then spontaneous IPSCs (sIPSCs) were measured in this neuronal pathway. Whole-cell patch-clamp electrophysiology of brain slices containing the dlBNST revealed that the frequency of sIPSCs significantly increased in VTA-projecting dlBNST neurons 4 weeks after surgery. Next, the role of corticotropin-releasing factor (CRF) signaling within the dlBNST in the increased sIPSCs was examined. CRF increased the frequency of sIPSCs in VTA-projecting dlBNST neurons in sham-operated controls, but not in chronic pain rats. By contrast, NBI27914, a CRF type 1 receptor antagonist, decreased the frequency of sIPSCs in VTA-projecting dlBNST neurons in the chronic pain rats, but not in the control animals. In addition, histological analyses revealed the increased expression of CRF mRNA in the dlBNST. Finally, bilateral injections of NBI27914 into the dlBNST of chronic pain rats activated mesolimbic dopaminergic neurons and induced conditioned place preference. Together, these results suggest that the mesolimbic dopaminergic system is tonically suppressed during chronic pain by enhanced CRF signaling within the dlBNST via increased inhibitory inputs to VTA-projecting dlBNST neurons.SIGNIFICANCE STATEMENT The comorbidity of chronic pain and depression has long been recognized. Although dysfunction of the mesolimbic dopaminergic system has been implicated in both chronic pain and depression, the underlying mechanisms remain to be elucidated. Here, we show that the inhibitory inputs to the neuronal pathway from the dorsolateral bed nucleus of the stria terminalis (dlBNST) to the ventral tegmental area increase during chronic pain. This neuroplastic change is mediated by enhanced corticotropin-releasing factor signaling within the dlBNST that leads to tonic suppression of the mesolimbic dopaminergic system, which may be involved in the depressive mood and anhedonia under the chronic pain condition.


Subject(s)
Chronic Pain/metabolism , Corticotropin-Releasing Hormone/metabolism , Dopaminergic Neurons/metabolism , Septal Nuclei/metabolism , Signal Transduction/physiology , Ventral Tegmental Area/metabolism , Aniline Compounds/pharmacology , Animals , Disease Models, Animal , Dopaminergic Neurons/drug effects , Male , Patch-Clamp Techniques , Pyrimidines/pharmacology , Rats , Rats, Sprague-Dawley , Septal Nuclei/drug effects , Signal Transduction/drug effects , Ventral Tegmental Area/drug effects
3.
Biol Psychiatry ; 86(3): 230-239, 2019 08 01.
Article in English | MEDLINE | ID: mdl-30635130

ABSTRACT

BACKGROUND: A method that promotes the retrieval of lost long-term memories has not been well established. Histamine in the central nervous system is implicated in learning and memory, and treatment with antihistamines impairs learning and memory. Because histamine H3 receptor inverse agonists upregulate histamine release, the inverse agonists may enhance learning and memory. However, whether the inverse agonists promote the retrieval of forgotten long-term memory has not yet been determined. METHODS: Here, we employed multidisciplinary methods, including mouse behavior, calcium imaging, and chemogenetic manipulation, to examine whether and how the histamine H3 receptor inverse agonists, thioperamide and betahistine, promote the retrieval of a forgotten long-term object memory in mice. In addition, we conducted a randomized double-blind, placebo-controlled crossover trial in healthy adult participants to investigate whether betahistine treatment promotes memory retrieval in humans. RESULTS: The treatment of H3 receptor inverse agonists induced the recall of forgotten memories even 1 week and 1 month after training in mice. The memory recovery was mediated by the disinhibition of histamine release in the perirhinal cortex, which activated the histamine H2 receptor. Histamine depolarized perirhinal cortex neurons, enhanced their spontaneous activity, and facilitated the reactivation of behaviorally activated neuronal ensembles. A human clinical trial revealed that treatment of H3 receptor inverse agonists is specifically more effective for items that are more difficult to remember and subjects with poorer performance. CONCLUSIONS: These results highlight a novel interaction between the central histamine signaling and memory engrams.


Subject(s)
Histamine Agonists/pharmacology , Memory Disorders/drug therapy , Mental Recall/drug effects , Perirhinal Cortex/drug effects , Adult , Animals , Betahistine , Cognition/drug effects , Double-Blind Method , Female , Humans , Male , Mice , Mice, Inbred C57BL , Object Attachment , Piperidines , Stochastic Processes , Young Adult
4.
Medicines (Basel) ; 6(1)2018 Dec 25.
Article in English | MEDLINE | ID: mdl-30585249

ABSTRACT

The present article reviews the research progress of three major polyphenols (tannins, flavonoids and lignin carbohydrate complexes), chromone (backbone structure of flavonoids) and herbal extracts. Chemical modified chromone derivatives showed highly specific toxicity against human oral squamous cell carcinoma cell lines, with much lower toxicity against human oral keratinocytes, as compared with various anticancer drugs. QSAR analysis suggests the possible correlation between their tumor-specificity and three-dimensional molecular shape. Condensed tannins in the tea extracts inactivated the glucosyltransferase enzymes, involved in the biofilm formation. Lignin-carbohydrate complexes (prepared by alkaline extraction and acid-precipitation) and crude alkaline extract of the leaves of Sasa species (SE, available as an over-the-counter drug) showed much higher anti-HIV activity, than tannins, flavonoids and Japanese traditional medicine (Kampo). Long-term treatment with SE and several Kampo medicines showed an anti-inflammatory and anti-oxidant effects in small size of clinical trials. Although the anti-periodontitis activity of synthetic angiotensin II blockers has been suggested in many papers, natural angiotensin II blockers has not yet been tested for their possible anti-periodontitis activity. There should be still many unknown substances that are useful for treating the oral diseases in the natural kingdom.

5.
Eur J Neurosci ; 48(9): 3052-3061, 2018 11.
Article in English | MEDLINE | ID: mdl-30240530

ABSTRACT

The bed nucleus of the stria terminalis (BNST) and the central amygdala (CeA) comprise a forebrain unit that has been described as the "extended amygdala". These two nuclei send dense projections to each other and have been implicated in the regulation of negative emotional states, including anxiety and fear. The present study employed an optogenetic technique to examine whether stimulation of CeA-projecting dorsolateral BNST (dlBNST) neuron terminals would influence anxiety-like behaviors in male Sprague-Dawley rats. Photostimulation of CeA-projecting dlBNST neuron terminals produced anxiogenic effects in an elevated plus maze test. This finding is inconsistent with previous reports showing that optogenetic stimulation of BNST neurons projecting to the lateral hypothalamus (LH) and ventral tegmental area (VTA) produces anxiolytic rather than anxiogenic effects. To address this issue, electrophysiological analyses were conducted to characterize dlBNST neurons projecting to the CeA, LH, and VTA. dlBNST neurons can be electrophysiologically classified into three distinct cell types (types I-III) according to their responses to depolarizing and hyperpolarizing current injections. Whole-cell patch-clamp recordings revealed that more than 60% of the CeA-projecting dlBNST neurons were type II, whereas approximately 80% of the LH- and VTA-projecting dlBNST neurons were type III. These electrophysiological results will help elucidate the mechanisms underlying the heterogeneity of BNST neurons during the regulation of anxiety-like behaviors.


Subject(s)
Anxiety/metabolism , Central Amygdaloid Nucleus/metabolism , Nerve Net/metabolism , Septal Nuclei/metabolism , Animals , Anxiety/psychology , Central Amygdaloid Nucleus/chemistry , Male , Maze Learning/physiology , Nerve Net/chemistry , Optogenetics/methods , Rats , Rats, Sprague-Dawley , Septal Nuclei/chemistry
7.
Food Chem ; 237: 749-755, 2017 Dec 15.
Article in English | MEDLINE | ID: mdl-28764062

ABSTRACT

The effect of electrostatic atomized water particle (EAWP) treatment on quality maintenance during postharvest storage was determined to clarify a physiological role of reactive oxygen species in broccoli florets. The EAWP treatment suppressed the floret yellowing of broccoli during storage. The hydrogen peroxide content increased immediately after EAWP treatment but then remained at a low level during storage. Catalase (CAT) activity increased rapidly with EAWP treatment and was higher than that in the control for the first 4days of storage. The BoCAT gene expression level with EAWP treatment also showed a high trend as compared to the control. The gene expressions of BoRboh, BoCu/ZnSOD, and BoAPX showed almost the same tendency with or without EAWP treatment. These findings suggested that hydrogen peroxide formed by EAWP treatment and regulated by CAT could be relevant to maintaining the quality of broccoli.


Subject(s)
Brassica , Hydrogen Peroxide , Reactive Oxygen Species , Static Electricity
8.
PLoS One ; 12(8): e0181784, 2017.
Article in English | MEDLINE | ID: mdl-28800607

ABSTRACT

The genus Allium is a rich source of steroidal saponins, and its medicinal properties have been attributed to these bioactive compounds. The saponin compounds with diverse structures play a pivotal role in Allium's defense mechanism. Despite numerous studies on the occurrence and chemical structure of steroidal saponins, their biosynthetic pathway in Allium species is poorly understood. The monosomic addition lines (MALs) of the Japanese bunching onion (A. fistulosum, FF) with an extra chromosome from the shallot (A. cepa Aggregatum group, AA) are powerful genetic resources that enable us to understand many physiological traits of Allium. In the present study, we were able to isolate and identify Alliospiroside A saponin compound in A. fistulosum with extra chromosome 2A from shallot (FF2A) and its role in the defense mechanism against Fusarium pathogens. Furthermore, to gain molecular insight into the Allium saponin biosynthesis pathway, high-throughput RNA-Seq of the root, bulb, and leaf of AA, MALs, and FF was carried out using Illumina's HiSeq 2500 platform. An open access Allium Transcript Database (Allium TDB, http://alliumtdb.kazusa.or.jp) was generated based on RNA-Seq data. The resulting assembled transcripts were functionally annotated, revealing 50 unigenes involved in saponin biosynthesis. Differential gene expression (DGE) analyses of AA and MALs as compared with FF (as a control) revealed a strong up-regulation of the saponin downstream pathway, including cytochrome P450, glycosyltransferase, and beta-glucosidase in chromosome 2A. An understanding of the saponin compounds and biosynthesis-related genes would facilitate the development of plants with unique saponin content and, subsequently, improved disease resistance.


Subject(s)
Allium/genetics , Saponins/biosynthesis , Sequence Analysis, RNA/methods , Steroids/biosynthesis , Transcriptome/genetics , Biological Assay , Biosynthetic Pathways/genetics , Gene Expression Profiling , Gene Expression Regulation, Plant , Genes, Plant , Models, Biological
9.
Genes Genet Syst ; 91(3): 161-173, 2016 Nov 26.
Article in English | MEDLINE | ID: mdl-27074981

ABSTRACT

The aim of this study was to obtain primary information about the global diversity of garlic (Allium sativum L.) by evaluating morphological, physiological and isozyme variation. A total of 107 garlic accessions collected worldwide were grown in Yamaguchi, Japan. Five morphological traits (bulb weight, bulb diameter, number of cloves per bulb, number of bulbils and scape length) and one physiological trait (bolting period) of the collected garlic showed wide variation. Meanwhile, a total of 140 garlic accessions, including the 107 mentioned above, were characterized by leucine aminopeptidase (LAP) and phosphoglucoisomerase (PGI) isozyme analyses; they clearly showed polymorphisms in putative isozyme loci (Lap-1, Lap-2 and Pgi-1). Allelic frequencies were estimated in each group of accessions categorized by their geographical origin, and the observed (Ho) and expected (He) heterozygosities were calculated. The allelic frequencies differed between groups. A principal component analysis based on morpho-physiological data indicated a grouping of the garlic accessions into Central Asian and Northern Mediterranean groups as well as others. We discuss the roles of artificial and natural selection that may have caused differentiation in these traits, on the assumption that ancestral domesticated garlic populations have adapted in various regions using standing variation or mutations that accumulated during expansion, and have evolved along with human-preferred traits over a long history of cultivation.


Subject(s)
Garlic/genetics , Genetic Variation , Isoenzymes/genetics , Leucyl Aminopeptidase/genetics , Garlic/enzymology , Gene Frequency , Heterozygote , Humans , Japan , Mutation , Phenotype
10.
Genome ; 58(4): 135-42, 2015 Apr.
Article in English | MEDLINE | ID: mdl-26218854

ABSTRACT

Bunching onion (Allium fistulosum L.; 2n = 16), bulb onion (Allium cepa L. Common onion group), and shallot (Allium cepa L. Aggregatum group) cultivars were inoculated with rust fungus, Puccinia allii, isolated from bunching onion. Bulb onions and shallots are highly resistant to rust, suggesting they would serve as useful resources for breeding rust resistant bunching onions. To identify the A. cepa chromosome(s) related to rust resistance, a complete set of eight A. fistulosum - shallot monosomic alien addition lines (MAALs) were inoculated with P. allii. At the seedling stage, FF+1A showed a high level of resistance in controlled-environment experiments, suggesting that the genes related to rust resistance could be located on shallot chromosome 1A. While MAAL, multi-chromosome addition line, and hypoallotriploid adult plants did not exhibit strong resistance to rust. In contrast to the high resistance of shallot, the addition line FF+1A+5A showed reproducibly high levels of rust resistance.


Subject(s)
Basidiomycota/physiology , Chromosomes, Plant/genetics , Disease Resistance/genetics , Onions/genetics , Plant Diseases/immunology , Shallots/genetics , Basidiomycota/immunology , Breeding , Onions/immunology , Onions/microbiology , Plant Diseases/microbiology , Plant Leaves/genetics , Plant Leaves/immunology , Plant Leaves/microbiology , Seedlings/genetics , Seedlings/immunology , Seedlings/microbiology , Shallots/immunology , Shallots/microbiology
11.
Biosci Biotechnol Biochem ; 78(7): 1112-22, 2014.
Article in English | MEDLINE | ID: mdl-25229844

ABSTRACT

To investigate the involvement of Allium roylei metabolites in the plant's defenses, a comprehensive analysis of the content of cysteine sulfoxides, flavonols, polyphenols, ascorbic acid, and saponins was carried out in the various organs of this species. Metabolomics high performance liquid chromatography (HPLC), spectral-based analysis, and histochemcial studies have given important insight to the validity of saponins as a key component involved in plant protection. The root-basal stem, bulb, and leaf extracts exhibited 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity with inhibition concentration (IC(50)) ranging from 0.649 to 0.757 mg/mL. The antimicrobial properties of the saponin and flavonoid crude extracts were evaluated. The saponin extracts demonstrated significant antifungal activity depending on the applied concentration, and the growth inhibition rate of the tested fungal pathogens ranged from 1.07 to 47.76%. No appreciable antibacterial activity was recorded in the same sample.


Subject(s)
Allium/metabolism , Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Ascorbic Acid/metabolism , Biological Transport , Flavonoids/metabolism , Flavonoids/pharmacology , Free Radical Scavengers/metabolism , Free Radical Scavengers/pharmacology , Organ Specificity , Polyphenols/metabolism , Saponins/metabolism , Saponins/pharmacology , Sulfoxides/metabolism
12.
Am J Ophthalmol ; 158(5): 983-93, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25036881

ABSTRACT

PURPOSE: To evaluate the therapeutic benefits of tear-exchangeable, limbal, rigid contact lenses (limbal CLs) in patients with Stevens-Johnson syndrome- or toxic epidermal necrolysis-associated ocular sequelae. DESIGN: Noncomparative, retrospective, interventional case series. METHODS: We enrolled 53 eyes of 42 patients (mean age, 51.8 ± 13.9 years; mean follow-up, 25.7 ± 15.7 months) with Stevens-Johnson syndrome- or toxic epidermal necrolysis-associated ocular sequelae and divided them into 3 groups according to the best-corrected visual acuity (BCVA) before limbal CL fitting: (1) BCVA worse than 20/2000 (11 eyes), (2) BCVA ranging from 20/200 to 20/2000 (31 eyes), and (3) BCVA of 20/200 or better (11 eyes). The BCVA and the 25-item National Eye Institute Visual Function Questionnaire (NEI VFQ-25) composite score before fitting and after 3 months of limbal CL use were evaluated. The change in BCVA (in logarithm of the minimal angle of resolution [logMAR] units) and 25-item National Eye Institute Visual Function Questionnaire composite score change were compared among the 3 groups. RESULTS: Best-corrected visual acuity improved from 1.61 to 0.86 logMAR at 3 months after fitting CL use. Improvement in BCVA in groups 1, 2, and 3 was 0.95 logMAR, 0.82 logMAR, and 0.37 logMAR, respectively. The mean 25-item National Eye Institute Visual Function Questionnaire composite score for the 11 subscales improved from 37.6 ± 16.0 to 58.4 ± 17.4 (P = .000001). All 11 subscores, except that for driving ability, improved significantly. The general vision subscore improved most in group 3, yet the general health subscore improved most in group 1. No serious adverse events attributable to limbal CL use occurred. CONCLUSIONS: The tear-exchangeable limbal CL is safe and effective for the improvement of vision and quality of life in Stevens-Johnson syndrome or toxic epidermal necrolysis patients with severe ocular sequelae.


Subject(s)
Contact Lenses , Corneal Diseases/therapy , Limbus Corneae/pathology , Stevens-Johnson Syndrome/therapy , Tears/physiology , Adult , Aged , Corneal Diseases/physiopathology , Equipment Design , Female , Follow-Up Studies , Humans , Male , Middle Aged , Quality of Life , Retrospective Studies , Stevens-Johnson Syndrome/physiopathology , Surveys and Questionnaires , Time Factors , Visual Acuity
13.
Food Chem ; 165: 224-31, 2014 Dec 15.
Article in English | MEDLINE | ID: mdl-25038670

ABSTRACT

Chlorophyll (Chl)-degrading peroxidase (POX) was purified from green and green-yellow broccoli florets to elucidate the physiological role and cell localisation of Chl-degrading POX. Effect of UV-B treatment on Chl-degrading POX activity was also determined. The Chl-degrading POX activity was separated by an ion exchange chromatography into 3 fractions, Chl-POX1, Chl-POX2 and Chl-POX3. The activities of Chl-POX2 and Chl-POX3, but especially that of Chl-POX3, were found in green-yellow florets and were clearly suppressed by UV-B irradiation. The protein molecular sizes of cPOX1 and cPOX3 purified from Chl-POX1 and POX3 were 43 and 34kDa, respectively. The cPOX1 was not located within chloroplast fraction, whereas the cPOX3 was found in intact chloroplasts extracted from senescing broccoli florets. We propose that cPOX3 is a chloroplast POX and the activity was suppressed by UV-B treatment, suggesting that cPOX3 might be involved in Chl degradation in stored broccoli florets.


Subject(s)
Brassica/chemistry , Chlorophyll/chemistry , Chloroplasts/metabolism , Peroxidase/metabolism , Oxidoreductases , Ultraviolet Rays
14.
Food Chem ; 156: 160-4, 2014 Aug 01.
Article in English | MEDLINE | ID: mdl-24629952

ABSTRACT

The effect of electrostatic atomized water particles (EAWP) on degreening of green sour citrus fruit during storage was determined. Superoxide anion and hydroxyl radicals included in EAWP were present on the surface of the fruit peel after the treatment. Hydrogen peroxide was formed from EAWP in an aqueous solution, which could indicate that a hydroxyl radical of EAWP turns to hydrogen peroxide in the fruit flavedo as well as in the aqueous solution. EAWP treatment effectively suppressed the degreening of green yuzu and Nagato-yuzukichi fruits during storage at 20°C. The enhancement in K+ ion leakage of both EAWP-treated fruits reduced in comparison with the control. In spite of EAWP treatment, total peroxide level in both fruits showed almost no changes during storage, suggesting that hydrogen peroxide formed by EAWP treatment could stimulate the activation of hydrogen peroxide scavenging system and control degreening of these fruits during storage.


Subject(s)
Citrus/chemistry , Fruit/chemistry , Reactive Oxygen Species , Static Electricity , Taste , Water
15.
Biosci Biotechnol Biochem ; 77(12): 2486-8, 2013.
Article in English | MEDLINE | ID: mdl-24317054

ABSTRACT

We measured the antioxidant contents and antioxidative activities in eight Allium fistulosum-shallot monosomic addition lines (MAL; FF+1A-FF+8A). The high antioxidative activity lines (FF+2A and FF+6A) showed high polyphenol accumulation. These additional chromosomes (2A and 6A) would therefore have anonymous genes related to the upregulation of polyphenol production, the antioxidative activities consequently being increased in these MALs.


Subject(s)
Allium/metabolism , Free Radical Scavengers/metabolism , Allium/genetics , Biphenyl Compounds/metabolism , Chromosomes, Plant/genetics , Picrates/metabolism
16.
Biomed Rep ; 1(3): 355-358, 2013 May.
Article in English | MEDLINE | ID: mdl-24648948

ABSTRACT

Kamaboko is a traditional type of processed seafood made from fish jelly paste that is unique to Japan. We supplemented Kamaboko with Japanese bunching onion (Allium fistulosum L.) with an alien monosome from shallot (Allium cepa L. Aggregatum group) and we measured in vitro the oxygen radical absorbance capacity (ORAC) value, an index of antioxidant activity. We also evaluated the results of sensory testing. The ORAC value of plain Kamaboko was 166±14 µmol trolox equivalent (TE)/100 g fresh weight (FW). The values of the edible Alliaceae powder, i.e., Japanese bunching onion (JBO, genome FF, 2n=2x=16) and the alien addition line of JBO carrying the 6A chromosome from shallot (FF+6A, 2n=2x+1=17), were 6,659±238 and 14,096±635 µmol TE/100 g dry weight (DW). We hypothesized that the 6A chromosome encoded the enhancement of polyphenol production. Subsequently, we created Kamaboko containing 4.8% JBO powder or 4.8% FF+6A powder. The ORAC value of each modified Kamaboko product was increased to 376±24 µmol TE/100 g FW for the JBO powder and to 460±16 µmol TE/100 g FW for the FF+6A powder, respectively. We next created Kamaboko containing 9.0% JBO powder or 9.0% FF+6A powder and the ORAC values of the respective modified Kamaboko products was increased to 671±16 and 740±21 µmol TE/100 g FW, i.e., 4.1- and 4.5-times the value of plain Kamaboko. Consequently, taking into consideration the sensory evaluation regarding taste and appearance as well, the use of Kamaboko supplemented with 4.8% FF+6A powder is recommended.

17.
Genome ; 55(11): 797-807, 2012 Nov.
Article in English | MEDLINE | ID: mdl-23199574

ABSTRACT

This study was carried out to evaluate the antifungal effect of Allium cepa Aggregatum group (shallot) metabolites on Fusarium oxysporum and to determine the shallot chromosome(s) related to Fusarium wilt resistance using a complete set of eight Allium fistulosum - shallot monosomic addition lines. The antifungal effects of hexane, butanol, and water extraction fractions from bulbs of shallot on 35 isolates of F. oxysporum were examined using the disc diffusion method. Only hexane and butanol fractions showed high antifungal activity. Shallot showed no symptom of disease after inoculation with F. oxysporum f. sp. cepae. The phenolic content of the roots and the saponin content of root exudates of inoculated shallot increased to much higher levels than those of the control at 3 days after inoculation. Application of freeze-dried shallot root exudates to seeds of A. fistulosum soaked in a spore suspension of F. oxysporum resulted in protection of seedlings against infection. Among eight monosomic addition lines and A. fistulosum, FF+2A showed the highest resistance to Fusarium wilt. This monosomic addition line also showed a specific saponin band derived from shallot on the thin layer chromatography profile of saponins in the eight monosomic addition lines. The chromosome 2A of shallot might possess some of the genes related to Fusarium wilt resistance.


Subject(s)
Allium/chemistry , Allium/genetics , Chromosomes, Plant/genetics , Fusarium/drug effects , Plant Diseases/immunology , Plant Extracts/pharmacology , Allium/immunology , Allium/microbiology , Chromatography, Thin Layer , Disk Diffusion Antimicrobial Tests , Fusarium/pathogenicity , Mycelium , Phenols/analysis , Phenols/isolation & purification , Phenols/pharmacology , Plant Diseases/microbiology , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Exudates/chemistry , Plant Exudates/isolation & purification , Plant Exudates/pharmacology , Plant Roots/chemistry , Plant Roots/genetics , Plant Roots/immunology , Plant Roots/microbiology , Saponins/analysis , Saponins/isolation & purification , Saponins/pharmacology , Seedlings/chemistry , Seedlings/genetics , Seedlings/immunology , Seedlings/microbiology , Seeds/chemistry , Seeds/genetics , Seeds/immunology , Seeds/microbiology , Spores, Fungal
18.
G3 (Bethesda) ; 2(6): 643-51, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22690373

ABSTRACT

Lachrymatory factor synthase (LFS) catalyzes the formation of lachrymatory factor, one of the most distinctive traits of bulb onion (Allium cepa L.). Therefore, we used LFS as a model for a functional gene in a huge genome, and we examined the chromosomal organization of LFS in A. cepa by multiple approaches. The first-level analysis completed the chromosomal assignment of LFS gene to chromosome 5 of A. cepa via the use of a complete set of A. fistulosum-shallot (A. cepa L. Aggregatum group) monosomic addition lines. Subsequent use of an F(2) mapping population from the interspecific cross A. cepa × A. roylei confirmed the assignment of an LFS locus to this chromosome. Sequence comparison of two BAC clones bearing LFS genes, LFS amplicons from diverse germplasm, and expressed sequences from a doubled haploid line revealed variation consistent with duplicated LFS genes. Furthermore, the BAC-FISH study using the two BAC clones as a probe showed that LFS genes are localized in the proximal region of the long arm of the chromosome. These results suggested that LFS in A. cepa is transcribed from at least two loci and that they are localized on chromosome 5.

19.
Theor Appl Genet ; 124(7): 1241-57, 2012 May.
Article in English | MEDLINE | ID: mdl-22234606

ABSTRACT

To produce alien monosomic addition lines (AMALs) of Allium cepa (genomes CC, 2n = 2x = 16) carrying extrachromosomes from Allium roylei (RR, 2n = 2x = 16), reciprocal backcrossing of allotriploids (2n = 24, CCR) with diploids (2n = 16, CC) and selfing of a single allotriploid were carried out. The chromosome numbers in the BC(2)F(1) and BC(1)F(2) progenies ranged from 16 to 32. Forty-eight plants were recorded to possess 2n = 17 among a total of 169 plants in observation. Through the analyses of isozymes, expressed sequence tag (EST) markers, and karyotypes, all eight possible types of A. cepa-A. roylei monosomic addition lines (CC+1R-CC+8R) could be identified. Seven types of representative AMALs (without CC+2R) were used for the GISH analysis of somatic chromosomes. Except for CC+6R, all AMALs showed an entire (unrecombined) extrachromosome from A. roylei in the integral diploid background of A. cepa. A single recombination between A. cepa and A. roylei was observed on the extrachromosome in the remaining type. All alloplasmic AMALs possessing A. roylei cytoplasm showed high or complete pollen sterility. Only the autoplasmic CC+4R with A. cepa cytoplasm possessed relatively high pollen fertility. The bulbs of CC+4R displayed the distinct ovoid shape that discriminates them from spherical or oval ones in other AMALs. Downy mildew screening in the field showed higher resistance in A. roylei, a hypo-allotriploid (CCR-nR, 2n = 23), and an allotriploid (CCR, 2n = 24). Meanwhile, no complete resistance was found in some AMALs examined. This was the first trial toward the establishment of a complete set of A. cepa-A. roylei monosomic additions.


Subject(s)
Allium/genetics , Chromosomes, Plant/genetics , Crosses, Genetic , Genes, Plant , Hybridization, Genetic , Monosomy , Breeding , Endangered Species , Karyotyping
20.
Genes Genet Syst ; 84(1): 43-55, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19420800

ABSTRACT

To develop the bunching onion (Allium fistulosum L.; genomes, FF) chromosome-specific genetic markers for identifying extra chromosomes, eight shallot (A. cepa L. Aggregatum group; genomes, AA)--A. fistulosum monosomic addition plants (AA+nF) and 62 shallot--A. fistulosum single-alien deletion plants (AAF-nF) were analyzed by 23 different chromosome-specific genetic markers of shallot. The eight monosomic addition plants consisted of one AA+2F, two AA+6F, and five AA+8F. Of the 62 single-alien deletion plants, 60 could be identified as six different single-alien deletion lines (AAF-1F, -3F, -4F, -6F, -7F, and -8F) out of the eight possible types. Several single-alien deletion lines were classified on the basis of leaf and bulb characteristics. AAF-8F had the largest number of expanded leaves of five deletion plants. AAF-7F grew most vigorously, as expressed by its long leaf blade and biggest bulb size. AAF-4F had very small bulbs. AAF-7F and AAF-8F had different bulbs from those of shallot as well as other types of single-alien deletion lines in skin and outer scale color. Regarding the sugar content of the bulb tissues, the single-alien deletion lines showed higher fructan content than shallot. Moreover, shallot could not produce fructan with degree of polymerization (DP) 12 or higher, although the single-alien deletion lines showed DP 20 or higher. The content of S-alk(en)yl-L-cysteine sulfoxide (ACSO) in the single-alien deletion lines was significantly lower than that in shallot. These results indicated that chromosomes from A. fistulosum might carry anonymous factors to increase the highly polymerized fructan production and inhibit the synthesis of ACSO in shallot bulbs. Accordingly, alien chromosomes from A. fistulosum in shallot would contribute to modify the quality of shallot bulbs.


Subject(s)
Chromosomes, Plant/genetics , Genes, Plant/physiology , Onions/genetics , Shallots/genetics , Chimera/genetics , Chimera/metabolism , Chromosomes, Plant/metabolism , Fructans/biosynthesis , Fructans/genetics , Onions/metabolism , Shallots/metabolism
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