Prevalence and Molecular Characterization Coat Protein Gene of Tobacco streak virus Causing Peanut Stem Necrosis Disease in Coastal and Rayalaseema Districts of Andhra Pradesh, South-India

2021 ◽  
Author(s):  
K. Saratbabu ◽  
K. Vemana ◽  
A.K. Patibanda ◽  
B. Sreekanth ◽  
V. Srinivasa Rao
Keyword(s):  
Coat Protein ◽  
Molecular Characterization ◽  
Andhra Pradesh ◽  
Disease Incidence ◽  
Tobacco Streak Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Streak Virus ◽  
Rt Pcr ◽  
Cp Gene ◽  
Stem Necrosis

Background: Peanut stem necrosis disease (PSND) caused by Tobacco streak virus (TSV) is a major constraint for groundnut production in Andhra Pradesh (A.P.). However, studies on prevalence and spread of the disease confined to only few districts of A.P. with this background current study focused on incidence and spread of the disease in entire state of A.P. Further an isolate of TSV occurring in A.P. characterized on the basis of genetic features by comparing with other TSV isolates originated from different hosts and locations from world.Methods: Roving survey was conducted during kharif 2017-18 in groundnut growing districts of Andhra Pradesh (A.P.) for peanut stem necrosis disease incidence. Groundnut plants showing PSND symptoms were collected and tested with direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA). Groundnut samples found positive by ELISA once again tested by reverse transcription polymerase chain reaction (RT-PCR). The representative TSV-GN-INDVP groundnut isolate from Prakasham district was maintained on cowpea seedlings by standard sap inoculation method in glasshouse for further molecular characterization. The Phylogenetic tree for coat protein (CP) gene was constructed using aligned sequences with 1000 bootstrap replicates following neighbor-joining phylogeny.Result: Thirty-eight (52.7%) of seventy-two groundnut samples collected from different locations in A.P were given positive reaction to TSV by DAC-ELISA. For the first time, PSND incidence observed in coastal districts (Krishna, Guntur, Sri Pottisriramulu Nellore, Prakasham) of A.P. Maximum PSND incidence recorded from Bathalapalli (22.2%) and the minimum incidence in Mulakalacheruvu (4.1%). The coat protein (CP) gene of TSV-GN-INDVP groundnut isolate was amplified by RT-PCR and it shared maximum per cent nucleotide identity (97.51-98.62%) with TSV isolates from groundnut and other different crops reported in India. All Indian isolates cluster together irrespective of crop and location based on the phylogenetic analysis.

scholarly journals First Report of Tobacco streak virus Infecting Guizotia abyssinica from India

Plant Disease ◽  
2007 ◽  
Vol 91 (3) ◽  
pp. 330-330 ◽  
Author(s):  
N. Arun Kumar ◽  
M. Lakshmi Narasu ◽  
U. B. Zehr ◽  
K. S. Ravi
Keyword(s):  
Amino Acid ◽  
Coat Protein ◽  
The United States ◽  
Tobacco Streak Virus ◽  
Streak Virus ◽  
Vegetable Crops ◽  
Guizotia Abyssinica ◽  
Oil Seed ◽  
Cp Gene ◽  
Seed Crops

Guizotia abyssinica (L.f.) Cass. (niger), an important oil seed crop grown in India, is used in foods, paints, soaps, and as an illuminant. During a survey conducted in 2004 to monitor Tobacco streak virus (TSV) in Helianthus annuus L. (sunflower) and Arachis hypogaea L. (groundnut), typical symptoms of leaf and petiole necrosis were observed in niger plants from Karnataka State, India. The field-collected samples reacted with TSV-specific polyclonal antiserum in direct antigen coated (DAC)-ELISA. Indicator host species were mechanically inoculated with extracts from symptomatic leaves and grown under greenhouse conditions. The inoculations resulted in local necrotic lesions on Vigna unguiculata cv. C-152 (cowpea), Gomphrena globosa, and Nicotiana tabacum cv. Xanthi (tobacco) at 3 to 4 days postinoculation (dpi) and systemic mosaic mottling on sunflower and G. globosa at 7 to 9 dpi. To identify the virus at the molecular level, total RNA was isolated (RNeasy kit, Qiagen Inc., Chatsworth, CA) from the virus-inoculated cowpea leaf and used for reverse transcription-PCR using TSV CP (coat protein) specific primers (2). The resulting ~720-bp amplicon corresponding to the CP gene of TSV was cloned into pGem-T vector (Promega, Madison, WI) and sequenced. The resulting sequence of the TSV-niger isolate (TSV-NG) comprised 717 nucleotides encoding 238 amino acid residues of the viral coat protein (GenBank Accession No. DQ864458). Comparison of the sequence with those of other TSV CP gene indicated 98.5 to 99.3% nucleotide and 97.9 to 99.6% amino acid sequence identity with TSV isolates from India (1,2; GenBank Accession Nos. AF505073, AY061930, AY061929, AF515823, AF515824, and AF515825). The sequence of TSV-NG had 89.5 and 80.0% amino acid identity with TSV-WC, type strain from the United States (GenBank Accession No. X00435) and TSV-BR, isolate from Brazil (GenBank Accession No. AY354406), respectively. On the basis of symptoms, transmission, and serological and molecular data, the causal agent of necrosis in niger was identified as a strain of TSV widely prevalent in other oil seed and vegetable crops in India. The new report of Tobacco streak virus infecting niger from India, indicated the expansion of host range among oil seed crops. References: (1) A. I. Bhat et al. Indian J Biotechnol. 1:350, 2002. (2) K. S. Ravi et al. Plant Pathol. 50:800, 2001.

scholarly journals Identification of Tobacco streak virus in Cranberry and the Association of TSV with Berry Scarring

Plant Disease ◽  
2016 ◽  
Vol 100 (4) ◽  
pp. 696-703 ◽  
Author(s):  
L. D. Wells-Hansen ◽  
J. J. Polashock ◽  
N. Vorsa ◽  
B. E. L. Lockhart ◽  
P. S. McManus
Keyword(s):  
New Jersey ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Management Strategies ◽  
Tobacco Streak Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Streak Virus ◽  
Transmission Electron ◽  
Gene Sequence Analysis

Cranberry plants bearing disfigured, scarred fruit were reported by growers in the major cranberry-growing region of central Wisconsin in July 2012. Plants bearing scarred fruit have since been observed in Massachusetts and New Jersey. Three complementary methods provided evidence of Tobacco streak virus (TSV) in symptomatic plants: (i) leaves and scarred berries tested positive for TSV by double-antibody sandwich enzyme-linked immunosorbent assay; (ii) quasi-isometric particles approximately 33 nm in diameter were extracted from leaves of symptomatic plants and visualized using transmission electron microscopy; and (iii) coat protein gene sequence analysis revealed 94 to 99% nucleotide similarity with reference TSV sequences. In newer cultivars, 99% of uprights with scarred berries tested positive for TSV. In older cultivars, 31% of uprights with scarred berries tested positive for TSV and the remaining 69% of uprights with scarred berries tested positive for Blueberry shock virus. TSV overwintered in cranberry plants, and leaves, pollen, and fruit tested positive for TSV the year following symptom occurrence. Attempts to inoculate cranberry using infected pollen or sap as inoculum failed, but several herbaceous hosts tested TSV positive following mechanical inoculation. Phylogenetic analysis of the coat protein gene of 26 TSV isolates from various cultivars of cranberry in Wisconsin, New Jersey, and Massachusetts revealed diversity. This work provides information that will be useful in understanding the epidemiology of TSV in cranberry and in the development of management strategies.

scholarly journals Biological and molecular characterization of an isolate of Tobacco streak virus obtained from soybeans in Brazil

2005 ◽  
Vol 30 (4) ◽  
pp. 366-373 ◽  
Author(s):  
Álvaro M. R. Almeida ◽  
Junichi Sakai ◽  
Kaoru Hanada ◽  
Thalita G. Oliveira ◽  
Priscila Belintani ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Molecular Characterization ◽  
Sucrose Gradient ◽  
Open Reading Frame ◽  
Spherical Particles ◽  
Tobacco Streak Virus ◽  
Streak Virus ◽  
Reading Frame ◽  
Cp Gene

A virus was isolated from soybean (Glycine max) plants with symptoms of dwarfing and bud blight in Wenceslau Braz County, Paraná, Brazil. The host range and properties resembled those of Tobacco streak virus (TSV). The purified virus showed three peaks in a frozen sucrose gradient. Antiserum was produced and the virus was serologically related to TSV. Electron microscopy detected 28 nm spherical particles. Coat protein (CP) had a Mr of 29.880 Da. A fragment of 1028 nt was amplified, cloned and sequenced. One open reading frame with 717 nt was identified and associated to the CP. The CP gene shared 83% identity with the sequence of TSV CP from white clover (Trifolium repens) (GenBank CAA25133). This is the first report of the biological and molecular characterization of TSV isolated from soybeans. It is proposed that this isolate be considered a strain of TSV named TSV-BR.

scholarly journals Sources of Resistance to Tobacco streak virus in Wild Arachis (Fabaceae: Papilionoidae) Germplasm

Plant Disease ◽  
2007 ◽  
Vol 91 (12) ◽  
pp. 1585-1590 ◽  
Author(s):  
G. Kalyani ◽  
A. S. Reddy ◽  
P. Lava Kumar ◽  
R. D. V. J. Prasada Rao ◽  
R. Aruna ◽  
...  
Keyword(s):  
Systemic Infection ◽  
Tobacco Streak Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Streak Virus ◽  
Sources Of Resistance ◽  
Potential Threat ◽  
Southern States ◽  
Severe Reduction ◽  
Stem Necrosis ◽  
Peanut Cultivars

Stem necrosis disease caused by Tobacco streak virus (TSV), first recognized in 2000, has emerged as a potential threat to peanut (Arachis hypogaea) in southern states of India. The virus induces severe necrosis of shoots leading to death of the plant, and plants that survive are malformed, with severe reduction in pod yield. All the currently grown peanut cultivars in India are highly susceptible to the virus. Therefore, wild relatives of peanut were evaluated to identify potential sources of resistance to TSV infection. In all, 56 germplasm accessions from 20 wild Arachis spp. in four sections (Arachis, Erectoides, Procumbente, and Rhizomatosae), along with susceptible peanut cultivars (JL 24 and K 1375), were evaluated for resistance to TSV under greenhouse conditions using mechanical sap inoculations. Systemic virus infection, determined by enzyme-linked immunosorbent assay (ELISA), in the test accessions ranged between 0 and 100%. Twenty-four accessions in section Arachis that had 0 to 35% systemically infected plants were retested, and systemic infection was not detected in eight of these accessions in repeated trials in the greenhouse. These are International Crops Research Institute for the Semi-Arid Tropics groundnut (ICG) accession nos. 8139, 8195, 8200, 8203, 8205, and 11550 belonging to A. duranensis; ICG 8144 belonging to A. villosa; and ICG 13210 belonging to A. stenosperma. Even though the resistant accessions had 0 to 100% TSV infection in inoculated leaves, TSV was not detected in the subsequently emerged leaves. This is the first report of TSV resistance in Arachis spp. The eight TSV resistant accessions are cross compatible with A. hypogaea for utilization in breeding for stem necrosis disease resistance.

scholarly journals Identification, Characterization, and Molecular Detection of Alfalfa mosaic virus in Potato

2006 ◽  
Vol 96 (11) ◽  
pp. 1237-1242 ◽  
Author(s):  
H. Xu ◽  
J. Nie
Keyword(s):  
Mosaic Virus ◽  
Gene Sequence ◽  
Alfalfa Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polymorphism Analysis ◽  
Rt Pcr ◽  
Potato Leaf ◽  
Simple Method ◽  
Cp Gene ◽  
Nucleotide Sequence Alignment

Alfalfa mosaic virus (AMV) was detected in potato fields in several provinces in Canada and characterized by bioassay, enzyme-linked immunosorbent assay, and reverse-transcription polymerase chain reaction (RT-PCR). The identity of eight Canadian potato AMV isolates was confirmed by sequence analysis of their coat protein (CP) gene. Sequence and phylogenetic analysis indicated that these eight AMV potato isolates fell into one strain group, whereas a slight difference between Ca175 and the other Canadian AMV isolates was revealed. The Canadian AMV isolates, except Ca175, clustered together among other strains based on alignment of the CP gene sequence. To detect the virus, a pair of primers, AMV-F and AMV-R, specific to the AMV CP gene, was designed based on the nucleotide sequence alignment of known AMV strains. Evaluations showed that RT-PCR using this primer set was specific and sensitive for detecting AMV in potato leaf and tuber samples. AMV RNAs were easily detected in composite samples of 400 to 800 potato leaves or 200 to 400 tubers. Restriction analysis of PCR amplicons with SacI was a simple method for the confirmation of PCR tests. Thus, RT-PCR followed by restriction fragment length polymorphism analysis may be a useful approach for screening potato samples on a large scale for the presence of AMV.

Survey for Yellow Mosaic Disease Occurring on Blackgram [Vigna mungo (L.) Hepper] and Weed Hosts in Four Major Blackgram Growing Districts of Andhra Pradesh and Characterization of Associated Viruses using Coat Protein Gene

2021 ◽  
Author(s):  
Vallabhaneni Tilak Chowdary ◽  
V. Manoj Kumar ◽  
P. Kishore Varma ◽  
B. Sreekanth ◽  
V. Srinivasa Rao
Keyword(s):  
Coat Protein ◽  
Andhra Pradesh ◽  
Disease Incidence ◽  
Pcr Amplification ◽  
Mosaic Disease ◽  
Yellow Mosaic Virus ◽  
Ageratum Conyzoides ◽  
Yellow Mosaic Disease ◽  
Weed Plants ◽  
Weed Hosts

Background: Yellow mosaic disease (YMD) caused by Yellow mosaic virus is one of the major constraints in the pulse production in Andhra Pradesh (A.P.) due to fast evolution of strains, like Mungbean yellow mosaic India virus (MYMIV). Keeping this in view, a survey was undertaken in the major blackgram growing districts of A.P. to know the YMD incidence in blackgram and weed hosts and were characterized based on genetic features by comparing with other YMV isolates from different hosts and locations across the world. Methods: Roving survey was conducted during rabi 2019-20 in major blackgram growing districts of A.P. viz., Krishna, Guntur, West Godavari and Prakasam districts for YMD incidence. Blackgram plants showing characteristic symptoms were collected as representative samples from each mandal along with the suspected weed plants and were subjected to amplification using coat protein (CP) specific primers followed by molecular characterization. Phylogenetic tree for coat protein (CP) gene was constructed using aligned sequences with 1000 bootstrap replicates following neighbor-joining phylogeny. Result: Out of the four districts surveyed, the highest disease incidence was recorded at Machavaram village of Prakasam district (43.22%), whereas least disease incidence was recorded at Chinaganjam village of Praksam district (2.4%). Six weeds viz., Ageratum conyzoides, Amaranthus viridis, Parthenium hysterophporus, Vigna trilobata, Abelmoscus moschatus, Desmodium laxiflorum have showed positive result in PCR amplification with MYMIV specific coat protein primers. Four isolates from blackgram samples and two from weed plants shared 94.85 to 99.58% nucleotide identity among themselves.

scholarly journals Molecular characterization of the Cassava brown streak virus coat protein

2001 ◽  
Vol 50 (4) ◽  
pp. 527-534 ◽  
Author(s):  
W. A. Monger ◽  
S. Seal ◽  
A. M. Isaac ◽  
G. D. Foster
Keyword(s):  
Coat Protein ◽  
Molecular Characterization ◽  
Streak Virus ◽  
Virus Coat Protein ◽  
Cassava Brown Streak Virus ◽  
Virus Coat ◽  
Brown Streak

scholarly journals First Report of Rice stripe necrosis virus Infecting Rice in Burkina Faso

Plant Disease ◽  
2014 ◽  
Vol 98 (10) ◽  
pp. 1451-1451 ◽  
Author(s):  
D. Sereme ◽  
B. J. Neya ◽  
M. Bangratz ◽  
C. Brugidou ◽  
I. Ouedraogo
Keyword(s):  
Burkina Faso ◽  
Disease Incidence ◽  
Polyclonal Antiserum ◽  
Rt Pcr ◽  
Necrosis Virus ◽  
Polymyxa Graminis ◽  
Cp Gene ◽  
Pcr Product ◽  
Colombian Isolate ◽  
Western Burkina Faso

Rice stripe necrosis virus (RSNV) was first described in 1977 as a new virus infecting rice in Cote d'Ivoire (3) and was subsequently observed in Liberia, Nigeria, and Sierra Leone (2). RSNV is a soil-borne virus transmitted by the fungus Polymyxa graminis (1) and belongs to the genus Benyvirus (4). During a survey carried out in April of 2013, severe symptoms characterized by seedling death, severe plant malformation, and foliar striping were observed on rice plants in an experimental field of INERA at Banfora located in western Burkina Faso. Disease incidence in the field was estimated to be 80 ± 5%. The symptoms of disease were successfully transmitted to the susceptible rice (Oryza sativa) cultivar IR64 by soil transmission experiments (1). RSNV was detected by ELISA using a polyclonal antiserum (1), kindly provided by Dr. Denis Fargette, IRD, Montpellier, France. Total nucleic acid was extracted with TRIzol reagent (Invitrogen) from IR64 and field infected samples. The presence of the virus was confirmed by RT-PCR using primers 5′-CATCTTGTCGAGATGAG-3′ and 5′-GCGTTGTCTTTATCAGTG-3′ for specific sequences flanking the RNA2 CP gene. The RT-PCR product was directly sequenced and the sequence was deposited in GenBank (Accession No. LK023710). Sequence analysis showed that the CP gene of the RSNV isolate from Burkina Faso shared the highest nucleotide sequence identity (97.6%) with the known RSNV CP gene sequence from the Colombian isolate (EU099845) available in GenBank, confirming the presence of RSNV in the rice crops in Burkina Faso. To our knowledge, this is the first confirmed report of RSNV in Burkina Faso. Further studies are needed to determine its incidence and spread in the country. Detection of RSNV in Burkina Faso signals the urgent need for adoption of appropriate measures to restrict the spread and impact of this virus within Africa. References: (1) C. Fauquet and J. C. Thouvenel. Proc. Acad. Sci. Ser. D 296:575, 1983. (2) C. Fauquet et al. Develop. Appl. Biol. 2:71, 1988. (3) D. Louvel and J.-M. Bidaux. Agronomie Tropicale 32:257, 1977. (4) I. Lozano and F. Morales. Eur. J. Plant Pathol. 124:673, 2009.

scholarly journals First Report of Sharka Disease Caused by Plum Pox Virus in Lithuania

Plant Disease ◽  
1998 ◽  
Vol 82 (12) ◽  
pp. 1405-1405 ◽  
Author(s):  
J. Staniulis ◽  
J. Stankiene ◽  
K. Sasnauskas ◽  
A. Dargeviciute
Keyword(s):  
Coat Protein ◽  
Plant Protection ◽  
Virus Disease ◽  
Pcr Amplification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
First Report ◽  
Das Elisa ◽  
Sharka Disease

Plum pox (sharka) disease caused by plum pox potyvirus (PPV) is considered the most important virus disease of stone fruit trees in Europe and the Mediterranean region. Nearly all those countries that produce stone fruits are affected (3). The causal virus of the disease is a European Plant Protection Organization A2 quarantine pathogen. Symptoms of leaf mottling, diffuse chlorotic spots, rings, and vein banding of varied intensity characteristic for plum pox virus infection were observed in the plum (Prunus domestica) orchard tree collection of the Lithuanian Institute of Horticulture in Babtai in 1996. Presence of this virus in the diseased trees was confirmed by double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) with kits from BIOREBA (Reinach, Switzerland) and by polyclonal antibodies raised against a Moldavian isolate of PPV courtesy of T. D. Verderevskaya (Institute of Horticulture, Kishinev, Moldova). ELISAs with both sources of antiserum were positive for presence of PPV. Electron microscopy revealed the presence of potyvirus-like particles averaging 770 nm in extracts of mechanically inoculated plants of Chenopodium foetidum (chlorotic LL [local lesions]) and Pisum sativum cvs. Rainiai and Citron (mottling). For molecular diagnosis and characterization of this isolate, PPV-971, reverse transcription-polymerase chain reaction (RT-PCR) was employed. Total RNA from the leaves of infected pea was isolated as described (2). High molecular weight RNA selectively precipitated with 2 M lithium chloride was used for RT-PCR amplification of the coat protein encoding sequence by use of specific primers complementary to 5′ and 3′ parts of PPV coat protein L1 (GenBank accession no. X81081). Amino acid sequence comparison with GenBank data indicated 98.2% similarity with coat protein of PPV potyvirus isolated by E. Mais et al. (accession no. X81083) and 97.3% with PPV strain Rankovic (1).The specific DNA fragment, corresponding to predicted coat protein sequence size, was cloned into Escherichia coli pUC57 for DNA sequencing. Expression of the cloned sequence in bacteria and yeast expression systems is under investigation. The presence of PPV in plum trees in the 9-year-old collection at Babtai was confirmed by DAS-ELISA in 1997 and again in 1998. PPV was then detected in 20% of symptomatic trees of three cultivars. The Lithuanian PPV isolate reacted positively with “universal” Mab.5b and with a Mab (Mab.4DG5) specific for PPV-D. No reaction was observed with Mabs specific for PPV-M (Mab.AL), PPV-C (Mab.AC and Mab.TUV), and PPV-El Amar (Mab.EA24). PPV-971 seems to be a typical member of the less aggressive Dideron strain cluster of PPV (D. Boscia, personal communication). This is the first report of PPV in Lithuania and confirms the necessity for continuing the precautionary measures established in this country for indexing of nursery plum trees used for graft propagation. References: (1) S. Lain et al. Virus Res. 13:157, 1989. (2) J. Logemann et al. Anal. Biochem. 163:16, 1987. (3) M. Nemeth. OEPP/EPPO Bull. 24:525, 1994.

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