Monitoring current season potato leafroll virus movement with an immunosorbent direct tissue blotting assay

A direct tissue blotting assay (DTBA) was used to track the movement of potato leafroll virus (PLRV) from newly infected foliage to the tubers. Plant and tuber characteristics were recorded to assess plant growth stage at inoculation and PLRV effect on yield. Russet Burbank potatoes were planted at different times in 1991 and 1992 to provide plants of different maturities which were then inoculated using PLRV carrying aphids. Aphids were allowed to feed two to three days after which an insecticide was applied. Stems and tubers were tested periodically for PLRV with DTBA after inoculation. Indexed tubers were grown out and ELISA tested in the greenhouse the following winter to confirm results of summer serological tests. Plant age affected percentages more consistently than did inoculation date. When plants approximately 43 days from planting were inoculated at different dates, early inoculation produced a higher percentage of infected plants. Conversely, when plants approximately 62 days from planting were inoculated at different dates, late inoculation resulted in a higher percentage of infected plants. However, early inoculation of young plants resulted in the highest infection percentages. Tuber size and yield were negatively affected by higher percentages of leafroll regardless of the stage of growth at inoculation. DTBA is best used for detecting PLRV in foliage of plants grown from infected tubers (i.e. secondary PLRV). DTBA is less accurate for detecting primary PLRV.     

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.     

Effect of time and date of inoculation,plant age,and temperature on translocation of potato leafroll virus into potato tubers

Movement of potato leafroll virus (PLRV) to tubers following placement of viruliferous aphids on foliage was studied on the medium-maturing cultivar, Katahdin and the late-maturing cultivar, Russet Burbank. Inoculation was begun on August 20 and continued at three-day intervals until mid-September. There was no trend of increasing or decreasing numbers of leafroll-infected tubers from early to late inoculation. Several variables were examined to determine their effects on the incidence of PLRV-infected tubers. Multiple regression analyses showed that inoculation date, maximum daily temperature, minimum daily temperature, plant age, and length of time from inoculation to harvest explained 38% of PLR incidence in tubers of cv. Katahdin. Length of time from inoculation to harvest and minimum daily temperature explained 40% of PLR incidence in tubers of the cv. Russet Burbank.     

Detection of potato leafroll virus by visual inspection,direct tissue blotting and ELISA techniques

Greenhouse-grown Russet Burbank and Russet Norkotah plants were tested for potato leafroll virus (PLRV) by visual examination, enzyme-linked immunosorbent assay (ELISA) and direct tissue-blotting assay (DTBA). Visibly infected and healthy plants were obtained from excised eyes of tubers submitted for winter certification tests. Each plant was sectioned into leaf, petiole, stem, root, seed piece, and tuber portions. Tissues were blotted on a nitrocellulose membrane for DTBA and then homogenized for use in a double-antibody sandwich ELISA system. Agreement between the two serological detection methods and with visual readings was high for petioles and stems, but lower for leaf, tuber and root tissues. Comparison of DTBA with ELISA and with visual plant symptoms suggest that DTBA can be used with the same accuracy as ELISA for detecting PLRV in stems and petioles     

Potato leafroll virus spread in differentially resistant potato cultivars under varying aphid densities

An action threshold of 3-10 green peach aphid,Myzus persicae (Sulzer), apterae per 100 lower leaves is recommended for use in Minnesota to prevent further spread of potato leafroll virus (PLRV) in potato,Solarium tuberosum L. This threshold was first developed and validated using the PLRV susceptible cultivar Russet Burbank. Here we report experiments to determine if higher aphid densities could be tolerated in PLRV resistant cultivars,i.e., Kennebec (moderately resistant) or Cascade (highly resistant), without an increase in PLRV infection. Insecticidal sprays were applied to plots when predetermined target aphid densities, based on number of apterae per 100 leaves, were reached: 3, 10, 30, and 100 (Russet Burbank); 10, 30, 100, and 300 (Kennebec); and 30, 100, 300, and 1000 (Cascade). The response variable was the average percentage of PLRV infected plants. Overall mean cumulative aphid-days and percent PLRV infection were 617 and 23.5% for Russet Burbank, 1,296 and 10.2% for Kennebec, and 4,816 and 9.5% for Cascade. For each cultivar, the highest target aphid density tolerated without an increase in PLRV spread was determined by comparing PLRV infection in plots sprayed on predetermined thresholds to PLRV infection in plots where aphids were rigorously controlled. This maximum density was 10 apterae per 100 leaves for Russet Burbank and 300 apterae per 100 leaves for Cascade. Results using Kennebec were ambiguous, but Kennebec was always more resistant to PLRV than Russet Burbank. Excised leaflet tests showed that the cultivars did not differ in resistance to green peach aphid. It appears that action thresholds based on green peach aphid apterae can be different depending upon the inherent PLRV-resistance of the cultivar.     

Western russet: A new potato variety with excellent fresh market and frozen-fried processing quality and field resistance to common scab and PVYo

Western Russet, designated experimentally as A7961-1, and resulting from the cross A68113-4 x Bel-Rus, was released in 2004 by the USDA/ARS and the agricultural experiment stations of Idaho, Oregon, and Washington. The foliage of Western Russet is spreading, with medium-sized olive-green leaves and abundant white flowers. The tubers are tan, with medium russet skin, oblong shape, a slightly prominent eyebrow, white flesh, and distinctly visible pith. Western Russet was compared with Russet Burbank in trials across the Pacific Northwest for yield, quality, and disease response. In general, Western Russet produced lower total yields than Russet Burbank, but on average and depending on location, similar U.S. No. 1 yields. When observed for defect problems, Western Russet exhibited resistance to second growth, growth cracks, shatter bruise, stem-end discoloration, and heat necrosis, moderate resistance to hollow heart, and moderate susceptibility to blackspot bruise. In product quality tests, Western Russet was rated similar to Russet Burbank for french fry and dehydrated potato flake quality and slightly inferior for baked potato quality. In replicated evaluations, Western Russet was found to have good field resistance to common scab and PVY^o, and moderate resistance to verticillium wilt and tuber net necrosis caused by PLRV. It demonstrated susceptibility to late blight, foliar PLRV, dry rot, soft rot, and tuber early blight. Biochemical analysis of Western Russet tubers showed markedly higher vitamin C content than those of Russet Burbank. Tuber glycoalkaloid concentration in tubers produced in a 1988 trial was 1.0 mg 100 g^?1.     

Summit Russet: A new russet potato variety with good fresh market and frozen processing qualities

Summit Russet (A84118-3), the result of the cross A77236-6 x TND329-1Russ, was released in 2003 by the USDA/ARS and the Agricultural Experiment Stations of Idaho, Oregon, and Washington. The vines of Summit Russet are erect, with very thick stems, large yellowish-green leaves and abundant white flowers. The tubers are tan, with medium russet skin, long-flattened shape, a slightly prominent eyebrow, white flesh, and moderately prominent pith. Tuber dormancy is very long. Summit Russet was compared with Russet Burbank in trials across the Pacific Northwest for yield, quality, and disease response. In general, Summit Russet produced similar or lower total yields than did Russet Burbank, but higher U.S. No. 1 yields. In Idaho variety trials, Summit Russet exhibited resistance to second growth, growth cracks, blackspot bruise, and stem-end discoloration, and moderate susceptibility to hollow heart and shatter bruise. In comprehensive product quality evaluations, Summit Russet was rated superior to Russet Burbank for french fry quality and similar for baked potato quality. French fry quality was retained after long-term storage. In replicated disease trials, Summit Russet was found to have good resistance to common scab, early blight tuber rot, and Verticillium wilt, and moderate resistance to Fusarium dry rot and tuber net necrosis caused by PLRV. It demonstrated susceptibility to late blight, foliar PLRV, PVX, PVY^o, and bacterial soft rot. Biochemical analysis of Summit Russet tubers showed them to be higher in total solids and sucrose, and lower in dextrose than those of Russet Burbank. Tuber glycoalkaloid concentration as measured from tubers produced in 1994–1996 trials was 6.7 mg 100 g¹.     

Effect of transplant container volume and growing season length on field performance of micropropagated potatoes

Micropropagation is a tissue culture technique adapted for the rapid multiplication of disease-free seed stocks. Procedures for propagating potatoes in the laboratory and acclimating plantlets in the greenhouse are available, but information on cultural practices for maximizing tuber yield of plantlets when transplanted to the field is lacking. Centennial Russet and Russet Burbank plantlets were transferred from culture jars to three sizes of transplant containers for establishment under greenhouse conditions before transplanting to the field. Length of field growing season was varied by using two transplant dates and two vine kill dates. Survival of field transplanted plantlets was above 95 percent in both 1983 and 1984. Cultural practices significantly affected the tuber yield of plantlets of both cultivars. Total yield and yield of tubers larger than 35 mm in diameter increased with increasing transplant container volume. Transplant container volume had no effect on the yield of tubers less than 35 mm in diameter. Highest total yields and tuber production per plantlet for both Centennial Russet and Russet Burbank were obtained from the longest growing season (early transplant date with late vine kill). Yield of Russet Burbank plantlets increased more in response to a longer growing season than did Centennial Russet.     

Association of viral,bacterial and fungal pathogens with vascular discoloration of Russet Burbank potato tubers

Russet Burbank potatoes from the 1979 and 1980 crop years, collected from Chicago, IL repack warehouses and retail markets, were sampled for vascular discoloration. The amount of discoloration varied among sampling months and states of origin and decreased from 7.7% in 1979 to 1.6% in 1980. Highest levels of vascular discoloration were detected in December and January samples. Vascular discolored and non-discolored (control) tubers were assayed for the presence of potato leafroll, potato virus X, and beet western yellows viruses by the enzyme-linked immunosorbent assay (ELISA). Tubers were also assayed for 2 subspecies ofErwinia carotovora and forVerticillium albo-atrum andV. dahliae. Potato leafroll virus was detected in 31 of 831 vascular discolored tubers.V. albo-atrum was detected in 1 of 180 discolored tubers. Beet western yellows virus was not detected in discolored or non-discolored tubers. Two subspecies ofE. carotovora and potato virus X were equally common in discolored and non-discolored tubers.     

Russet Nugget: A fresh market and processing potato cultivar with resistance to common scab

Russet Nugget is an oblong, smooth, high yielding potato with fresh market and processing qualities. It emerges quickly, produces an erect, large, vigorous vine with a profusion of white flowers, and is late maturing. Russet Nugget is very resistant to common scab, and moderately resistant to leafroll net necrosis,Vertcillium wilt, and early blight (tuber and foliage). In the San Luis Valley of Colorado, yield of Russet Nugget is comparable to Russet Burbank, with a significantly greater percentage of U.S. No. 1 tubers. In Texas, total yield is comparable to Norgold 40, with a slightly lower percentage of U.S. No. 1 tubers. Tubers of Russet Nugget have high specific gravity and vitamin C content.     

Heat inactivation of leafroll virus in potato tuber tissues

Heat inactivation of leafroll virus in tuber tissues of three potato varieties (Russet Burbank, Katahdin, and Mohawk) was studied. Russet Burbank did not tolerate high constant temperatures and a low proportion of tubers and eye-pieces survived the treatments. On the other hand, Russet Burbank eye-pieces survived, with few exceptions, treatment at 40 C for four hours alternating with room temperaure (16–20 C) for 20 hours daily for as long as eight weeks. Inactivation of the virus was complete after six weeks of this treatment. Results obtained with the Katahdin and Mohawk varieties in similar tests were variable, and this possibly may be attributed to the higher room temperature (25–30 C) prevailing during these experiments.     

Transmission of potato leafroll virus to or from tuber slices by aphids feeding through a membrane

Summary Eye-bearing slices, cut from healthy potato tubers and placed between Parafilm membranes, were inoculated with potato leafroll virus (PLRV) byMyzus persicae. PLRV was detected by ELISA and by transmission tests in tuber slices and in plants grown from the slices of the susceptible cv. Désirée, but not in those of the resistant cv. Arkula. These results suggest that PLRV replication and transport within tuber phloem is controlled by specific mechanisms of resistance.M. persicae was also able to acquire and transmit PLRV toPl floridana from slices cut from tubers of infected plants. The aphids effectively transmitted PLRV from slices cut from the sprouting rose end but they failed to transmit it from slices cut from the heel end of tubers.     

Anatomy and compositional variations within potatoes. IV. Total solids distribution in different cultivars

Distribution of cortical, perimedullary, and pith tissues and total solids content were determined on both a fresh and a dry weight basis for six potato cultivars grown concurrently in three locations in California. At one location, tubers harvested 80 days from planting were 1.0 to 2.5% lower in total solids than tubers harvested at later dates. The high solids cortical tissues ranged from 39 to 51% of whole tubers on a dry basis, and the amount appeared to be related to tuber shape. White Rose and Red LaSoda showed greater difference in per cent total solids between cortical and perimedullary tissues than did Kennebec, Lenape,³ Norchip, and Russet Burbank. The high total solids of the cortical tissues is particularly significant to processing losses. Any method of peeling that removes one half the linear depth of the cortical tissues can result in a loss of 25% or more of total tuber solids, depending upon cultivar and tuber shape. Tuber development and distribution of total solids and other components were previously described for Red Pontiac, Russet Burbank, and White Rose tubers obtained from the retail market (6,9,10,11). It was particularly noted that the high-solids cortical tissues, although averaging only about 6mm in depth, comprised between 40 and 50% of both fresh and dry tuber weights (6,11). Because this sort of distribution of solids is especially significant to peeling loss, possible differences in solids among cultivars and at different harvest dates were investigated.     

Potato leafroll virus purification and antiserum preparation for enzyme-linked immunosorbent assays

Potato leafroll virus (PLRV) was purified from potato foliage and stems with an average yield of 0.14 mg of PLRV/kg of potato. Modifications of an existing purification procedure are reported. Five low dosage (38-118 μg of PLRV) intravenous injections were used to produce a PLRV antiserum for use in enzyme-linked immunosorbent assays (ELISA) from tubers. PLRV was readily detected in ELISA testing of potato tubers and leaves and inPhysalis floridana Rybd. Non-specific reactions were low with all tissues. In parallel tests, a Canadian antiserum produced higher nonspecific reactions with tuber and leaf tissue. The results indicated that the use of low dosage-intravenous injections might be necessary methodology for producing PLRV antiserum for use in ELISA diagnostic tests with tuber tissue where high non-specific reactions have been reported.     

GemStar russet: A potato variety with high yield, good culinary quality, excellent fresh market appearance, and resistance to common scab

GemStar Russet, derived from the cross Gem Russet x A8341-5, was released in 2004 by the USDA/ARS and the agricultural experiment stations of Idaho, Oregon, and Washington. The foliage of GemStar Russet is dark yellowish-green, spreading, with large leaves and abundant white flowers. The tubers are brown, medium to heavily russeted, oblong, with a slightly prominent eyebrow, white flesh, and indistinct pith. GemStar Russet was compared with Russet Burbank and Russet Norkotah in trials across the Pacific Northwest for yield, quality, and disease response. Except for locations in southern Idaho, in general, GemStar Russet produced slightly lower total yield than Russet Burbank, but much higher U.S. No. 1 yield. When compared with Russet Norkotah, GemStar Russet produced similar total and U.S. No. 1 yields in early harvest trials but higher total and U.S. No. 1 yields in late-harvest trials. When observed for defect problems, GemStar Russet exhibited resistance to second growth, growth cracks, and stemend discoloration, moderate resistance to blackspot and shatter bruising, but a high level of susceptibility to hollow heart. In product quality tests, GemStar Russet was rated superior to Russet Burbank for french fry quality and comparable for baked potato quality. GemStar Russet was found to be immune to PVX, resistant to common scab and powdery scab, moderately resistant to Verticillium wilt, tuber net necrosis caused by PLRV, and corky ringspot. It demonstrated susceptibility to late blight, PLRV, dry rot, soft rot, and ringrot and extreme susceptibility to PVY^o. Biochemical analysis of GemStar Russet tubers showed them to be higher in protein and much higher in vitamin C than those of Russet Burbank or Russet Norkotah. Three-year average for tuber glycoalkaloid concentration was 1.5 mg 100 g^?1.     

An approach to field screening potato genotypes for potato leaf roll virus resistance

Eight potato cultivars and two advanced breeder selections were assessed for field resistance to the potato leaf roll virus (PLRV) following field exposures in which PLRV-infected Russet Burbank plants were used as inoculum sources within treatments. This screening protocol provided consistent PLRV resistance ratings despite year-to-year variation in PLRV pressure. Secondary disease incidence based on enzyme-linked immunosorbent assay (ELISA) of foliage from tuber progeny ranged from 0–87% in 1990 and 0–67% in 1991, and was consistent with reported PLRV resistance ratings for eight of ten genotypes. Agreement between visual assessment and ELISA on plants from harvested tubers was 94% in 1990 and 83% in 1991, for all genotypes. However, agreement data were inconsistent from year-to-year, with the exception of three genotypes. In both years, current season infection, based on ELISA of foliage, was detected in less than two percent of the plants and, was inadequate as a measure of secondary PLRV incidence. Green peach aphid (GPA) populations did not differ among genotypes at sampling times during the season, but the PLRV concentration in GPA colonizing Russet Burbank plots was significantly higher than in GPA colonizing any other genotype.     

Enzyme-linked immunosorbent assay (ELISA) for the detection of potato leafroll virus and potato virus Y in potato tubers after artificial break of dormancy

Summary Conditions necessary for the detection of potato leafroll virus (PLRV) and potato virus Y (PVY) in tubers from primary and secondary infected plants were investigated. Tubers were analysed before and after breaking dormancy by rindite treatment. PLRV was reliably detected indormant tubers whereas PVY was readily detected only when tubers had been rindite-treated and held for two to three weeks at 22°C and high humidity in the dark. PLRV occurred in higher concentration at the heel end than at the rose end of infected tubers and the concentration remained nearly unchanged during the experimental period of 35 days, whereas PVY was found to be more concentrated at the rose end and was rapidly accumulating in the tubers after the break of dormancy. In dormant tubers PVY concentration dropped during storage at 22°C. The use of ELISA for tuber indexing is discussed.     

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.     

First report ofPythium sylvaticum causing potato tuber rot

Leak is a post-harvest disease of potato tubers manifested by a wet, spongy rot that can quickly lead to complete tuber decay. In North America, leak is most commonly caused byPythium ultimum var.ultimum Trow, although a number of otherPythium spp. have been found to be pathogenic to potato tubers. We report that an isolate identified by DNA sequencing asP. sylvaticum Campbell & Hendrix was pathogenic to potato tubers. The organism produced a dark, hollow cavity in tubers that was much drier in texture than typical symptoms of leak. Symptomology was similar in tubers of the cultivars Yukon Gold, Green Mountain, and Russet Burbank. Studies to complete Koch’s postulates confirmed thatP. sylvaticum could be reisolated from diseased tissue and produced the same disease symptoms when isolates were subsequently re-inoculated into healthy tubers. This is the first report ofP. sylvaticum causing rot in potato tubers.     

Tuber initiation and development in irrigated and non-irrigated potatoes

Tuber initiation and development are processes basic to potato production and are particularly critical in areas with short growing seasons. It is important to know how and to what extent management decisions affect these processes in order to maximize the yield of marketable tubers. A two-year field study, conducted in southcentral Alaska, examined top growth, tuber initiation, and tuber development in eight potato cultivars grown with and without irrigation. Plants of the cultivars Allagash Russet, Bake-King, Green Mountain, Kennebec, Lemhi Russet, Russet Burbank, Shepody, and Superior were harvested weekly throughout the growing season, and top dry weight, numbers of tubers, and individual tuber fresh weights were recorded. Top dry weight was reduced by moisture stress shortly after emergence in 1993, and about one month following emergence in 1994, when early-season soil moisture was greater. The weight of tubers was similarly affected within approximately 5 wk of emergence in 1993 and 6 weeks in 1994. Tuber weight at harvest was increased two-to three-fold by irrigation in all cultivars. The number of tubers each plant set was affected by irrigation in most, but not all, cultivars. Some varieties (Lemhi Russet in 1994, Allagash Russet both years) set more tubers than were maintained through the growing season. Tuber remnants found during sample collection indicated that tuber reabsorption had occurred. Irrigated Green Mountain had more than one tuber initiation period during the season, whereas other varieties such as Shepody maintained a relatively constant number of tubers following initial tuber set. Tuber size distribution at the end of the growing season showed that larger tubers were favored by irrigation.