scholarly journals Use of Posttranscription Gene Silencing in Squash to Induce Resistance against the Egyptian Isolate of theSquash Leaf Curl Virus

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Omnia Taha ◽  
Inas Farouk ◽  
Abdelhadi Abdallah ◽  
Naglaa A. Abdallah
Keyword(s):  
Gene Silencing ◽  
Real Time ◽  
Real Time Pcr ◽  
Protein Gene ◽  
Bipartite Begomovirus ◽  
Squash Leaf Curl Virus ◽  
Induce Resistance ◽  
Biotype B ◽  
Leaf Curl Virus ◽  
Leaf Curl

Squash leaf curl virus(SqLCV) is a bipartite begomovirus affecting squash plants. It is transmitted by whiteflyBemisia tabacibiotype B causing severe leaf curling, vein banding, and molting ending by stunting. In this study full-length genomic clone of SqLCV Egyptian isolated and posttranscriptional gene silencing (PTGS) has been induced to develop virus resistance. The Noubaria SqLCV has more than 95% homology with Jordon, Israel, Lebanon, Palestine, and Cairo isolates. Two genes fragment from SqLCV introduced in sense and antisense orientations using pFGC5049 vector to be expressed as hairpin RNA. The first fragment was 348 bp from replication associated protein gene (Rep). The second fragment was 879 bp representing the full sequence of the movement protein gene (BC1). Using real-time PCR, a silencing record of 97% has been recorded toRep/TrAPconstruct; as a result it has prevented the appearance of viral symptoms in most tested plants up to two months after infection, while construct containing theBC1gene scored a reduction in the accumulation of viral genome expression as appearing in real-time PCR results 4.6-fold giving a silencing of 79%, which had a positive effect on symptoms development in most tested plants.

TaqMan real-time PCR for detection and quantitation of squash leaf curl virus in cucurbits

2012 ◽  
Vol 179 (2) ◽  
pp. 367-372 ◽  
Author(s):  
Cheng-Ping Kuan ◽  
Hung-Chang Huang ◽  
Chia-Che Chang ◽  
Yi-Lin Lu
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

scholarly journals Cucurbit leaf curl virus, a New Whitefly Transmitted Geminivirus in Arizona, Texas, and Mexico

Plant Disease ◽  
2000 ◽  
Vol 84 (7) ◽  
pp. 809-809 ◽  
Author(s):  
J. K. Brown ◽  
A. M. Idris ◽  
M. W. Olsen ◽  
M. E. Miller ◽  
T. Isakeit ◽  
...  
Keyword(s):  
Binding Site ◽  
Field Isolate ◽  
Bipartite Begomovirus ◽  
Degenerate Primers ◽  
Field Isolates ◽  
Squash Leaf Curl Virus ◽  
Cp Gene ◽  
Honeydew Melon ◽  
Leaf Curl Virus ◽  
Leaf Curl

In 1998 to 1999, geminivirus-like symptoms were observed in whitefly-infested pumpkin, honeydew melon, and muskmelon in Arizona and Texas and in Coahuilla, Mexico (MX), respectively. Plants exhibited leaf curl and/or mottling, reminiscent of symptoms caused by Squash leaf curl virus (SLCV-WAZ) described from Arizona in 1981 (2). The isolate from Arizona pumpkin fields was experimentally transmitted to pumpkin seedlings by the “B type” of Bemisia tabaci (Genn.), and symptoms were indistinguishable from those observed in infected fields. Samples from AZ, MX, and TX were assessed for begomovirus presence by polymerase chain reaction (PCR) using degenerate primers that amplify a contiguous fragment containing the viral coat protein (Cp) gene and common region (CR) of the A component (CR-A) (~2,100 bp) and a fragment containing the CR of the B component (CR-B) (~1,100 bp). One to four isolates from each location were examined by PCR using both primer pairs, and at least three amplicons per isolate were cloned and their sequences determined. Alignment of viral Cp nucleotide (nt) sequences revealed that AZ [AF256199], MX, and TX field isolates shared 98.7 to 100% sequence identity, but were only 84.5 to 85.6% identical to the Cp gene of SLCV-extended (SLCV-E) [M38183] and SLCV-restricted (SLCV-R) (S. G. Lazarowitz, unpublished), respectively, suggesting a new, previously undescribed begomoviral species (3). Further, the Cp nt sequence of the three field isolates was 6 nt shorter than SLCV-E, SLCV-WAZ [AF256203], and SLCV-R Cp sequences. The CR-A [AF256200] and CR-B [AF256201] sequences (179 nt, each) of field isolates, including the theoretical Rep binding element, GGTGT, were 100% identical. Although the Rep binding site is identical among field isolates, SLCV-E, SLCV-R, and SLCV-WAZ, the field isolate CR sequence shared only 64.2, 67.5, and 66.9% overall identity with CR-A SLCV-E, SLCV-R [M63155], and SLCV-WAZ [AF256202], respectively. Prior to 1998 to 1999, SLCV-WAZ was the only New World begomovirus of cucurbits known to infect both melon (Cucumis) and pumpkin (Cucurbita) (1). Therefore, SLCV was initially suspected as the causal agent. However, here we provide evidence for a new, previously undescribed bipartite begomovirus of cucurbits in AZ, MX, and TX that is herein provisionally designated Cucurbit leaf curl virus (CuLCV). Prediction of its closest begomovirus relatives by Cp nt sequence and Rep binding site comparisons suggest that CuLCV is a new member of the SLCV lineage, also containing Bean calico mosaic virus, Cabbage leaf curl virus, SLCV-E, and Texas pepper virus-TAM. References: (1) J. K. Brown and M. R. Nelson. Phytopathology 74:1136, 1984. (2) J. K. Brown and M. R. Nelson. Ann. Appl. Biol. 115:243, 1986. (3) M. A. Mayo and C. R. Pringle. J. Gen. Virol. 97:649, 1998.

scholarly journals Squash Leaf Curl Virus: A New World Bipartite Begomovirus Threatening Squash Production in Oman

Plant Disease ◽  
2020 ◽  
Vol 104 (9) ◽  
pp. 2533-2533
Author(s):  
M. S. Shahid ◽  
H. Al-Sulaimani ◽  
A. M. Al-Sadi
Keyword(s):  
New World ◽  
Bipartite Begomovirus ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

scholarly journals Identification of the Begomoviruses Squash Leaf Curl Virus and Watermelon Chlorotic Stunt Virus in Various Plant Samples in North America

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 810
Author(s):  
Rafaela S. Fontenele ◽  
Amulya Bhaskara ◽  
Ilaria N. Cobb ◽  
Lucas C. Majure ◽  
Andrew M. Salywon ◽  
...  
Keyword(s):  
North America ◽  
High Throughput Sequencing ◽  
Essential Feature ◽  
Native Vegetation ◽  
Bipartite Begomovirus ◽  
Squash Leaf Curl Virus ◽  
The Usa ◽  
Leaf Curl Virus ◽  
Watermelon Chlorotic Stunt Virus ◽  
Leaf Curl

Geminiviruses are a group of plant-infecting viruses with single-stranded DNA genomes. Within this family, viruses in the genus Begomovirus are known to have a worldwide distribution causing a range of severe diseases in a multitude of dicotyledonous plant species. Begomoviruses are transmitted by the whitefly Bemisia tabaci, and their ssDNA genomes can be either monopartite or bipartite. As part of a viral survey, various plants including those in the families Alliaceae, Amaranthaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Malvaceae, Oleaceae and Solanaceae were sampled and screened for begomoviruses using both a high-throughput sequencing and a begomovirus-specific primer pair approach. Based on the sequences derived using these approaches, the full-length genome of various begomoviruses were amplified from plants using abutting primers. Squash leaf curl virus (SLCV) and watermelon chlorotic stunt virus (WCSV) were identified in Cactaceae (n = 25), Solanaceae (n = 7), Cucurbitaceae (n = 2) and Lamiaceae (n = 1) samples. WCSV is an Old World bipartite begomovirus that has only recently been discovered infecting watermelons in the Americas. Our discovery of WCSV in the USA is the first indication that it has reached this country and indicates that this virus might be widespread throughout North America. Phylogenetic analysis suggests WCSV was introduced to the New World twice. The detection of begomoviruses in cactus plants suggests possible spillover events from agricultural areas into native vegetation. Since WCSV and SLCV have previously been found in mixed infections, pseudo-recombination infection experiments were conducted. We demonstrate that WCSV DNA-B is successfully trans-replicated by SLCV DNA-A despite very low degree of similarity between the replication-associated iterative sequences present in their common region, an essential feature for binding of the replication associated protein. This study highlights the importance of viral surveys for the detection of spillover events into native vegetation, but also suggests the need for more surveillance of WCSV in the USA, as this virus is a serious threat to watermelon cultivation in the Middle East.

scholarly journals Identifikasi Begomovirus yang Berasosiasi dengan Penyakit Kuning pada Mentimun di Jawa Barat dan Bali

2016 ◽  
Vol 7 (1) ◽  
pp. 9 ◽  
Author(s):  
Rizki Haerunisa ◽  
Gede Suastika ◽  
Tri Asmira Damayanti
Keyword(s):  
Nicotiana Benthamiana ◽  
Protein Gene ◽  
Tomato Leaf ◽  
New Delhi ◽  
West Java ◽  
Ageratum Yellow Vein Virus ◽  
Squash Leaf Curl Virus ◽  
Tomato Leaf Curl ◽  
Leaf Curl Virus ◽  
Leaf Curl

<p>ABSTRACT<br />A survey conducted from several cucumber cultivation area of West Java and Bali found some plants showing yellow mosaic, vein banding, and stunting symptoms, caused by Begomovirus infection. This study aimed to detect and determine incidence of Begomovirus on cucumber plants, and analyze variation of Begomovirus coat protein gene in West Java dan Bali. Leaf samples from 50 plants were taken randomly from each location in Sumedang, Karawang, Sukabumi (West Java) and Tabanan, Gianyar, Klungkung (Bali). Disease incidence was determined based on serological assay using specific antiserum of Tomato leaf curl New Delhi virus (ToLCNDV) dan Squash leaf curl virus (SLCV). Incidence of ToLCNDV and SLCV were 28-100% and 30-80%, respectively. PCR using Begomovirus degenerate primers successfully amplified coat protein gene about ± 550bp. There were three Begomovirus species associated with yellowing disease on cucumber plant i.e Squash leaf curl China virus (SLCCNV), ToLCNDV, and Ageratum yellow vein virus (AYVV). Based on nucleotide sequences analysis, it was found that isolate SLCCNV had highest nucleotide homology with SLCCNV isolate Malaysia (EF197940) about 94.5%, and was considered as a strain “China”, whereas ToLCNDV has highest nucleotide similarity with ToLCNDV isolate Indonesia (AB613825) about 99.4% and was considered as a strain “Indonesia”. The AYVV sequences showed highest nucleotide AYVV isolate Nicotiana benthamiana from Indonesia (AB100305) about 92.1%.</p><p>Keywords: Cucumis sativus, Dot immunobinding assay, Squash leaf curl China virus, Tomato leaf curl New Delhi virus</p><p>ABSTRAK<br />Survei yang dilakukan di beberapa pertanaman mentimun di Jawa Barat dan Bali menemukan gejala mosaik kuning, daun keriting, penebalan tulang daun, dan kerdil akibat infeksi Begomovirus. Penelitian ini bertujuan untuk mendeteksi dan menghitung insidensi penyakit yang disebabkan oleh Begomovirus pada tanaman mentimun, serta menganalisis keragaman gen protein selubung Begomovirus di Jawa Barat dan Bali. Sampel daun diambil secara acak sebanyak 50 tanaman dari tiap lokasi pertanaman mentimun di Sumedang, Karawang, dan Sukabumi (Jawa Barat), Tabanan, Gianyar, dan Klungkung (Bali). Deteksi Begomovirus dan insidensi penyakit ditentukan dengan uji serologi DIBA menggunakan antiserum Tomato leaf curl New Delhi virus (ToLCNDV) dan Squash leaf curl virus (SLCV). Insidensi ToLCNDV dan SLCV berturut-turut berkisar 28-80% dan 30-80%. Deteksi dengan teknik PCR menggunakan primer universal Begomovirus berhasil mengamplifikasi gen protein selubung berukuran ± 550 pb. Hasil perunutan nukleotida menunjukkan terdapat tiga spesies Begomovirus yang menginfeksi tanaman mentimun di Jawa Barat dan Bali, yaitu Squash leaf curl China virus (SLCCNV), ToLCNDV, dan Ageratum yellow vein virus (AYVV). Isolat SLCCNV Bali memiliki kesamaan nukleotida dan asam amino terhadap isolat SLCCNV dari negara lainnya berkisar antara 89.8-94.5% dan 94.2-96.3%, dan dikelompokkan ke dalam strain “Cina”. ToLCNDV isolat Jawa Barat dan Bali memiliki kesamaan nukleotida dan asam amino berkisar antara 92.8-99.4% dan 97.3-99.4% dengan isolat ToLCNDV dari negara lainnya, tergolong ke dalam strain “Indonesia”. Gen protein selubung AYVV Bali memiliki kesamaan nukleotida dan asam amino berkisar antara 89.5-92.1% dan 94.7-95.2%, dengan kesamaan tertinggi dengan isolat AYVV asal Indonesia yang menginfeksi Nicotiana benthamiana.</p><p>Kata kunci: Cucumis sativus, Dot immunobinding assay, Squash leaf curl China virus, Tomato leaf curl New Delhi virus.</p>

scholarly journals A New Tomato-Infecting Begomovirus in Barbados

Plant Disease ◽  
2000 ◽  
Vol 84 (12) ◽  
pp. 1342-1342 ◽  
Author(s):  
M. E. Roye ◽  
N. M. Henry ◽  
P. D. Burrell ◽  
W. A. McLaughlin ◽  
M. K. Nakhla ◽  
...  
Keyword(s):  
Plant Species ◽  
Western Hemisphere ◽  
Pairwise Comparisons ◽  
Bipartite Begomovirus ◽  
Weak Signals ◽  
Tomato Plants ◽  
Leaf Curling ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

In September 1998, tomato plants in Barbados exhibited symptoms of severe leaf curling without marginal chlorosis. These symptoms were often associated with an increase in whitefly (Bemisia tabaci) populations. DNA was extracted from leaf tissue from symptomatic tomato plants. Polymerase chain reaction (PCR) was performed with DNA-A degenerate primer pair PAC1v1978/PAV1c715, which amplifies part of the rep gene, the cp gene, and the common region (CR), and with DNA-B primer pair PBC1v2039/PBV1c800, which amplifies part of the bc1 and bv1 genes and the CR (2). The amplified PCR fragments of DNA-A and DNA-B were 1.3 and 1.4 kb, respectively, which are the expected sizes from bipartite, whitefly-transmitted geminiviruses of the Western Hemisphere (2). DNA sequence of the cloned fragments of DNA-A and DNA-B are available as GenBank No. AF213013 and AF213014, respectively. The 181 nucleotides of the CR of DNA-A had a nucleotide identity of 96% with the CR of DNA-B, which indicates that this is a bipartite begomovirus. Pairwise comparisons using DNASTAR (DNASTAR, Madison, WI) of the sequenced part of DNA-A was most similar to Cabbage leaf curl virus (CaLCuV, 69%, U65529) and Squash leaf curl virus extended host range isolate (SqLCV-E, 64%, M38183), and <59% to 13 other bipartite Western Hemisphere geminiviruses and Tomato yellow leaf curl virus from Israel (X15656). Pairwise comparisons of the DNA-B fragment sequence was 59 and 55% similar to CaLCuV (U65530) and SqLCV-E (M38182), respectively. Phylogenetic analysis of DNA-A of the major groups of Western Hemisphere begomoviruses placed the Barbados tomato-infecting geminivirus in the cluster with CaLCuV and SqLCV-E (1), while DNA-B analysis placed it with CaLCuV. The DNA-A amplified fragment was used as a probe at high stringency with the dot blot hybridization assay using the Genius II labeling and detection kit (Boeringer Mannheim) to detect this geminivirus in tomato and several other plant species, which had typical geminiviral symptoms. Strong hybridization signals were obtained for all 23 tomato plants with symptoms, weak signals were observed for two of three muskmelon and two of seven watermelon plants, all with leaf curling symptoms. No hybridization signals were observed for peppers with leaf curling symptoms and two weed species, Macroptilium lathyroides and Rhynchosia minima, with golden mosaic symptoms or with the symptomless plant species used as negative controls. The weak signals observed from watermelon and muskmelon samples indicated the presence of low virus titer or geminiviruses distinct from this tomato virus. The presence of viral DNA in these two plant species was confirmed by PCR with degenerate primers described above. Resulting database searches of sequences in the GenBank revealed that the Barbados tomato virus appears to be a previously unreported virus. This new virus is given the provisional name Tomato leaf curl Barbados virus (ToLCBBV). References: (1) J. C. Faria et al. Phytopathology 84:321, 1994. (2) M. R. Rojas et al. Plant Dis. 77:340, 1993.

scholarly journals Ultra-rapid Real-time PCR for the Detection of Tomato yellow leaf curl virus

2012 ◽  
Vol 18 (4) ◽  
pp. 298-303
Author(s):  
Tack-Soo Kim ◽  
Seung-Kook Choi ◽  
Min-Jung Ko ◽  
Minho Lee ◽  
Hyung Seok Choi ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Tomato Yellow Leaf ◽  
Yellow Leaf ◽  
Leaf Curl Virus ◽  
Leaf Curl

scholarly journals Resistanceto Cucurbit Leaf Crumple Virus in Melon

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1108E-1109 ◽  
Author(s):  
James D. McCreight ◽  
Hsing-Yeh Liu ◽  
Thomas A. Turini
Keyword(s):  
Sweet Potato ◽  
Cucumis Melo ◽  
Field Data ◽  
Field Tests ◽  
Squash Leaf Curl Virus ◽  
Biotype B ◽  
Leaf Curl Virus ◽  
Cucurbit Leaf Crumple Virus ◽  
Greenhouse Tests ◽  
Leaf Curl

Cucurbit leaf crumple geminivirus (CuLCrV) is transmitted by sweet-potato whitefly (Bemisia tabaci) biotype B (SPWF-B) and occurs on cucurbits in Arizona, California, Texas, and Mexico. This virus is identical to Cucurbit leaf curl virus, and their symptoms are similar to Squash leaf curl virus on squash (Cucurbita sp.) and Melonleaf curl virus on melon (Cucumis melo L.). Melon has been reported to be either susceptible to CuLCrV, or to have the ability to recover from infection. Twenty-three melon cultigens were inoculated with CuLCrV in greenhouse tests using SPWF-B. Eighteen of the cultigens tested were highly susceptible to CuLCrV (≥60% infected plants) and generally exhibited pronounced CuLCrV symptoms: `Amarillo', `Edisto 47', `Esteem', `Fuyu 3', `Impac', `Moscatel Grande', `Negro', `Perlita', PI 234607, PI 236355, PI 414723, `PMR 5', `Seminole', `Sol Dorado', `Sol Real', `Top Mark', `Vedrantais', and WMR 29. Five cultigens were resistant to CuLCrV (<40% infected plants that exhibited restricted, mild symptoms): MR-1, PI 124111, PI 124112, PI 179901, and PI 313970. Symptoms abated with time in both groups although infected plants remained positive for the virus. Ten of the cultigens (`Edisto 47', `Fuyu 3', `Impac', MR-1, PI 124112, PI 313970, PI 414723, `PMR 5', `Top Mark', and WMR 29) were included in field tests in 2003 and 2004 that were naturally infected with CuLCrV. With the exception of PI 414723, the greenhouse and field data were consistent for reaction to CuLCrV.

scholarly journals Emergence of a New Cucurbit-Infecting Begomovirus Species Capable of Forming Viable Reassortants with Related Viruses in the Squash leaf curl virus Cluster

2002 ◽  
Vol 92 (7) ◽  
pp. 734-742 ◽  
Author(s):  
J. K. Brown ◽  
A. M. Idris ◽  
C. Alteri ◽  
Drake C. Stenger
Keyword(s):  
Nucleotide Sequence ◽  
Host Range ◽  
Nucleotide Sequence Identity ◽  
Begomovirus Species ◽  
Progeny Virus ◽  
Bipartite Begomovirus ◽  
Sequence Identity ◽  
Squash Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

Cucurbit leaf curl virus (CuLCV), a whitefly-transmitted geminivirus previously partially characterized from the southwestern United States and northern Mexico, was identified as a distinct bipartite begomovirus species. This virus has near sequence identity with the previously partially characterized Cucurbit leaf crumple virus from California. Experimental and natural host range studies indicated that CuLCV has a relatively broad host range within the family Cucurbitaceae and also infects bean and tobacco. The genome of an Arizona isolate, designated CuLCV-AZ, was cloned and completely sequenced. Cloned CuLCV-AZ DNA A and B components were infectious by biolistic inoculation to pumpkin and progeny virus was transmissible by the whitefly vector, Bemisia tabaci, thereby completing Koch's postulates. CuLCV-AZ DNA A shared highest nucleotide sequence identity with Squash leaf curl virus-R (SLCV-R), SLCV-E, and Bean calico mosaic virus (BCaMV) at 84, 83, and 80%, respectively. The CuLCV DNA B component shared highest nucleotide sequence identity with BCaMV, SLCV-R, and SLCV-E at 71, 70, and 68%, respectively. The cis-acting begomovirus replication specificity element, GGTGTCCTGGTG, in the CuLCV-AZ origin of replication is identical to that of SLCV-R, SLCV-E, and BCaMV, suggesting that reassortants among components of CuLCV-AZ and these begomoviruses may be possible. Reassortment experiments in pumpkin demonstrated that both reassortants of CuLCV-AZ and SLCV-E A and B components were viable. However, for CuLCV-AZ and SLCV-R, only one reassortant (SLCV-R DNA A/CuLCV-AZ DNA B) was viable on pumpkin, even though the cognate component pairs of both viruses infect pumpkin. These results demonstrate that reassortment among sympatric begomovirus species infecting cucurbits are possible, and that, if generated in nature, could result in begomoviruses bearing distinct biological properties.

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