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.

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

Translocation and rotation of microtubules caused by multiple species of Chlamydomonas inner-arm dynein

1992 ◽  
Vol 103 (3) ◽  
pp. 653-664 ◽  
Author(s):  
O. Kagami ◽  
R. Kamiya
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Light Chains ◽  
Maximal Rate ◽  
Heavy Chains ◽  
Torque Generation ◽  
Multiple Species ◽  
Monoq Column

Dynein was extracted from outer arm-less axonemes of the mutant oda1 and fractionated by high-pressure liquid chromatography on a MonoQ column into seven distinct subspecies (named a-g). Each subspecies contained one or two heavy chains and several medium-sized and light chains; by vanadate/UV-induced photocleavage and SDS- polyacrylamide gel electrophoresis, eight distinct heavy chains were identified. Analysis of the mutant axonemes indicated that the subspecies f (containing two heavy chains) is missing in the inner-arm mutant ida1 and the subspecies a, c and d are missing in the mutant ida4. Six subspecies (all but f) supported microtubule translocation with the maximal rate ranging from 2 to 12 micrometre s-1 and the apparent Km for ATP ranging from about 10 to 100 micromolar. All the subspecies translocated microtubules with the plus end leading, indicating that all the inner-arm dyneins are minus end-directed motors. Five subspecies (all but b and f) displayed microtubule rotation during translocation at rates of up to about 10 Hz. Unexpectedly, the Km values for ATP for translocation and rotation did not always agree; because of this, the pitch of the movement was variable with some subspecies. These observations indicate that axonemes are equipped with several inner-arm subspecies and that torque generation is a feature common to many of them.

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.

Screening Method for Vitamins A and D in Fortified Skim Milk, Chocolate Milk, and Vitamin D Liquid Concentrates

1979 ◽  
Vol 62 (6) ◽  
pp. 1358-1360
Author(s):  
Susan K Henderson ◽  
Lucia A Mclean
Keyword(s):  
Vitamin D ◽  
High Pressure ◽  
Liquid Chromatography ◽  
Vitamin A ◽  
Mobile Phase ◽  
Screening Method ◽  
Skim Milk ◽  
Chocolate Milk ◽  
Reverse Phase ◽  
Milk Chocolate

Abstract Vitamin A was determined in fortified chocolate milk and skim milk; vitamin D was determined in fortified chocolate milk, skim milk, and vitamin D concentrates, using reverse phase high pressure liquid chromatography (HPLC). The sample is saponified, extracted with hexane, and chromatographed in an HPLC system on a 10 μm Vydac TP reverse phase C18 column, using acetonitrile-methanol (9+1) as the mobile phase. For 6 replicates, the recoveries of vitamins A and D, using this procedure, were 99 and 98%, respectively.

The Determination of Urinary Oxypurines as Markers of Gastrointestinal Tumors

1987 ◽  
Vol 73 (3) ◽  
pp. 289-294 ◽  
Author(s):  
Marco Lorenzi ◽  
Daniela Vannoni ◽  
Roberto Leoncini ◽  
Ranieri Caldarone ◽  
Enrico Marinello
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Urinary Excretion ◽  
Tumor Marker ◽  
Cancer Patients ◽  
Plasma Levels ◽  
Colorectal Tumor ◽  
Reverse Phase ◽  
Tumor Bearing

Plasma levels and urinary excretion of oxypurines – hypoxanthine and xanthine – were evaluated by reverse-phase high-pressure liquid chromatography in 13 patients affected by gastric tumors and in 19 colorectal tumor-bearing patients. Preliminary results indicate higher values of urinary xanthine and an increase in the xanthine/hypoxanthine ratio in cancer patients. The increase was not generalized to all subjects, and did not appear related either to the stage of the disease or to CEA values. The limits within which the determination of urinary oxypurines can be employed as a tumor marker are discussed.

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