Comparative Epidemiology of Three Virus Diseases on Zucchini Squash

Zucchini yellow mosaic virus (ZYMV), Papaya ringspot virus – type W (PRSV‐W) and Zucchini lethal chlorosis virus (ZLCV) cause important diseases on zucchini squash crops in Brazil. ZYMV and PRSV‐W belong to the genus Potyvirus and are transmitted by aphids, whereas ZLCV belongs to Tospovirus and is transmitted by the thrips Frankliniella zucchini. These three viruses may occur simultaneously in the field, and the epidemiology of the corresponding diseases may be determined by interactions among viruses, hosts and vectors. In this work, the progress of the diseases caused by these viruses was studied over a temporal and geographic range for three planting seasons (PS). For the lethal chlorosis (ZLCV), a monomolecular model was found to be the best fit for the data, though only during the third PS. For data collected during the first two PS, the Gompertz model was found to fit the data best. The spatial distribution of disease indicated disease aggregation at the end of the crop cycle. For the yellow mosaic (ZYMV), the model that best fit in the 1st PS was the logistic and in the 2nd and 3rd PS was monomolecular. The spatial pattern of the disease was random when the disease incidence was low but aggregated when the disease incidence was high. The common mosaic (PRSV‐W) showed the lowest incidence in all three PS. An exponential model was the best fit for data collected during all PS, and the spatial pattern of the disease was random. Interactions among the three viruses apparently did not result in changes in the epidemiology of the diseases. Removal of sources of inoculum and planting at an unfavourable time for reproduction of virus vectors are the two main measures recommended for the control of these diseases. The use of insecticide is indicated only for the control of the F. zucchini.     

Molecular evidence of occurrence of Tomato Leaf Curl New Delhi Virus infecting cucurbits in several states in India

Abstract

Tomato Leaf Curl New Delhi Virus (ToLCNDV) is an important begomovirus constraint to the production of cucurbitaceous and other crops in India and its subcontinent. In this study first time, symptomatic samples of different cucurbits were collected from Telangana, Uttar Pradesh (Varanasi) and Madhya Pradesh location in India. The symptomatic samples were associated with begomovirus infection was confirmed by PCR amplification using specific universal primers Deng 540/541. Thirteen out of fifteen samples were amplified with newly designed bipartite specific primers JKR 58?F and JKR 59?R. In eleven out of thirteen bipartite samples contain betasatellite, confirmed by PCR amplification using specific primer CLB36F and CLB37R. The amplified PCR product with JKR58F and 59R of bitter gourd sample collected from Telangana was sequenced. The sequence was share 97.53% similarity with ToLCNDV infecting bottle gourd in Haryana in India (FN645905). Phylogenic analysis revealed that ToLCNDV infecting cucurbits are different, from ToLCNDV infecting tomato.     

Molecular characterization of pathogenic Fusarium species in cucurbit plants from Kermanshah province, Iran

Fusarium is one of the important phytopathogenic genera of microfungi causing serious losses on cucurbit plants in Kermanshah province, the largest area of cucurbits plantation in Iran. Therefore, the objectives in this study were to isolate and identify disease-causing Fusarium spp. from infected cucurbit plants, to ascertain their pathogenicity, and to determine their phylogenetic relationships. A total of 100 Fusarium isolates were obtained from diseased cucurbit plants collected from fields in different geographic regions in Kermanshah province, Iran. According to morphological characters, all isolates were identified as Fusarium oxysporum, Fusarium proliferatum, Fusarium equiseti, Fusarium semitectum and Fusarium solani. All isolates of the five Fusarium spp. were evaluated for their pathogenicity on healthy cucumber (Cucumis sativus) and honeydew melon (Cucumis melo) seedlings in the glasshouse. F. oxysporum caused damping-off in 20–35 days on both cucurbit seedlings tested. Typical stem rot symptoms were observed within 15 days after inoculation with F. solani on both seedlings. Based on the internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) restriction fragment length polymorphism (RFLP) analysis, the five Fusarium species were divided into two major groups. In particular, isolates belonging to the F. solani species complex (FSSC) were separated into two RFLP types. Grouping among Fusarium strains derived from restriction analysis was in agreement with criteria used in morphological classification. Therefore, the PCR-ITS-RFLP method provides a simple and rapid procedure for the differentiation of Fusarium strains at species level. This is the first report on identification and pathogenicity of major plant pathogenic Fusarium spp. causing root and stem rot on cucurbits in Iran.     

Assessing indigenous and local knowledge of farmers about pollination services in cucurbit agro-ecosystem of Punjab,Pakistan

Being the ultimate beneficiary of ecosystem services provided by on-farm agricultural biodiversity, the participation of farmers in its sustainable utilization and conservation is crucial. How much aware they are with the significance and conservation of agricultural biodiversity in order to improve their crop yield remains unclear, especially from the developing courtiers. Pollination is one of such ecosystem services, enormously contributed by the wild bees. In the present study, we have investigated the knowledge of farmers about bees and pollination in general in three districts i.e. Multan, Bahawalpur and Khanewal of southern Punjab, Pakistan. Some 300 farmers (100 cucurbit growers in each district using convenient sampling method) were interviewed using a semi-structured questionnaire. Respondents were first presented with a box of insect specimens and then were asked to identify bees among those. Those who identified correctly were asked to state about their nesting sites. Only 11% of the respondents could correctly identify the bees and half of them could report something about nesting sites. A majority (63%) of the farmers was unable to tell fertilization requirements in cucurbits, 59% could not distinguish female flower from the male flower and 64% could not state any benefit of bees. However, upon briefing about the significance of bee pollinators, 58% of the farmers showed eagerness to conserve bees at their farms. Keeping in view the inadequacies of farmers' knowledge about wild bees and pollination in general, the present study also gives some policy recommendations.     

Hormonal regulation of sex expression in Momordica charantia

Sex expression in a monoecious cucurbit, Momordica charantia L. (long–fruited variety 'Karela') can be regulated by application of gibberellin A₃ (GA₃), indole-3-acetic acid (IAA) and 3-hydroxymethyl oxindole (HMO), an oxidation product of IAA, all of which promote female flowering. Both IAA and HMO accelerated ethylene evolution in the seedlings of this plant. While a low concentration of Ethrel promotes flowering, the effect was reversed with increased concentrations. Surprisingly, GA₃ was the most effective growth regulator in increasing, femaleness. In untreated plants, levels of endogenous GA-like substance increased progressively up to the age of 60 days, at which the ratio of male to female flowers was minimum.     

Proteomics and metabolomics analyses reveal the cucurbit sieve tube system as a complex metabolic space

The plant vascular system, and specifically the phloem, plays a pivotal role in allocation of fixed carbon to developing sink organs. Although the processes involved in loading and unloading of sugars and amino acids are well characterized, little information is available regarding the nature of other metabolites in the sieve tube system (STS) at specific sites along the pathway. Here, we elucidate spatial features of metabolite composition mapped with phloem enzymes along the cucurbit STS. Phloem sap (PS) was collected from the loading (source), unloading (apical sink region) and shoot–root junction regions of cucumber, watermelon and pumpkin. Our PS analyses revealed significant differences in the metabolic and proteomic profiles both along the source–sink pathway and between the STSs of these three cucurbits. In addition, metabolite profiles established for PS and vascular tissue indicated the presence of distinct compositions, consistent with the operation of the STS as a unique symplasmic domain. In this regard, at various locations along the STS we could map metabolites and their related enzymes to specific metabolic pathways. These findings are discussed with regard to the function of the STS as a unique and highly complex metabolic space within the plant vascular system.     

Transferability of Cucurbita SSR markers for genetic diversity assessment of Turkish bottle gourd (Lagenaria siceraria) genetic resources

The genetic diversity present in crop landraces represents a valuable genetic resource for breeding and genetic studies. Bottle gourd (Lagenaria siceraria) landraces in Turkey are highly genetically diverse. However, the limited genomic resources available for this crop hinder the molecular characterization of Turkish bottle gourd germplasm for its adequate conservation and management. Therefore, we evaluated the efficacy of 40 SSR markers from major cucurbit crops (Cucurbita pepo L. and Cucurbita moschata L.) in 30 bottle gourd landraces, together with 16 SRAP primer combinations. In addition, we compared the genetic relationship between bottle gourd and 31 other cucurbit accessions (11 Cucurbita maxima, 3 C. moschata, 5 C. pepo subsp. ovifera, 10 C. pepo and 2 Luffa cylindrica). Twenty-seven Cucurbita SSR markers showed transferability to bottle gourd. SSR markers amplified 59 alleles, in bottle gourd genome with an average of 1.64 alleles per locus. Together, SSR and SRAP markers amplified 453 fragments across the 61 accessions, and clearly discriminated L. siceraria and L. cylindrica from the other cucurbit species. Genetic diversity analysis separated edible cucurbit from ornamentals, while population structure analysis classified L. siceraria in two subpopulations defined by fruit shape, rather than geographical origin. The results indicated that the genomic resources available for Cucurbita species are valuable to study and preserve the genetic diversity of bottle gourd in Turkey.     

Fruit fly (Diptera: Tephritidae) diversity in cucurbit fields and surrounding forest areas of Himachal Pradesh,a north-western Himalayan state of India

During the course of studies, Bactrocera (Bactrocera) latifrons (Hendel), B. (B.) nigrofemoralis White and Tsuruta, Dacus (Callantra) longicornis Wiedemann, Dacus (Callantra) sphaeroidalis (Bezzi), Cyrtostola limbata (Hendel) and Pliomelaena udhampurensis Agarwal and Kapoor were recorded for the first time in Himachal Pradesh in a cucurbit ecosystem. Apart from these, other species viz. Bactrocera tau, Bactrocera cucurbitae, Bactrocera dorsalis, Bactrocera zonata, Bactrocera scutellaris, Bactrocera diversa and Dioxyna sororcula (Wiedemann) were also identified. Distribution records of B. (B.) dorsalis (Hendel), B. (B.) zonata (Saunders), Bactrocera (Hemigymnodacus) diversa (Coquillett), B. (Zeugodacus) cucurbitae (Coquillett), B. (Z.) scutellaris (Bezzi) and B. (Z.) tau (Walker) has been described.     

Laboratory evaluation of certain natural compounds against the melon ladybird beetle,Epilachna chrysomelina F. attacking cucurbit plants

Neem Azal-T/S, Neudosan and Spruzit flüssig at different concentrations were evaluated against the second and fourth instar larvae of E. chrysomelina under laboratory conditions. In case of acute effects, the highest mortality was recorded when larvae were offered squash leaves treated with the highest concentration (2%) of Neem Azal-T/S, and the lowest mortality was, however recorded after treating them with the lowest concentration (0.25%). On the other hand, the second instar larvae were more susceptible to any compound at any concentration than the fourth instar one. One per cent Neudosan formulation showed the highest toxicity to both the 2nd and 4th instar larvae. As the concentration of this compound decreased the mortality percentage of treated larvae decreased to reach the minimum at the lowest concentration (0.25%). The same trend could be applied for Spruzit flüssig compound. When Neem Azal-T/S was compared with Neudosan, the later at any concentration was more toxic than the former. Generally, Neudosan and Spruzit flüssig seemed to be active against E. chrysomelina during the first three days after treatment, then, deterioration occurred among this compound. In case of latent effects, the duration of the treated fourth instar larvae of E. chrysomelina with any tested compound, was prolonged. An increase in the concentration caused prolongation of this period to reach the longest at the highest concentration. This prolongation, was more pronounced after Neem Azal-T/S treatment followed by Neudosan and Spruzit flüssig. The same trend could be applied for the duration of the pupal stage as being affected by the different concentrations of the tested compounds which were applied to the fourth instar larvae. However, adults of E. Chrysomelina treated as fourth instar larvae with high concentrations of Neem Azal-T/S showed some adult malformations. The sex ratios of adults produced from fourth instar larvae treated with different concentrations of the tested natural compounds were about 1?:?1, except for two cases; 1% Neudosan where the number of females were double the number of males and 0.5% Spruzit flüssig where the number of males were double the number of females. An increase in the concentration of any tested compound caused an obvious decrease in the percentage of emerged adults. The least emergence percentage was recorded in the case of Spruzit flüssig, followed by Neudosan and Neem Azal-T/S. Treating the fourth instar larvae with the different concentrations of the tested natural compounds significantly shortened the longevity of produced adults and decreased the fecundity.     

Inoculation of cucumber,melon and zucchini varieties with Tomato leaf curl New Delhi virus and evaluation of infection using different detection methods

The disease caused by Tomato leaf curl New Delhi virus (ToLCNDV), which is naturally transmitted by the whitefly Bemisia tabaci, causes important economic losses in cucurbit crops. The availability of simple and efficient inoculation protocols and detection methods is necessary for screening varieties and germplasm collections as well as for breeding populations. We evaluated the infectivity of ToLCNDV inocula prepared using three different buffers for mechanical sap inoculation in a susceptible variety of zucchini. We found that inoculum prepared with buffer III, which contains polyvinylpyrrolidone, is highly efficient for mechanical inoculation, with 100% of plants displaying severe symptoms 21 days post‐inoculation. Using this buffer, we mechanically inoculated 19 commercial varieties of cucurbit crops (six of cucumber, six of melon and seven of zucchini), evaluated the evolution of symptoms and diagnosed infection using nine different ToLCNDV detection methods (four based on serology, four based on molecular hybridization and one based on PCR detection). The results revealed that all varieties are susceptible, although cucumber varieties display less severe symptoms than those of melon or zucchini. All detection methods were highly efficient (more than 85% of plants testing positive) in melon and zucchini, but in cucumber, the percentage of positive plants detected with serology and molecular hybridization methods ranged from 20.4% with Squash leaf curl virus (SLCV) antiserum, to 78.5% with DNA extract hybridization. Overall, the best detection results were obtained with PCR, with 92.6%, 92.4% and 98.4% cucumber, melon and zucchini plants, respectively, testing positive. When considering the overall results in the three crops, the best serology and molecular hybridization methods were those using Watermelon chlorotic stunt virus (WmCSV) antiserum and DNA extract, respectively. The inoculation methodology developed and the information on detection methods are of great relevance for the selection and breeding of varieties of cucurbit crops that are tolerant or resistant to ToLCNDV.