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1.
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.  相似文献   

2.
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.  相似文献   

3.
The Green Mountain cultivar was used in field tests to determine the effects of inoculating potato plants at various times with the potato leafroll virus (PLRV) on development of internal necrosis of tuber tissue. Viruliferous apterae of the green peach aphid,Myzus persicae (Sulz.), were placed on each stem in all hills to be inoculated in each 3.0 m single-row plot. Planting and inoculation dates were varied in all field experiments and, in one, several vine-killing dates were also included. All harvested tubers were stored for approximately four months at 10°C to enhance development of internal necrosis prior to examination. Similar but smaller greenhouse studies involving both apterous and alate green peach aphids were also conducted using Green Mountain, Irish Cobbler, and Russet Burbank cultivars. All results showed that as inoculation was delayed relative to plant development, more net necrosis (NN) occurred. Conversely, when plants were inoculated early, stem-end browning (SEB) rather than NN predominated. A high percentage of naturally occurring SEB tubers (cv. Russet Burbank) were found by ELISA to contain PLRV. Plants produced by these tubers only rarely developed leafroll symptoms. These findings suggest a previously unsuspected causal relationship between SEB and PLRV. Implications of this apparent relationship on the epidemiology of potato leafroll in Maine are discussed.  相似文献   

4.
Development of alternative serological techniques to ELISA for detection of potato viruses offers advantages for monitoring virus incidence and for seed potato certification systems. Several trials showed that multiplex tissue print immunoassay (TPIA) and dot blot immunoassay (DBIA) might represent fast, practical, and sensitive alternatives for the detection of: Potato leaf roll virus (PLRV), Potato virus S (PVS), Potato virus X (PVX) and Potato virus Y (PVY), from green and/or tuber tissues. In TPIA, the specific precipitation patterns in infected tissues of leaf petioles or stem cross sections, observed with each virus, allowed identification of the specific virus or mixed infections in a single multiplex assay. For detection of PVY in green tissues, DBIA was shown to be over 50 times more sensitive than ELISA. TPIA and ELISA from the tuber stem end or from eyes might be used for rapid detection of PVY and PVS in seed potato tubers without prior germination. PVS was evenly distributed in potato tuber tissue, while PVY was localized in the vascular tissue beneath the epidermis, with irregular distribution along the periphery of the potato tuber. For laboratories in developing countries lacking time and facilities for tests based on tuber germination, monitoring for PVS and PVY using TPIA in tuber tissue may be a suitable alternative to ELISA.  相似文献   

5.
This report describes a simple, rapid and inexpensive procedure for sampling large numbers of dormant tubers for analysis of potato leafroll luteovirus (PLRV) infection. The procedure uses a common electric drill to simultaneously remove and macerate tuber-eye samples for detection of PLRV by the enzyme-linked immunosorbant assay (ELISA) and the polymerase chain reaction (PCR). By using these sampling and analysis approaches, 19 of 20 different PLRV isolates were detected in dormant tubers from plants with primary infections. Results from the dormant tuber analysis, were verified by planting the tubers and testing leaf tissue by ELISA and PCR. Similar sampling and testing done on healthy dormant tubers and sprouts from the tubers consistently gave negative results as expected.  相似文献   

6.
Potato varieties Russet Burbank and Ranger Russet were transformed with a cDNA version of the 23 kD coat protein cistron (CP) of the potato leafroll virus (PLRV) using anAgrobacterium-mediated procedure. Clones were assayed for presence of thenpt- II and CP genes by Southern analysis, for expression of CP mRNA by Northern analysis, and for presence of PLRV coat protein in uninoculated and aphid inoculated plants by ELISA and Western blot analysis in uninoculated plants. Two putative Russet Burbank transformants were escapes, lacking eithernptll or CP, while one putative Ranger Russet transformant possessed thenpt- II gene but not the CP gene. In Russet Burbank and Ranger Russet, some transformants had statistically lower virus titer. The lowered titer was consistent throughout assays at three times during primary infection and one assay of secondary infection. The ranking of virus titer across all tests was statistically consistent. Transformants with the lowest secondary titers had virus contents 15 and 31 % of the titers of untransformed controls for Russet Burbank and Ranger Russet, respectively. The virus titer of the two Russet Burbank escapes and the Ranger Russet with only thenpt- II gene did not differ significantly from their respective untransformed controls.  相似文献   

7.
To evaluate four diagnostic methods for potato leaf roll virus (PLRV), an antiserum was prepared against a virus preparation purified from infectedDatura stramonium L. by an exudation method. The antiserum had a titer of 1:64 in microprecipitin tests. In a procedure developed subsequently PLRV was purified by a method that involved grinding liquid nitrogen frozen stems, petioles, and veins from different solanaceous hosts. A chloroform and n-butanol clarification was done followed by two cycles of differential centrifugation. On sucrose gradients, these preparations formed one band containing spherical particles 25 nm in diameter. In Ouchterlony double-diffusion plates, the antiserum reacted positively with virus preparations from frozenPhysalis, Datura or potato tissue partially purified by two cycles of differential centrifugation. No visible reaction was obtained between the antiserum and the virus preparation from density gradient centrifugation. Antiserum neutralized infectivity of 40–50% of the virions in a partially purified preparation. No virus peak was detected in the scanning patterns obtained after a mixture of antiserum and virus was centrifuged on a sucrose density gradient. A few virus particles were seen attached to serologically specific grids. Methods to diagnose PLRV in infected potato tubers were then compared. In the first, aphids were used to acquire the virus from tuber sprouts. They transmitted the virus to 100% of thePhysalis test plants. Results of the second method, visual inspection of symptoms on potato plants, were similar to those of the first method. Sarkar’s electron microscopic method and immuno-specific grids were less efficient than aphid transmission to diagnose PLRV in tuber sprouts. The low virus concentration in infected tissue made these last two methods unreliable.  相似文献   

8.
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.  相似文献   

9.
Collectotrichum coccodes, the pathogen causing black dot disease of potato, was investigated to determine its potential for causing symptoms on the aerial parts of potato plant, premature vine death, and reduction in yield under greenhouse and field conditions. Spray inoculations with conidial suspensions (106 colony forming units/ml) ofC. coccodes caused sunken, dark, necrotic lesions on stems, leaves and petioles, vein and leaf necrosis, dieback and premature vine death under growth chamber and greenhouse conditions. Field inoculations in 1986 and 1987 provided evidence for an increased incidence of premature vine death in seven potato genotypes (Russet Burbank, Norgold Russet, Superior, Butte, NDA 8694-3, A68113-4 and A66107-51). Late season application of metribuzin at 0.84 kg/ha significantly increased the incidence of stems with black dot symptoms in cv. Russet Burbank at one location. Field inoculations in 1987 at two locations resulted in yield reduction in the late-maturing cv. Russet Burbank (6.3 and 6.5 t/ha) and clone A68113-4 (5.0 t/ha), but not in the early maturing cv. Norgold Russet. Similar results were obtained in greenhouse studies following soil inoculations withC. coccodes: yield reduction was observed in cv. Russet Burbank but not in cvs. Norgold Russet, Superior and Katahdin. Colonization of stem ends of tubers byC. coccodes from spray inoculated field plots was higher than in tubers from uninoculated plots at one of the two locations. These results point to a much broader potential ofC. coccodes as a potato pathogen, than hitherto documented. Colletotrichum coccodes (Wallr.) Hughes (syn.C. atramentarium (Berk. & Br. Taub.) was described in 1825 (21) and has a wide host range (5). Potato and tomato are the more economically important hosts. This fungus has been variously referred to as an unspecialized, minor pathogen or secondary invader (18), weak pathogen (22), specialized parasite (5), or superficial colonizer and widespread epiphyte (19). The symptoms produced by this pathogen on potato have been described as black dot (10, 14) on stems, stolons, tubers and roots, sometimes associated with leaf scorch or wilt (24), rubbery tuber wilt (1), and tuber skin necrosis (17). The fungus is commonly associated with senescent, decaying root and stem tissue, especially at the end of the season. In Idaho, instances of premature death of potato without typical symptoms of Verticillium wilt, but accompanied by a high incidence of stems with black dot symptoms, have been observed in potato crops grown on fields fumigated with metam sodium. Additional observations by growers have also implicated the possible interaction of the herbicide metribuzin with the incidence of black dot. Although it was observed by some workers to accelerate leaf senescence (12, 22), Schmiedeknecht (21) concluded thatC. coccodes was unable to infect the green leaves of potato plant. Even thoughVerticillium dahliae is generally considered the major factor in potato early dying (20), several workers (7, 11, 13, 19, 22) have investigated the possible role ofC. coccodes in causing premature death of potato (8). However, these studies examinedC. coccodes only as a soilborne, root and tuber pathogen of potato; its effects on shoot portions (aerial parts) of the plant were not considered. In addition to causing anthracnose of fruits of tomato, pepper and squash (18), recent reports indicate thatC. coccodes causes foliage blights on eastern black nightshade (2) and velvetleaf (25). The objectives of this study were: (1) to examine the potential ofC. coccodes to cause symptoms on aerial parts of potato plant, (2) to investigate the effect of spray inoculation in the field withC. coccodes on incidence of premature vine death, tuber colonization and yield of potato, and (3) to document the effect of soil inoculation withC. coccodes alone or in combination withV. dahliae on potato varieties under greenhouse conditions.  相似文献   

10.
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, PVYo, 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.  相似文献   

11.
Potato (Solanum tuberosum L. cv. Russet Burbank) tuber periderm resistance to water loss was monitored during the harvest season. Periderm resistance of tubers from water-stressed plants was compared with tubers from unstressed plants throughout a six-week harvest period, in order to determine the effect of plant condition on tuber resistance to water loss. Tubers stored for ten days at 4°C immediately following harvest were also monitored to determine if storage enhanced periderm resistance and if any difference occurred between tubers from stressed vines compared with those from unstressed vines. During the harvest season, tuber resistance to water loss was positively correlated with plant water stress, as measured by leaf water potential and leaf diffusive resistance. Storage of tubers after harvest also caused an increase in periderm resistance. The commercial practice of vine killing, therefore, may have a positive effect on reducing weight loss in storage by enhancing the resistance of tuber periderm to water loss.  相似文献   

12.
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.  相似文献   

13.
The response of Russet Burbank grown at five plant densities (4.0 to 11.1 × 104 plants ha?1), to level of fertilization was studied in field experiments at Guelph, Canada in the 1972 and 1973 growing seasons. Growth analysis techniques were used to establish growth patterns of Russet Burbank at three of the above plant densities (4.0,6.3 and 11.1 × 104plants ha?1), while the effect on root development of rate of fertilization was observed in controlled environments. In both seasons total tuber yield was not influenced by plant density, while marketable yield showed a negative response to increased density. The interaction between plant density and level of fertilization did not approach significance in either season. Due to greater axillary branching at the lower plant densities, leaf area index and photosynthetically active radiation measured at two levels in the canopy, were similar at all densities. Fertilizer levels used in the field showed no negative effect on root growth in experiments conducted in controlled environments. Marked variation in the number of mainstems and thus the number of tubers produced per plant between seasons indicated the importance of using the mainstem as the basic population unit.  相似文献   

14.
Potato cultivars Norland, Kennebec, and Russet Burbank were inoculated with a Minnesota isolate of potato virus S (PVS) in the field. The incidence of secondary infection, measured by enzyme-linked immunosorbent assay (ELISA), indicated that significantly more PVS infection occurred in Russet Burbank than Norland or Kennebec, and that Norland had significantly more infection than Kennebec (P≤ 0.01). Mature plant resistance was not observed. Rapid reinfection of PVS-free seedlots resulted from inoculum point-sources when routine cultivation practices were followed. Reinfection rates were significantly greater for seedlots grown at Grand Forks, ND, when compared with seedlots grown at Becker, MN, during both years of the study (P≤ 0.01). After two years in the field, reinfection rates for Norland (71.8%) and Russet Burbank (73.0%) did not differ significantly, but were significantly greater than the reinfection rate for Kennebec (29.5%) (P≤ 0.01). In greenhouse-grown plants, PVS moved out of rub-inoculated leaves within 24 hr, however, 13 and 20 days were required before PVS was detected with ELISA in foliage above and below the inoculated leaf, respectively. The frequency of PVS detection was significantly greater in foliage above the inoculated leaf compared to foliage below the inoculated leaf (P≤ 0.05). Translocation of PVS from inoculated leaves to daughter tubers occurred within 13 days for Russet Burbank and Norland and within 20 days for Kennebec.  相似文献   

15.
A potato leafroll virus (PLRV) isolate has been successfully transmitted to and recovered from two wild crucifers,Sisymbrium altissimum L. (Jim Hill or tumble mustard) andCapsella. bursa-pastoris (L.) Medic. (shepherd’s purse) by the green peach aphid (GPA),Myzus persicae (Sulzer). Virus antigen in both plant species was found to be higher in root tissue than in foliar tissue, based on enzyme-linked immunosorbent assay (ELISA) determinations.C. bursa-pastoris was apparently a relatively poorer source of inoculum for the GPA thanS. altissimum. Using two geographically-separated biotypes ofC. bursa-pastoris, a Washington biotype was found to contain higher antigen titer in both leaf and root tissue than a California biotype, as determined by ELISA. Field studies demonstrated that both weed species can serve as overwintering sources of PLRV  相似文献   

16.
Field performance comparisons between ex vitro plantlet-derived (EVPD) plants and seed tuber-derived (STD) plants ofRusset Burbank were conducted for two consecutive years under a short growing season in Quebec. The EVPD plants produced more potential seed tubers than STD plants although total tuber biomass of EVPD plants was less than that of STD plants. The frequency and weight distributions of tubers formed by the two types of propagates were different. The EVPD plants produced significantly more small sized tubers and fewer large sized tubers than the STD plants. Average tuber weight was less for EVPD than STD plants for each of five size categories. No significant differences were found in the fresh and dry weight of plants, leaves and stems at harvest time. However, plant height and growth habit differed between the two types of propagates. The EVPD plants had a unique appearance. They produced a single stem with extensive axillary bud development. The EVPD plants were apparently more susceptible to early water stress in the field than STD plants, probably due to an earlier production of stolons. No differences were found between ex vitro plantlet- or microtuber-derived plants, in plant growth or yield characteristics.  相似文献   

17.
Using enzyme-linked immunosorbent assay (ELISA) and dotimmunobinding assay (DIBA) for potato viruses A (PVA), M (PVM), S (PVS), X (PVX), YN (PVYN), YO (PVYO) and leafroll (PLRV) and nucleic acid spot hybridization (NASH) for potato spindle tuber viroid (PSTVd), virus and viroid were detected reliably from single leaf discs (6 mm) of tissue-culture plantlets. Leaf discs taken from leaf positions (1 to 8) (bottom to top) can be used for detection of all viruses except PLRV where the lower leaves had higher concentrations of virus than the leaves from the upper part of the plantlet. Virus cultures were maintained for 1 to 4 years in several potato cultivars. The levels of virus remained reproducible except for PVM concentration, which was found to be very low in cv. Green Mountain. Using densitometry software, the DIBA spots were quantified and results were comparable to A405 values obtained by ELISA. PSTVd concentration as measured by densitometry from spots of NASH indicated no loss of viroid over 1–4 years in tissue culture in two potato cultivars.  相似文献   

18.
以马铃薯晚熟品种“下寨６５”为试验材料,研究了离体诱导微型薯直播田间繁殖块茎繁殖脱毒田间１～５代种薯的质量和效率。结果表明,生育阶段比同品种网室繁殖脱毒小薯同代脱毒种薯略迟;生长势、株高、茎粗、主茎分枝数、单株叶面积、根系数、匍匐茎数和匍匐茎长度等生长发育指标,ＰＶＸ、ＰＶＹ、ＰＬＲＶ和ＰＶＡ等重要病毒害发病率及病情指数,干腐病和软腐病发病率,晚疫病发病率和叶面枯死率相同;未发现环腐病和黑经病。单薯重量平均４５．１ｇ,随田间种植代数增加而增大;单株结薯数、单位面积结薯数逐代减小,脱毒田间１～４代单株产量和单位面积产量逐代增加,繁殖效率与脱毒小薯相同,故离体诱导微型薯田间繁殖块茎繁殖脱毒种薯是经济有效且切实可行的。  相似文献   

19.
对母本抗PLRV、父本免疫PVX、PVY按Line×tester (12× 3)设计获得的 36个家系 ,通过 1998～ 2 0 0 0年大田暴露试验 ,进行PLRV感染率、平均单栋结薯数量和重量的配合力及遗传分析 ,结果表明 :PLRV感染率的遗传力为 71 6 8% ,特殊配合力负值最高为 - 6 85 ,母本一般配合力负值最高为 - 14 87,父本一般配合负值最高为 - 3 13,选择一般配合力负值较高母本和特殊配合力负值较高的组合是获得高抗PLRV后代的前提 ;单株结薯数量的遗传力为 5 0 4 8% ,组合特殊配合力最高为 2 1,母本一般配合力最高为 2 8,父本一般配合力最高为 0 3,特殊配合力较高的组合在一般配合力高的和差的亲本组合中出现频率较高 ,父本在块茎数量的遗传中起重要作用。单株结薯重量的遗传力为 75 19% ,特殊配合力最高为 0 5 8,母本一般配合力最高为 0 4 2 ,而父本最高为0 33,产量的亲本选配应以一般配合力为主  相似文献   

20.
Detailed growth analyses were conducted with Russet Burbank potato to provide validation and parameterization data for a computer growth model being developed cooperatively at the University of Idaho and University of California, Davis. At each of two locations (Aberdeen and Kimberly, Idaho) plants were sampled seven times at about two-week intervals. Plants were separated into individual leaf blades, petioles, internodes, tubers, and roots. Leaf area indices and leaf elevation angles were determined in 20 cm strata. Plants at Kimberly emerged approximately 10 days prior to those at Aberdeen. At both locations, the increase in total dry weight was curvilinear for 30–40 days after emergence and then was linear until the end of the study. The maximum top weight per hill was slightly higher at Aberdeen, but near-maximum values occurred for much longer at Kimberly. Total leaf area at Aberdeen reached a maximum of approximately 4.0, while the maximum at Kimberly was 5.5. Root dry weight reached a maximum much earlier at Kimberly and the change in root dry weight paralleled that of tops in both places. Tuber initiation occurred approximately 19 June at both locations. Following an initial increase, tuber number/hill decreased for several weeks. Final tuber numbers per hill were approximately six at Aberdeen and 12 at Kimberly. Dry weight per tuber showed a curvilinear increase for 20–30 days and then was constant at 0.68 and 0.37 g/tuber.day at Aberdeen and Kimberly, respectively. Bulking rates were 17 and 21 g/m2.day. Final yields were 39 T/ha at Aberdeen and 49 T/ha at Kimberly. The higher final tuber yield at Kimberly was the result of a higher rate of bulking and a longer leaf area duration.  相似文献   

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