Post-Transcriptional Gene Silencing of the p23 Silencing Suppressor of Citrus tristeza virus Confers Resistance to the Virus in Transgenic Mexican Lime

2006 ◽  
Vol 60 (2) ◽  
pp. 153-165 ◽  
Author(s):  
Carmen Fagoaga ◽  
Carmelo López ◽  
Alfonso Hermoso de Mendoza ◽  
Pedro Moreno ◽  
Luis Navarro ◽  
...  
Plant Disease ◽  
2008 ◽  
Vol 92 (10) ◽  
pp. 1468-1468
Author(s):  
R. Abou Kubaa ◽  
K. Djelouah ◽  
A. M. D'Onghia ◽  
R. Addante ◽  
M. Jamal

During the spring of 2006, the main Syrian citrus-growing areas of Lattakia (Jableh, Aledyye, Eseelya, Siano, and Hresoon provinces) and Tartous (Almintar, Aljammase, Karto, Majdaloonelbahr, Yahmour, Amreet, Althawra, and Safita provinces) were surveyed to assess the presence of Citrus tristeza virus (CTV). Eight nurseries (approximately 130 plants per nursery), two budwood source fields (approximately 230 trees per field), and 19 groves (approximately 60 trees per grove) containing the main citrus varieties were visually inspected and sampled for serological assays. The hierarchical sampling method was carried out in each selected grove (2). Infected samples were collected from two nurseries, two budwood source fields, and six groves. Stems and leaf petioles from nursery trees and flower explants from the groves were collected and analyzed for CTV by direct tissue blot immunoassay (DTBIA) with the commercial kit from Plantprint (Valencia, Spain). Of 2,653 samples tested, 89 (4%) CTV-infected plants were detected. Five citrus varieties were found to be infected and Meyer lemon (Citrus limon ‘Meyer’) had the highest incidence at 16%. Numerous sweet orange varieties (Citrus sinensis L.) were found to be highly infected in the field, but only the Washington navel sweet orange was found to be infected in the nurseries. No clear CTV symptoms were observed during the survey. Samples that were positive for CTV by DTBIA were also positive by biological indexing on Mexican lime (C. aurantifolia) and immunocapture-reverse transcription-PCR as described by Nolasco et al. (3). Coat protein gene sequences obtained from five selected clones of a Syrian CTV isolate (GenBank Accession No. EU626555) showed more than 99 and 98% nucleotide sequence identity to a Jordanian CTV isolate (GenBank Accession No. AY550252) and the VT isolate (GenBank Accession No. U56902), respectively. Almost all infected samples induced moderate vein clearing symptoms when grafted to Mexican lime. Symptoms of vein clearing, leaf cupping, stunting, and stem pitting on Mexican lime were induced by graft transmission of CTV from one Valencia sample from the Tartous area. The viral inoculum is widely and randomly distributed in commercial groves, especially in the southern Tartous area and in some nurseries. To our knowledge, this is the first report of CTV in Syria. However, CTV was reported from the neighboring citrus-growing countries of Lebanon, Turkey, and Jordan (1), and the severe seedling yellows strain is present in this area, which poses a potential threat to Syrian citriculture. References: (1) G. H. Anfoka et al. Phytopathol. Mediterr. 44:17, 2005. (2) G. Hughes and T. R. Gottwald, Phytopathology 88:715, 1998. (3) G. Nolasco et al. Eur. J. Plant Pathol. 108:293, 2002.


2011 ◽  
Vol 12 (9) ◽  
pp. 898-910 ◽  
Author(s):  
CARMEN FAGOAGA ◽  
GIOVANNI PENSABENE-BELLAVIA ◽  
PEDRO MORENO ◽  
LUÍS NAVARRO ◽  
RICARDO FLORES ◽  
...  

2019 ◽  
Vol 13 (2) ◽  
pp. 179-192 ◽  
Author(s):  
Nabil Killiny ◽  
Yasser Nehela ◽  
Faraj Hijaz ◽  
Sulley K. Ben-Mahmoud ◽  
Subhas Hajeri ◽  
...  

Plant Disease ◽  
2005 ◽  
Vol 89 (6) ◽  
pp. 575-580 ◽  
Author(s):  
C.M. Herron ◽  
T.E. Mirkov ◽  
N. N. Solís-Gracia ◽  
C.J. Kahlke ◽  
M. Skaria ◽  
...  

Citrus tristeza virus (CTV) isolates collected from the Lower Rio Grande Valley in south Texas and east Texas were characterized using citrus indicators and molecular methods. The citrus indicators were Mexican lime (Citrus aurantifolia), sour orange (C. aurantium), sweet orange (C. sinensis) grafted to sour orange, Duncan grapefruit (C. × paradisi), and Madam Vinous sweet orange, with some CTV isolates additionally indexed using the Texas commercial grapefruit cvs. Rio Red and Star Ruby, and Marrs and N-33 sweet orange. Severity ratings used 11 biotype groups or cumulative mean relative indices. Molecular characterization was carried out using poly- and monoclonal antibodies, seven strain-specific probes and single-stranded conformational polymorphism, and all were based on the CTV major coat protein or gene. All Texas CTV isolates produced vein clearing symptoms on inoculated Mexican lime plants. Over half of the CTV isolates tested were placed in biotype groups IX and X (causing decline of sweet orange on sour orange, seedling yellows on sour orange and grapefruit seedlings, and stem pitting of grapefruit or sweet orange), and one isolate was in biotype I (mild).


Virology ◽  
2010 ◽  
Vol 405 (2) ◽  
pp. 300-308 ◽  
Author(s):  
Muhammad Shah Nawaz-ul-Rehman ◽  
Nazia Nahid ◽  
Shahid Mansoor ◽  
Rob W. Briddon ◽  
Claude M. Fauquet

2005 ◽  
Vol 18 (5) ◽  
pp. 435-445 ◽  
Author(s):  
Carmen Fagoaga ◽  
Carmelo López ◽  
Pedro Moreno ◽  
Luis Navarro ◽  
Ricardo Flores ◽  
...  

Ectopic expression of the p23 gene from a severe (T36) strain of Citrus tristeza virus (CTV) induces viral-like symptoms in Mexican lime. Here, we report that expressing the same gene from a mild strain induced similar symptoms that correlated with accumulation of p23 protein irrespective of the source strain. CTV inoculation of transgenic limes showing CTV-like leaf symptoms and high p23 accumulation did not modify symptoms initially, with the virus titer being as in inoculated nontransgenic controls; however, at later stages, symptoms became attenuated. Transformation with p23-T36 of CTV-susceptible sweet and sour orange and CTV-resistant trifoliate orange also led to CTV-like leaf symptoms that did not develop when plants were transformed with a truncated p23 version. In transgenic citrus species and relatives other than Mexican lime, p23 was barely detectable, although symptom intensity correlated with levels of p23 transcripts. The lower accumulation of p23 in sweet and sour orange compared with Mexican lime also was observed in nontransgenic plants inoculated with CTV, suggesting that minimal p23 levels cause deleterious effects in the first two species. Conversely, transgenic expression of p23 in CTV nonhost Nicotiana spp. led to accumulation of p23 without phenotypic aberrations, indicating that p23 interferes with plant development only in citrus species and relatives.


2013 ◽  
Vol 88 (2) ◽  
pp. 1394-1397 ◽  
Author(s):  
P. Serra ◽  
S. M. Bani Hashemian ◽  
C. Fagoaga ◽  
J. Romero ◽  
S. Ruiz-Ruiz ◽  
...  

Viruses ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 782 ◽  
Author(s):  
Zhuoran Li ◽  
Yizhong He ◽  
Tao Luo ◽  
Xi Zhang ◽  
Haoliang Wan ◽  
...  

The severe strain of citrus tristeza virus (CTV) causes quick decline of citrus trees. However, the CTV mild strain causes no symptoms and commonly presents in citrus trees. Viral suppressor of RNA silencing (VSR) plays an important role in the successful invasion of viruses into plants. For CTV, VSR has mostly been studied in severe strains. In this study, the N4 mild strain in China was sequenced and found to have high sequence identity with the T30 strain. Furthermore, we verified the functions of three VSRs in the N4 strain, and p23 was found to be the most effective in terms of local silencing suppressor activity among the three CTV VSRs and localized to both nucleus and plasmodesmata, which is similar to CTV T36 strain. Several conserved amino acids were identified in p23. Mutation of E95A/V96A and M99A/L100AA impaired p23 protein stability. Consequently, these two mutants lost most of its suppressor activity and their protein levels could not be rescued by co-expressing p19. Q93A and R143A/E144A abolished p23 suppressor activity only and their protein levels increased to wild type level when co-expressed with p19. This work may facilitate a better understanding of the pathogenic mechanism of CTV mild strains.


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