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At least 253 records · Page 14Linked to original sources

A polygalacturonase from citrus leaf explants: role in abscission.

The relationship between polygalacturonase activity and abscission of citrus leaf explants was studied. Determination of polygalacturonase activity in citrus tissues requires concentration of the enzyme, use of a proper assay method, and inhibition of an oxidase present in the extracts which oxidizes the reaction products of the polygalacturonase. The polygalacturonase from citrus leaf explants is an exopolygalacturonase and appears to be a soluble enzyme.Polygalacturonase activity increased during abscission of citrus leaf explants and was localized in the separation layer. Ethylene accelerated the increase in polygalacturonase activity, but its effect was evident only after at least an 8-hour lag period. 2,4-Dichlorophenoxyacetic acid and cycloheximide inhibited abscission and polygalacturonase activity. It is concluded that polygalacturonase, in addition to cellulase, plays a role in abscission.

Journal Article↗

Accumulation of Free Proline in Citrus Leaves during Cold Hardening of Young Trees in Controlled Temperature Regimes.

Free proline increased in leaves of orange (Citrus sinensis [L.] Osb. cv. Valencia) and grapefruit (Citrus paradisi Macfad. cv. Star Ruby) trees on a wide range of citrus rootstocks during cold hardening. Increases in sugars accompanied proline accumulation. During cold hardening, the rate of proline accumulation was greater in old than in young leaves. In leaves of grapefruit trees kept in the dark during cold hardening, neither proline nor sugars increased and the degree of cold hardiness was less than in trees exposed to light. Like sugar accumulations, proline accumulation does not reflect specific degrees of cold hardiness in citrus cultivars.

Journal Article↗

Genomic Relatedness of Xanthomonas campestris Strains Causing Diseases of Citrus.

Xanthomonas campestris strains that cause disease in citrus were compared by restriction endonuclease analysis of DNA fragments separated by pulsed-field gel electrophoresis and by DNA reassociation. Strains of X. campestris pv. citrumelo, which cause citrus bacterial spot, were, on average, 88% related to each other by DNA reassociation, although these strains exhibited diverse restriction digest patterns. In contrast, strains of X. campestris pv. citri groups A and B, which cause canker A and canker B, respectively, had relatively homogeneous restriction digest patterns. The groups of strains causing these three different citrus diseases were examined by DNA reassociation and were found to be from 55 to 63% related to one another. Several pathovars of X. campestris, previously shown to cause weakly aggressive symptoms on citrus, ranged from 83 to 90% similar to X. campestris pv. citrumelo by DNA reassociation. The type strain of X. campestris pv. campestris ranged from 30 to 40% similar in DNA reassociation experiments to strains of X. campestris pv. citrumelo and X. campestris pv. citri groups A and B. Whereas DNA reassociation quantified the difference between relatively unrelated groups of bacterial strains, restriction endonuclease analysis distinguished between closely related strains.

Journal Article↗

Distribution of TGG repeat-related sequences in 'Trovita' orange (Citrus sinensis Osbeck) chromosomes

The clone pAS"C" is a sequence related to M13 phage minisatellite sequences isolated from bovine DNA. It contains TGG repeats, and related sequences are abundant in the Citrus genome. Fluorescent in situ hybridization (FISH) signals detected using this clone as a probe showed that the TGG repeat-related sequences are found on all the Citrus chromosomes, but are not found in Citrus-specific CMA+/DAPI- heterochromatic regions located at the extreme ends of each chromosome. These data suggest that the TGG-repeated sequences are evolutionarily conserved and that the CMA+/DAPI- heterochromatic regions were added to the chromosome ends at a recent stage in Citrus evolution.

Journal Article↗

CgMYC2 directly activates jasmonate-induced naringin biosynthesis in Citrus grandis 'Tomentosa'.

CgMYC2 links jasmonate signaling to naringin biosynthesis by binding G-box motifs and activating flavonoid-pathway promoters in Citrus grandis 'Tomentosa' Naringin, the predominant bitter-flavanone glycoside in Citrus grandis 'Tomentosa', has well-characterized biosynthetic enzymes, yet the transcriptional regulators coupling hormonal signals to pathway activation remain poorly understood. We demonstrate that CgMYC2, a jasmonate-responsive bHLH transcription factor, functions as a central activator of naringin biosynthesis. Exogenous methyl jasmonate (MeJA) treatment increased naringin content 3.45-fold in seedlings, coinciding with a rapid 6.6-fold induction of CgMYC2 that preceded the peak transcription of five core biosynthetic genes (CgPAL5, CgCHS, CgFNS, Cg7GlcT, and Cg1,2RhaT). Physical interaction between CgMYC2 and the JAZ protein CgJAZ3 was confirmed by pull-down and Co-IP assays, placing CgMYC2 within the canonical jasmonate signaling cascade. Y1H confirmed CgMYC2 binding to the Cg1,2RhaT promoter, EMSA demonstrated direct G-box-dependent binding to all five pathway promoters, and dual-luciferase assays showed transactivation of all five promoters, with the strongest activation for CgCHS. As complementary chromatin-level support, a single-sample CUT&Tag profile revealed G-box-enriched CgMYC2-associated chromatin regions across jasmonate-responsive and secondary-metabolic loci. Virus-induced gene silencing (VIGS) of CgMYC2 reduced naringin content by ~21% and suppressed biosynthetic gene expression, supporting its positive contribution. Furthermore, heterologous overexpression in tomato activated the flavonoid pathway and elevated 16 flavonoid compounds, consistent with evolutionary conservation of the MYC2-G-box regulatory logic. These findings establish CgMYC2 as a central, JA-responsive activator bridging jasmonate perception and naringin biosynthesis, providing a molecular framework for the targeted improvement of bitter-flavonoid traits in citrus.

Citrus↗

Amplification of DNA of Xanthomonas axonopodis pv. citri from historic citrus canker herbarium specimens.

Herbaria are important resources for the study of the origins and dispersal of plant pathogens, particularly bacterial plant pathogens that incite local lesions in which large numbers of pathogen genomes are concentrated. Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus bacterial canker disease, is a notable example of such a pathogen. The appearance of novel strains of the pathogen in Florida and elsewhere make it increasingly important to understand the relationships among strains of this pathogen. USDA-ARS at Beltsville, Maryland maintains approximately 700 herbarium specimens with citrus canker disease lesions up to 90 years old, originally collected from all over the world, and so is an important resource for phytogeographic studies of this bacterium. Unfortunately, DNA in herbarium specimens is degraded and may contain high levels of inhibitors of PCR. In this study, we compared a total of 23 DNA isolation techniques in combination with 31 novel primer pairs in order to develop an efficient protocol for the analysis of Xac DNA in herbarium specimens. We identified the most reliable extraction method, identified in terms of successful amplification by our panel of 31 primer pairs. We also identified the most robust primer pairs, identified as successful in the largest number of extracts prepared by different methods. We amplified Xac genomic sequences up to 542 bp long from herbarium samples up to 89 years old. Primers varied in effectiveness, with some primer pairs amplifying Xac DNA from a 1/10,000 dilution of extract from a single lesion from a citrus canker herbarium specimen. Our methodology will be useful to identify pathogens and perform molecular analyses of bacterial and possibly fungal genomes from herbarium specimens.

Biological Specimen Banks↗

Capillary electrophoresis and high-performance liquid chromatography determination of polyglutamyl 5-methyltetrahydrofolate forms in citrus products.

The 5-methyltetrahydrofolate (5mTHF) polyglutamates in citrus products were analyzed by capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC). Folate species were purified from citrus products and concentrated from 2- to 100-fold using combined folate-affinity chromatography and C18 extraction. Seven polyglutamyl 5mTHFs were found in most not-from-concentrate (NFC) orange juices (OJ) in total amounts of approximately 1 nmol/mL, with varying distributions of individual polyglutamates. Folate amounts and distributions were also measured in orange fractions, single-strength OJ from concentrate, NFC grapefruit juice, and citrus peel molasses. Models containing ascorbic acid had folate thermal degradation rates one-seventh that of models without ascorbic acid. Pasteurization studies demonstrated that folate loss was <2% for commercial OJ pasteurization conditions (i.e., 93 degrees C for 5 s, 88 degrees C for 15 s, and 82 degrees C for 30 s). Both methods were precise, reproducible, and potentially faster than traditional analytical procedures requiring enzymatic deconjugation and microbial assays.

Beverages↗

Moderate shade can increase net gas exchange and reduce photoinhibition in citrus leaves.

Daily variations in net gas exchange, chlorophyll a fluorescence and water relations of mature, sun-acclimated grapefruit (Citrus paradisi Macfady.) and orange (Citrus sinensis L. Osbeck) leaves were determined in tree canopies either shaded with 50% shade screens or left unshaded (sunlit). Mean daily maximum photosynthetic photon flux density (PPFD) under shade varied from 500 to 700 micromol m-2 s-1 and was sufficient to achieve maximum net CO2 assimilation rates (A CO2). Responses of grapefruit and orange leaves to shading were remarkably similar. At midday, on bright clear days, the temperatures of sunlit leaves were 2-6 degrees C above air temperature and 1-4 degrees C above the temperatures of shaded leaves. Although midday depressions of stomatal conductance (gs) and A CO2 were observed in both sunlit and shaded leaves, shaded leaves had lower leaf-to-air vapor pressure differences (D) along with higher gs, A CO2 and leaf water-use efficiency than sunlit leaves. Estimated stomatal limitation to A CO2 was generally less than 25% and did not differ between shaded and sunlit leaves. Leaf intercellular CO2 partial pressure was not altered by shade treatment and did not change substantially with increasing D. Radiation and high temperature stress-induced non-stomatal limitation to A CO2 in sunlit leaves was greater than 40%. Reversible photoinhibition of photosystem II efficiency was more pronounced in sunlit than in shaded leaves. Thus, non-stomatal factors play a major role in regulating A CO2 of citrus leaves during radiation and high temperature stress.

Carbon Dioxide↗

High-efficiency genome-editing, transgene evaluation, and antimicrobial efficacy testing using Citrus medica L. hairy roots.

Huanglongbing (HLB) disease, associated with the fastidious bacterium Candidatus Liberibacter asiaticus (CLas), has a significant impact on citrus production worldwide. Conventional biochemical and genetic evaluation studies to identify potential disease resistance strategies have been mainly hindered due to the inability to culture CLas in a defined medium and the general recalcitrance of Citrus cultivars (grapefruits and oranges) to Agrobacterium-mediated plant transformation. We previously demonstrated the utility of plant hairy roots to co-cultivate CLas. In this study, we developed a hairy root transformation system using citron (Citrus medica L.), which is highly amenable to Rhizobium-mediated hairy root transformation. The explant survival and hairy root transformation efficiencies were up to 100% and 73%, respectively, and transgenic roots can be attained in as little as 30-60&#x2009;days. We demonstrate the utility of this citron-based hairy root transformation for rapid CRISPR/Cas9-mediated gene editing, transgene evaluation, and antimicrobial efficacy testing. The citron-based hairy root transformation system will significantly help the research community to speed-track the assessment of potential HLB disease resistance strategies.

Citrus↗

In vitro availability of minerals of some tropical and citrus fruits as influenced by antinutritional factors.

Four tropical fruits and three citrus fruits were analyzed for moisture, ash, antinutritional factors (phytate, oxalate, and polyphenols) contents, and total and available minerals. Moisture contents ranged from 6.0 to 83.17% for tropical fruits while that of citrus fruits ranged from 88.20 to 89.50%. Ash contents ranged from 2.56 to 4.50% and from 3.83 to 4.83%, for tropical and citrus fruits, respectively. All fruits contained no oxalate while phytate and polyphenols ranged from 48.1 to 134.1 mg/ 100 g and from 0.115 to 0.34%, respectively. For all fruits major minerals contents ranged from 7.7 to 433.3 while trace ones ranged from 0.116 to 1.91 mg/100 g. In vitro availabilities of major minerals (% of total) varied from 11.1 to 86.2% while for minor ones it ranged from 13 to 72.5%.

Biological Availability↗

Determination of flavonoids in citrus juices by micro-HPLC-ESI/MS.

Flavonoids are a large class of naturally occurring aromatic secondary plant metabolites. They constitute one of the most characteristic classes found in nature and more than 4000 flavonoids have been identified and divided into several subclasses. Flavonoids have several effects on human health, mainly related to their antioxidant activity. As a result of the increased interest in the biologically active compounds in food, many research studies have investigated the detection and quantification of antioxidants. Citrus plants are of great interest since they accumulate large amounts of flavonoids and fruit and juices of these species are consumed in large quantities. Despite the extensive research performed on Citrus flavonoids, many compounds still remain unidentified. In this work, flavonoids of five citrus juices: lemon, grapefruit, bergamot, orange, and mandarin were analyzed by microHPLC coupled on-line with an MS detector equipped with an ESI source operating in negative mode. The use of microcolumn HPLC greatly enhanced detection performance. In total, 20 flavonoids of two different subclasses (flavanone and flavone glycosides) were identified with the help of retention data and MS spectral information.

Beverages↗

Spatial and temporal distribution of imidacloprid and thiamethoxam in citrus and impact on Homalodisca coagulata populations.

Titers of two systemic neonicotinoid insecticides in citrus trees were measured in conjunction with conventional evaluations of their impact on glassy-winged sharpshooter (Homalodisca coagulata (Say); GWSS) populations. Xylem fluid samples were collected at regular intervals and from multiple locations within field-grown citrus trees to determine imidacloprid and thiamethoxam concentrations using commercial ELISA kits. Uptake profiles varied considerably with peak mean titers of imidacloprid occurring 6-8 weeks after application compared with 2 weeks for thiamethoxam. The persistence of each compound also varied as near-peak levels of imidacloprid were sustained for another 6-10 weeks before gradually declining. In contrast, thiamethoxam titers declined more rapidly after the initial peak, possibly reflecting an application rate only one-quarter of that used for imidacloprid. Within-tree distributions were more similar for the two compounds, with no significant effect due to height of the sample (upper or lower half) or to the quadrant location within the tree, with the exception of one quadrant in the thiamethoxam-treated trees. Substantial reductions in GWSS nymphs and adults were observed in imidacloprid-treated trees during the 2001 trial and were sustained for 4-5 months after treatment. Treatment effects on nymphs were not as well pronounced in the 2002 trial, when overall GWSS infestations were much reduced from the previous year. However, consistently lower adult infestations were still observed in 2002 for both treatments compared with untreated trees. Information on the spatial and temporal profiles in citrus trees was obtained for both compounds to complement field impact data and improve understanding of their pest management potential.

Animals↗

PCR detection of the two 'Candidatus' Liberobacter species associated with greening disease of citrus.

Greening is a severe and widespread disease of citrus in two main citrus growing areas of the world, Asia and Africa. It is caused by an uncultured phloem restricted bacterium that we have recently characterized from the sequence of its 16S ribosomal DNA. The bacterium is a new "Candidatus' genus, Liberobacter, in the alpha subdivision of the proteobacteria, and two 'Candidatus species' have been recognized: Liberobacter asiaticum and Liberobacter africanum. In this paper we describe a PCR method to detect the two liberobacter species in citrus trees by amplification of a 1160 bp fragment of their 16S rDNA. Distinction between the two species has also been achieved by Xbal digestion of the amplicons.

Citrus↗

Characterization of citrus tristeza virus subgenomic RNAs in infected tissue.

Citrus tristeza virus (CTV) specific RNAs extracted from infected citrus tissue were analyzed by Northern blot hybridization. RNAs were characterized by size and identified using cDNA probes specific to nine open reading frames (ORFs) identified by the analysis of sequence obtained from cDNA clones of the T36 isolate of CTV. Sequence specific cDNA probes identified the genomic RNA as well as subgenomic RNAs representing the p33, p65, p61, p27, p25, p18, p13, p20, and p23 ORFs in extracts of total or double-stranded RNA (dsRNA) isolated from infected tissue. A probe derived from the 3' terminal ORF (p23) hybridized to each of these subgenomic RNAs, indicating that the RNAs are 3' coterminal. The relative amounts of the different subgenomic RNAs varied widely. The RNAs for the p20 and p23 ORFs were the most abundant and surpassed the amount of the p25 or capsid protein specific subgenomic RNA. The number and sizes of the CTV subgenomic RNAs were the same in total RNA and dsRNA preparations. Propagation of T36 in seven different citrus hosts did not alter the pattern of subgenomic RNAs.

Citrus↗

Polyphenol antioxidants in citrus juices: in vitro and in vivo studies relevant to heart disease.

It is well known that eating fruits and vegetables lowers the risk of chronic diseases such as heart disease and cancer. The question of what is/are the active ingredient(s) is still unresolved. The initial hypothesis was that the antioxidant vitamins were responsible. However, recently the polyphenols have been investigated since they have been found to have beneficial properties such as being strong antioxidants. We measured the polyphenol content of citrus juices by an oxidation-reduction colorimetric method (Folin) using catechin as the standard. The order was tangerine juice > grapefruit juice > orange juice. The antioxidant contribution of ascorbic acid was measured by the difference in Folin reactive content following removal by ascorbate oxidase. Ascorbate contributed 56 to 77% of the antioxidant content of orange juice, 46% of the single tangerine juice measured, and 66 to 77% of grapefruit juices. Polyphenol quality in the juices was analyzed by using the inhibition of lower density lipoprotein oxidation promoted by cupric ion, an in vitro model of heart disease. Quality decreased in the following order: orange juice > grapefruit juice > tangerinejuice. In orange juice polyphenols accounted for 84-85% of antioxidant quality. The pure polyphenol hesperidin, which is common in juices, ascorbic acid, and the citrus juices, were not able to bind with LDL+VLDL and protect it from oxidation. In a hamster model of atherosclerosis, the juices were able to significantly inhibit atherosclerosis and lowered cholesterol and triglycerides. Ascorbic acid alone in the dose provided by the juices was found to have the same effect on atherosclerosis. However, the polyphenols in the citrus

Adult↗

Inhibition of growth and aflatoxin production of Aspergillus parasiticus by citrus oils.

Aspergillus parasiticus was inoculated into grapefruit juice and a glucose-yeast extract medium; both contained 500-7000 ppm of citrus oils that were incorporated into the media by sonication. Orange and lemon oil were more inhibitory to mold growth and aflatoxin production than was dlimonene, the main constituent of the two peel oils. After 7 days at 28 degrees C, 2000 ppm of lemon and 3000 ppm of orange oil in grapefruit juice afforded maximum suppression of mold growth and toxin formation. When the glucose-yeast extract medium was used, 3000 ppm of either oil were needed to achieve the same result. After 4 days at 28 degrees C, orange oil at 3500 ppm in either medium markedly inhibited mold growth (as evidenced by dry weight of mold mycelium) and aflatoxin production (only 14 and 1% of the amount normally produced in the juice and artificial medium, respectively). Higher concentrations of orange oil further reduced mold growth and aflatoxin production and also delayed the onset of sporulation, if it occurred. Although aflatoxin was detected in all samples, only 0.2 to 0.5% of the amount found in controls (without the citrus oil) was present when the medium contained 7000 ppm orange oil. The mold consistently grew, albeit very poorly, on the glass at the liquid-atmosphere interface even when the substrate contained a large amount of citrus oil.

Aflatoxins↗

[The antioxidant activity of citrus fruit peels].

The antioxidant properties of freeze-dried citrus fruit peels (orange, lemon, grapefruit) and methanolic extracts from the peel were studied. Freeze-dried orange peel showed the highest, lemon peel somewhat less and grapefruit peel the lowest but still remarkable antioxidant activity. This could be significantly improved by preparing methanolic extracts of the peels. Comparative examinations and autoxidation studies with the flavanon glycosides hesperidin and naringin as well as with their aglycones hesperetin and naringenin showed that the former are mainly responsible for the antioxidative activity of the citrus peel and extracts. In order to compare their antioxidative activity with that of the commercially available natural antioxidants alpha-tocopherol and ascorbylpalmitate, the freeze-dried citrus peels and their methanolic extracts should be used in higher concentrations, in consideration of their peculiar properties and complex natural composition. Furthermore, aspects of the correlation between antioxidant activity and molecular structure of the flavanones were discussed.

Antioxidants↗

Alteration of O,O-dimethyl S-[alpha-(carboethoxy)benzyl] phosphorodithioate (phenthoate) in citrus, water, and upon exposure to air and sunlight.

The fate of 32P- and 14C-labeled O,O-dimethyl S-[alpha-(carboethoxy)benzyl] phosphorodithioate (phenthoate) was determined in citrus, water, and upon exposure to air and sunlight. The products recovered from citrus and glass plates exposed to sunlight were unchanged phenthoate, phenthoate oxon, demethyl phenthoate, mandelic acid, bis-[alpha-(carboethoxy)benzyl] disulfide, O,O-dimethyl phosphorothioic and phosphorodithioic acid. Similar products generally were found in citrus leaf and fruit extracts. Phenthoate was fairly stable in phosphate buffered water with a half-life of approximately 12 days at pH 8.0. The major hydrolysis products were phenthoate acid, demethyl phenthoate and demethyl phenthoate oxon.

Air↗