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1.
Characterization of Citrus tristeza virus isolates in northern Iran   总被引：1，自引：0，他引：1  
The biological and molecular properties of four Citrus tristeza virus (CTV) isolates isolated from infected Satsuma trees imported from Japan, and growing in citrus groves in northern Iran (Mahdasht orchards, Mazandaran Province), were investigated. CTV-infected samples were collected from sweet orange trees and grafted onto Alemow (Citrus macrophylla Wester) seedlings. On indicator plants, these isolates produced various symptoms including vein clearing and stem pitting on Mexican lime, Alemow, and Citrus hystrix, and yellowing and stunting on sour orange and grapefruit seedlings. Citrus samples were also surveyed for CTV using serological tests. The coat protein (CP) gene of these isolates was amplified using specific primers, yielding an amplicon of 672 bp for all isolates. Sequence analysis showed 98%–99% sequence homology of Iranian isolates with the Californian CTV severe stem-pitting isolate SY568 and 97%–98% homology with the Japanese seedling yellows isolate NUagA. The Iranian isolates were compared by restriction fragment length polymorphism (RFLP) analysis of the CP amplicon for further classification.  相似文献   

2.
The complete nucleotide sequence of a seedling-yellows-inducing isolate NUagA of Citrus tristeza virus (CTV) was determined. It consisted of 19302 nucleotides and contained 12 open reading frames (ORF) organized identically to those of previously sequenced isolates. This genome is the largest among the CTV genome sequenced so far ; it is 6 nucleotides (nt), 76 nt, 43 nt, and 53 nt longer than that of T36 (quick decline, Florida), VT (seedling yellows, Israel), T385 (mild, Spain), and SY568 (stem pitting, California), respectively. Sequence comparison of NUagA and the other isolates revealed approximately 90% identities throughout the 3′ half of the genome. The 5′ half of the genome was only about 70% identical to that of T36 but still high at about 90% to those of VT, SY568, and T385. Comparison of amino acid sequences on ORF1a encoding polyproteins, the most variable region, reflects the CTV isolate relationship ; NUagA is closely related to VT, SY568, and T385, but distantly related to T36. Received 29 May 2000/ Accepted in revised form 16 November 2000  相似文献   

3.
The nucleotide sequences for the minor coat protein (CPm) gene and its deduced amino acid sequences for two aphid-transmissible and two nontransmissible isolates of Citrus tristeza virus (CTV) from symptomless orchard trees of Miyagawa satsuma [Citrus unshiu (Macf.) Marc.] on trifoliate orange [Poncirus trifoliate (L.) Raf.] and declining Washington navel [C. sinensis (L.) Osb.] trees on sour orange (C. aurantium L.) rootstocks were analyzed and compared with those of highly transmissible CTV strains available in GenBank. The isolates produced severe symptoms on indicator plants and their aphid transmissibility was assayed through acquisition by A. gossypii of CTV and subsequent inoculation feeding on young Mexican lime seedlings. The CPm gene nucleotides and coded amino acid sequences were very similar among the nontransmissible isolates and among the transmissible. Five of 73 nucleotide substitutions that existed between CPm gene nucleotide sequence of nontransmissible and transmissible isolates caused changes in the deduced amino acid sequences of the nontransmissible isolates. Two nucleotide substitutions yielded new amino acids with similar properties. However, the three remaining mutations led to substitution of new amino acids with a different charge and polarity at positions 14, 238 and 239. The last two mutations occurred at the C-terminal region of the CPm, which is implicated in the formation of a salt bridge that helps to maintain the protein’s tertiary structure. Amino acid substitutions can affect aphid transmission efficiency by altering the conformation of the proteins or masking motifs involved in the interaction between CPm and aphid stylets.  相似文献   

4.
柑橘衰退病毒含量对其蚜传效率的影响   总被引：1，自引：0，他引：1  
为明确毒源植株和蚜虫中柑橘衰退病毒(Citrus tristeza virus,CTV)的发生情况与蚜虫传播CTV能力的关系,将褐色橘蚜、棉蚜、橘二叉蚜和绣线菊蚜置于分别感染了4个CTV分离株的锦橙上取食24 h后,运用巢式RT-PCR和实时RT-PCR检测蚜虫和锦橙的带毒情况,并分析蚜虫的传毒能力。结果显示,蚜虫中CTV的平均带毒率为0.76~0.84,其中棉蚜的最高,其次为绣线菊蚜、褐色橘蚜和橘二叉蚜。锦橙中各CTV分离株的含量差异不大,与蚜虫传毒效率间无显著相关性;也未发现蚜虫带毒率与其传播CTV能力之间存在相关性。蚜虫体内CTV的含量为5.36×103±2.33×103~2.01×106±3.67×105拷贝,褐色橘蚜中含量最高,其次为棉蚜、橘二叉蚜和绣线菊蚜;且高蚜传能力CTV分离株在褐色橘蚜体内的含量远高于低蚜传能力分离株。表明蚜虫体内CTV的含量可能与蚜虫传毒能力有关。  相似文献   

5.
A Citrus tristeza virus (CTV) isolate (L192GR) naturally occurring in lemon trees of more than 100 years old in Greece was fully characterized. Virus‐derived small interfering RNAs, induced by Dicer processing of dsRNAs formed during RNA virus replication, were isolated and used as targets for sequencing. Next‐generation high‐throughput sequencing using the Ion Torrent platform was performed. A total of 432 632 sequences, 94·05% of which corresponded to L192GR, were determined. Subsequent bioinformatics analysis enabled the determination of the full‐length 19 251 nt genome of the L192GR isolate (GenBank no. KC262793 ). Comparative analysis of complete genomes revealed molecular homology with CTV‐VT isolate FS2‐2 from Florida (GenBank no. EU937519 ) with 98·2% nucleotide sequence identity. Recombination events were detected in L192GR and they probably contribute to its unique characteristics. Specifically, although most isolates of the CTV‐VT group induce the seedling yellows syndrome and react positively with the monoclonal antibody MCA13, which is typically associated with severe CTV isolates, the MCA13‐positive L192GR gave very mild or even no symptoms in the seedling yellows indicator plants. Furthermore, experimental aphid transmissibility studies revealed a poor transmission efficiency of 20%. This is the first report of a CTV isolate originating from a lemon tree being fully characterized at biological, serological and molecular levels. The present study further confirms that, when the goal is the risk assessment associated with a new pathogen or isolate in a particular area, molecular data have to be combined with the biological properties of the pathogen.  相似文献   

6.
Darjeeling and Sikkim hills of India are well known for production of mandarin orange (Citrus reticulata). The recent spread of Citrus tristeza virus (CTV) has threatened the citrus cultivation in this region. During a survey in Darjeeling and Sikkim hills, pomelo trees were recorded as CTV free. Since pomelo trees did not show any disease appearance, a study was undertaken to ascertain whether they are resistant to CTV infection or resistant to aphid feeding or both. Toxoptera citricida, the most efficient aphid vector and which is abundantly present in this region, did not feed on pomelo when other Citrus species such as mandarin, kagzi lime and rough lemon were available. Additionally, CTV isolates of Darjeeling and Sikkim hills were not transmissible to pomelo either by T. citricida or grafting. We report for the first time that pomelo is resistant to isolates of CTV present in this region.  相似文献   

7.
ABSTRACT A total of 14 Spanish isolates of Citrus tristeza virus (CTV) and 1 isolate from Japan were transmitted by Aphis gossypii, and the subisolates obtained were compared with the source isolates for symptom expression and double-stranded RNA (dsRNA) pattern. Of the 14 Spanish isolates, 9 showed altered dsRNA patterns after aphid transmission but only minor variations in the intensity of symptoms induced on Mexican lime. Northern blot hybridization with complementary DNA (cDNA) probes corresponding to both the 5' and the 3' termini of the CTV genomic RNA (gRNA) showed that the dsRNA bands that could be used to discriminate between the dsRNA pattern of the source and the aphid-transmitted isolates were the replicative forms of defective RNAs (D-RNAs). Conversely, the Japanese isolate and two subisolates obtained from it by aphid transmission had the same dsRNA pattern, but one of the subisolates induced milder symptoms in several hosts. Dot-blot hybridization with cDNA probes representing several regions of the gRNA showed that most of the aphid-transmitted isolates differed from the corresponding source isolate by their hybridization pattern. Our results indicate that aphid transmission often sorts the populations of gRNA variants and D-RNAs present in CTV isolates.  相似文献   

8.
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9.
The population of genomic RNA sequence variants of citrus tristeza virus (CTV) isolates was characterized by single-strand conformation polymorphism (SSCP) analysis of complementary DNA (cDNA) of the genes p18 and p20. Comparison of field and aphid-transmitted isolates showed that aphid transmission frequently altered the single-strand conformation polymorphism (SSCP) pattern of both genes, indicating changes in the population of genomic RNA variants. SSCP analysis of the cDNA of RNA extracted from small pieces of tissue sampled at different sites of the same plant sometimes yielded different patterns, indicating uneven distribution of the genomic RNA variants within the infected plant. Different SSCP patterns were also obtained when the RNA extracted from individual aphids probing in the same infected leaf was used as a template. Uneven distribution of the genomic RNA variants within the infected plant and sorting of some of these variants by individual aphids probably contribute to changes observed in the CTV population following aphid transmission.  相似文献   

10.
A large‐scale survey of Citrus tristeza virus (CTV) was carried out from 2016 to 2018 in the Chlef Valley, one of the main citrus growing areas in Algeria. In this study a total of 1680 citrus trees from 93 commercial orchards were sampled. The collected samples were tested by direct tissue blot immunoassay analysis and by the double antibody sandwich enzyme‐linked immunosorbent assay technique, and 54 trees were identified as being infected with CTV. This result confirmed that 54 trees were infected by the virus, corresponding to an infection rate of 3.21% throughout the studied area. Five of these local CTV sources were chosen for further molecular investigations to determine the genotype associated with the CTV isolates now spreading in the Chlef area. Characterization with multiple molecular markers showed the presence of the T30 and VT genotypes. This result allowed confirmation of the presence of a virulent strain belonging to the VT genotype. The other CTV isolates were similar to those from the Mitidja region, which showed 99% nucleotide identity with the Spanish mild CTV isolate. This early finding of a strain belonging to the VT genotype is an issue for Algerian citrus producers and needs rapid actions to be taken by the National phytosanitary services, extending the surveillance to other citrus production regions and uprooting the infected trees.  相似文献   

11.
Citrus tristeza virus (CTV) represents one of the major threats to citrus production worldwide. In the East Adriatic region, CTV symptoms are mostly absent due to traditional citrus grafting on trifoliate orange (Poncirus trifoliata), a CTV-tolerant rootstock. Therefore, the virus has been continuously spreading by the propagation of infected material. The genetic variability of CTV was studied on nineteen citrus samples, collected from orchards in the coastal region of Croatia, Montenegro and Albania, that previously tested positive by ELISA and immunocapture RT-PCR. Single-strand conformation polymorphism of the amplified coat protein gene demonstrated the presence of different CTV variants in each amplicon, while sequence analysis of cloned CP gene variants confirmed their clustering into six out of the seven phylogenetic groups so far delineated. Four of these groups include sequences of severe quick decline, seedling yellows and stem-pitting (SP) isolates, thought to be found only rarely in the Mediterranean region. Regardless of the lack of symptoms in the field, CTV isolates from the East Adriatic displayed high genetic variability and pathogenic potential, additionally confirmed by biological characterisation. The high percentage of mixed infections suggest the potential for further diversification and a greater risk of severe variants spreading into new areas.  相似文献   

12.
The effect of Citrus tristeza virus (CTV) infection on photosynthetic activity and antioxidant metabolism was analysed in plants of the highly susceptible citrus genotype Mexican lime (Citrus aurantifolia). Two virus isolates differing in their virulence (the severe T318 and the mild T385) were used in the experiments. CTV infection caused a reduction in photosynthetic capacity in infected plants. This limitation was mainly due to a reduction in the carboxylative efficiency whereas the limitation of CO2 diffusion through the stoma had lower impact. The virus did not damage the antennae and did not reduce the efficiency of light harvesting complexes. Oxidative damage occurred in infected plants, as evidenced by the increase in malondialdehyde levels. Indeed, CTV infection caused an increase in ascorbate peroxidase activity in new shoots developed in infected plants during the 2 years of the experiment. Data suggest that the H2O2 removal machinery was not damaged as a result of stress but the defence mechanism was overwhelmed with time due to the continuing pressure of biotic stress.  相似文献   

13.
 柑橘衰退病毒(Citrus tristeza virus,CTV),柑橘碎叶病毒(Citrus tatter\|leaf virus,CTLV),柑橘裂皮病类病毒(Citrus exocortis viroid,CEVd)和柑橘黄龙病(Huanglongbing, HLB)亚洲种病原(Candidatus liberobacter asiaticus)是重要的柑橘嫁接传播病原。本文建立了同时检测HLB病菌、CTV、CEVd 和CTLV 4种柑橘嫁接病原的一步法、双温多重PCR检测技术体系,同时在体系中设置内参基因。应用该体系快速评价了4种嫁接传播病原在田间侵染情况,结果表明28个田间样品CTV、CEVd、CTLV和HLB感染率分别为89.3 %、17.9 %、10.7 %和28.6 %,接近半数样品为混合感染。并且将该方法应用于快速评价茎尖嫁接苗病毒的脱除情况。  相似文献   

14.
Citrus tristeza virus (CTV) is the causal agent of the most important virus disease of citrus. Numerous CTV isolates differing in biological and molecular characteristics have been reported worldwide. Recently, CTV was detected in Italy in several citrus crops from three separate areas: (1) Cassibile, province of Syracuse; (2) Massafra, province of Taranto; and (3) Belpasso, province of Catania. CTV isolates from Massafra and Cassibile were mild, whereas isolates from Belpasso induced severe symptoms. To study the genetic variation of CTV populations of these areas, 150 samples per area were examined by single strand conformation polymorphism (SSCP) and nucleotide sequence analysis of CTV gene p20. All isolates from the same area showed the same SSCP pattern whereas for each area a different SSCP pattern was obtained. The Massafra and the Cassibile isolates had a nucleotide identity higher than 99% with a mild isolate from Spain and about 92% with the Belpasso isolates, which were similar (identity higher than 99%) to severe isolates from California and Japan. These results suggest at least two independent introductions of CTV in Italy, probably by import of CTV-infected budwoods. Within each area, the virus population was homogeneous suggesting diffusion of CTV by aphid transmission. The GenBank accession numbers of the sequences reported in this paper are: AY262000, AY263360 and AY263361 corresponding to gene p20 of CTV isolates from Massafra, Cassibile and Belpasso (Italy), respectively.  相似文献   

15.

The resistance of ashgourd to the melon-cottonaphid, Aphis gossypii (Glover), and the incidence of aphid-transmitted viral diseases was investigated following recording of aphid numbers and trichome (plant hair) numbers per cm2 of leaf on four parts of the cucurbit, ashgourd, representing four genotypes (Local Sylhet, Local Round, High Female and CQ-10-90). The Local Sylhet genotype was found to have the highest trichome density compared with those of Local Round, High Female, and CQ-10-90 ashgourd genotypes, and was least infested by Aphis gossypii. The percentage of virus-infected plants was also found to be lowest on the Local Sylhet genotype. Aphid numbers were lowest on the terminal and young leaves of ashgourd compared with mature and senescent leaves, due to the higher trichome density in the former leaf categories. Trichome density was found to have a significant negative influence on the number of aphids. Trichomes in ashgourd play a major role by providing a first line defence in reducing aphid infestation, with a subsequent beneficial effect of reducing the incidence of virus.  相似文献   

16.
S. Korkmaz 《Phytoparasitica》2002,30(4):420-428
Biological properties and dsRNA patterns of one Cyprus and three Turkish isolates of citrus tristeza virus (CTV) were investigated. In addition, CTV antigen concentration and effect of tissue sampling time from naturally infected Shamouti sweet orange trees grown in the field of Icel Province, Turkey, were also determined. The Cyprus isolate showed vein clearing symptoms on grapefruit, ‘Madam Vinous’ and Mexican lime and stem pitting symptoms on Mexican lime. The three Turkish isolates showed only vein clearing symptoms on Mexican lime. All four isolates showed a full-length major double-stranded RNA (dsRNA) band of 13.3 × 106 Da mol. wt in extracts from infected Madam Vinous sweet orange trees, and major or minor dsRNA bands with 2.0. 0.8 and 0.5 × 106 mol.wt. All seven different citrus varieties inoculated with the Igdir (D) strain contained full-length dsRNA. The additional two dsRNA of 0.8 and 0.5 × 106 mol.wt were also detected as clearly as full-length dsRNA in these hosts, but were weaker inCitrus exelsa and ‘Interdonat’ lemon. Madam Vinous, rough lemon and Mexican lime were the best hosts for dsRNA analysis. ELISA values were highest in April (OD405nm =0.476), decreased steadily until August, and then increased gradually through December. ELISA values were lowest in July and August (OD405nm =0.157 and 0.141, respectively). dsRNA recovery from a field tree infected with isolate Igdir D was good in March, April and May and poor in January and February. No dsRNA band was detected in August or September. http://www.phytoparasitica.org posting July 9, 2002.  相似文献   

17.
An island wide survey for Citrus tristeza virus (CTV) in citrus orchards across Jamaica (13 regions) was conducted over 2 years. Trees (1, 885) showing virus-like symptoms as well as asymptomatic trees were randomly sampled for testing by ELISA and 55 samples from the 6 major citrus growing regions were graft inoculated on indicator plants. Most samples (74%) reacted to polyclonal antibodies against CTV in ELISA, while 20% were positive in tests using monoclonal antibodies specific to severe CTV strains. Samples collected from the 6 major citrus growing regions produced vein clearing and stem pitting symptoms on Mexican lime indicator plants (87%). In addition, stem pitting symptoms were induced on Duncan grapefruit, sweet orange, sour orange or sweet orange grafted on sour orange. Nucleotide sequencing of the coat protein gene sequences isolated from these samples indicated high identities (88 to 95.5%) among the Jamaican isolates and previously reported stem pitting strains from Central and North America and Eurasia (88 to 100%). The results suggest a shared ancestry with isolates from other geographical locations, rather than geographical speciation, and presumably separate CTV introductions into Jamaica.  相似文献   

18.
Reactions that occur when a plant is subjected to Citrus tristeza virus (CTV) infection often result in triggering of numerous defence mechanisms to fight the infection. The reactions vary according to virus strain, host genotype, time of exposure to the infection and environmental conditions. To date, no study has examined in detail the consequences of 10‐year exposure to CTV infection on the biochemical and physiological status of susceptible Mexican lime plants (Citrus aurantifolia). To understand the reaction of such plants, changes in nutrient status, total proteins, enzyme activity involved in scavenging of reactive oxygen species, photosynthetic and transpiration rates, chlorophyll content, membrane permeability and water content were analysed in plants infected with different CTV isolates and in healthy plants. The activity of superoxide dismutase and polyphenol oxidase significantly decreased in the infected leaves, and membrane permeability was lower in the infected plants. Macro‐ and micronutrient elements were significantly changed: concentrations of leaf nitrogen, zinc, magnesium and iron were elevated but potassium concentration depressed in comparison to noninfected control leaves. Levels of other analysed nutrient elements, enzymes, photosynthesis and stomatal conductance, chlorophyll content and relative water content were unchanged. Clear physiological changes were found among infected and noninfected control plants but none between plants infected with different CTV isolates. The data suggest that some of the defence mechanisms investigated here were suppressed due to the continuous and long‐term pressure of biotic stress.  相似文献   

19.
The phloem‐sucking psyllid Cacopsylla picta plays an important role in transmitting the bacterium ‘Candidatus Phytoplasma mali’, the agent associated with apple proliferation disease. The psyllid can ingest ‘Ca. Phytoplasma mali’ from infected apple trees and spread the bacterium by subsequently feeding on uninfected trees. Until now, this has been the most important method of ‘Ca. Phytoplasma mali’ transmission. The aim of this study was to investigate whether infected C. picta are able to transmit ‘Ca. Phytoplasma mali’ directly to their progeny. This method of transmission would allow the bacteria to bypass a time‐consuming reproductive cycle in the host plant. Furthermore, this would cause a high number of infected F1 individuals in the vector population. To address this question, eggs, nymphs and adults derived from infected overwintering adults of C. picta were reared on non‐infected apple saplings and subsequently tested for the presence of ‘Ca. Phytoplasma mali’. In this study it was shown for the first time that infected C. picta individuals transmit ‘Ca. Phytoplasma mali’ to their eggs, nymphs and F1 adults, thus providing the basis for a more detailed understanding of ‘Ca. Phytoplasma mali’ transmission by C. picta.  相似文献   

20.
The metabolism of the chloronicotinyl insecticide imidacloprid is strongly influenced by the method of application. Whilst in foliar application most of the residues on the leaf surface display unchanged parent compound, most of the imidacloprid administered to plants by soil application or seed treatment is metabolized more or less completely, depending on plant species and time. The present study revealed that certain metabolites of imidacloprid which have been described in crop plants are highly active against aphid pests in different types of bioassays. Some of these metabolites showed a high oral activity against the green peach aphid (Myzus persicae), and the cotton aphid (Aphis gossypii). The aphicidal potency of the metabolites investigated was weaker in aphid dip tests than in oral ingestion bioassays using artificial double membranes. The most active plant metabolite was the imidazoline derivative of imidacloprid. The LC50 values of this metabolite for M. persicae and A. gossypii in oral ingestion bioassays were in the lower ppb-range, i.e. 0·0044 and 0·0068 mg litre-1, respectively. Most of the other reported metabolites showed much weaker activity. Compared to imidacloprid, the imidazoline derivative showed superior affinity to housefly (Musca domestica) head nicotinic acetylcholine receptors, while all other metabolites were less specific than imidacloprid. It seems possible that, after seed treatment or soil application, a few of the biologically active metabolites arising are acting in concert with remaining levels of the parent compound imidacloprid, thus providing good control and long-lasting residual activity against plant-sucking pests in certain crops. © 1998 SCI.  相似文献   

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