Detection and relationships of cotton leaf curl virus and allied whitefly-transmitted geminiviruses occurring in Pakistan

A stock culture of cotton leaf curl virus from Pakistan (CLCuV-PK), was transmitted by whiteflies (Bemisia tabaci) to seven plant species, including French bean, okra, tobacco and tomato, and caused vein thickening and leaf curl symptoms. It was readily detected in triple antibody sandwich ELISA (TAS-ELIS A) by 11 out of 31 monoclonal antibodies raised against the particles of three other geminiviruses: African cassava mosaic, Indian cassava mosaic and okra leaf curl viruses. Reaction strength was enhanced when the tissue extraction fluid contained sodium sulphite. Minor variations in epitope profile were found among virus isolates from cotton (Gossypium hirsutum) collected from different districts in Pakistan over a 5-year period. These epitope profiles were distinguishable from that of cotton leaf curl virus from G. barbadense in southern India but indistinguishable from the profiles of viruses causing yellow vein disease of okra in India or Pakistan, or leaf curl of okra {Abelmoschus esculentus), Hibiscus tiliaceus, radish or sunflower in Pakistan, suggesting that these plants are putative natural hosts of CLCuV-PK. The viruses in cotton, and in okra with leaf curl or yellow vein symptoms, were also detected by PCR with three pairs of CLCuV-PK-specific primers. Five additional whitefly-transmitted geminiviruses were found among isolates from 11 other naturally-infected species in Pakistan, and were distinguished by their epitope profiles. These viruses were associated, respectively, with tobacco leaf curl, squash yellow blotch, tomato yellow leaf curl, watermelon leaf crinkle and soybean yellow mosaic diseases. The first four of these viruses were detected readily by PCR with geminivirus general primers but only weakly, if at all, with two pairs of CLCuV-PK-specific primers. Pakistani crops are infected with a range of distinguishable but relatively closely related whitefly-transmitted geminiviruses, some of which resemble those found in India.     

Epidemiology of tobacco leaf curl virus in India

The incidence of disease caused by tobacco leaf curl geminivirus (TbLCV) in ten tobacco growing areas of India ranged from 1.2% to 77%. The highest incidence of disease was observed in Andhra Pradesh (77%) followed by Gujarat (59%), Karnataka (17%), Bihar (11.6%) and West Bengal (5.4%). Under field conditions, an average of 32 adult whiteflies (Bemisia tabaci) per plant were recorded in Andhra Pradesh followed by Gujarat (20), Karnataka (12), Bihar (8) and West Bengal (5). In sequential sowings at Bangalore, all the plants were infected within 90 days in plots planted from February to June. Infection in plots planted later was progressively less. There was a positive correlation between whitefly catches and the final incidence of leaf curl disease in plantings. TbLCV was transmitted by Bemisia tabaci to 35 plant species, including Beta vulgaris, Capsicum annuum, Carica papaya, Cymopsis tetragonoloba, Lycopersicon esculentum, Sesamum indicum, Phaseolus vulgaris and Petunia hybrida. Forty five TbLCV isolates from different parts of India induced four distinct types of symptoms on tobacco cultivars Samsun and Anand 119. Group 1 isolates caused severe curling and cup-shaped enations; group II isolates induced pale green leaves, pit-like depressions and thorny enations: group III isolates caused leathery leaves, narrow and tiny protruding enations between the veins, and group IV isolates induced irregular thickening and swelling of veins and green flap-like enations on veins. Nylon net covers protected tobacco seedlings in nursery beds for 45 days. Ricinus communis and Helianthus annuus sown around the tobacco nursery bed as barrier crops attracted adult whiteflies and decreased the number found on tobacco.     

Host range, vector relations and serological relationships of cotton leaf curl virus from southern India

In 1989 to 1991, leaf curl disease was observed in cotton (Gossypium bar-badense cv. Local) grown in kitchen gardens in five districts in Karnataka State, India, and in 1994 it was recorded in G. hirsutum cv. Sharada in two districts. Symptoms consist of leaf curling, vein thickening, leaf enations, and stunting and distortion of plants. The disease is caused by cotton leaf curl virus (CLCuV-K), which was transmitted by the whitefly Bemisia tabaci to 24 plant species in six families. Hosts include bean (Phaseolus vulgaris), pepper, tobacco, tomato and several weeds, almost all of which developed leaf curl, with or without vein thickening. CLCuV-K was transmitted from cotton to cotton by adult B. tabaci after an acquisition access period of 1 h, could be inoculated in 5 min, had a minimum latent period of 8 h and was retained by viruliferous insects for up to 9 days. Female B. tabaci transmitted more frequently than males. CLCuV-K is a whitefly-transmitted geminivirus. It reacted with two out of 17 monoclonal antibodies (MAbs) raised to African cassava mosaic virus and five out of 10 MAbs raised to Indian cassava mosaic virus. CLCuV-K isolates from different locations in Karnataka had similar epitope profiles. As judged by these profiles, CLCuV-K is closely related to Indian tomato leaf curl virus from Karnataka, is distinguishable from several other whitefly-transmitted geminiviruses found in India and is still more distantly related to those, including cotton leaf crumple virus from the USA, found in other continents. CLCuV-K infected all cultivars tested of G. barbadense and one of six cultivars of G. hirsutum but none of G. arboreum or G. herbaceum.     

Ipomoea crinkle leaf curl caused by a whitefly-transmitted gemini-like virus

A whitefly-transmitted infectious agent, associated with geminate particles, induced distinct symptoms on several Ipomoea species, but not on I. batatas cv. Georgia Jet. The virus was transmitted by Bemisia argentifolii in a persistent manner and by grafting, but not mechanically. No transmission to species outside Ipomoea was obtained. Extracts from infected Ipomoea plants hybridised with a bean yellow mosaic virus riboprobe and a tomato yellow leaf curl virus riboprobe, although not so strongly as hybridisation of these riboprobes with extracts from plants infected with the homologous viruses. Based on host range, we consider this virus to be distinct from sweet potato leaf curl virus reported from the Far East, and propose it be named “Ipomoea crinkle leaf curl virus” (ICLCV).     

Complete nucleotide sequence and the genome organization of tobacco leaf curl geminivirus from Japan

The genomic DNA of tobacco leaf curl geminivirus (TLCV) from tomato plants with leaf curl disease in Japan has been sequenced. The single circular DNA molecule comprises 2,761 nucleotides. TLCV DNA contains six open reading frames (ORFs) capable of encoding proteins with a molecular weight greater than 10 K. In total nucleotide sequence comparisons with other geminiviruses, TLCV was most closely related to tomato leaf curl virus from Taiwan (TwToLCV) (76% identity), tomato leaf curl virus from Bangalore (ToLCV-Ba) (74%) and agerantum yellow vein virus (AYVV) (74%), all possessing a monopartite genome. The significant but relatively low sequence similarity in the genomic DNA between TLCV and other geminiviruses suggests it is a distinct geminivirus in genus Begomovirus.     

Tomato yellow leaf curl virus DNA in callus cultures derived from infected tomato leaves

Callus cultures were induced from leaves of a tomato plant infected with tomato yellow leaf curl virus (TYLCV) and analyzed for viral DNA presence during successive subcultures. No TYLCV DNA was detected in calli sampled after eight months of culture. Considerable differences in the presence of TYLCV DNA were found within sectors of a callus culture and between different callus cultures, throughout the entire eight months period. Infected calli which were cultured at sub-optimal temperature (15°C) retained the viral DNA longer than at 25 °C. The results suggested that TYLCV disappearance during callus culture was due to a disruption of some of the cell-to-cell connections, resulting in islands of infected cells in the midst of uninfected tissue and/or to the competition between the rate of cell division and that of viral DNA replication.Abbreviations BA benzyladenine - CMV cucumber mosaic virus - NAA naphthaleneacetic acid - TMV tobacco mosaic virus - TYLCV tomato yellow leaf curl virus     

Chinese tomato yellow leaf curl virus--a new species of geminivirus

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Replication of tomato yellow leaf curl virus (TYLCV) DNA in agroinoculated leaf discs from selected tomato genotypes

The leaf disc agroinoculation system was applied to study tomato yellow leaf curl virus (TYLCV) replication in explants from susceptible and resistant tomato genotypes. This system was also evaluated as a potential selection tool in breeding programmes for TYLCV resistance. Leaf discs were incubated with a head-to-tail dimer of the TYLCV genome cloned into the Ti plasmid ofAgrobacterium tumefaciens. In leaf discs from susceptible cultivars (Lycopersicon esculentum) TYLCV single-stranded genomic DNA and its double-stranded DNA forms appeared within 2–5 days after inoculation. Whiteflies (Bemisia tabaci) efficiently transmitted the TYLCV disease to tomato test plants following acquisition feeding on agroinoculated tomato leaf discs. This indicates that infective viral particles have been produced and have reached the phloem cells of the explant where they can be acquired by the insects. Plants regenerated from agroinfected leaf discs of sensitive tomato cultivars exhibited disease symptoms and contained TYLCV DNA concentrations similar to those present in field-infected tomato plants, indicating that TYLCV can move out from the leaf disc into the regenerating plant. Leaf discs from accessions of the wild tomato species immune to whitefly-mediated inoculation,L. chilense LA1969 andL. hirsutum LA1777, did not support TYLCV DNA replication. Leaf discs from plants tolerant to TYLCV issued from breeding programmes behaved like leaf discs from susceptible cultivars.The Hebrew University of Jerusalem, Faculty of Agriculture, Department of Field and Vegetable Crops     

Evaluation of Gossypium species for resistance to cotton leaf curl Burewala virus

Cotton leaf curl disease (CLCuD), caused by cotton leaf curl Burewala virus (CLCuBV), has emerged as a major threat to cotton production in Pakistan. Resistance to CLCuBV was evaluated in cultivated and wild cotton genotypes representing six Gossypium species by visual symptom scoring and virus assessment using PCR tests. Considerable variation in responses was observed when using whitefly and graft transmission to inoculate Gossypium genotypes with CLCuBV in field and greenhouse experiments. Under field evaluation, all cultivated genotypes of Gossypium hirsutum and three genotypes of G. barbadense were susceptible. Eleven genotypes that represented six wild and cultivated Gossypium species were considered to be highly resistant as they were free from infection. Similar results were obtained when these genotypes were tested using whitefly transmission. To verify these findings, 132 cultivated and wild genotypes were tested by graft inoculation. All G. hirsutum genotypes (116 cultivated, 1 wild, 1 transgenic Coker-312 and 1 non-transgenic Coker-312), three G. barbadense genotypes and one G. thurberi genotype were highly susceptible and exhibited symptoms 9–12 days after grafting. Four genotypes of G. arboreum and one genotype of G. anomalum did not express symptoms but had a detectable level of virus. One genotype of G. herbaceum and three wild genotypes of G. hirsutum showed mild symptoms (severity indexes of 1–2) and exhibited delayed disease development. These genotypes were classified as moderately resistant to resistant. Resistant genotypes that were identified in this study will be useful sources for exploitation of breeding programmes aimed at developing CLCuBV-resistant varieties and increasing genetic diversity.     

Prediction of cotton leaf curl virus disease and its management through resistant germplasm and bio-products

Cotton leaf curl virus disease reduces the cotton yield significantly every year and is transmitted by Bemisia tabaci. The study was designed to evaluate 15 varieties/lines against the disease. Multiple regression analysis was performed based on a-biotic environmental variables (maximum air temperature, minimum air temperature, relative humidity and rainfall) to predict disease incidence and its vector (Bemisia tabaci). Two bio-products were evaluated against the whitefly population to control the disease. Out of 15 cotton varieties/lines, no one was found highly resistant against the disease. Five varieties/lines (BT BT-980, BT-457, KIRAN, BT-666 and SLH-BT-6) exhibited moderately resistant response. Maximum air temperature (34–35.5 °C), minimum temperature (25.75–26.25 °C), relative humidity (64.14–66%), rainfall (1–2 mm) and wind speed (5.50–5.75 Kmh^?1) favoured the disease development. Maximum whitefly population was favoured by maximum air temperature from 34–35.5 °C, 25.8–26.2 °C minimum air temperature, 64.14–66% relative humidity, 1–2 mm from rainfall and 5.50–5.75 Kmh^?1 wind speed. Datura stramonium was found more effective as compared to Aviara (Homoeopathic) but not from the positive control (Acetamiprid).     

DGP1, a drought-induced guard cell-specific promoter and its function analysis in tobacco plants

Stomatal pores on the surface of leaf are gate-ways of gas exchange between plants and environment. For example, plants can take in CO2 via photosynthe-sis and lose water by transpiration. It was estimated that plants account for around 65% fresh water use every year, which was mainly lost through stomata[1]. So they attracted much more attention to increase the ability of drought resistance by regulating stomatal movement. Then a tentative plan came to our brains, is it possible for us to as…     

Cloning of the promoter region of plasma membrane aquaporin BnPIP1 from Brassica napus and its functional analysis

Aquaporins make water-selective channels in plants, facilitating the permeation of water through membranes and adjusting water fast transport during seed germination, cell elongation, stoma movement, fertilization and responses to environmental stresses. They belong to the MIP (major intrinsic protein) family with molecular weight of 2629 kD and are characterized by six membrane-spanning a-helixes connected by five loops and short N-terminal and C-terminal domains in the cytoplasm[13]. The p…