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PubMed2002

Evaluation of the genetic structure of Xylella fastidiosa populations from different Citrus sinensis varieties.

Xylella fastidiosa was isolated from sweet orange plants (Citrus sinensis) grown in two orchards in the northwest region of the Brazilian state of São Paulo. One orchard was part of a germ plasm field plot used for studies of citrus variegated chlorosis resistance, while the other was an orchard of C. sinensis cv. Pêra clones. These two collections of strains were genotypically characterized by using random amplified polymorphic DNA (RAPD) and variable number of tandem repeat (VNTR) markers. The genetic diversity (H(T)) values of X. fastidiosa were similar for both sets of strains; however, H(T)(RAPD) values were substantially lower than H(T)(VNTR) values. The analysis of six strains per plant allowed us to identify up to three RAPD and five VNTR multilocus haplotypes colonizing one plant. Molecular analysis of variance was used to determine the extent to which population structure explained the genetic variation observed. The genetic variation observed in the X. fastidiosa strains was not related to or dependent on the different sweet orange varieties from which they had been obtained. A significant amount of the observed genetic variation could be explained by the variation between strains from different plants within the orchards and by the variation between strains within each plant. It appears, therefore, that the existence of different sweet orange varieties does not play a role in the population structure of X. fastidiosa. The consequences of these results for the management of sweet orange breeding strategies for citrus variegate chlorosis resistance are also discussed.

Bacterial Typing Techniques↗
PubMed2000

Sequences of Citrus tristeza virus separated in time and space are essentially identical.

The first Citrus tristeza virus (CTV) genomes completely sequenced (19.3-kb positive-sense RNA), from four biologically distinct isolates, are unexpectedly divergent in nucleotide sequence (up to 60% divergence). Understanding of whether these large sequence differences resulted from recent evolution is important for the design of disease management strategies, particularly the use of genetically engineered mild (essentially symptomless)-strain cross protection and RNA-mediated transgenic resistance. The complete sequence of a mild isolate (T30) which has been endemic in Florida for about a century was found to be nearly identical to the genomic sequence of a mild isolate (T385) from Spain. Moreover, samples of sequences of other isolates from distinct geographic locations, maintained in different citrus hosts and also separated in time (B252 from Taiwan, B272 from Colombia, and B354 from California), were nearly identical to the T30 sequence. The sequence differences between these isolates were within or near the range of variability of the T30 population. A possible explanation for these results is that the parents of isolates T30, T385, B252, B272, and B354 have a common origin, probably Asia, and have changed little since they were dispersed throughout the world by the movement of citrus. Considering that the nucleotide divergence among the other known CTV genomes is much greater than those expected for strains of the same virus, the remarkable similarity of these five isolates indicates a high degree of evolutionary stasis in some CTV populations.

Base Sequence↗
PubMed2005

Identification and genomic characterization of a new virus (Tymoviridae family) associated with citrus sudden death disease.

Citrus sudden death (CSD) is a new disease that has killed approximately 1 million orange trees in Brazil. Here we report the identification of a new virus associated with the disease. RNAs isolated from CSD-affected and nonaffected trees were used to construct cDNA libraries. A set of viral sequences present exclusively in libraries of CSD-affected trees was used to obtain the complete genome sequence of the new virus. Phylogenetic analysis revealed that this virus is a new member of the genus Marafivirus. Antibodies raised against the putative viral coat proteins allowed detection of viral antigens of expected sizes in affected plants. Electron microscopy of purified virus confirmed the presence of typical isometric Marafivirus particles. The screening of 773 affected and nonaffected citrus trees for the presence of the virus showed a 99.7% correlation between disease symptoms and the presence of the virus. We also detected the virus in aphids feeding on affected trees. These results suggest that this virus is likely to be the causative agent of CSD. The virus was named Citrus sudden death-associated virus.

Amino Acid Sequence↗
PubMed1987

Diversity of citrus tristeza virus isolates indicated by dsRNA analysis.

One major dsRNA of molecular weight (MW) 13.3 X 10(6) and two others (MW 1.9 X 10(6) and 0.8 X 10(6] were routinely detected by polyacrylamide gel electrophoresis in extracts from sweet orange (Citrus sinensis) or citron (Citrus medica) infected with each of 66 isolates of citrus tristeza virus (CTV). Several additional dsRNA were also commonly detected, usually as weakly stained bands in reproducible positions in gels, but some were very prominent, e.g., a dsRNA of MW 1.7 X 10(6) associated with a seedling yellows isolate (sy-1). No dsRNA was detected in equivalent extracts from noninoculated sweet orange and citron. End-labeled [32P] probes were made from purified full-length viral RNA or polyacrylamide gel-purified full-length dsRNA of a nonseedling yellows (nsy-1) and a seedling yellows (sy-1) isolate of CTV. Each of the four probes was able to hybridize to all major and most minor dsRNAs of both isolates in composite polyacrylamide/agrarose gels, including the 1.7 X 10(6) dsRNA specific to the seedling yellows isolate, and could readily detect CTV nucleic acid sequences in extracts from bark of infected sweet orange plants spotted onto nitrocellulose membranes. One dsRNA (MW 0.5 X 10(6] was very prominent in some isolates and much less so, or undetectable, in other isolates and 66 isolates have been screened for the presence of this dsRNA. There was a strong correlation between inability to detect the 0.5 X 10(6) dsRNA and the designation of an isolate as neither a seedling yellows type nor a stem pitting isolate of grapefruit; these properties were typical for isolates of CTV from southern California.

Citrus↗
PubMed2000

Suspected citrus pulp toxicosis in dairy cattle.

Thirteen lactating dairy cows from a herd of 650 died over a 6-week period. Most animals were down in milk production at 1 milking and were found dead at the next milking. Two cows had elevated heart rate and enlarged mandibular lymph nodes. Two others had azotemia, elevated heart rate, hyperglycemia, and weight loss. Necropsy of 10 cows revealed hemorrhages on the intestinal serosa and epicardium, lymphadenopathy, interstitial nephritis, small intestinal hemorrhage, and interstitial pneumonia. Histopathology showed lymphocytic to lymphogranulomatous inflammation in the heart, spleen, kidney, lymph nodes, liver, lung, pancreas, and adrenal gland. Phlebitis was present in 2 livers. The lesions resembled those of hairy vetch toxicosis, but no vetch was being fed. Similar lesions have been reported with the feeding of citrus pulp. Citrus pulp was being fed to the lactating cows and had been added to the diet 6 weeks before the first death. The syndrome resolved with elimination of citrus pulp from the diet.

Animals↗
PubMed2001

Differential inhibition of human cancer cell proliferation by citrus limonoids.

Limonoids have been shown to inhibit the growth of estrogen receptor-negative and -positive human breast cancer cells in culture. The primary objective of this study was to test the antiproliferative activity of limonoids (obacunone 17 beta-D-glucopyranoside, nomilinic acid 17 beta-D-glucopyranoside, limonin, nomilin, and a limonoid glucoside mixture), found in high concentrations in mandarin (Citrus reticulata Blanco), against a series of human cancer cell lines. The human cancer cell lines included leukemia (HL-60), ovary (SKOV-3), cervix (HeLa), stomach (NCI-SNU-1), liver (Hep G2), and breast (MCF-7). The growth-inhibitory effects of the four limonoids and the limonoid glucoside mixture against MCF-7 cells were significant, and the antiproliferative activity of the different citrus limonoids was also dose and time dependent. No significant effects were observed on growth of the other cancer cell lines treated with the four individual limonoids at 100 micrograms/ml. At 100 micrograms/ml, the limonoid glucoside mixture demonstrated a partial inhibitory effect on SKOV-3 cancer cells. With use of flow cytometry, it was found that all the limonoid samples could induce apoptosis in MCF-7 cells at relatively high concentrations (100 micrograms/ml). Considering the high concentration needed to induce apoptosis, it is unlikely that this is the primary mechanism of action for the cytotoxic effects seen with limonoids in this study. Further work is needed in this area to establish the mechanism of action of citrus limonoids on human breast cancer cells.

Apoptosis↗
PubMed2004

Screening of furanocoumarin derivatives in citrus fruits by enzyme-linked immunosorbent assay.

This paper reports a sensitive and specific enzyme-linked immunosorbent assay for screening of furanocoumarin derivatives as cytochrome P450 3A4 inhibitors in citrus fruits. Anti-6',7'-dihydroxybergamottin antibody was obtained by immunizing rabbits with 6',7'-dihydroxybergamottin conjugated with bovine serum albumin using the N-succinimidyl ester method. An enzyme marker was similarly prepared by coupling 6',7'-dihydroxybergamottin with beta-D-galactosidase. The enzyme-linked immunosorbent assay is capable of detecting as little as 800 pg/ml of 6',7'-dihydroxybergamottin and 4 ng/ml of bergamottin. Cross-reactivity data showed that the antibody well recognizes both the furanocoumarin and 6,7-dihydroxy-3,7-dimethyloct-2-enyloxy moieties of the 6',7'-dihydroxybergamottin, and is thus specific to the structure of furanocoumarin derivatives containing geranyloxy side chain as the cytochrome P450 3A4 inhibitors in grapefruit juice. The antibody was, therefore, used for screening a large number of citrus fruits for furanocoumarin derivatives such as 6',7'-dihydroxybergamottin. Fifteen citrus fruits were examined and significant reactivity was observed in 8 of these: red pummelo, sweetie, melogold, banpeiyu pummelo, hassaku, sour orange, lime and natsudaidai. This enzyme-linked immunosorbent assay may be a powerful tool for screening for furanocoumarin derivatives as cytochrome P450 3A4 inhibitors in grapefruit juice.

Beverages↗
PubMed2005

[Chemical study of citrus plants in the search for cancer chemopreventive agents].

The constituents of Citrus plants were investigated to develop useful agents that are effective in cancer chemoprevention. We examined the roots and bark of the roots of various Citrus plants, resulting in the isolation of many novel compounds. Their structures were determined using spectroscopic methods, especially 2D-NMR spectra. The following new compounds of constituents were found: dimeric coumarins, dimeric acridone alkaloids, and acridone-coumarin dimers. Auraptene and nobiletin are known as useful constituents of the peel of Citrus plants for cancer chemoprevention. However, we found that 3,5,6,7,8,3',4'-heptamethoxyflavone (HPT) has both antitumor promotion and initiation activities and is more effective than auraptene and nobiletin. We synthesized pentaallyl quercetin (QPA), a useful antitumor compound that has the additional effects of a P-glycoprotein (P-gp) inhibitor. The inhibitory effects of QPA on P-gp were more effective than those of the typical P-gp inhibitors cyclosporin A and verapamil. Both HPT and QPA have antitumor promotion activity and are potential candidates for effective multidrug resistance agents in cancer chemotherapy.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2001

Cloning and expression of CitPDS1, a gene encoding phytoene desaturase in citrus.

A cDNA clone encoding phytoene desaturase (PDS) was isolated from citrus (Citrus unshiu Marc.). The transcript of the isolated PDS (CitPDS1) was not detected by conventional RNA gel-blot analysis; instead, it was detected by a sensitive reverse transcription-PCR (RT-PCR). The CitPDS1 transcript in the juice sacs/segment epidermis (edible part) was at a low level in the young fruit, and it increased toward maturation like citrus phytoene synthase (CitPSY1). In the peel, in contrast to CitPSY1, the transcript of which was induced toward maturation, the level of the CitPDS1 transcript remained constant after an increase in July, indicating non-coordinate regulation of CitPDS1 and CitPSY1 in the peel.

Amino Acid Sequence↗
PubMedDate not supplied

Diversity of flies (Diptera: Tephritidae and Lonchaeidae) in organic citrus orchards in the vale do Rio Caí, Rio Grande do Sul, southern Brazil.

This study was carried out in the Counties of Montenegro and Pareci Novo located in the region of the Vale do Rio Cai, Rio Grande do Sul, Southern Brazil, aiming to determine the fruit fly species of Tephritidae and Lonchaeidae that occur in organic orchards of sweet orange [Citrus sinensis (L.) Osb.] cultivar Céu, and Murcott tangor (Citrus reticulata Blanco x C. sinensis), during the fruit ripening stages in 2003 and 2004. Eight McPhail traps baited with integral grape juice diluted to 25% were installed in four orchards, two in each citrus species. The traps were checked weekly, when the baits were changed, the flies separated and preserved in 70% ethyl alcohol. Fruits were also sampled from the orchards, placed in containers with damp soil and closed with a mesh. The tephritid flies represented 86.2% of all captured flies in the four orchards during both years. Five Tephritidae species were captured from traps: Anastrepha fraterculus (Wiedemann), Anastrepha grandis (Macquart), Anastrepha pseudoparallela (Loew), Anastrepha dissimilis Stone and Ceratitis capitata (Wiedemann). The captured species of Lonchaeidae were: Neosilba zadolicha McAlpine & Steyskal, Neosilba n.sp.3, Neosilba sp. and Lonchaea sp. Anastrepha fraterculus was found in 99% of the fruit samples, both in 'Céu' orange and 'Murcott' tangor, and Neosilba n.sp.3 were only obtained from 'Murcott' tangor fruits.

Animals↗
PubMed2006

Reproductive performance of the mite Brevipalpus phoenicis (Geijskes, 1939) on citrus and coffee, using life table parameters.

The flat-mite Brevipalpus phoenicis (Geijskes, 1939) (Acari: Tenuipalpidae) is considered important in citrus (Citrus spp.) and coffee plants (Coffea spp.) in Brazil, and is known as the leprosis and coffee ring spot mite, as being a vector of the Citrus Leprosis Rhabdovirus - CitLV and Coffee Ring Spot Virus - CoRSV. The objective of this work is to find out about the reproductive success of B. phoenicis on citric fruits and coffee leaves by fertility life table parameters and its biology. The experiments were carried out in laboratory conditions at 25 +/- 2 degrees C, 70 +/- 10% of relative humidity and 14 h of photophase. The lengths of embryonic and post-embryonic periods were different due to the host where the mite was reared. B. phoenicis showed better development and higher survival and fecundity in citric fruits than coffee leaves. The intrinsic rate of the population increase (r(m)) was 0.128 and 0.090 - females/female/day on citric fruits and coffee leaves, respectively. The citric fruits were more appropriate for the development of B. phoenicis than coffee leaves.

Animals↗
PubMed2006

Reduced oviposition by Diaprepes abbreviatus (Coleoptera: Curculionidae) and growth enhancement of citrus by Surround particle film.

Regularly applied sprays of a particle film, Surround WP, greatly enhanced the growth of citrus trees on a poorly drained Winder soil at Fort Pierce, FL. After 3 yr of applications every 3 or 4 wk, Surround-treated trees had at least 5 times the mass, 6 times the canopy volume, and approximately 4 times the cross-sectional area of the tree stems at the graft union compared with untreated trees. The larger Surround-treated trees attracted a higher number of adult weevil Diaprepes abbreviatus (L.) and to a lesser extent citrus root weevil, Pachnaeus litus (Germar), per tree, but there was an equivalent number of egg masses per tree compared with the control trees. The number of egg masses per female weevil oviposited on Surround-treated trees was significantly less than either the control trees or trees treated biannually with an entomopathogenic nematode, BioVector. The number of larvae per tree recovered from the roots of excavated trees was greater from trees treated with Surround once every 3 wk compared with control trees. The data suggest that Surround particle film greatly enhanced the growth of citrus trees grown in a poorly drained soil. The reduction in oviposition by D. abbreviatus was insufficient to significantly reduce the number of root weevil larvae per tree feeding on the roots. However, the more vigorous trees resulting from Surround applications may be more resistant or tolerant to root weevil feeding.

Analysis of Variance↗
PubMed2002

Sampling methods as abundance indices for adult Diaprepes abbreviatus (Coleoptera: Curculionidae) in citrus.

Beat sampling and two type of traps, cup traps and Tedders traps, were evaluated as sampling methods to detect and estimate population densities of adult Diaprepes abbreviatus L. weevils newly colonizing young citrus trees. The study was conducted over a 65-wk period across a 0.25-ha area of 80 citrus trees [Citrus sinensis (L.) Osbeck] (1.2-1.5 m tall). Beat samples were taken weekly to determine the number of trees infested and number of new adult weevils per tree. Sixteen of the 80 trees studied were each monitored weekly using one of the following trapping methods: cup traps in trees, cup traps on a stake in the ground within the tree drip line, cup traps on a stake in the ground outside of the drip line, Tedders traps on the ground within the drip line, and Tedders traps on the ground outside of the drip line. Weevils collected each week from trees and traps were removed from the study site. Based on the coefficients associated with Taylor's power law, the optimum numbers of trees to sample for an SEM equal to 25% of the mean estimate decreased from 50 trees at a mean of 0.5 new weevils per tree to 30 trees at a mean of 0.8 new weevils per tree. A significant relationship was found between the weekly mean number of new weevils per tree and the proportion of trees infested, a binomial relationship that could be further explored in the search for a sampling program for adult D. abbreviatus. Regression analyses indicated that three of the trapping methods served at least as weak indices of the presence and abundance of new weevils: cup traps in trees, Tedders traps inside the dripline and Tedders traps outside the dripline. Cup traps in trees and Tedders traps inside the dripline captured the most weevils and most frequently detected weevils. Although relatively inefficient as abundance indices of populations of new weevils, these two trapping methods appeared to have some value with respect to signaling when weevils first appeared in trees during the spring.

Animals↗
PubMed2002

Is there meaningful plant resistance to diaprepes abbreviatus (Coleoptera: Curculionidae) in citrus rootstock germplasm?

Host plant resistance to the Diaprepes root weevil, Diaprepes abbreviatus (L.) was assessed for seedlings of 54 Poncirus trifoliata (L.) Raf. selections and two families of progeny from crosses between Citrus and P. trifoliata. Weight gain was consistently lower when larvae were reared in pots containing the progeny of Citrus reticulata Blanco 'Sunki' x P. trifoliata 'Flying Dragon' compared with larvae reared on progeny of 'Pearl' (C. reticulata x C. paradisi Macf.) x 'Flying Dragon'. This is the first evidence of genetic control of resistance to the Diaprepes root weevil within sexually compatible citrus rootstock germplasm. There was a significant positive correlation between percentage root loss and larval weight gain within the resistant progeny, indicating a possible antixenotic effect. Two varieties of P. trifoliata were identified as more resistant than 'Flying Dragon' based on larval weight gain.

Animals↗
PubMed2004

Sucrose octanoate toxicity to brown citrus aphid (Homoptera: Aphididae) and the parasitoid Lysiphlebus testaceipes (Hymenoptera: Aphidiidae).

We report the toxicological effects of a new biorational, synthetic sucrose octanoate (AVA Chemical Ventures L.L.C., Portsmouth, NH), on brown citrus aphid, Toxoptera citricida (Kirkaldy), nymphs and adults and to its native parasitoid Lysiphlebus testaceipes (Cresson). Sucrose octanoate topically applied was equally toxic to brown citrus aphid adults and nymphs with LC50 and LC90 values ranging from 356 to 514 and 1029 to 1420 ppm, respectively. Mortalities of both stages did not differ significantly over time during the 3-24-h sampling period. Dry residues of sucrose octanoate exhibited similar levels of toxicity to both nymphs and adults. Mortality ranged from 60 to 70% at 6,000 ppm 4 h after exposure. L. testaceipes was not harmed by treatments as high as 4,000 ppm of sucrose octanoate as long as the parasitoid had mummified before treatment. Based on these results, sucrose octanoate would be a useful biorational in citrus integrated pest management programs.

Animals↗
PubMed2006

Evaluation of mating disruption for control of lightbrown apple moth (Lepidoptera: Tortricidae) in citrus.

Three large-scale mating disruption (MD) trials were conducted from 2001 to 2004 in an organic citrus orchard in inland southeastern Australia to evaluate the effectiveness of the MD dispenser Isomate LBAM Plus in controlling lightbrown apple moth, Epiphyas postvittana (Walker), in citrus. At the application rate of 364-728 dispensers per ha, the dispensers reduced pheromone trapping of E. postvittana to almost undetectable levels for approximately 6 mo in the treated area. During this period, most sentinel females in the treated area failed to mate. Infestation by E. postvittana in the treated area was reduced by >50%. If distributed in citrus orchards in late winter (August), the dispensers can be expected to remain effective until next February in southeastern Australia, covering the period when most fruit scarring caused by its larvae occurs.

Animals↗
PubMed2006

Application Fourier transform near infrared spectrometer in rapid estimation of soluble solids content of intact citrus fruits.

Nondestructive method of measuring soluble solids content (SSC) of citrus fruits was developed using Fourier transform near infrared reflectance (FT-NIR) measurements collected through optics fiber. The models describing the relationship between SSC and the NIR spectra of citrus fruits were developed and evaluated. Different spectra correction algorithms (standard normal variate (SNV), multiplicative signal correction (MSC)) were used in this study. The relationship between laboratory SSC and FT-NIR spectra of citrus fruits was analyzed via principle component regression (PCR) and partial least squares (PLS) regression method. Models based on the different spectral ranges were compared in this research. The first derivative and second derivative were applied to all spectra to reduce the effects of sample size, light scattering, instrument noise, etc. Different baseline correction methods were applied to improve the spectral data quality. Among them the second derivative method after baseline correction produced best noise removing capability and yielded optimal calibration models. A total of 170 NIR spectra were acquired; 135 NIR spectra were used to develop the calibration model; the remaining spectra were used to validate the model. The developed PLS model describing the relationship between SSC and NIR reflectance spectra could predict SSC of 35 samples with correlation coefficient of 0.995 and RMSEP of 0.79 degrees Brix.

Biochemistry↗
PubMedDate not supplied

Soil temperature, nitrogen concentration, and residence time affect nitrogen uptake efficiency in citrus.

We try to elucidate which environmental and soil factors control nitrogen uptake efficiency in citrus. Effects of residence time and nitrogen (N) concentration (three 500-mL applications of 7 mg N L(-1), representative of reclaimed water used for citrus irrigation in central Florida, or one 150-mL application of 70 mg N L(-1)) on nitrogen uptake efficiency (NUE) of young citrus seedlings were studied. Increasing residence times from 2 to 8 h increased NUE from 36 to 82% and from 17 to 34% for high and low application frequencies, respectively. We developed a model to predict N uptake based on root density, N concentration, and soil temperature (Ts). Assuming a base temperature (Tb) of 10 degrees C, N uptake temperature sum (UTS) = sigma(Ts - Tb)/24 (degrees CdN, degree day units of N uptake). To eliminate the risk of N leaching for young seedlings, minimum uptake periods of 5 and 16 degrees CdN were required at initial soil N concentrations of 0.9 and 2.5 mg N L(-1), respectively. After correcting for differences in root length, this information was then used to predict the effect of irrigation practices on N uptake from reclaimed water for mature trees. Applying 2500 mm yr(-1) vs. 400 mm yr(-1) reclaimed water reduced the NUE of N in this water from 100 to 63% during the summer and from 100 to 28% during the winter. Reductions in NUE at higher irrigation rates appeared to be related to N displacement below the root zone prior to complete N uptake.

Citrus↗