Zucchini Yellow Mosaic Potyvirus Research Articles

Softening of Processed Plant Virus Infected Cucumis sativus L. Fruits

Texture softening of pickled cucumbers does not meet consumers’ quality expectations and leads to economic losses. The factor(s) triggering this phenomenon is still unknown. We investigated the importance of plant viruses such as Cucumber green mottle mosaic tobamovirus (CGMMV) and Zucchini yellow mosaic potyvirus (ZYMV) in the context of softening of pickles. Cucumber plants (Cucumis sativus) were infected by mechanical inoculation, grown under greenhouse conditions and tested positive for the viral infection by ELISA. The severity of virus infection was reflected in yield and symptom expression. Histological and morphological alterations were observed. All fruits were pasteurized, separately stored in jars and subjected to texture measurements after four, six and 12 months. CGMMV-infections were asymptomatic or caused mild symptoms on leaves and fruit, and texture quality was comparable to control. At the same time, fruits of ZYMV-infected plants showed severe symptoms like deformations and discoloration, as well as a reduction in firmness and crunchiness after pasteurization. In addition, histological alterations were detected in such fruits, possibly causing textural changes. We conclude that plant viruses could have a considerable influence on the firmness and crunchiness of pickled cucumbers after pasteurization. It is possible that the severity of symptom expression has an influence on texture properties.

Incidence and disease control of Zucchini yellow mosaic potyvirus

Zucchini yellow mosaic potyvirus (ZYMV) was first identified in northern Italy. It likes other species of the family Potyuiridue. ZYMV has been recorded in many countries since 1981. The efficient intercontinental spread of the virus can be explained by international trading of infected seeds. Since coat protein (CP) analysis has become a primary method for taxonomic assignment of potyviruses the aims were to characterize this genomic region of ZYMV originating from virus-infected cucurbitaceous seedlings. Virus infection in cucurbits is typically associated with mosaic symptoms on leaves and lumpy, distorted fruit. The range of symptoms produced by each virus can overlap and plants are commonly infected by more than one virus at once. The viruses are spread by many species of aphids moving through or within a crop. Control options include: destroying old cucurbit crops as soon as harvesting is completed destroying weeds and volunteer cucurbits, within and around crops as these harbor the viruses and/or the aphids separating new crops from maturing crops as these will have high levels of virus infection avoiding overlapping crops of cucurbits.

Detection of Some Virus Diseases of Edible Seed Squash (Cucurbita pepo L.) in Nevşehir Province, Turkey

Squash is a member of Cucurbitaceae family. It is grown for fresh consuming and its seeds are used as a snack in Turkey like some Mediterranean countries and Germany, Hungary, Austria and China. Cucurbita pepo L. is mostly used for cultivating squash seeds in Turkey. Also, a small amount of seeds obtain from Cucurbita moschata Duch (Butternut squash or pumpkin). Virus diseases are one of the most destructive diseases on squash which is grown for seeds in Nevsehir province. In this study, it was aimed to determine the virus infections in major squash growing areas in Nevsehir province. Totally 134 plant samples with common virus symptoms like mosaic, curling, blistering, mottling, distortion, shoestring, stunting and vine decline were collected from squash plants during 2018. In this study. Double-Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS-ELISA) method is used for identifying the virus infections on the plant samples. According to the results of the DAS-ELISA, 97.76 % of plant samples were infected with Zucchini yellow mosaic potyvirus (ZYMV), Watermelon mosaic potyvirus-2 (WMV), Cucurbit aphid borne yellow virus (CABYV), Cucumber mosaic cucumovirus (CMV), Papaya ringspot potyvirus -watermelon strain (PRSV-W) and Squash mosaic comovirus (SqMV). WMV was predominant in the research area with the ratio of 89.55 %. ZYMV was the second important virus disease in the surveyed area and it was detected on the samples at the ratio of 57.46 %. Also, mixed infections of those virus infections were detected commonly in squash. Especially, ZYMV+WMV (40.29 %) and WMV+ZYMV+PRSV-W (8.20 %) mixed infections were common.

Natural infection of squash fruits (Cucurbita pepo) by Zucchini Yellow Mosaic potyvirus (ZYMV) in Alexandria governorate

Natural infection of squash fruits (Cucurbita pepo) by Zucchini Yellow Mosaic potyvirus (ZYMV) in Alexandria governorate

Occurrence and Characterization of Coat Protein Gene of Zucchini yellow mosaic potyvirus (ZYMV) Isolate Infecting Pumpkin (Cucurbita pepo L.) in Bingol Province (Turkey)

Zucchini yellow
 mosaic potyvirus
 (ZYMV) belongs to family Potyviridae, which causes serious economic losses in
 many cucurbits from worldwide. In 2018 (September), leaf samples of pumpkin
 exhibiting typical viral symptoms including mosaic, blistering and wrinkling
 and healthy pumpkin leaf samples were collected from Bingöl and screened by
 Reverse- Transcription Polymerase Chain Reaction (RT-PCR) against ZYMV
 infection. Tested leaf samples were reacted positive resulting in an expected
 about 840 bp DNA fragments of partial coat protein (CP) gene of ZYMV. ZYMV-CP
 gene was further inserted into a pGEM-T Easy prokaryotic cloning vector and
 their partial nucleotide sequences and deduced amino acid were ascertained. The
 provided ZYMV- CP gene sequence consisted of 837 nucleotides in length coded
 for 279 amino acid residues of approximately 31.2 kDa. The isolate was
 denominated as ZYMV-Bingol and registered with MK689858 accession number in the
 NCBI. The sequence of ZYMV- Bingol CP gene were aligned with 21 isolates
 deposited in GenBank from different geographical location and its phylogenetic
 relationships were determined. Molecular analysis of the ZYMV CP gene sequence
 indicated the highest similarity with 100% Turkish isolate (JF317296) and the
 lowest with 91.64% of Korea isolate (AF062518), at nucleotide level. Moreover,
 phylogenetic analyses revealed that ZYMV Bingöl isolate is clustered with the
 Turkish- Adana isolate (JF317296) and Pakistan isolate (AB127936). ZYMV has
 been reported for the first time in the pumpkin plant from Bingöl province of
 Turkey by this study.

Yozgat İli Çerezlik Kabak (Cucurbita pepo L.) Ekim Alanlarında Görülen Virüslerin Belirlenmesi

Cucurbits (Cucurbitaceae family) consist of 119 genera and 825 species and are cultivated worldwide, especially in tropical and subtropical regions. In the world, squash (Cucurbita pepo L.), watermelon (Citrullus lanatus (Thunb) Matsum&Nakai), cucumber (Cucumis sativus L.), and melon (C. melo L.) are the most cultivated cucurbit species as vegetable crops. Squash is grown for fresh consuming, as well as its seeds are used like a snack in Turkey. Viruses are one of the most destructive pathogen groups on squash which is grown for seeds in Yozgat province. In this study, it was aimed to determine the virus infections in economically important squash planting fields in the province. Totally 140 plant samples which show widespread virus symptoms such as mottling, shoe-string, mosaic, stunting, curling, vine decline, distortion, and blistering were collected from plants in 2017. In this study, DAS-ELISA method is used for identifying the virus infections on the plant samples. According to the results of the DAS-ELISA 84.28 % of plant samples were infected with Cucumber mosaic cucumovirus (CMV), Watermelon mosaic potyvirus (WMV), Papaya ring spot potyvirus-watermelon strain (PRSV-W), Zucchini yellow mosaic potyvirus (ZYMV), and Squash mosaic comovirus (SqMV). The prevalent virus was WMV with the ratio of 57.85 %. It was followed by ZYMV which was detected on the samples at the ratio of 53.57 %. Besides, double and multiple virus infections were determined commonly in the samples. Especially, ZYMV+WMV mixed infections were common. The infection of Cucumber green mottle mosaic tobamovirus (CGMMV) wasn’t determined in the survey area.

Aksaray İli Çerezlik Kabak (Cucurbita pepo L.) Ekim Alanlarında Görülen Bazı Virüs Hastalıkları

Cucurbits (the Cucurbitaceae family) consist of 119 genera and 825 species and are cultivated worldwide, especially in tropical and subtropical regions. Squash (Cucurbita pepo L.), a member of this family, is one of the most cultivated and important vegetable species worldwide. As well as, squash is grown for fresh consuming; its seeds are used as a snack in some Mediterranean countries, Austria, Germany, China and Hungary like in Turkey. One of the most destructive diseases on seed squash is virus diseases in Aksaray province. In this study, it was aimed to determine the virus infections in main squash planting areas in Aksaray. For that purpose, totally 153 plant samples which showing widespread virus symptoms such as curling, mosaic, blistering, mottling, shoe-string, distortion, and stunting were collected from seed squash plants in 2014 and tested by DAS-ELISA method. According to the results of the DAS-ELISA 84.96 % of the samples were infected with Cucumber mosaic cucumovirus (CMV), Papaya ring spot potyvirus - watermelon strain (PRSV-W), Zucchini yellow mosaic potyvirus (ZYMV), Squash mosaic comovirus (SqMV) and Watermelon mosaic potyvirus - 2 (WMV-2). The prevalent virus was ZYMV with the ratio of 66.01 % and followed by WMV-2 with 57.51 %. Besides, double and multiple virus infections were determined commonly in the samples such as, ZYMV+WMV-2, ZYMV+CMV, WMV-2+CMV, ZYMV+SqMV, ZYMV+WMV-2+PRSV-W, and ZYMV+WMV-2+SqMV. Especially, ZYMV+WMV-2 mixed infections were the most common. The infection of Cucumber green mottle mosaic tobamovirus (CGMMV) wasn’t determined in the survey area.

Viral Diseases of Crops in Nepal

Viral diseases are the important diseases next to the fungal and bacterial in Nepal. The increase in incidence and severity of viral diseases and emergence of new viral diseases causes the significant yield losses of different crops in Nepal. But the research and studies on plant viral diseases are limited. Most of the studies were focused in viral diseases of rice (Rice tungro virus and Rice dwarf virus), tomato (Yellow leaf curl virus) and potato (PVX and PVY). Maize leaf fleck virus and mosaic caused by Maize mosaic virus were recorded as minor disease of maize. Citrus Tristeza Virus is an important virus of citrus fruit in Nepal while Papaya ringspot potyvirus, Ageratum yellow vein virus (AYVV), Tomato leaf curlJava betasatellite and Sida yellow vein Chinaalphasatellite were recorded from the papaya fruit. The Cucumber mosaic virus (CMV) and Zucchini yellow mosaic potyvirus (ZYMV) are the viral diseases of cucurbitaceous crop reported in Nepal. Mungbean yellow mosaic India virus (MYMIV) found to infect the many crops Limabean, Kidney bean, blackgram and Mungbean. Bean common mosaic necrosis virus in sweet bean, Pea leaf distortion virus (PLDV), Cowpea aphid‐borne mosaic potyvirus (CABMV), Peanut bud necrosis virus (PBNV) in groundnut, Cucumber mosaic virus (CMV). Chili veinal mottle potyvirus (CVMV) and Tomatoyellow leaf curl gemini virus (TYLCV) were only reported and no any further works have been carried out. The 3 virus diseases Soyabean mosaic (SMV), Soybean yellow mosaic virus and Bud blight tobacco ring spot virus (TRSV) were found in soybean.Int. J. Appl. Sci. Biotechnol. Vol 6(2): 75-80

IDENTIFIKASI PENYEBAB PENYAKIT DAUN KERITING KUNING PADA TANAMAN MENTIMUN

ABSTRACTIdentification of the causal agent of yellow leaf curl disease on cucumbers. Yellow leaf curl disease has been reported to cause serious diseases and yield losses on tobacco, chilli pepper, and tomato plants in Java. Similar symptoms were observed recently on cucumber plants from several growing areas in West Java (Bogor), Central Java (Tegal and Sukoharjo), and Yogyakarta (Sleman). Symptom variations including mosaic, chlorotic spotting, leaf curling, blistering, vein banding, reduction and distortion of leaf and fruit were observed. Serological detection using Enzyme Linked Immunosorbent Assay (ELISA) showed infection of several viruses. Antibodies specific to Squash mosaic comovirus (SqMV), Zucchini yellow mosaic potyvirus (ZyMV), dan Cucumber mosaic cucumovirus (CMV) were reacted positively with field samples. No serological reactions were observed with antibodies to Tobacco ringspot potyvirus (TRSV) and Watermelon mosaic potyvirus (WMV). Molecular detection approach based on Polymerase Chain Reaction was undergone using universal primers for Geminivirus, pAL1v1978 and pAR1c715. DNA fragment 1600 bp in size, was successfully amplified from leaf samples originated from Tegal, Sleman, Bogor, and Sukoharjo. Further identification by nucleotide sequencing indicated that virus isolates causing yellow leaf curl disease on cucumber have highest homology (95.7% to 98.6%) with Tomato leaf curl New Delhi virus-[Cucumber:Indonesia] (AB613825) from Klaten, Central Java, Indonesia.

A High Level of Transgenic Viral Small RNA Is Associated with Broad Potyvirus Resistance in Cucurbits

Gene-silencing has been used to develop resistance against many plant viruses but little is known about the transgenic small-interfering RNA (t-siRNA) that confers this resistance. Transgenic cucumber and melon lines harboring a hairpin construct of the Zucchini yellow mosaic potyvirus (ZYMV) HC-Pro gene accumulated different levels of t-siRNA (6 to 44% of total siRNA) and exhibited resistance to systemic ZYMV infection. Resistance to Watermelon mosaic potyvirus and Papaya ring spot potyvirus-W was also observed in a cucumber line that accumulated high levels of t-siRNA (44% of total siRNA) and displayed significantly increased levels of RNA-dependent RNA (RDR)1 and Argonaute 1, as compared with the other transgenic and nontransformed plants. The majority of the t-siRNA sequences were 21 to 22 nucleotides in length and sense strand biased. The t-siRNA were not uniformly distributed throughout the transgene but concentrated in "hot spots" in a pattern resembling that of the viral siRNA peaks observed in ZYMV-infected cucumber and melon. Mutations in ZYMV at the loci associated with the siRNA peaks did not break this resistance, indicating that hot spot t-siRNA may not be essential for resistance. This study shows that resistance based on gene-silencing can be effective against related viruses and is probably correlated with t-siRNA accumulation and increased expression of RDR1.

Viruses of vegetable crops in Albania

Field surveys were carried out in the main vegetable‐growing areas of Western and Central Albania to evaluate the sanitary status of open‐field and protected cultivations of capsicum, tomato, potato, watermelon, cucumber, courgette, aubergine, lettuce, cabbage, chicory, leek and celery. The following viruses were detected: Alfalfa mosaic alfamovirus (AMV), Cucumber mosaic cucumovirus (CMV), Potato Y potyvirus (PVY), Tomato spotted wilt tospovirus (TSWV) and Watermelon mosaic potyvirus 2 (WMV‐2). The virus found most frequently was CMV and all the isolates identified were of subgroup IA. AMV was also detected in several areas and all isolates were of subgroup II, suggesting a French origin. Finally, at the time of this survey, TSWV infections appeared to be moderately relevant and absent in protected crops. This is the first documented record of AMV, CMV, PVY, TSWV and WMV‐2 in Albania. No infection by Tomato yellow leaf curl begomovirus (TYLCV) or Zucchini yellow mosaic potyvirus (ZYMV) was detected during this survey.

Cucurbits under protected cultivation

Cucurbits under protected cultivation

Virus Disease Problems on Field Cucumber in Hungary with Some International Aspects

Viruses caused severe yield losses of field cucumber in Hungary last years. Field surveys have been carried out on field cucumber to evaluate virus infection in the region of Gyor-Moson-Sopron county. Virus infection was determined on the basis of symptoms, biotest and with DAS-ELISA serological method for the presence of 15 viruses. The virus vector aphid flight was monitored by Moericke yellow water pan. On the basis of field surveys 100% virus infection has been observed in cucumber fields. Virus symptoms were various, depending on varieties, environmental factors, viruses and strains. Out of the viruses investigated only three [Cucumber mosaic cucumovirus (CMV) (DTL serotype); Zucchini yellow mosaic potyvirus (ZYMV) and Watermelon mosaic 2 potyvirus (WMV-2)] have been occurred on cucumber samples. Biological tests confirmed the results of DAS-ELISA. There was much difference, regarding the frequency of viruses. ZYMV was dominant in 1998, while CMV was dominant in 1999. The proportion of the complex infection was very high. Till now, besides CMV other viruses cannot be detected from weeds in cucumber ecosystems. The peak of aphid flight was in the middle of June and later a secondary peak was observed at the beginning of July, which coincided with the appearance of the first virus symptoms. Regarding, that cucumber production occurs in fields at the same place year by year, soil borne virus vectors may play important role in virus infection. To reduce virus infection we can suggest using of light summer oils, which prevents virus transmission by aphids. It is concluded that team work of virologists, pathologists, growers, technologists and the breeders is necessary to solve virus problems of field cucumber in Hungary.

Biological and serological characterization of viruses of summer squash crops in Yugoslavia

A survey on summer squash open field crops was carried out during 2000 and 2001 in order to identify the major viruses infecting these crops in different localities. Plants showed different types of symptoms: mild mosaic, chlorotic spotting, distinctive mosaic, blistering of leaf lamina leaf yellowing, deformation of leaf lamina, knobbed fruits and stunting of plants. The symptoms were very variable but showed the viral nature of the investigated summer squash diseases. The collected samples were tested by bioassay and by two serological methods ELISA and EBIA using cucumber mosaic cucumovirus (CMV), zucchini yellow mosaic potyvirus (ZYMV), watermelon mosaic potyvirus 2 (WMV-2), zucchini yellow flack potyvirus (ZYFV) watermelon mosaic potyvirus 1 (WMV-1), squash mosaic comovirus (SqMV) and cucurbit aphid-borne yellows polerovirus (CABYV) polyclonal antisera. In all tested samples single or mixed infection with ZYMV, CMV and WMV-2 was detected. The most prevalent virus infecting summer squash was ZYMV. This is the first report of ZYMV, the most destructive virus infecting cucurbits, in Yugoslavia. It was also proven that the identified viruses are transmissible by Aphis gossypii in a non-persistent manner, but possible role of seed in virus transmission was not confirmed.

Identification of viruses infecting pumpkins (Cucurbita pepo L) in Yugoslavia

This study was carried out in order to identify the major viruses infecting pumpkins (Cucurbita pepo)grown in Serbia. Leaf samples from virus-infected pumpkin plants were collected in mid-July 2001. Naked-seeded and hulled oil pumpkins, patty pan, zucchini and summer squash from three different locations were included (Table 1). Virus-infected plants showed different symptoms (Table 2 and Figures 1-4). Due to the great variability of the symptoms, the causal viruses could not be fully and precisely determined by visual examination only. The infected samples were tested by the biotest, as well as by two serological methods, ELISA and EBIA. Polyclonal antibodies raised against cucumber mosaic cucumovirus (CMV), zucchini yellow mosaic potyvirus (ZYMV), watermelon mosaic potyvirus 1 (WMV-1), watermelon mosaic potyvirus 2 (WMV-2) and squash mosaic comovirus (SqMV) were used. In each of the 50 collected samples one or two viruses were detected (Tables 3 and 4). The most prevalent viruses infecting pumpkins were ZYMV (62%) and CMV (58%). WMV-2 was extremely rare.

A nonviral peptide can replace the entire N terminus of zucchini yellow mosaic potyvirus coat protein and permits viral systemic infection.

Systematic deletion and peptide tagging of the amino-terminal domain (NT, ~43 amino acids) of an attenuated zucchini yellow mosaic potyvirus (ZYMV-AGII) coat protein (CP) were used to elucidate its role in viral systemic infection. Deletion mutants truncated by 8, 13, and 33 amino acid residues from the CP-NT 5' end were systemically infectious and produced symptoms similar to those of the AGII virus. Tagging these deletion mutants with either human c-Myc (Myc) or hexahistidine peptides maintained viral infectivity. Similarly, addition of these peptides to the intact AGII CP-NT did not affect viral life cycle. To determine which parts, if any, of the CP-NT are essential for viral systemic infection, a series of Myc-tagged mutants with 8 to 43 amino acids removed from the CP-NT were constructed. All Myc-tagged CP-NT deletion mutants, including those from which virtually all the viral CP-NT had been eliminated, were able to encapsidate and cause systemic infection. Furthermore, chimeric viruses with deletions of up to 33 amino acids from CP-NT produced symptoms indistinguishable from those caused by the parental AGII virus. In contrast to CP-NT Myc fusion, addition of the foot-and-mouth disease virus (FMDV) immunogenic epitope to AGII CP-NT did not permit systemic infection. However, fusion of the Myc peptide to the N terminus of the FMDV peptide restored the capability of the virus to spread systemically. We have demonstrated that all CP-NT fused peptides were exposed on the virion surface, masking natural CP immunogenic determinants. Our findings demonstrate that CP-NT is not essential for ZYMV spread and that it can be replaced by an appropriate foreign peptide while maintaining systemic infectivity.

Spread of zucchini yellow mosaic potyvirus in squash in Hungary

The temporal and spatial distribution of zucchini yellow mosaic potyvirus (ZYMV) was studied in a 3000‐m2 zucchini squash field. The first infected plants were found 4 weeks after the field was exposed to virus source plants. The infection increased to nearly 74% by the end of the study. Alate aphids were active from the beginning of the study and 43 species were trapped in the field. Flights of vector species Acyrthosiphon pisum and Myzus persicae peaked during the fourth week which resulted in high virus incidence 4 weeks later. There was a significant correlation between the number of vectors caught in yellow pan traps and the number of infected plants in the field. In laboratory studies evaluating 11 aphid species, Aphis pomi de Geer was identified as a new vector species of ZYMV. Although this aphid was not caught in our field studies, it may be an important contributor in other areas where cucurbits are grown in close proximity to apple or other hosts of this aphid.

Replacement of the coat protein gene of plum pox potyvirus with that of zucchini yellow mosaic potyvirus: characterization of the hybrid potyvirus

Infectious hybrid virus was generated by replacing part of the coat protein gene of plum pox potyvirus with that of the zucchini yellow mosaic potyvirus. This viable hybrid contains 84.5% of zucchini yellow mosaic potyvirus coat protein gene while the rest of the sequence was derived from plum pox potyvirus. Changing the coat protein gene between these two viruses had no effect on the experimental host range. Pathogenicity, stability and replication capacity of the hybrid virus were nearly identical to the parent viruses.

Engineering zucchini yellow mosaic potyvirus as a non-pathogenic vector for expression of heterologous proteins in cucurbits

Plant virus vectors provide an attractive biotechnological tool for the transient expression of foreign genes in whole plants. As yet there has been no use of recombinant viruses for the improvement of commercial crops. This is mainly because the viruses used to create vectors usually cause significant yield loss and can be transmitted in the field. A novel attenuated zucchini yellow mosaic potyvirus (AG) was used for the development of an environmentally safe non-pathogenic virus vector. The suitability of AG as an expression vector in plants was tested by analysis of two infectious viral constructs, each containing a distinct gene insertion site. Introduction of a foreign viral coat protein gene into AG genome between the P1 and HC-Pro genes, resulted in no expression in planta . In contrast, the same gene was stably expressed when inserted between NIb and CP genes, suggesting that this site is more suitable for a gene vector. Virus-mediated expression of reporter genes was observed in squash and cucumber leaves, stems, roots and edible fruit. Furthermore, AG stably expressed human interferon-alpha 2, an important human anti-viral drug, without affecting plant development and yield. Interferon biological activity was measured in cucumber and squash fruit. Together, these data corroborate a biotechnological utility of AG as a non-pathogenic vector for the expression of a foreign gene, as a benefit trait, in cucurbits and their edible fruit.

Cucurbit protoplast isolation for the study of plant virus replication

A cucurbit protoplast isolation protocol was established for the study of plant virus replication in vivo. This protocol is applicable to both cucumber and squash leaf tissue with significant increases in yields of viable protoplasts suitable for electroporation, compared to other published methods. A combination of Cellulase RS, Macerozyme R10 and mannitol was used as digestion enzymes and osmoticum. An average of 1.7×10 7 protoplasts per gram of fresh leaf tissue were obtained from cucumber cultivar Bet-α. Both cucumber cultivar Shimson and squash cultivar First Taste produced an average yield of 6.0×10 6 protoplasts per g of fresh leaf tissue. Electroporation of 10 μg of Zucchini yellow mosaic potyvirus (ZYMV-S) RNA into the protoplasts resulted in virus replication and synthesis of coat protein (CP). SDS-PAGE and immunoblotting were used to detect the CP 48 h post-electroporation. This protocol is highly reproducible and will assist researchers who require cucurbit protoplasts to study virus replication.