MOLECULAR CHARACTERIZATION OF TURNIP MOSAIC POTYVIRUS (TuMV) INFECTING RADISH (Raphanus sativus L.)
KrishiKosh
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Title |
MOLECULAR CHARACTERIZATION OF TURNIP MOSAIC POTYVIRUS (TuMV) INFECTING RADISH (Raphanus sativus L.)
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Creator |
PARMAR, NEHANJALI
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Contributor |
BHARDWAJ, S.V.
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Subject |
extraction, dna, fractionation, biological phenomena, livestock, alcohols, organochlorine compounds, area, planting, organic acid salts
Radish (Raphanus sativus L.), Turnip mosaic potyvirus |
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Description |
ABSTARCT Radish (Raphanus sativus L.) is an edible root crop of family Brassicaceae which is grown worldwide especially in Asia. Turnip mosaic potyvirus (TuMV; genus: Potyvirus, family: Potyviridae) is considered as one of the most important viruses in the world that infect field-grown vegetables and displays a large natural as well as experimental host range. TuMV was found to be prevalent in different regions of India. In all, nine isolates were collected on the basis of symptoms from areas comprising Himachal Pradesh (Mandi, Solan, Shimla and Kinnaur), Chandigarh, Punjab (Ludhiana), Haryana (Karnal), New Delhi (West Patel Nagar) and Rajasthan (Bharatpur). During serological detection, ELISA tests were conducted. All seven isolates except from New Delhi and Rajasthan reacted positively with monoclonal antibodies against TuMV. These studies were further confirmed through RT-PCR using specific primers for coat protein (CP) gene as a molecular detection procedure. A cDNA of approximately 1000 bp was amplified from all the seven TuMV Indian isolates. The RTPCR products were subsequently cloned and sequenced. The sequenced product of all the seven TuMV Indian isolates (IND1-IND7) was approximately 986 bp which comprised of 54 bp of the 3´ end of nuclear inclusion b (NIb) gene, the whole CP gene and 65 bp of the 3´ untranslated (UTR) region. CP gene of all the seven Indian isolates of TuMV was 867 bp long, encoding 288 amino acid residues which were submitted to NCBI. Accession numbers JQ246074 to JQ246080 and AFE55681 to AFE55687 were assigned to seven TuMV Indian isolates IND1 to IND7 CP gene nucleotide and amino acid sequences, respectively. Conserved motif DAG (Asp-Ala-Gly) and NAG (Asn-Ala-Gly), which has been reported to be important for potyvirus transmission by aphids, were found at positions 6-8 and 56-58 aa residues, respectively in the seven TuMV Indian isolates CP gene sequences. Another conserved motif, GDD (Gly-Asp-Asp) which has been identified as a hallmark of RNA dependent RNA polymerase was observed at 158-160 aa position in all the seven CP gene sequences of Indian isolates of TuMV. Percent homology of CP gene of seven Indian isolates among themselves and with other TuMV isolates retrieved from NCBI database was within the range of 87-99% and 92-100% at nucleotide and amino acid level, respectively. Phylogenetic analysis based upon nucleotide and amino acid sequences using UPGMA, NJ, MP and ME methods inferred classification of seven TuMV Indian isolates, tentatively into basal- BR group, due to its occurrence nearest to those isolates of TuMV which have been earlier classified to this group. Conserved domain for TuMV CP gene was observed at 51-287aa position in all the seven test Indian isolates. Computational predictions for various restriction enzymes were also carried out. Alpha helix consensus secondary structure was predicted and found to dominate in all the seven protein sequences of CP gene of Indian isolates of TuMV |
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Date |
2016-06-17T14:43:24Z
2016-06-17T14:43:24Z 2012 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/67547
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Language |
en
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Format |
application/pdf
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