Eradication of potato virus X from potato by ribavirin treatment of cultured potato shoot tips

Ribavirin treatment of cultured potato shoot tips was tested as a means of eradicating potato virus X (PVX) from two potato cultivars. Shoot tips were cultured on liquid medium containing 0, 1, 10 or 100μg/ml ribavirin. Cultures were evaluated periodically for viability, and scored for vigor on a relative growth scale. Developed plantlets were assayed for PVX by transmission tests toGomphrena globosa. Ribavirin treatment was phytotoxic at all tested concentrations, and lethal to all cultivars treated at 100 μg/ml. Treatment also delayed plantlet development by up to 2 months at 1 and 10 μg/ml as compared with nontreated controls. PVX assays indicated that 80 and 83% of the plantlets were free of PVX following treatment with 10 μg/ml for cultivars Russet Burbank and Red McClure, respectively. Five and 6% of the plantlets developed from the 1 μg/ml treatment were PVX-free, whereas 0 and 2% of the controls were PVX-free for the same cultivars. Six to 8 months were required to develop plants from shoot-tip cultures treated with 10 μg/ml ribavirin.     

Reinfection of potato seed stocks with potato virus S and potato virus X in Wisconsin

Reinfection of potato seed stocks with the potato viruses S (PVS) and X (PVX) varied with cultivar, virus, and grower. Rapid recontamination was observed for the cultivars Norgold Russet and Ontario with PVS and for the cultivars LaChipper, Norchip, and Norgold Russet with PVX. Recontamination was low for the cultivars LaChipper and Monona with PVS and for the cultivars Kennebec, Monona, Norland, and Superior with PVX. Survey results suggest that PVS and/or PVX can be eliminated from cultivars which appear to possess field resistance to infection, but that further evaluation of cultivars which are very susceptible to reinfection will be necessary.     

Enzyme-linked immunosorbent assay with single or combined antisera for viruses S and X in potato tubers and plants

Enzyme-linked immunosorbent assay (ELISA), with single or combined antisera was effective for diagnosing potato virus S (PVS), potato virus X (PVX) or both viruses in plants grown in the greenhouse or field. In dormant tubers, stolon, middle and apical end composite sampling with or without eyes and sprouted tubers produced reliable positive assays for PVX. Only tuber pieces with sprouts resulted in consistently reliable assays for PVS. Composite sampling of potato foliage was effective for detecting one PVX infected plant in a total of 100 Kennebec, Norland, or Russet Burbank plants. There were some false negative results and greater variability in composite PVS assays, but on average, one PVS infected plant can be detected in composites of 10 Kennebec, Norland, or Russet Burbank plants. Sodium diethyldithiocarbamate (0.01M NaDIECA) in phosphate buffered saline + 0.5% Tween (PBS-T) added to plant extracts enhanced specific reactions for either virus. Onceor twiceused enzyme conjugate was effective in ELISA of either virus from potato foliage.     

A Reexamination of the Effectiveness of Ribavirin on Eradication of Viruses in Potato Plantlets in vitro Using ELISA and Quantitative RT-PCR

The potato plantlets singly infected by PVA, PLRV, PVS, PVX and PVY and mix-infected by PVM, PVS and PVY were cultured on MS medium with different concentration of ribavirin. The effects of ribavirin on growth of the plantlets and efficiency of virus elimination were investigated. Results showed that the plant height and fresh weight obviously decreased with increase of ribavirin concentration from 0 mg/L to 150 mg/L, and most of the plantlets could not survive when the concentration reached 200 mg/L. According to the ELISA tests, ribavirin was more efficient for eradicating PVA, PVM, PVS and PVX than PVY and PLRV, and healthy plantlets could be obtained with high frequency (up to 100 %) by culturing with 75?~?150 mg/L ribavirin after 2?~?3 subcultures. Whereas, only 33?~?66 % PVY and PLRV infected plantlets were found to be virus-free after 3 subcultures with 75?~?150 mg/L ribavirin. The results of quantitative RT-PCR (qPCR) indicated that ribavirin could obviously reduce virus content in the plantlets. Except PLRV was detected positive after 3 subcultures with ribavirin, the healthy seedlings were obtained from infected stocks at the first or end of propagation and no viruses could be detected at the post-eradication stage. No apparent difference of genetic variation resulted from ribavirin treatment was found by SSR analysis between the control and the treated plantlets. All of these results above proved that ribavirin treatment in vitro was an effective method to eliminate viruses in the propagation of potato.     

Length of dormancy and sprouting characteristics of ten potato cultivars

The length of dormancy and sprouting characteristics of ten cultivars (Bison, Kennebec, Norchip, Norgold Russet, Norland, Red Pontiac, Russet Burbank, Viking, Dakchip, and ND8891-3), were compared. Russet Burbank had the longest dormancy period and the slowest rate of sprouting after dormancy ended. Viking and Norgold Russet had the most sprouts and Kennebec the fewest. Dakchip had the shortest dormancy period, the greatest weight of sprouts and the greatest weight loss. Dormancy was shortest in the warmest (20°C) storage. Storage at 2°C for six to nine weeks increased sprouting after dormancy ended compared with continuous 10 or 20°C storage.     

Daniel-Navan: A white-skinned,medium-late potato cultivar suitable for regions with a hot climate

Daniel-Navan is a new potato cultivar suitable for the spring crop (between February and July) in the Negev region of Israel, and having a high yielding potential with excellent tuber appearance. The cultivar is resistant to heat stress, Verticillium wilt (V. dahliae) and early blight (Alternaria solani). Tubers of Daniel-Navan are oval, white-skinned, with creamy-white flesh, free of deformities, and growth cracks. It has a high dry matter content (22.7%) and boils and bakes well. The marketable yield of Daniel-Navan obtained at maturity consistently surpasses that of Desiree, the control. Daniel-Navan is a main crop cultivar bred and selected by the Horticultural and Plant Breeding Station, Northern Ireland, and the Agricultural Research Organization, Gilat Experiment Station, Israel. Owing to its high yielding performance and tuber quality, this cultivar could be suitable for use in regions with a hot climate.     

Tertiary treatment of potato processing waste by biological activated carbon process

A biological activated carbon process was investigated for its feasibility as a tertiary treatment process for potato processing wastewaters. Bench-scale continuous complete-mix activated sludge reactors and biological activated carbon units were operated to evaluate the removal kinetics of organic materials and to determine the design parameters for a full-scale treatment unit. Raw wastewater contained 4300 mg/1, which was reduced to 130 to 340 mg/1 following the complete-mix activated sludge treatment and to 30 to 90 mg/1 after the biological activated carbon treatment.     

Objective methods for testing potato flake quality as influenced by free starch and surfactants

A method was developed which uses changes in rates of retrogradation measured by a Brabender Amylograph to assess the complexing ability of surfactants for free starch in potato flakes during cooling. The rate of retrogradation was linearly related to surfactant concentration. A Blue Value Index method, adapted from use with cereal flours for determination of starch damage, was used to monitor the amount of free starch created by the required particle size reduction for Amylograph analysis. The change in Blue Value Index was also used to monitor the ability of surfactants to complex free starch and thereby influence potato flake quality. Distilled monoglyceride appeared to be more effective than sodiumor calcium stearoyl–2-lactylate. However, the combination of 0.3% distilled monoglyceride with 0.2% sodium stearoyl–2-lactylate was nearly as effective as 0.5% distilled monoglyceride in terms of free starch complexing ability. Instron back extrusion tests for mealiness of reconstituted flakes were found to be influenced by the level of free starch; therefore, back extrusion data should be accompanied by Blue Value Index data.     

Use of enzyme-linked immunosorbent assay to detect potato leafroll virus in field grown potato,cv. Russet Burbank

Use of enzyme-linked immunosorbent assay (ELISA) in detecting potato leafroll infections in field grown potato, cv. Russet Burbank, was studied from 1986 to 1988 at Rosemount, Minnesota. The objective was to determine relative reliability of current season foliage ELISA, tuber tissue ELISA, and tuber progeny foliage ELISA. Serological tests were most accurate when foliage of tuber progenies was tested. ELISA underestimated total leafroll infection when current season foliage from the inoculated plant was used, in those plants inoculated during late tuber bulking stage. Current season foliage ELISA tests using newly expanded terminal leaflets were more reliable than were tests using older leaflets. Leafroll infection was detected in the current season foliage and tuber progenies (tuber tissue as well as tuber progeny foliage) of some plants seven days after inoculation. Most current season foliage infections were detected by day 14–28 depending on year. Differences among years were most likely caused by variation in quality of virus source plants and numbers of vectors used in inoculation. ELISA tests on tuber tissue were almost as effective as ELISA tests on tuber progeny foliage in detecting potato leafroll 20 days after inoculation, but ELISA on tuber tissue substantially underestimated infection if plants were sampled earlier. Maximum percent tuber infection occurred 20 days or more after inoculation. Movement of the virus from the inoculated stem to other stems decreased with increased plant age at inoculation. Percent infected tubers declined with increased plant age at inoculation. Action thresholds developed for aphids in managing potato leafroll virus should take into account the temporal change in percent infected tubers.     

Changes in the nutrient composition of potatoes during home preparation: III. Minerals

The effect of several cooking methods on the mineral content of three major U.S. potato varieties was assessed. Significant losses of minerals in potato flesh were minimal and were unaccountably associated with the Norchip variety grown in the Red River Valley. The Irish potato was found to contribute significantly to the U.S. Recommended Dietary Allowance for copper, iron, magnesium, and phosphorus in the diet of man.     

The Use of Mineral Oil in Potato Protection: Dynamics in the Plant and Effect on Potato Virus Y Spread

The focus of this study was to evaluate two mineral oils (Superior 70 and Vazyl-Y) in reducing the seasonal spread of Potato Virus Y (PVY). Three concentrations of oil (0, 5, and 10 L ha^?1 of Superior 70; 0, 7.5, and 15 L ha^?1 of Vazyl-Y) and three spray regimes for both oils (every 3–4, 7, and 10–11 days) were tested. Two weeks after top-kill, two tubers from each of 49 plants free of virus at emergence were harvested from treatment plots, sprouted, and tested for PVY with enzyme-linked immunosorbent assay (ELISA). Results revealed that in the case of Superior 70, PVY spread in mineral oil treated plots ranged from 2.1 to 12.2 %, while in the control plots it ranged from 20.4 to 37.7 % across three cultivars. In the case of Vazyl-Y, PVY spread in mineral oil treated plots ranged from 2.1 to 26.5 %, while in the control plots it ranged from 49.9 to 85.7 % across three cultivars. These data show that there was a significant reduction in PVY due to spray of mineral oils. In addition, mineral oil was quantified in plants from the Superior 70 treated and the control plots to understand the dynamics of mineral oil during the season. While there was little to no oil measured in the leaves at the early stages of plant growth, a considerable amount of mineral oil was detected close to plant maturity. A basic model of the concentration of oil in the treated foliage was formulated to confirm our understanding of the factors at play. The model could explain from 50 to 90 % of the variation in oil content observed in the field. Plant growth and size are important factors affecting oil content in mineral oil treated foliage.     

Relationship of virus concentration with the field resistance to potato virus Y in potatoes

The concentration of potato virus Y (PVY) was determined, using ELISA values (A405 nm), in twenty-six potato cultivars belonging to five resistance groups, grown in the field and in the greenhouse. On the basis of virus concentration, potato cultivars of group A, B, and C did not differ significantly and constitute the most susceptible group; those of group D and E differ significantly with each other and with group A, B, C, and constitute moderate and highly resistant groups, respectively. In the second year of infection, virus concentration was higher in each group, irrespective of resistance level. Thus, the infected plants of resistant groups, in a second year of growth, could be as rich sources of virus to aphids as plants from susceptible groups.     

A rapid and simple method for determination of potato chloroplast DNA type

A procedure for restriction enzyme analysis of the potato chloroplast DNA (ctDNA) is described. The advantages of this method are: 1) rapid determination of ctDNA type, 2) no ultracentrifugation, and 3) low cost of analyses. This method makes it easy to distinguish the ctDNA types of wild and cultivated potato accessions.     

A method for large scale laboratory inoculation of potato tubers with late blight fungus

As part of a project to investigate the possibility that teratogenic factors are formed in potato tubers infected with the late blight fungus,Phytophthora infestans, a method was devised which allowed quick, efficient inoculation of large numbers of potato tubers. Tubers were placed in specially constructed trays, wounded by puncturing the periderm with flower “pin frogs”, and sprayed with a zoospore suspensionof P. infestans prepared from cultures grown on rye agar at 20C (68F) for 10 days. Approximately 95% of Katahdin and 93% of Russet Sebago tubers inoculated by this method exhibited typical late blight lesions after two weeks incubation at 20C, 80–90% RH. Eight tons of potatoes were processed in this manner.     

Detection of potato viruses X and S in tissue culture plantlets

The latex agglutination test (LAT) and enzyme-linked immunosorbent assay (ELISA) were evaluated in separate studies for their ability to detect potato virus X (PVX) and potato virus S (PVS) in tissue culture plantlets. Healthy and infected clonal lines of several potato cultivars were used. LAT was unsatisfactory because only low levels of agglutination were obtained with infected samples, and because variable and inconsistent results were obtained with both healthy and infected clones. ELISA, however, consistently gave high spectrophotometric readings and intense visual reactions for infected but not for healthy clones. The results indicate that ELISA can be used to detect PVX and PVS in tissue culture plantlets, and in programs where tissue culture is employed, early detection and elimination of infected plantlets is possible.     

Kinetin,thermotherapy, and tissue culture to eliminate potato virus (PVX) in potato

PVX infected plantlets from two potato cultivars grownin vitro with 0.3, 3, 30 or 300 ppm kinetin were exposed to temperatures of 28 or 35 C. After 3 wk, axillary buds were isolated and grown aseptically in organogenic media, followed by PVX testing by ELISA. The serological test was also run on whole plantlets at the end of the kinetin-temperature exposure. No donor plants exposed to 28 C nor the plantlets derived from their buds gave an ELISA (-) reaction, regardless of the kinetin content of the media or that of the cultivar. At 35 C the virus was suppressed to undetectable levels in several whole plantlets. In the cultivar Alpha, 2 out of 6 resulting plantlets after isolation of buds were virus-free in the presence of 3.0 mg/1 kinetin during the treatments. From Atzimba, about 15–40% of the regenerated plants were ELISA (-), without any relationship to the cytokinin content in the media. Heat had a stronger influence on virus elimination than kinetin and the results varied with the cultivar.     

Purification of potato virus A and its detection in potato by enzyme-linked immunosorbent assay (ELISA)

Potato virus A (PVA) was purified fromNicandra physaloides by a simple method that omitted organic solvent clarification and consisted of differential centrifugation followed by equilibrium centrifugation in CsCl. An antiserum was produced that specifically detected PVA in potato leaf sap using either the SDS-agar test or enzyme-linked immunosorbent assay (ELISA). No heterlogous reaction of the antiserum with potato virus Y was detected. Purified PVA was highly infectious; it had an A 258/280 nm absorbance ratio of 1.28. The particles had a normal intact appearance in the electron microscope. Detection of PVA in potato sprouts and foliage by ELISA was compared with the local lesion assay onPhysalis angulata L. plants. ELISA was superior over an indicator plant test when sprout tissue was used. PVA antiserum reacted similarly with mild and crinkle isolates.     

Shoot formation in potato tuber discs in tissue culture

Tuber discs of potato (Solanum tuberosum) growing on a modified Murashige and Skoog medium produced callus and embryoid bodies but failed to form shoots. However, if 0.4 ppm of 6-benzylaminopurine was added to the basal medium, shoots became visible seven weeks after inoculation.     

Anatomical evidence for the existence of roots on potato tubers and stolons

Roots on potato tubers and stolons displayed the normal root anatomy which consisted of a central vascular cylinder surrounded by endodermis with Casparian strips, the cortex and epidermis. Tuber roots appear to initiate from the parenchyma cells adjacent to the vascular tissue. Shoot tips were similar to normal apical meristems. These observations support our research demonstrating the growth of functional roots from potato tubers and stolons.