A Partial Characterization of Acyrthosiphon Pisum (Harris) Strains Efficient and Inefficient in the Transmission of Pea Enation Mosaic Virus PDF Download

Author: Emmanuel Chukwuemeka Igbokwe
Publisher:
ISBN:
Category : Aphids
Languages : en
Pages : 140

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Author: J. Victor French
Publisher:
ISBN:
Category : Mosaic diseases
Languages : en
Pages : 186

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Author: James Hsi-cho Tsai
Publisher:
ISBN:
Category : Beetles
Languages : en
Pages : 148

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Author: Lowell Raymond Nault
Publisher:
ISBN:
Category : Mosaic diseases
Languages : en
Pages : 116

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Author: James Hsi-cho Tsai
Publisher:
ISBN:
Category : Pea aphid
Languages : en
Pages : 138

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Author: Kerry F. Harris
Publisher:
ISBN:
Category : Virus diseases of plants
Languages : en
Pages : 320

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Author: Paul D. Kohnen
Publisher:
ISBN:
Category : Peas
Languages : en
Pages : 192

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Book Description
Biological, serological, and molecular characteristics of the P4 pathotype of pea seedborne mosaic potyvirus (PSbMV) were investigated. The characterization allowed P4 to be differentiated from other pathotypes of the same virus. P4 was distinguished from P1 on the basis of host symptomatology. It induced varied, but often distinctive symptoms in systemically-infected pea (Pisum sativum) cultivars. P4 inoculation of indicator hosts, notably Chenopodium album, resulted in the formation of local lesions. Pathotype P4 was seed transmitted at low frequencies relative to P1. In ten selected pea cultivars, seed transmission of P4 never exceeded 0.7%, while P1 was transmitted at frequencies as high as 33%. In mixed infections with P1, pathotype P4 was seed transmitted at 0- 2% as determined by P4-specific indirect ELISA. However, limited polymerase chain reaction (PCR) data suggest that P4 may be transmitted with P1 at higher rates than indicated by ELISA data. P4 was also aphid transmitted at relatively low rates. Pea aphids (Acyrthosiphon pisum) allowed 3 or 5 min acquisition access periods (AAP's) transmitted P4 at a maximum rate of 16% (3 aphids/plant). P4 transmission required short (1-7 min) AAP's and exhibited no bimodal character within tested parameters. P4 could be serologically distinguished from the other pathotypes using monoclonal and polyclonal antibodies. Tests with cross-reactive P4 antiserum suggest that P4 is more closely related to P2 than P1. P4-specific antiserum was derived by cross-absorbing diluted antiserum with P1- infected pea sap to eliminate P1- reactive antibodies. P1 and P4 RNA was detected and differentiated using pathotype-specific oligonucleotide primers in conjunction with PCR. Viral RNA sequences were amplified in pure preparations as well as crude pea tissue (leaf, root, pollen, and seed) extracts.

Author: Ya-Chu Judy Kao
Publisher:
ISBN:
Category : Pea aphid
Languages : en
Pages : 258

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Author: Wa-lee Lim
Publisher:
ISBN:
Category : Peas
Languages : en
Pages : 300

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Author: Jody Jellison Goodell
Publisher:
ISBN:
Category : Lentils
Languages : en
Pages : 250

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Book Description
The lentil strain of pea seedborne mosaic virus, PSbMV-L, was distinguished from the standard isolates of PSbMV by a natural inoculum reservoir in lentil seed and by separate resistance genes in Pisum and Lens. PSbMV-L was intermediate in symptom severity between the standard and New Zealand isolates of PSbMV. Seed transmission rates up to 16.7% were observed for PSbMV-L in seed increase plots of infected lentil accessions, but the virus was not detected in commercial lentil plantings. The virus was nonpersistently transmitted by Acyrthosiphon pisum. Maximum rates of aphid transmission occurred with acquisition access periods of 3 to 5 minutes and ranged from 80 to 100%. Aphid transmission rates decreased with increasing temperature. Apparent trimodal transmission of PSbMV-L was observed. PSbMV-L is characterized by intrinsic particle instability. Partial purification was obtained by extracting infected lentil leaves in a neutral buffer, clarification with 7.5% chloroform, centrifugation through PEG (6 %)- sucrose (35%) cushions and equilibrium centrifugation in CsC1 gradients. The absorbance ratio 260/280 was 1.15. PSbMV-SL25 and PSbMV-L were indistinguishable on the basis of protein subunit molecular weight and could not be differentiated using classical serological techniques. The strains could be separated by quantitative ELISA using polyclonal sera produced in rabbit or chicken. Attempts to achieve greater antibody specificity were made with spleen cells from previously immunized mice that were fused with myoloma cells to create antibody-secreting hybridomas. The resultant colonies were screened using a modified ELISA against PSbMV-SL25 and PSbMV-L for the presence of specific antibody production. Selected colonies were expanded. The molecular weight of PSbMV-L RNA was estimated at 3.5 million. DNA complementary to PSbMV-C4 RNA was prepared using the random primer method of Taylor et al. Hybridization kinetics with PSbMV-L, PSbMV-SL25 and BYMV-Gil RNA were studied using Sl-nuclease assays. The lentil strain showed significantly lower levels of hybridization than the homologous control. Heterologous hybridization was severely decreased when the stringency of the assay conditions was increased.