Characterization of Chickpea Chlorotic Dwarf Virus (CpCDV) Associated with Chickpea Stunt Disease

2021 ◽  
Author(s):  
L. Manjunatha ◽  
N. Srinivasa ◽  
T. Basavaraja ◽  
M.C. Keerthi
Keyword(s):  
Coat Protein ◽  
Infected Plant ◽  
Pcr Amplification ◽  
Uttar Pradesh ◽  
Coat Protein Sequence ◽  
Causal Agent ◽  
Dwarf Virus ◽  
Limiting Factors ◽  
Specific Pcr ◽  
Yellow Orange

Background: Stunt disease is becoming the major yield limiting factors for the chickpea production and its occurrence has been reported form different states of India. The symptoms of stunt disease caused by chickpea chlorotic dwarf virus are difficult to distinguish Mastrevirus-infected plant from other disease-causing pathogens. Therefore, it’s an imperative for precise detection of causal agent of the disease for development of management strategy against chickpea stunt.Methods: Survey for the incidence of stunt disease with most characteristic symptoms of leaf reddening and yellow orange typical to Mastrevirus infection was conducted in chickpea fields. The causal agent of the stunt was characterized and described through conventional and virus-specific PCR-based diagnostic technique.Result: The study revealed that maximum of 60% of the chickpea stunt was observed in three districts of Uttar Pradesh with an average incidence of 12.90%. The PCR amplification using CpCDV-specific primers encoding coat protein resulted in an expected amplicon size of 350bp. The comparison of the partial coat protein sequence of virus revealed that maximum homology of 98.70% with previously identified chickpea chlorotic dwarf virus (CpCDV) strains, indicating that CpCDV associated with the chickpea stunt. Based on molecular characterization, chickpea stunt disease caused by Chickpea chlorotic dwarf virus (ssDNA), belongs to the genus Mastrevirus which is also responsible for the lentil stunt disease.

scholarly journals RAPDs are Useful for Genetic Analysis of Xylella fastidiosa and for Development of Strain-specific PCR Primers

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 783D-783
Author(s):  
Margaret R. Pooler ◽  
John S. Hartung
Keyword(s):  
Xylella Fastidiosa ◽  
Genetic Relationships ◽  
Pcr Amplification ◽  
Pcr Primers ◽  
Causal Agent ◽  
Plant Diseases ◽  
Citrus Variegated Chlorosis ◽  
Specific Pcr ◽  
Horticultural Plant ◽  
Similarity Relationships

Xylella fastidiosa is a fastidious gram-negative, xylem-limited leafhopper-transmitted bacterium that has proven to be the causal agent of many economically important horticultural plant diseases, including Pierce's disease of grapevine and citrus variegated chlorosis. Genetic relationships among 11 X. fastidiosa strains isolated from mulberry, almond, ragweed, grape, plum, elm, and citrus were determined using randomized amplified polymorphic DNA (RAPD). Twenty-two 10-base primers amplified a total of 77 discrete polymorphic bands. Phenetic analysis based on a similarity matrix corresponded well with previous reports on RFLP-based similarity relationships, indicating that RAPD-PCR amplification products can be used as a reliable indicator of genetic distance in X. fastidiosa. RAPD products have been cloned and sequenced, and pairs of 21-nucleotide PCR primers have been developed that detect X. fastidiosa in general and the causal agent of citrus variegated chlorosis specifically.

Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

1996 ◽  
Vol 75 (05) ◽  
pp. 757-759 ◽  
Author(s):  
Rainer Blasczyk ◽  
Markus Ritter ◽  
Christian Thiede ◽  
Jenny Wehling ◽  
Günter Hintz ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Pcr Amplification ◽  
Factor V ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Specific Amplification ◽  
Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

scholarly journals Molecular characterization of chickpea chlorotic dwarf virus and peanut witches’ broom phytoplasma associated with chickpea stunt disease and identification of new host crops and leafhopper vectors in India

3 Biotech ◽  
2021 ◽  
Vol 11 (3) ◽  
Author(s):  
Madem Gurivi Reddy ◽  
Virendra Kumar Baranwal ◽  
Doddachowdappa Sagar ◽  
Govind Pratap Rao
Keyword(s):  
Sequence Comparison ◽  
Disease Incidence ◽  
Uttar Pradesh ◽  
Rflp Analysis ◽  
Dwarf Virus ◽  
Rrna Gene ◽  
New Delhi ◽  
New Host ◽  
Severe Stunting ◽  
Virtual Rflp

AbstractAn investigation was carried out to identify and characterize the phytoplasma and viruses associated with the chickpea varieties showing severe stunting, leaf reddening, yellowing and phyllody symptoms during the summer season of 2018–2019 and 2019–2020 in eight states of India. The average disease incidence was recorded from 3 to 32% in different states. The presence of chickpea chlorotic dwarf virus (CpCDV) was confirmed in thirty-seven chickpea samples by amplification of CpCDV coat protein gene and sequence comparison analysis. No record of association of luteovirus, polerovirus and cucumovirus could be detected in any of the symptomatic chickpea samples by RT-PCR assay. Brassica nigra, B. juncea, Lens culinaris, two weeds (Heteropogan contartus, Aeschynomene virginica) and one leafhopper (Amarasca biguttula) were identified as new putative hosts for CpCDV. Association of peanut witches’ broom phytoplasma was confirmed in twenty-eight chickpea samples, Sesamum indicum, five weeds hosts and two leafhopper species (Exitianus indicus, Empoasca motti) using nested PCR assays with primer pairs P1/P7 and R16F2n/R16Rn. The results of phytoplasma association in plants and leafhopper samples were further validated by using five multilocus genes (secA, rp, imp, tuf and secY) specific primers. Sequence comparison, phylogenetic and virtual RFLP analysis of 16S rRNA gene and five multilocus genes confirmed the identity of association of 16SrII-C and 16SrII-D subgroups of phytoplasmas strain with chickpea samples collected from Andhra Pradesh (AP), Telangana, Karnataka, Madhya Pradesh, Uttar Pradesh and New Delhi. Mixed infection of phytoplasma (16SrII-D) and CpCDV was also detected in symptomatic chickpea samples from AP and Telangana. The reports of association of 16SrII-C subgroup phytoplasma in chickpea and 16SrII-D subgroup phytoplasma in C. sparsiflora and C. roseus are the new host records in world and from India, respectively.

scholarly journals First Report of Raspberry bushy dwarf virus in Rubus multibracteatus from China

Plant Disease ◽  
2003 ◽  
Vol 87 (5) ◽  
pp. 603-603 ◽  
Author(s):  
C. J. Chamberlain ◽  
J. Kraus ◽  
P. D. Kohnen ◽  
C. E. Finn ◽  
R. R. Martin
Keyword(s):  
Amino Acid ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Dwarf Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Black Raspberry ◽  
First Report ◽  
Raspberry Bushy Dwarf Virus ◽  
Plant Pathol

Raspberry bushy dwarf virus (RBDV), genus Idaeovirus, has been reported in commercial Rubus spp. from North and South America, Europe, Australia, New Zealand, and South Africa. Infection can cause reduced vigor and drupelet abortion leading to crumbly fruit and reduced yields (3,4). In recent years, Rubus germplasm in the form of seed, was obtained on several collection trips to The People's Republic of China to increase the diversity of Rubus spp. in the USDA-ARS National Clonal Germplasm Repository, (Corvallis, OR). Before planting in the field, seedlings were tested for the presence of RBDV, Tomato ringspot virus, and Tobacco streak virus using triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) (antiserum produced by R. R. Martin). One symptomless plant of R. multibracteatus H. Lev. & Vaniot (PI 618457 in USDA-ARS GRIN database), from Guizhou province in China, tested positive for RBDV (RBDV-China). After mechanical transmission on Chenopodium quinoa Willd., this isolate produced typical symptoms of RBDV (3). To determine if RBDV-China was a contaminant during the handling of the plants, or if the source was a seedborne virus, the coat protein gene was sequenced and compared to published sequences of RBDV. RNA was extracted from leaves of R. multibracteatus and subjected to reverse transcription-polymerase chain reaction (RT-PCR) using primers that flank the coat protein gene. Products from four separate PCR reactions were sequenced directly or were cloned into the plasmid vector pCR 2.1 (Invitrogen, Carlsbad, CA) and then sequenced. The coding sequence of the coat protein gene of RBDV-China was 87.5% (722/825) identical to that isolated from black raspberry (Genbank Accession No. s55890). The predicted amino acid sequences were 91.6% (251/274) identical. Previously, a maximum of five amino acid differences had been observed in the coat proteins of different RBDV strains (1). The 23 differences observed between RBDV-China and the isolate from black raspberry (s55890) confirm that the RBDV in R. multibracteatus is not a greenhouse contaminant but is indeed a unique strain of RBDV. In addition, monoclonal antibodies (MAbs) to RBDV (2) were tested against RBDV-China. In these tests, MAb D1 did not detect RBDV-China, whereas MAb R2 and R5 were able to detect the strain. This is the first strain of RBDV that has been clearly differentiated by MAbs using standard TAS-ELISA tests. Although RBDV is common in commercial Rubus spp. worldwide, to our knowledge, this is the first report of RBDV in R. multibracteatus, and the first report of RBDV from China. The effects of this new strain of RBDV could be more or less severe, or have a different host range than previously studied strains. It is more divergent from the type isolate than any other strain that has been studied to date. Phylogenetic analysis of coat protein genes of RBDV may be useful in understanding the evolution and spread of this virus. References: (1) A. T. Jones et al. Eur. J. Plant Pathol. 106:623, 2000. (2) R. R. Martin. Can. J. Plant. Pathol. 6:264, 1984. (3) A. F. Murant. Raspberry Bushy Dwarf. Page 229 in: Virus Diseases of Small Fruits. R. H. Converse, ed. U.S. Dep. Agric. Agric. Handb. 631, 1987. (4) B. Strik and R. R. Martin. Plant Dis. 87:294, 2003.

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.

Onion yellow dwarf virus. [Distribution map].

2018 ◽  
Author(s):  
Keyword(s):  
New York ◽  
Far East ◽  
Uttar Pradesh ◽  
Yellow Dwarf ◽  
Dwarf Virus ◽  
Onion Yellow Dwarf Virus ◽  
Distribution Map ◽  
Yellow Dwarf Virus ◽  
Virus Distribution ◽  
Distribution In Europe

Abstract A new distribution map is provided for Onion yellow dwarf virus. Potyviridae: Potyvirus. Hosts: onion (Allium cepa) and garlic (Allium sativum). Information is given on the geographical distribution in Europe (Austria, Croatia, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Italy, mainland Italy, Sicily, Lithuania, Moldova, Netherlands, Poland, Portugal, Romania, Russia, Far East, Siberia, Serbia, Slovenia, Spain, UK, England and Wales and Ukraine), Asia (China, Henan, Hubei, Hunan, Jiangsu, Shandong, Yunnan, Zhejiang, India, Delhi, Gujarat, Haryana, Madhya Pradesh, Maharashtra, Indian Punjab, Rajasthan, Uttar Pradesh, Indonesia, Java, Iran, Israel, Japan, Pakistan, Taiwan, Turkey, Vietnam and Yemen), Africa (Egypt, Morocco, Nigeria and Sudan), North America (Canada, British Columbia, Nova Scotia, Ontario, Mexico, USA, California, Colorado, Florida, Idaho, Iowa, Minnesota, Nevada, New York, Oregon, Texas, Vermont, Washington and West Virginia), Central America and Caribbean (Cuba), South America (Argentina, Brazil, Bahia, Goias, Minas Gerais, Parana, Rio Grande do Sul, Santa Catarina, Sao Paulo, Chile, Ecuador and Urugay) and Oceania (Australia and New Zealand).

scholarly journals Identification and Characterization of Janthinobacterium svalbardensis F19, a Novel Low-C/N-Tolerant Denitrifying Bacterium

2019 ◽  
Vol 9 (9) ◽  
pp. 1937 ◽  
Author(s):  
Yinyan Chen ◽  
Peng Jin ◽  
Zhiwen Cui ◽  
Tao Xu ◽  
Ruojin Zhao ◽  
...  
Keyword(s):  
Nitrogen Removal ◽  
Removal Rate ◽  
Pcr Amplification ◽  
Domestic Wastewater ◽  
Heterotrophic Nitrification ◽  
Aerobic Denitrification ◽  
Nitrite Accumulation ◽  
Specific Pcr ◽  
Domestic Wastewater Treatment ◽  
Initial Ph

Herein, we isolated Janthinobacterium svalbardensis F19 from sludge sediment. Strain F19 can simultaneously execute heterotrophic nitrification and aerobic denitrification under aerobic conditions. The organism exhibited efficient nitrogen removal at a C/N ratio of 2:1, with an average removal rate of 0.88 mg/L/h, without nitrite accumulation. At a C/N ratio of 2, an initial pH of 10.0, a culturing temperature of 25 °C, and sodium acetate as the carbon source, the removal efficiencies of ammonium, nitrate, nitrite, and hydroxylamine were 96.44%, 92.32%, 97.46%, and 96.69%, respectively. The maximum removal rates for domestic wastewater treatment for ammonia and total nitrogen were 98.22% and 92.49%, respectively. Gene-specific PCR amplification further confirmed the presence of napA, hao, and nirS genes, which may contribute to the heterotrophic nitrification and aerobic denitrification capacity of strain F19. These results indicate that this bacterium has potential for efficient nitrogen removal at low C/N ratios from domestic wastewater.

scholarly journals Detection of maternal cells in human fetal blood during the third trimester of pregnancy using allele‐specific PCR amplification

1997 ◽  
Vol 98 (3) ◽  
pp. 767-771 ◽  
Author(s):  
THIERRY PETIT ◽  
MARC DOMMERGUES ◽  
GÉRARD SOCIÉ ◽  
YVES DUMEZ ◽  
ELIANE GLUCKMAN ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Fetal Blood ◽  
Third Trimester ◽  
The Third ◽  
Specific Pcr ◽  
Allele Specific ◽  
Allele Specific Pcr
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