Maintenance,symptoms and distribution of potato viruses X,S, Y,A, M and leafroll in potato tissue culture plantlets

Maintaining potato viruses X, S, Y, A, M and leafroll in tissue culture plantlets is a convenient, cost and space effective alternative to the use of greenhouse plants. Of these six viruses, only certain strains of PVX induced symptoms in tissue culture plantlets. Nevertheless, all infected tissue culture plants were found to be more reliable than greenhouse grown plants as virus-infected controls in enzyme-linked immunosorbent assay (ELISA) testing. Another advantage of maintaining viruses in tissue culture plantlets was the elimination of contamination by other viruses or other pathogens. Leaves, stems, and roots of virus infected plantlets were tested separately for antigen levels by ELISA. In these tests, the stems and leaves of all but PVA infected tissue culture plants consistently gave positive ELISA values. In contrast, root tissue from PVY infected tissue culture plantlets was not reliable for PVY detection. In all cases, the viruses detected in the original source material were detected in the resulting tissue culture plantlets.     

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.     

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.     

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.     

A rapid method for determining blackspot susceptibility of potato clones

Two methods of determining susceptibility of potato clones to blackspot were compared: (1) bruising by weight dropping and (2) bruising by abrasive peeling. A highly significant positive correlation was obtained between the intensity of enzymatic discoloration following abrasive peeling and the amount of blackspot that developed by weight dropping (r=0.93). Abrasive peeling was more rapid than the weight-dropping method. Tuber samples were abraded 30 sec and the amount of enzymatic discoloration evaluated after 24 hr. The need for individually bruising and hand peeling of tubers was eliminated with this method. Because of the rapidity of the abrasive peeling method, it can be used effectively in potato breeding programs to screen large numbers of clones for blackspot susceptibility. Results indicate that tuber maturity affects enzymatic discoloration and blackspot susceptibility. Immature tubers, dug while the vines are still green, are more resistant to blackspot than mature tubers. Tuber maturity therefore must be considered when screening clones for susceptibility to blackspot.     

Improved procedures for the isolation and culture of protoplasts fromSolanum etuberosum

An efficient and convenient procedure for the isolation of protoplasts fromSolanum etuberosum is described. Excised leaves of plants grownin vitro were preconditioned for 3 days in the dark at 4 C. A 16 h enzyme digestion followed with purification by flotation in Babcock bottles produced a mean yield of 12.5 × 10⁶ protoplasts/g fresh weight. Protoplast viability as judged by morphology at the time of isolation was 89%. Liquid preculture for 1 day at 1 × 10⁶ protoplasts/ml was used prior to plating at 1 × 10⁵ protoplasts/ml and gave a mean survival of protoplasts of 43% after 8 days in culture.     

Pre and early post-emergence weed control with Paraquat in potatoes

Studies using Paraquat herbicide for early post-emergence control of broadleaved and grass weeds in Katahdin and Russet Burbank potatoes were conducted in Maine during four growing seasons. All rates and times of application of Paraquat gave good commercial control of grass and broadleaf weeds when compared to Premerge and Dowpon treatment as checks. Paraquat applied to Katahdins 2 weeks after ground crack reduced the yield of tubers but did not significantly affect specific gravity. Yield and specific gravity of Russet Burbank was reduced by Paraquat applied one and 2 weeks after ground crack. Paraquat can be used effectively for weed control in Katahdin up to one week after ground crack without crop damage. In Russet Burbank it appeared that application at ground crack was about as late as Paraquat could be applied without affecting yield or specific gravity of tubers.     

Albinism of potato spindle tuber viroid-infected Rutgers tomato in continuous light

Potato spindle tuber viroid (PSTV) caused albinism in the new growth of Rutgers tomato under continuous light. The albinism response was obtained with 4 isolates causing severe symptoms in tomato and with 1 mild isolate which is normally symptomless. Rutgers tomato inoculated with the 4 severe strains of PSTV developed typical ‘bunchy top’ symptoms in 12 to 16-hr day lengths. Symptoms developed slower and much less distinctly on plants incubated in 6-hr days. New growth in PSTV-infected plants was green in short days (12-hr illumination) and chlorotic to white in continuous light. Infectivities of extracts from white portions of plants were from 3 to 10 times greater than those from green portions of infected plants. Albino symptoms also developed in other tomato varieties when inoculated with PSTV and kept under continuous light. Albinism was most pronounced under continuous light at 30°C, somewhat less intense at 24°, and at 16°, new growth of infected leaves and stems remained green. Albinism did not develop in otherSolanum species which are symptomless hosts of PSTV nor in seedling potato plants grown in continuous light.     

The effect of nitrogen on the coumarin-induced tuberization of potato axillary shoots culturedin vitro

Coumarin-induced tuberization was studied on axillary shoots excised from etiolated potato sprouts each of which was cultured in vitro. Sprouts grown on culture medium with high nitrogen concentrations produced axillary shoots which elongated and would not readily tuberize. Sprouts cultured on medium low in nitrogen produced axillary shoots which tuberized with limited elongation. Carbohydrate concentration of the culture medium for the excised axillary shoot could significantly modify the nitrogen effect of the potato sprout medium. Axillary shoots growing on high nitrogen medium did not tuberize except in cases where the carbohydrate concentration was increased.     

Laboratory testing of potato tubers for multigenic resistance to late blight

Resistance in the tubers of potato clones with various levels of foliar multigenic resistance toPhytophthora infestans was measured to detect possible correlations between tuber and foliar resistance. A highly significant correlation was found using wound-healed tuber tissues. High levels of tuber resistance were detected in most foliar-resistant clones when inoculations with race 1,2,3,4 were made after 24–48 hours of wound periderm formation at 20 C. The resistance of wound-healed tissues increased as the time between wounding and inoculation increased. This increase in resistance was significantly greater for clones with foliar resistance than for clones with susceptible foliage.     

Effect of temperature on the starch content of potato callus tissue

Starch content was determined in Lemhi Russet callus grown on MS medium for 2 and 4 weeks at 25, 15, and 5°C with 50 μM 2,4-D or 10 μM picloram as the only plant growth regulator. A treatment of 5°C for 4 weeks followed by 25°C for 2 and 4 weeks was included to simulate reconditioning. Callus starch content ranged from 0.01 to 0.08% on a wet weight basis and 0.5 to 4.0% on a dry weight basis. Starch content was highest after 2 weeks at 25 and 15°C and decreased with time in a similar manner at both 25 and 15°C. Starch content decreased at 5°C, then increased significantly during simulated reconditioning at 25°C. Starch content was higher in callus grown with 50 μM 2,4-D than in callus grown with 10 μM picloram. After staining the callus with I₂-KI solution, starch granule formation was observed in the callus grown with 50 μM 2,4-D, while little granule formation was observed in the callus grown with 10 μM picloram.     

A method for estimating the loss in tuber yield caused by late blight of potato

A method was developed for estimating tuber yield losses caused by late blight of potato (Phytophthora infestans) (Mont.) de Bary. The method is based on yield and disease data collected for the varieties Green Mountain and Katahdin grown in eastern Canada during the period 1953–1970. The percentage yield loss can be calculated for any disease progress curve. The estimated loss was within 5% of the actual loss in nine cases out of ten.     

Determination of total glycoalkaloids in potato tubers using a modified titration method

A new method for determining total glycoalkaloid content of potatoes (TGA) was developed by extensively modifying the comprehensive titration procedure of Fitzpatrick and Osman (1). This modified titration method eliminates the need to hydrolyze the glycoalkaloids and to purify the resulting aglycones by partitioning them into an organic solvent such as benzene or methylene chloride. Instead the glycoalkaloids are precipitated using ammonium hydroxide with the precipitate being dissolved in a tetrahydrofuran-water-acetonitrile (50:30:20) mixture. An aliquot is evaporated and quantitated using the nonaqueous titration procedure developed by Fitzpatrick and Osman (1). A comparison between this modified method and a high-performance liquid Chromatographic (HPLC) procedure was made using three commercial potato varieties and five experimental selections. Results from the two methods compared favorably.     

Mechanical and rheological properties of calcium carbonate-filled ethylene propylene diene elastomer reinforced by metallic acrylate salt

High performance ethylene propylene diene methylene elastomer (EPDM)/calcium carbonate (CaCO₃)/aluminum triacrylate (ALTA) hybrids have been prepared by a melt compounding process. The mechanical properties of the peroxide cured EPDM/CaCO₃/ALTA vulcanizates were investigated by tensile, hardness, resilience, abrasion and fatigue life tests. The results showed that the ALTA can greatly improve the modulus at 100 %, tensile strength, abrasion resistance and fatigue life of the EPDM/CaCO₃/ALTA vulcanizates, while retaining their high elongation at break. ALTA as a reactive filler had accelerating effect on the vulcanization reaction of and increased the crosslink density of the EPDM/CaCO₃ composites. This phenomenon is due to increasing the ionic bonds arise from both homo-polymerization of ALTA and graft co-polymerization of it onto the EPDM. Scanning electron microscopy (SEM) revealed the presence of ALTA causes to the roughness of the fracture surface of CaCO₃-filled EPDM compounds demonstrating high interaction between the fillers and EPDM improved by introduction of ALTA. Dynamic mechanical analysis revealed that the glass transition temperature (T _g ) and dissipation peak (tan δ) of EPDM composites decreases with increasing ALTA content. Decrease in tan δ value and inward shifting of T _g were related to improved interaction of filler and EPDM.     

Bactericidal activity of zinc oxide particles embedded in poly(vinyl pyrrolidone) electrospun fibers

Composites of metal oxide nanoparticles incorporated into poly(vinyl pyrrolidone) electrospun fibers can be considered potential and low cost bactericidal agents. Homogeneous distribution of nanoparticles in the core of fibers and minimal aggregation induced by strong interaction with polymeric template are factors that contribute with diffusion of metal oxide nanoparticles from bulk to the fiber surface and subsequent release of nanoparticles to bacterial culture. Hybrid electrospun fibers were successfully applied against bacteria as a result of reasonable degree of interaction between zinc oxide and fibers as detected from FTIR analysis, DSC curves and SEM images.     

Phytophthora erythroseptica in Peru: Its identification and pathogenesis

EightPhytophthora isolates cultured from diseased potato plants and tubers in Peru were identified asPhytophthora erythroseptica Pethyb. var.erythroseptica. This pathogen was not previously reported from South America. The isolates were pathogenic to potato plants but they differed in virulence. Injuring roots prior to inoculation enhanced infection and disease development but all isolates infected noninjured roots. Continuous high soil moisture during incubation favored disease development. Under these conditions, disease developed at soil temperatures from 10 C to 30 C, but most rapidly at 25 C and 30 C. Plants grown 60 days before inoculation developed symptoms earlier and more rapidly than plants grown 15-, 30-, and 45 days. Four concentrations of mycelial-suspension inocula did not alter disease development, but zoospore inoculum induced symptoms and plant death more rapidly than mycelial suspensions at 25 C.     

Bison,a new red-skinned potato variety

Bison, a new red potato, was introduced by North Dakota State University. This new red variety has smooth tuber type and bright red skin color. Bison yields somewhat less than Norland and Red Pontiac but the advantage of Bison over these two varieties is its uniformity and bright red color. Bison is about medium in total solids and makes chips comparable in color to Norchip but lighter in color than Kennebec. Bison is resistant to race 0 of the late blight organismPhytophthora infestans, but susceptible to race 1–4.     

Variability for photoperiodic reaction among diploid and tetraploid potato clones from three taxonomic groups

The photoperiodic behaviour of 18 potato clones from three taxonomic groups (Andigena, Phureja, and Tuberosum diploid and tetraploid) was studied under controlled environments. Day lengths of 11, 13, 15, and 17 hours were provided with all other environmental conditions common. Six characters were studied: tuber formation, stem height, haulm weight, tuber number, tuber weight, and tuberization efficiency (tuber weight/haulm weight). All clones showed well defined critical daylength values at which tuberization was either absent or irregular. A wide variability for critical day length was observed among clones, the higher values corresponding to Tuberosum tetraploid and the lower values to the Phureja group. Stem height and haulm weight increased as photoperiod increased. Tuber number remained practically the same for all groups but Phureja which showed an inverse relationship. Tuber weight decreased as daylength increased for the groups Andigena, Phureja, and Tuberosum diploid which on the average showed a short day reaction for tuber yield. Tuberosum tetraploid did not show any significant change, behaving as a day neutral type. Tuberization efficiency was, in general, decreased as daylength increased suggesting a short day behaviour for all groups. A regression analysis of tuber yield and tuberization efficiency on photoperiod indicated that, in general, tetraploid genotypes were less sensitive to drastic increases in daylength than diploids.