scholarly journals The First Citrus tristeza virus Outbreak Found in a Relevant Citrus Producing Area of Sicily, Italy

Plant Disease ◽  
2003 ◽  
Vol 87 (3) ◽  
pp. 314-314 ◽  
Author(s):  
S. Davino ◽  
M. Davino ◽  
A. Sambade ◽  
M. Guardo ◽  
A. Caruso
Keyword(s):  
Citrus Tristeza Virus ◽  
Polyclonal Antibodies ◽  
Accurate Estimation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sour Orange ◽  
Citrus Industry ◽  
Aphid Vectors ◽  
History Of ◽  
Tristeza Virus ◽  
Citrus Tristeza

In the course of a survey to select superior old citrus lines in the area of Siracusa (Sicily, Italy), trees in several blocks of Fortune (Citrus reticulata Blanco), Nova (C. reticulata Blanco), Satsuma (C. unshiu (Macfad.) mandarins Marc.), and Marsh grapefruit (C. paradisi Macfad.) propagated on sour orange (C. aurantium L.) rootstock showed stunting, decline, dieback, and small-sized fruits. Stunting was particularly evident in grapefruit. Declined plants consistently showed pin-holing in the cambial face of sour orange bark below the bud union line, which is often associated with Citrus tristeza virus (CTV) infection. Young shoots from 600 Fortune, 300 Nova, 400 Satsuma, and 20 Marsh grapefruit plants showing decline were analyzed by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) (Loewe Phytodiagnostica Biochemica, Sauerlach, Germany) and by immunoprinting-ELISA (Agritest Srl Valenzano-Bari-Italy) using CTV specific polyclonal antibodies. All decline tree samples reacted positively with both techniques while healthy greenhouse controls were negative. Total RNA was extracted from 50 of those plants, 25 Fortune and 15 Nova mandarins, 5 Satsuma, and 5 Marsh grapefruit (Qiagen RNeasy Plant minikit, Qiagen S.P.A., Milan, Italy), and tested in reverse transcription-polymerase chain reaction (RT-PCR) using specific primers for genes p20 (forward 5′-CGA GCT TAC TTT AGT GTT A-3′ from CTV T36 genomic position 17767-17786 and reverse 5′-TAA TGT CAA ACT GAC CGC from CTV T36 position 18269-18286) and p23 (forward 5′-ACT AAC TTT AAT TCG AAC A-3′ from CTV T36 position 18347-18286 and reverse 5′-AAC TTA TTC CGT CCA CTT C-3′ from CTV T36 position 19026-19044) (2). In all cases, DNA fragments of the expected size were amplified. Equivalent samples from CTV-free greenhouse control plants did not react in ELISA and yielded no DNA after amplification with the same primers. When the history of the plants in the affected blocks was traced, it was found that all Fortune, Nova, satsuma and Marsh grapefruit trees had been propagated from budwood illegally imported from Spain 10 years before, suggesting the possibility that the imported buds were infected with CTV. The estimated number of infected plants in the area of Siracusa is approximately 10,000, and some evidence suggests that the virus might be spreading in the area (work in progress). Only scattered CTV-infected trees had been detected in Italy previously (1). To our knowledge, this is the first report of an important CTV outbreak in Italy. Additional surveys are being conducted to get a more accurate estimation of the CTV incidence, to determine if the virus is being dispersed by aphid vectors, and to biologically and molecularly characterize the virus strains present in the affected area. Presently, there are approximately 100,000 ha of citrus in Sicily, mostly grown on decline susceptible sour orange rootstock. The presence and potential spread of CTV is a major threat for this citrus industry. References: (1) M. Davino and G. Terranova. Frutticoltura 61:18, 1999. (2) A. Sambade et al. Plant Pathol. 51:257, 2002.

scholarly journals Cross-Protection of Grapefruit from Decline-Inducing Isolates of Citrus Tristeza Virus

Plant Disease ◽  
1999 ◽  
Vol 83 (11) ◽  
pp. 989-991 ◽  
Author(s):  
C. A. Powell ◽  
R. R. Pelosi ◽  
P. A. Rundell ◽  
E. Stover ◽  
M. Cohen
Keyword(s):  
Citrus Tristeza Virus ◽  
Natural Infection ◽  
Tree Height ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sour Orange ◽  
River Region ◽  
Indian River ◽  
Tristeza Virus ◽  
Better Than ◽  
Citrus Tristeza

The ability of three mild isolates of citrus tristeza virus (CTV) to prevent natural infection of 84 Ruby Red grapefruit on sour orange rootstock by aphid-transmitted, decline-inducing isolates of CTV was assessed by symptoms and verified by enzyme-linked immunosorbent assay (ELISA) after 16 years. Of 21 trees in each of four treatments protected by the DD 102 bb, Guettler HS, and DPI 1-12-5-X-E mild CTV isolates, 14, 10, and 14% were infected by severe isolates (MCA13 monoclonal antibody reactive) compared with 67% for unprotected control trees. The health of trees protected by the DD 102 bb CTV isolate was significantly better than that of unprotected control trees as measured by decline, tree ratings, and tree height. These data suggest that infection by certain mild isolates of CTV can cross-protect grapefruit trees on sour orange rootstock from decline-inducing isolates of CTV that are prevalent in the Indian River region of Florida.

scholarly journals Suppression of Decline-inducing Isolates of Citrus Tristeza Virus by Nondecline-inducing Isolates

HortScience ◽  
2003 ◽  
Vol 38 (1) ◽  
pp. 62-64 ◽  
Author(s):  
Charles A. Powell ◽  
Phyllis A. Rundell ◽  
Robert R. Pelosi
Keyword(s):  
Citrus Tristeza Virus ◽  
Sweet Orange ◽  
Minor Component ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sour Orange ◽  
Citrus Paradisi ◽  
Indicator Plants ◽  
Tristeza Virus ◽  
Swingle Citrumelo ◽  
Citrus Tristeza

Bark chips from six container-grown citrus trees, infected with nondecline-inducing citrus tristeza virus (CTV) isolates and maintained in a vector-free greenhouse for 10 years, 15 commercial grapefruit (Citrus paradisi Macf.) trees, and 16 commercial sweet orange [C. sinensis (L.) Osbeck] trees were used to inoculate three indicator plants each of `Madam Vinous' sweet orange [C. sinensis (L.) Osbeck], sour orange (C. aurantium L.), `Duncan' grapefruit (C. paradisi Macf.), `Mexican' lime [C. aurantifolia (Christm.)], Swingle citrumelo [C. paradisi Macf. × Poncirus trifoliota (L.) Raf.], and sour orange grafted with `Hamlin' sweet orange [C. sinensis (L.) Osbeck]. All plants providing bark chips had repeatedly tested positive by enzyme-linked immunosorbent assay (ELISA) for CTV [reacted with monoclonal antibody (MAb) 17G11], but tested negative for Florida decline-inducing isolates of CTV (did not react with MAb MCA13). After 6 months in vector-free greenhouses, all in oculated trees (except Swingle citrumelo, which is considered CTV resistant) were positive for CTV by 17G11 ELISA. In addition, some indicator plants inoculated from nine (two container, two commercial grapefruit, and five commercial orange trees) of the 37 bark chip source trees also were positive for decline-inducing CTV by MCA13 ELISA. Some of these positive indicators also showed vein-clearing symptoms characteristic of infection with a severe isolate of CTV. No control, noninoculated indicators in the same greenhouse, became infected with either decline-inducing or nondecline-inducing CTV. These results indicate that decline-inducing isolates of CTV can be present as a minor component of a mixture at levels undetectable by ELISA, and that these decline-inducing isolates can become detectable by ELISA and sometimes by symptoms when inoculated into indicator plants.

scholarly journals Breakdown of Cross-Protection of Grapefruit from Decline-Inducing Isolates of Citrus tristeza virus Following Introduction of the Brown Citrus Aphid

Plant Disease ◽  
2003 ◽  
Vol 87 (9) ◽  
pp. 1116-1118 ◽  
Author(s):  
C. A. Powell ◽  
R. R. Pelosi ◽  
P. A. Rundell ◽  
M. Cohen
Keyword(s):  
Immunosorbent Assay ◽  
Citrus Tristeza Virus ◽  
Cross Protection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Experimental Area ◽  
Toxoptera Citricida ◽  
The Cross ◽  
Tristeza Virus ◽  
Citrus Tristeza

A 21-year-old replicated field planting of 84 ‘Ruby Red’ grapefruit trees cross-protected with three mild isolates of Citrus tristeza virus (CTV) was assessed for decline-inducing and non-decline-inducing isolates of the virus 5 years after the brown citrus aphid (BrCA) (Toxoptera citricida Kirkaldy) first was established in the experimental area. Prior to the introduction of the BrCA, the cross-protecting mild isolates had significantly reduced detectable infection with decline-inducing isolates of CTV for 16 years (average infection of 13% in cross-protected trees compared with 67% in unprotected trees). After the introduction of the BrCA, infections with decline-inducing CTV (measured by enzyme-linked immunosorbent assay) were 57, 81, and 71% for trees protected with three mild isolates, respectively, compared with 95% in unprotected trees. These results suggest that the introduction of BrCA accelerated the breakdown of cross-protection against decline-inducing isolates of CTV in grapefruit.

scholarly journals In Situ Immunoassay (ISIA) of Field Grapefruit Trees Inoculated with Mild Isolates of Citrus tristeza virus Indicates Mixed Infections with Severe Isolates

Plant Disease ◽  
2002 ◽  
Vol 86 (5) ◽  
pp. 458-461 ◽  
Author(s):  
Youjian Lin ◽  
Phyllis A. Rundell ◽  
Charles A. Powell
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Citrus Tristeza Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mixed Infections ◽  
The Cross ◽  
Tristeza Virus ◽  
Mild Isolate ◽  
Citrus Tristeza

Ten grapefruit trees that had been inoculated with a mild isolate of Citrus tristeza virus (CTV) and maintained in the field for 18 years were found in a previous study to be declining and infected with severe isolates of CTV, or symptomless and infected with mild isolates of CTV, using enzyme-linked immunosorbent assay (ELISA). They were assayed with an in situ immunoassay (ISIA) procedure using monoclonal antibodies 17G11 (reacts with most Florida isolates of CTV) and MCA13 (reacts with severe, but not Florida mild isolates of CTV). All the grapefruit trees were 17G11 positive by ELISA and ISIA. The five trees that showed moderate decline symptoms were MCA13 positive by ELISA and ISIA. The five symptomless trees were MCA13 negative by ELISA. However, four of the five symptomless trees were MCA13 positive by ISIA, which showed that ISIA with MCA13 had greater sensitivity in detecting severe CTV isolates than ELISA. These results suggested that the cross-protected grapefruit trees, regardless of symptoms, were infected with both mild and severe isolates of CTV.

scholarly journals Citrus tristeza virus: a pathogen that changed the course of the citrus industry

2008 ◽  
Vol 9 (2) ◽  
pp. 251-268 ◽  
Author(s):  
PEDRO MORENO ◽  
SILVIA AMBRÓS ◽  
MARIA R. ALBIACH-MARTÍ ◽  
JOSÉ GUERRI ◽  
LEANDRO PEÑA
Keyword(s):  
Citrus Tristeza Virus ◽  
Citrus Industry ◽  
Tristeza Virus ◽  
Citrus Tristeza

scholarly journals Field Diagnosis of Citrus Tristeza Virus

EDIS ◽  
1969 ◽  
Vol 2005 (1) ◽  
Author(s):  
Stephen H. Futch ◽  
Ronald H. Brlansky
Keyword(s):  
Citrus Tristeza Virus ◽  
Cooperative Extension Service ◽  
Extension Service ◽  
Tree Decline ◽  
Sour Orange ◽  
University Of Florida ◽  
Nursery Trees ◽  
Citrus Macrophylla ◽  
Tristeza Virus ◽  
Citrus Tristeza

Citrus tristeza virus (CTV) is one of the most important pathogens affecting citrus worldwide. Tristeza was first reported in Florida in the 1950s. By the 1980s, it produced serious losses due to tree decline and death on sour orange and Citrus macrophylla rootstocks. Tree decline continues to be a problem today in groves that still have sour orange rootstock trees remaining. Due to CTV, few if any nursery trees are being propagated in Florida on sour orange, bittersweet or Citrus macrophylla rootstock. This document is HS996, one of a series of the Horticultural Sciences Department, Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida. Published February 2005. HS996/HS242: Field Diagnosis of Citrus Tristeza Virus (ufl.edu)

scholarly journals PRODUCTION OF POLYCLONAL ANTIBODY TO THE COAT PROTEIN OF CITRUS TRISTEZA VIRUS IN CHICKEN EGGS

2016 ◽  
Vol 4 (1) ◽  
pp. 18
Author(s):  
Nurhadi Nurhadi ◽  
Kamaruzaman Sijam ◽  
Inon Sulaiman
Keyword(s):  
Coat Protein ◽  
Polyclonal Antibody ◽  
Citrus Tristeza Virus ◽  
Sodium Dodecyl ◽  
Virus Titer ◽  
Healthy Plant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sds Page ◽  
Tristeza Virus ◽  
Citrus Tristeza

Citrus tristeza virus (CTV) is one of the most destructive diseases in many citrus growing areas of Indonesia. Effective strategies for controlling CTV depend on diagnostic procedure namely enzyme-linked immunosorbent assay (ELISA). Study aimed to purify the CTV antigen and produced its polyclonal antibody. Virion of the severe CTV isolate designated UPM/ T-002 was concentrated by polyethylene glycol (PEG) precipitation combined with low speed centrifugation. Semipurified antigen was further purified by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). The specific coat protein (CP) band of CTV with molecular weight of 25 kD was excised and eluted using elution buffer containing 0.25 M Tris-HCl pH 6.8 + 0.1% SDS, then used as antigen for injection into 6-month-old female of White Leghorn chicken. Results, showed than the specific polyclonal antibody raised against the 25-kDa CP had a titer of approximately 104, gave low background reaction with healthy plant sap and reacted specifically with CTV isolates. The reaction was equally strong for a severe, a moderate, a mild, and a symptomless isolate, suggesting a broad reaction range of this antibody toward different CTV isolates. Optimal virus titer can be obtained since virus loss during purification could be minimized and the highly purified antigen as an immunogen could be obtained by cutting out the CP band from SDS-PAGE gels. Large amount of highly titer of CTV antibody can be produced in chicken egg. The simplicity of the procedure makes it economically acceptable and technically adoptable because the antibody can be produced in basic laboratory.

scholarly journals PRODUCTION OF POLYCLONAL ANTIBODY TO THE COAT PROTEIN OF CITRUS TRISTEZA VIRUS IN CHICKEN EGGS

2016 ◽  
Vol 4 (1) ◽  
pp. 18
Author(s):  
Nurhadi Nurhadi ◽  
Kamaruzaman Sijam ◽  
Inon Sulaiman
Keyword(s):  
Coat Protein ◽  
Polyclonal Antibody ◽  
Citrus Tristeza Virus ◽  
Sodium Dodecyl ◽  
Virus Titer ◽  
Healthy Plant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sds Page ◽  
Tristeza Virus ◽  
Citrus Tristeza

Citrus tristeza virus (CTV) is one of the most destructive diseases in many citrus growing areas of Indonesia. Effective strategies for controlling CTV depend on diagnostic procedure namely enzyme-linked immunosorbent assay (ELISA). Study aimed to purify the CTV antigen and produced its polyclonal antibody. Virion of the severe CTV isolate designated UPM/ T-002 was concentrated by polyethylene glycol (PEG) precipitation combined with low speed centrifugation. Semipurified antigen was further purified by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). The specific coat protein (CP) band of CTV with molecular weight of 25 kD was excised and eluted using elution buffer containing 0.25 M Tris-HCl pH 6.8 + 0.1% SDS, then used as antigen for injection into 6-month-old female of White Leghorn chicken. Results, showed than the specific polyclonal antibody raised against the 25-kDa CP had a titer of approximately 104, gave low background reaction with healthy plant sap and reacted specifically with CTV isolates. The reaction was equally strong for a severe, a moderate, a mild, and a symptomless isolate, suggesting a broad reaction range of this antibody toward different CTV isolates. Optimal virus titer can be obtained since virus loss during purification could be minimized and the highly purified antigen as an immunogen could be obtained by cutting out the CP band from SDS-PAGE gels. Large amount of highly titer of CTV antibody can be produced in chicken egg. The simplicity of the procedure makes it economically acceptable and technically adoptable because the antibody can be produced in basic laboratory.

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