scholarly journals Detection of pepper leaf curl virus through PCR amplification and expression of its coat protein in Escherichia coli for antiserum production

2011 ◽  
Vol 10 (17) ◽  
pp. 3290-3295 ◽  
Author(s):  
Sinha D P ◽  
Saxena S ◽  
Kumar S ◽  
Singh M
Keyword(s):  
Escherichia Coli ◽  
Coat Protein ◽  
Pcr Amplification ◽  
Leaf Curl Virus ◽  
Leaf Curl

scholarly journals Serological Studies Using Polyclonal Antisera Prepared Against the Viral Coat Protein of Four Begomoviruses Expressed in Escherichia coli

Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1109-1114 ◽  
Author(s):  
A. M. Abouzid ◽  
J. Freitas-Astua ◽  
D. E. Purcifull ◽  
J. E. Polston ◽  
K. A. Beckham ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Coat Protein ◽  
Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thin Sections ◽  
Golden Yellow ◽  
Polyclonal Antisera ◽  
Leaf Curl Virus ◽  
Leaf Curl

Polyclonal rabbit antisera were produced to the coat protein of Bean golden mosaic virus Brazil isolate (BGMV), Cabbage leaf curl virus (CabLCV), Tomato yellow leaf curl virus (TYLCV), and Tomato mottle virus (ToMoV), all expressed in Escherichia coli by the pETh expression vector. The expressed coat protein of each virus was purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for use as an immunogen. The antisera to BGMV, CabLCV, TYLCV, and ToMoV reacted in indirect (plate-trapping) enzyme-linked immunosorbent assay (ELISA) with extracts from begomovirus-infected tissue. The antisera to BGMV, CabLCV, TYLCV, and ToMoV also reacted specifically with the test begomovirus antigens in leaf imprint blots and Western blots. The CabLCV and TYLCV antisera were used to detect Bean golden yellow mosaic virus antigens by immunogold labeling of thin sections of infected bean tissues. In tissue blot immunoassays, the TYLCV antiserum reacted well with TYLCV antigens but not with ToMoV antigens, while CabLCV antiserum reacted well with ToMoV antigens and weakly with TYLCV antigens. The results indicate that polyclonal antisera prepared to expressed begomovirus coat proteins were useful for the detection of begomoviruses in an array of assays.

Expression and functional characterisation of a soluble form of Tomato yellow leaf curl virus coat protein

2014 ◽  
Vol 70 (10) ◽  
pp. 1624-1631 ◽  
Author(s):  
Lan-Lan Wang ◽  
Xue-Mei Wei ◽  
Xiao-Dong Ye ◽  
Hong-Xing Xu ◽  
Xue-Ping Zhou ◽  
...  
Keyword(s):  
Coat Protein ◽  
Tomato Yellow Leaf ◽  
Soluble Form ◽  
Yellow Leaf ◽  
Virus Coat Protein ◽  
Functional Characterisation ◽  
Virus Coat ◽  
Leaf Curl Virus ◽  
Leaf Curl

Occurrence and genetic variability of partial coat protein gene of Sweet potato leaf curl virus (SPLCV) in Kenya

2017 ◽  
Vol 16 (45) ◽  
pp. 2112-2120 ◽  
Author(s):  
S. Maina ◽  
D. W. Miano ◽  
E. Mbogo ◽  
J. O. Amimo ◽  
J. Irungu ◽  
...  
Keyword(s):  
Coat Protein ◽  
Genetic Variability ◽  
Sweet Potato ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Potato Leaf ◽  
Leaf Curl Virus ◽  
Leaf Curl

Coat protein promoter from cotton leaf curl virus is not a tissue-specifically expressed promoter

2000 ◽  
Vol 45 (20) ◽  
pp. 1869-1874 ◽  
Author(s):  
Yingqiu Xie ◽  
Zhen Zhu ◽  
Yule Liu ◽  
Qian Wu ◽  
Honglin Xu
Keyword(s):  
Coat Protein ◽  
Cotton Leaf ◽  
Cotton Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

scholarly journals Detection of a Whitefly-Transmitted Geminivirus in Launaea spp. in India

Plant Disease ◽  
2000 ◽  
Vol 84 (5) ◽  
pp. 593-593
Author(s):  
S. K. Raj ◽  
G. Chandra ◽  
B. P. Singh
Keyword(s):  
Virus Transmission ◽  
Pcr Amplification ◽  
African Cassava Mosaic Virus ◽  
Ammonium Bromide ◽  
Pcr Product ◽  
Total Dna ◽  
Cetyl Trimethyl Ammonium ◽  
Tomato Leaf Curl ◽  
Leaf Curl Virus ◽  
Leaf Curl

Launaea acaulis (Roxb.) Babc. Aerr. and L. procumbeus (Roxb.) Ramayya & Rajgopal are weeds that grow near agricultural fields in India during the rainy season. Yellow net symptoms were observed in both species, but not in crops, in 1998 and 1999. Nicotiana tabacum cv. White Burley developed leaf curl symptoms after virus transmission from L. acaulis by whiteflies (Bemisia tabaci). Thus, the involvement of a geminivirus was suspected. Total DNA from infected and uninfected tobacco and Launaea plants was extracted by the cetyl trimethyl ammonium bromide method and precipitated by ethanol. Polymerase chain reaction (PCR), using DNA from infected and uninfected plants as a template and degenerate oli-gonucleotide primers designed from DNA-A of whitefly-transmitted African cassava mosaic virus (ACMV) (1) was performed. An expected ≈500-bp fragment was observed after agarose gel electrophoresis of the PCR product. Total DNA from infected plants gave positive signals and was homologous with the probe from cloned DNA-A of Indian tomato leaf curl virus (2) in high-stringency Southern hybridization tests. No PCR amplification and positive signals were obtained from DNA from uninfected plants. PCR amplification of the 500-bp fragment with primers from DNA-A of whitefly-transmitted ACMV resulted in positive signals, and homology with the probe indicated that L. acaulis and L. procumbens harbor whitefly-transmitted geminivirus infections. This is the first report of geminivirus infection in these Launaea spp. in India. References: (1) D. Deng et al. Ann. Appl. Biol. 125:327, 1994. (2) K. M. Srivastava, et al. J. Virol. Methods 51:297, 1985.

scholarly journals Engineering resistance against Tomato yellow leaf curl virus via the CRISPR/Cas9 system in tomato

2017 ◽  
Author(s):  
Manal Tashkandi ◽  
Zahir Ali ◽  
Fatimah Aljedaani ◽  
Ashwag Shami ◽  
Magdy M. Mahfouz
Keyword(s):  
Coat Protein ◽  
Virus Resistance ◽  
Nicotiana Benthamiana ◽  
Tomato Yellow Leaf ◽  
Yellow Leaf ◽  
Tomato Plants ◽  
Guide Rna ◽  
Multiple Generations ◽  
Leaf Curl Virus ◽  
Leaf Curl

AbstractCRISPR/Cas systems confer molecular immunity against phages and conjugative plasmids in prokaryotes. Recently, CRISPR/Cas9 systems have been used to confer interference against eukaryotic viruses. Here, we engineered Nicotiana benthamiana and tomato (Solanum lycopersicum) plants with the CRISPR/Cas9 system to confer immunity against the Tomato yellow leaf curl virus (TYLCV). Targeting the TYLCV genome with Cas9-single guide RNA at the sequences encoding the coat protein (CP) or replicase (Rep) resulted in efficient virus interference, as evidenced by low accumulation of the TYLCV DNA genome in the transgenic plants. The CRISPR/Cas9-based immunity remained active across multiple generations in the N. benthamiana and tomato plants. Together, our results confirmed the efficiency of the CRISPR/Cas9 system for stable engineering of TYLCV resistance in N. benthamiana and tomato, and opens the possibilities of engineering virus resistance against single and multiple infectious viruses in other crops.

scholarly journals Evolutionary and Molecular Aspects of Indian Tomato Leaf Curl Virus Coat Protein

2012 ◽  
Vol 2012 ◽  
pp. 1-12 ◽  
Author(s):  
Sivakumar Prasanth Kumar ◽  
Saumya K. Patel ◽  
Ravi G. Kapopara ◽  
Yogesh T. Jasrai ◽  
Himanshu A. Pandya
Keyword(s):  
Coat Protein ◽  
Viral Particle ◽  
Tomato Leaf ◽  
Host Cells ◽  
Coat Proteins ◽  
Tomato Leaf Curl Disease ◽  
Tomato Leaf Curl Virus ◽  
Tomato Leaf Curl ◽  
Leaf Curl Virus ◽  
Leaf Curl

Tomato leaf curl disease (ToLCD) is manifested by yellowing of leaf lamina with upward leaf curl, leaf distortion, shrinking of the leaf surface, and stunted plant growth caused by tomato leaf curl virus (ToLCV). In the present study, using computational methods we explored the evolutionary and molecular prospects of viral coat protein derived from an isolate of Vadodara district, Gujarat (ToLCGV-[Vad]), India. We found that the amino acids in coat protein required for systemic infection, viral particle formation, and insect transmission to host cells were conserved amongst Indian strains. Phylogenetic studies on Indian ToLCV coat proteins showed evolutionary compatibility with other viral taxa. Modeling of coat protein revealed a topology similar to characteristic Geminate viral particle consisting of antiparallel β-barrel motif with N-terminus α-helix. The molecular interaction of coat protein with the viral DNA required for encapsidation and nuclear shuttling was investigated through sequence- and structure-based approaches. We further emphasized the role of loops in coat protein structure as molecular recognition interface.

scholarly journals A Mixed Infection of Lettuce chlorosis virus, Papaya ringspot virus, and Tomato yellow leaf curl virus-IL Detected in a Texas Papaya Orchard Affected by a Virus-Like Disease Outbreak

Plant Disease ◽  
2017 ◽  
Vol 101 (7) ◽  
pp. 1094-1102 ◽  
Author(s):  
Olufemi J. Alabi ◽  
M. Al Rwahnih ◽  
J. L. Jifon ◽  
M. Sétamou ◽  
J. K. Brown ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Host Record ◽  
Tomato Yellow Leaf ◽  
Ringspot Virus ◽  
Yellow Leaf ◽  
Papaya Ringspot Virus ◽  
Perennial Crop ◽  
New Host ◽  
Leaf Curl Virus ◽  
Leaf Curl

Severe virus-like symptoms consisting of mosaic, distortion, yellowing, and brittleness were observed on papaya plants in a 20-ha orchard in South Texas during the 2014–15 growing season. Incidence of symptomatic plants increased from ∼40 to 100% within 6 months of the outbreak; the most severely affected plants were stunted, and fruit yield and quality were reduced compared with asymptomatic plants. The orchard papaya plant virome was explored using the Illumina NextSeq 500 platform and results were validated by Sanger DNA sequencing of complete viral genomes obtained by PCR amplification. The combined results revealed the presence of Papaya ringspot virus (PRSV; Potyvirus), Lettuce chlorosis virus (LCV; Crinivirus), and Tomato yellow leaf curl virus-IL (TYLCV-IL; Begomovirus). The RT-PCR analyses of leaves from 51 randomly sampled papaya plants indicated the presence of PRSV, LCV, and TYLCV-IL in 100, 39.2, and 15.7% of the samples, respectively. Plants infected with PRSV, in combination with LCV and/or TYLCV-IL, exhibited more severe symptoms compared with plants infected with PRSV alone. Furthermore, successful whitefly-mediated transmission of TYLCV-IL and LCV was accomplished by exposing virus-free papaya seedlings to viruliferous Bemisia tabaci (Genn.) under greenhouse conditions. The results of this study document a new host record for LCV and the first successful whitefly-mediated transmission of TYLCV-IL and LCV to papaya. As a perennial crop, infected papaya serving as an over-seasoning reservoir for TYLCV-IL and LCV, presents a new challenge to viral disease management in papaya orchards.

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