A SURVEY OF THE INCIDENCE OF POTATO SPINDLE TUBER VIROID IN PRINCE EDWARD ISLAND USING TWO TESTING METHODS

1988 ◽  
Vol 68 (4) ◽  
pp. 1229-1236 ◽  
Author(s):  
R. P. SINGH ◽  
T-L. DeHAAN ◽  
A. S. JASWAL
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Prince Edward Island ◽  
Potato Crop ◽  
Polyacrylamide Gel Electrophoresis ◽  
Leaf Disc ◽  
Potato Spindle Tuber Viroid ◽  
Polyacrylamide Gel ◽  
Nucleic Acid Hybridization ◽  
Dot Blot

Return-polyacrylamide gel electrophoresis (R-PAGE) and nucleic acid hybridization (dot-blot) were used for the detection of potato spindle tuber viroid (PSTV) from potato leaves. Both methods detected potato plants experimentally infected in the first season or those produced from infected tubers (secondarily infected). PSTV concentration was lower in the first-season infected plants than those in the second. Both methods detected PSTV in a single leaf disc from field-grown infected plants combined with 399 – 499 discs from field-grown healthy plants. The sensitivity of detection by R-PAGE was lower for certain cultivars and increased with the age of plants. About 85 000 leaf samples collected from 123 tablestock fields, 170 seed fields, and 63 cultivars from the Fox Island Elite Seed Farm in Prince Edward Island were found to be free from PSTV infection. Reasons for PSTV absence in the potato crop are discussed.Key words: Diagnosis, dot-blot, return polyacrylamide gel electrophoresis, nucleic acid hybridization, Solanum tuberosum, potato

scholarly journals Evidence for Association of a Viroid with Tapping Panel Dryness Syndrome of Rubber (Hevea brasiliensis)

Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1155-1155 ◽  
Author(s):  
P. Ramachandran ◽  
S. Mathur ◽  
L. Francis ◽  
A. Varma ◽  
J. Mathew ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Blot Hybridization ◽  
Potato Spindle Tuber Viroid ◽  
Low Molecular Weight ◽  
Total Nucleic Acid ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Low Salt ◽  
Tapping Panel Dryness

Tapping panel dryness (TPD) is one of the most destructive maladies affecting rubber plantations and is becoming a matter of serious concern. Reduced latex yield leading to total drying of the tapping panel is the obvious symptom. The cause of TPD syndrome is unknown but has been mostly attributed to abiotic causes. In India, the high yielding commercial clone RRII 105 is affected by TPD, leading to enormous losses. We have observed that TPD-affected trees show symptoms of bark scaling, cracking, drying, necrotic streaking, and browning of internal bark leading to the decay of internal tissues. Often prominent abnormal bulges on the lower part of tree trunks occur where the first panel begins to dry. Investigations on TPD-affected rubber samples did not reveal the association of fungus, bacterium, virus, or a protozoan. Total nucleic acid extracts purified from leaf and bark tissues of affected samples and analyzed by polyacrylamide gel electrophoresis under denaturing conditions of low salt and high temperature showed the presence of nucleic acids similar in electrophoretic mobility to low molecular weight (LMW) RNA, of ~359 nucleotides such as potato spindle tuber viroid (PSTVd). The LMW nucleic acid detected from TPD-affected samples was found to be RNA based on its sensitivity to RNase and insensitivity to DNase, phenol, and heat treatments. The LMW RNA was purified and cloned in a pUC 19-derived vector by using primers specific to PSTVd (1). The cloned DNA, when random labeled and used as probe reacted specifically to nucleic acid extracts from TPD-affected rubber trees but not from healthy tissue in dot-blot hybridization assays. Based on the above findings, a viroid etiology for TPD syndrome is proposed. Reference: (1) R. A. Owens, A. T. Candresse, and T. O. Diener. Virology 175:238, 1990.

Separation of potato spindle tuber viroid ribonucleic acid from Scopolia sinensis into three infectious forms and the purification and oligonucleotide pattern of fraction II RNA. Part IX

1976 ◽  
Vol 54 (7) ◽  
pp. 600-608 ◽  
Author(s):  
R. P. Singh ◽  
J. J. Michniewicz ◽  
S. A. Narang
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Cetyltrimethylammonium Bromide ◽  
Polyacrylamide Gel Electrophoresis ◽  
Potato Spindle Tuber Viroid ◽  
Ribonuclease A ◽  
Reverse Phase ◽  
Polynucleotide Kinase

The existence of three infectious forms of potato spindle tuber viroid (PSTV) RNA from Scopolia sinensis was demonstrated by fractionation with high salt, by reverse phase and high pressure liquid chromatography, and by polyacrylamide gel electrophoresis. Purification of fraction II was achieved by the following steps: extraction of nucleic acid with phenol, precipitation of the RNA with cetyltrimethylammonium bromide, fractionation of the RNA with lithium chloride and isopropanol, and finally gel electrophoresis. A procedure using reverse phase chromatography was developed to obtain 70–90% recovery of RNA from polyacrylamide gels. Purified PSTV fraction II RNA was digested with ribonuclease A and T1 and labelled with [γ-32P]ATP using polynucleotide kinase. The labelled digests were separated by the electrophoresis–homochromatography procedures of Sanger. About 20 and 30 spots were obtained with ribonuclease A and T1, respectively.

Ultrastructural localization of specific nucleoprotein fractions

1978 ◽  
Vol 36 (2) ◽  
pp. 204-205
Author(s):  
G. L. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultrastructural Localization ◽  
Polyacrylamide Gel ◽  
Acid Extraction ◽  
Acid Solutions ◽  
Low Salt ◽  
Liver Nuclei ◽  
Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

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