A Graft-Transmissible Stunting Factor in Citrus

Roistacher, C. N.; Calavan, E. C.; Blue, Rupert; Nauer, E. M.

doi:10.5070/c53g20f5dh

Cited by 17 publications

(30 citation statements)

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“…The recommended technique of Barbosa et al (2005) and Roistacher (1991) was followed during the study for the confirmation of CBLVd under controlled conditions. The infected budwood of grapefruit and sweet lime with characteristic symptoms of petiole necrosis, yellowing and leaf bending were collected from district Sahiwal.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of divergent isolates of Citrus bent leaf viroid (CBLVd) from citrus cultivars of Punjab, Pakistan

et al. 2023

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Citrus viroid infection is emerging as a serious threat because of its efficient systemic movement within the host plant and its quick spread due to contaminated pruning tools. A survey was conducted to investigate the primary distribution and molecular characterization of Citrus bent leaf viroid (CBLVd) and its variants in different citrus cultivars. A total of 154 symptomatic citrus samples were collected and detected by RT‒PCR with newly designed specific primers with the incidence of 36.33%. During biological indexing study on Etrog citron, expressions of reduced leaf size, yellowing with a light green pattern, and bending were observed. Amplified products were sequenced and analyzed using a nucleotide BLAST search, which showed 98% homology with other CBLVd isolates. The results of the phylogenetic tree analysis showed the presence of two main groups (A and B), with the predominant variants of CBLVd, i.e., CVd-I-LSS (Citrus viroid Low Sequence Similarity) sequences, clustering in subgroup A1 along with newly detected CVd-I-LSS from Palestinian sweet lime (Citrus limettioides), which has been identified as a new host of CVd-I-LSS in Pakistan. Further analysis of the sequences in subgroup A1 showed that the variant of CVd-I-LSS infecting citrus cultivars had a close relationship with isolates reported from China, Japan, and Iran, which may have resulted from the exchange of planting material. This study also unveiled the variability in nucleotide sequences of CBLVd, which made it unable to be detected by old primers. The results of this study indicate that the widespread presence of divergent variants of CBLVd is a major concern for the citrus industry in Pakistan and other countries where virulent isolates of CBLVd are prevalent. These findings suggest the need for future research on effective management and quarantine measures to stop the spread of CBLVd.

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Section: Methodsmentioning

confidence: 99%

Molecular characterization of divergent isolates of Citrus bent leaf viroid (CBLVd) from citrus cultivars of Punjab, Pakistan

et al. 2023

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“…(1). The collected samples were indexed biologically according to Roistacher (1991) and Roistacher et al, (2000) on indicator seedlings of Madam Vinous and Navelina sweet orange budded on rough lemon and kept under greenhouse conditions.…”

Section: Virus Sourcementioning

confidence: 99%

“…Bark scaling on infected branches and trunks is often accompanied by wood staining. Sour oranges, lemon, pummelo and rough lemon usually are not showing external symptoms (Roistacher, 1991). Two types of symptoms have been suggested, Psorosis A (PsA) and Psorosis B (PsB) (Fawcett and Klotz, 1939).…”

Section: Introductionmentioning

confidence: 99%

Elimination of Citrus psorosis virus using shoot tip grafting and electrotherapy techniques

Sayed

Saker²,

Shorbagy³

et al. 2022

Egyptian Journal of Agricultural Research

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During the spring of 2020, Citrus psorosis virus isolate was identified in citrus orchards in Qalyubia Governorate, displaying typical virus symptoms such as bark scaling lesions and ringspot on the leaves and fruits. Using chip buds from the donor plant grafted into Madam Vinous and Navelina sweet orange as indicator plants, the isolated virus was identified using biological indexing. DAS-ELISA and RT-PCR were used to test the plants that yielded positive results. RT-PCR employing virus-specific primers revealed single amplified fragments of the appropriate size (434 bp) in infected plants, while no bands in healthy plants were detected. Electrotherapy was used to eliminate the infection. Stem cuttings from infected plants were subjected to electric currents of 10, 20, and 40 mA for 10 and 20 minutes, respectively, before being grafted on the indicator plants. The treatment of 40 mA for 20 minutes was shown to be more effective in developing virus-free plants (85%). Also, for the elimination of Citrus psorosis virus, small shoot tip explants (0.1-0.2 mm) from virus-infected shoots were grafted onto a young rootstock of troyer citrange, growing in vitro on Murashige and Skoog medium with a success rate of 3%, after which the grafted plants were transferred to the greenhouse for acclimatization. The micrografted plants were successfully regrafted on potted sour orange seedlings in the greenhouse. DAS-ELISA was used to test all plants which survived for Citrus psorosis virus.

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“…The causal agent of CSD is Spiroplasma citri , a helical motile, wall-less, and cultivable microorganism that belongs to the class Mollicutes [ 3 ]. The pathogen is phloem limited, leafhopper transmitted [ 4 , 5 ], and also graft transmissible [ 6 , 7 , 8 ]. In addition to citrus species, the mollicute infects a wide range of non-rutaceous plant species such as carrot ( Daucus carota subsp.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Serological and Molecular Methods for Detection of Spiroplasma citri in Moroccan Citrus-Growing Areas

Sagouti

Rhallabi

Polizzi

et al. 2023

Plants

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Spiroplasma citri, a helical motile, wall-less, and cultivable microorganism of the class Mollicutes, is the agent of the citrus stubborn disease. There is currently a lack of data about the presence of this pathogen in Moroccan citrus orchards. This study aims to validate serological and molecular methods for routine S. citri diagnosis in Moroccan citrus groves. To provide an update on the present status of the outbreak of the pathogen in Moroccan citrus orchards, a survey of S. citri was conducted in the main citrus-growing regions of Morocco. A total of 575 leaf samples were collected from citrus trees with symptoms attributable to S. citri infection. Samples were collected during 2020 and 2021 from 23 citrus orchards. The presence of S. citri was tested in all samples using the double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Using this method, 57 samples were found to be infected with S. citri, 41 samples had doubtful results, and the remaining samples were negative. To corroborate the results of the DAS-ELISA test, 148 samples were chosen for additional molecular testing using conventional polymerase chain reaction (PCR) and real-time PCR (qPCR) based on specific primer pairs targeting three different genes (putative adhesion-like gene P58, putative adhesion gene P89, and spiralin gene). Using primers that target the putative adhesion-like gene P58, S. citri was detected by conventional and real-time PCR amplification from plant tissue with differing degrees of specificity. The results allowed us to determine the incidence of S. citri in all Moroccan citrus orchards, with a wide range of positive samples varying from 6.5% to 78%, and to show that molecular tests, particularly real-time PCR assays that target the putative adhesion-like gene P58, are the most sensitive for making an accurate diagnosis of S. citri. Indeed, the real-time PCR with P58-targeting primers yielded positive results from all positive and doubtful ELISA samples as well as some negative samples, with an OD value close to 1.5 × times healthy samples, thus demonstrating a high sensibility of this technique.

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A Graft-Transmissible Stunting Factor in Citrus

Cited by 17 publications

References 0 publications

Molecular characterization of divergent isolates of Citrus bent leaf viroid (CBLVd) from citrus cultivars of Punjab, Pakistan

Molecular characterization of divergent isolates of Citrus bent leaf viroid (CBLVd) from citrus cultivars of Punjab, Pakistan

Elimination of Citrus psorosis virus using shoot tip grafting and electrotherapy techniques

Comparison of Serological and Molecular Methods for Detection of Spiroplasma citri in Moroccan Citrus-Growing Areas

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