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The EFSA Panel on Plant Health conducted a pest categorisation of chickpea chlorotic dwarf virus (CpCDV) for the EU territory. The identity of CpCDV, a member of the genus Mastrevirus (family Geminiviridae) is established. Reliable detection and identification methods are available. The pathogen is not included in the EU Commission Implementing Regulation 2019/2072. CpCDV has been reported in Africa, Asia and Oceania. It has not been reported in the EU. CpCDV infects plant species in the family Fabaceae and several species of other families (Amaranthaceae, Brassicaceae, Caricaceae, Cucurbitaceae, Malvaceae and Solanaceae), including weeds. It may induce symptoms on its hosts, causing severe yield reduction. The virus is transmitted in a persistent, circulative and non‐propagative manner by the leafhopper species Orosius orientalis and O. albicinctus, which are not regulated. O. orientalis is known to be present in some EU member states. Plants for planting (other than seeds), parts of plants and cut flowers of CpCDV hosts and viruliferous leafhoppers were identified as the most relevant pathways for the entry of CpCDV into the EU. Cultivated and wild hosts of CpCDV are distributed across the EU. Would the pest enter and establish in the EU territory, impact on the production of cultivated hosts is expected. Phytosanitary measures are available to prevent entry and spread of the virus in the EU. CpCDV fulfils the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest. [ABSTRACT FROM AUTHOR]

A duplex PCR assay was standardized by optimizing PCR reaction constituents and cycles for the simultaneous detection of chickpea chlorotic dwarf virus (CpCDV) and a peanut witches' broom (PnWB) phytoplasma associated with the chickpea stunt disease. Coat protein gene and tuf gene specific primers for CpCDV and phytoplasmas were used. Different concentrations of the PCR components such as Taq polymerase, primers and PCR annealing temperature were standardized for the identification of the two agents by a duplex PCR assay. Expected amplicons of 590 bp for CpCDV and 1090 bp for phytoplasmas were consistently amplified from the symptomatic chickpea tissues. That resulted in equally efficient and sensitive in detecting single or mixed infection of CpCDV and PnWB phytoplasma in 148 symptomatic chickpea stunt samples collected in two states of India. The results indicate the robustness in the detection of pathogens present in chickpea showing stunt disease and for theoretical use in epidemiological studies that would help the appropriate disease management strategies. [ABSTRACT FROM AUTHOR]

A duplex PCR assay was standardized by optimizing PCR reaction constituents and cycles for the simultaneous detection of chickpea chlorotic dwarf virus (CpCDV) and a peanut witches' broom (PnWB) phytoplasma associated with the chickpea stunt disease. Coat protein gene and tuf gene specific primers for CpCDV and phytoplasmas were used. Different concentrations of the PCR components such as Taq polymerase, primers and PCR annealing temperature were standardized for the identification of the two agents by a duplex PCR assay. Expected amplicons of 590 bp for CpCDV and 1090 bp for phytoplasmas were consistently amplified from the symptomatic chickpea tissues. That resulted in equally efficient and sensitive in detecting single or mixed infection of CpCDV and PnWB phytoplasma in 148 symptomatic chickpea stunt samples collected in two states of India. The results indicate the robustness in the detection of pathogens present in chickpea showing stunt disease and for theoretical use in epidemiological studies that would help the appropriate disease management strategies. [ABSTRACT FROM AUTHOR]

In recent years, yield losses due to chickpea chlorotic dwarf virus (CpCDV, Mastrevirus, Geminiviridae) infection have been intensified in a large number of plants such as watermelon, okra and cotton and the virus has been reported around the world (Kanakala and Kuria 2018). In the current study, natural infection of watermelon with CpCDV was investigated. Watermelon samples showing leaf chlorosis, hardness and discoloration of the flesh with whitish stripes, deformation and increasing seed abortion of fruits (Fig. 1) were collected from watermelon-growing farms in Behbahan (Khuzestan Province) in 2018. Total DNA of the samples was extracted from leaf tissue using the cetyltrimethylammonium bromide (CTAB) method followed by the enrichment of the extracted circular DNA molecules by rolling circle amplification (RCA) method using phi 29 DNA polymerase (TempliPhi kit, GE Healthcare, USA). Polymerase chain reaction (PCR) tests using RCA product as DNA template and the primer pair CpCDV-752-F/CpCDV-1326-R (Askari et al. 2021) resulted in the amplification of an expected 575 bp DNA fragment (Fig. 2) indicating the infection of the all tested samples with CpCDV. The definitive infection of a PCR positive sample (W1) with CpCDV was confirmed by sequencing of PCR product. CpCDV has a broad host range and infects members of 11 plant families including several cucurbitaceous plants, such as cucumber, squash and watermelon (Kanakala and Kuria 2018; Zaagueri et al. 2017). The observed symptoms of the infected watermelon plants in this study were very similar to the symptoms of infected samples of the same plant, which was previously reported as “hard fruit syndrome” of watermelon in Tunisia, and the pathogenesis of the causal virus (CpCDV) was demonstrated (Zaagueri et al. 2017). The surveys of this study showed that the disease severity is remarkable in watermelon-growing farms in Behbahan and similar symptoms were also observed in some watermelon farms in Kerman and Hormozgan Provinces. Considering the wide host range of the virus, for example, several grain legumes, okra, tomato, spinach, pepper and cotton in neighboring countries of Iran and recent findings, it seems that CpCDV may has more natural hosts and a wide spread in Iran. However, further studied are required to confirm this hypothesis. This is the first report of the CpCDV infection of watermelon in Iran. [ABSTRACT FROM AUTHOR]

The legume crops such as chickpea and lentils are mainly cultivated in semi-arid tropical lands. Chickpea chlorotic dwarf virus (CpCDV) causes major losses to legumes throughout the world. Producing of specific antibody against this virus is crucial for surveys of disease in the fields and assessment of vial resistance in plant cultivars. Present article describes developing of specific antibody against the CpCDV virus by applying recombinant protein. In this study, coat protein of CpCDV was selected as a target for detection and preparation of polyclonal antibody. To achieve this aim CP gene encoding coat protein of CpCDV was initially PCR-amplified and inserted into bacterial expression vector. Expression of recombinant protein was performed in Bl21 strain of Escherichia coli. Purification was carried out under native conditions and the accuracy of recombinant protein production was confirmed by electrophoresis. The purified recombinant coat protein of CpCDV was used for immunization of rabbit. Purification of immunoglobulin molecules was performed by affinity chromatography using protein A column followed by conjugating of IgG to alkaline phosphatase enzyme. The capability of purified antibodies and conjugates for efficient detection of infected plants was assessed by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), western blotting and dot immunosorbent assay (DIBA). These results proved that prepared IgG and conjugate are able to distinguish with high efficiency CpCDV infected plants. To the best of our knowledge, this is the first report for production of anti-CpCDV antibodies raised through recombinant protein technology.

Chickpea chlorotic dwarf virus (CpCDV) a polyphagous mastrevirus, family Geminiviridae, is reported to infect several economically important hosts, among which tomato, in numerous world areas. NGS studies have recently linked the "hard fruit syndrome" of watermelon to CpCDV infection. In this study, we surveyed for CpCDV infection tomato fields in several Tunisian areas and we report this virus for the first time in tomato in this Mediterranean country. We detected CpCDV by PCR with specific primers and obtained a full-length sequence of a tomato-derived isolate, resulting identical to the Tunisian watermelon-derived isolate, and classifiable as CpCDV strain A. Following tomato agro-inoculation of the CpCDV infectious clone under controlled conditions, we checked infection by stem squash, dot blotting and PCR, and demonstrated the ability of CpCDV to replicate in tomato tissues. The alterations observed in fruits of CpCDV-infected tomatoes were similar to those previously described for watermelon, providing hints about the perturbation of fruit development mediated by this virus. [ABSTRACT FROM AUTHOR]

Cotton leaf curl disease in the Indian subcontinent is associated with several distinct begomoviruses that interact with a disease-specific DNA satellite named Cotton leaf curl Multan betasatellite (CLCuMB). However, we isolated the distinct strain (L) of leafhopper transmitted chickpea chlorotic dwarf virus, CpCDV-L (genus Mastrevirus, family Geminiviridae) from cotton plants affected by leaf curl disease in a small number of plants. The question as to whether CpCDV-L contributes to the development of disease symptoms such as leaf curling and enations remains to be answered. Standard methods were used to produce partial direct and tandem repeat constructs of CpCDVL for Agrobacterium-mediated inoculation in the binary vector pBIN19. The role of CpCDV-L in the induction of typical disease symptoms was studied by Agrobacteriummediated inoculation of the partial repeat construct into Nicotiana benthamiana. CpCDV-L induced downward leaf curling leading to cupping in N. benthamiana. [ABSTRACT FROM AUTHOR]

Chickpea chlorotic dwarf virus (CpCDV, genus Mastervirus, family Geminiviridae) is the most common viral disease of chickpea in Pakistan. Two aphid [Aphis craccivora Koch, Myzus persicae (Sulzer)], two leafhopper [Empoasca devastans Distant, Orosius albicinctus (Distant)] species and an unidentified brown leafhopper were collected in a chickpea field by hand and sweep nets for transmission studies of CpCDV. Transmission results showed that only the leafhopper O. albicinctus successfully transmitted the CpCDV from diseased to healthy chickpea plants. The presence of CpCDV in inoculated plants and the vector O. albicinctus were confirmed by DAS-ELISA test using specific polyclonal antibodies.

Chickpea stunt disease (CSD), caused by Chickpea chlorotic dwarf virus (CpCDV) is a threat to chickpea production leading to yield losses of 75–95%. Chickpea chlorotic dwarf virus is a monopartite, single-stranded circular DNA virus in the genus Mastrevirus and family Geminiviridae. It is transmitted by Orosius albicinctus in a circulative (persistent) and nonpropagative manner. Symptoms of CSD include very small leaves, intense discoloration (yellowing (kabuli type) and reddening (desi type)), and bushy stunted appearance of the plant. Presently, CpCDVs occurs in Africa, Asia, Australia, and the Middle East, causing extensive losses on economically important crops in in the families Fabaceae, Asteraceae, Amaranthaceae, Brassicaceae, Cucurbitaceae, Caricaceae, Chenopodiaceae, Leguminosae, Malvaceae, Pedaliaceae, and Solanaceae. High frequency of recombinations has played a significant role in the wide host range, diversification, and rapid evolution of CpCDVs. This review highlights the extensive research on the CpCDV genome diversity, host range, plant–virus–insect interactions, and RNA interference-based resistance of CpCDV, providing new insights into the host adaptation and virus evolution.

Chickpea stunt disease (CSD), caused by Chickpea chlorotic dwarf virus (CpCDV) is a threat to chickpea production leading to yield losses of 75–95%. Chickpea chlorotic dwarf virus is a monopartite, single-stranded circular DNA virus in the genus Mastrevirus and family Geminiviridae. It is transmitted by Orosius albicinctus in a circulative (persistent) and nonpropagative manner. Symptoms of CSD include very small leaves, intense discoloration (yellowing (kabuli type) and reddening (desi type)), and bushy stunted appearance of the plant. Presently, CpCDVs occurs in Africa, Asia, Australia, and the Middle East, causing extensive losses on economically important crops in in the families Fabaceae, Asteraceae, Amaranthaceae, Brassicaceae, Cucurbitaceae, Caricaceae, Chenopodiaceae, Leguminosae, Malvaceae, Pedaliaceae, and Solanaceae. High frequency of recombinations has played a significant role in the wide host range, diversification, and rapid evolution of CpCDVs. This review highlights the extensive research on the CpCDV genome diversity, host range, plant–virus–insect interactions, and RNA interference-based resistance of CpCDV, providing new insights into the host adaptation and virus evolution. [ABSTRACT FROM AUTHOR]