The extraction and partial purification of a rhamnosidase from Colorado potato beetle larvae (Leptinotarsa decemlineata (Say))

An enzyme which hydrolyzed a terminal rhamnose moiety from α-chaconine was extracted and partially purified from Colorado potato beetle (CPB) (Leptinotarsa decemlineata) larvae. This ammonium sulfate precipitated enzyme fraction resulted in nearly 97% metabolism of α-chaconine after a 144 hour incubation period at 42 C as determined by high performance liquid Chromatographic (HPLC) analysis. Approximately 81% of the metabolized α-chaconine was detected asβ ₂-chaconine. The isolated enzyme fraction from CPB larvae exhibited no metabolism of α-solanine. Metabolism was also followed over time by thin layer chromatography (tlc).     

Performance of Colorado potato beetle larvae,Leptinotarsa decemlineata (Say), reared on synthetic diets supplemented withSolanum glycoalkaloids

Glycoalkaloids are a class of secondary compounds (nitrogenous, steroidal glycosides), ubiquitously distributed throughout the Solanaceae. Numerous studies (in planta) have shown that certain glycoalkaloids, e.g., α-tomatine, solanocardenine, and leptine, have a negative impact on performance of the Colorado potato beetle,Leptinotarsa decemlineata (Say). However, the presence of other secondary compounds, as well as the nutritional status of the plants used, are additional variables in such experiments. By rearing Colorado potato beetle, from egg to prepupal stage, on a synthetic diet supplemented with the glycoalkaloids α-tomatine, α-chaconine, α-solanine, leptine I and the steroidal aglycone solanidine, we have been able to further establish the effects (ex planta) of these compounds on the Colorado potato beetle. Leptine I displayed a dose-dependent negative activity against the Colorado potato beetle (as measured by larval weight gain and tune to molt), when assayed at 0.31, 0.62 and 1.23 mM concentrations; however, by the fourth stadium no effect was found. When Colorado potato beetle were fed a higher concentration of leptine I (2.4 mM), there was a sustained effect in all stadia. At 2.4 mM, leptine I displayed a greater negative impact on Colorado potato beetle growth and development than did α-tomatine. α-Chaconine at 2.4 mM did not impair Colorado potato beetle performance relative to Colorado potato beetle reared on control diets. α-Chaconine plus α-solanine, at concentrations commonly found inSolanum tuberosum L. foliage (0.6 and 0.3 mM respectively), did not impair Colorado potato beetle performance. The steroidal aglycone solanidine (2.4 mM) had a significant negative impact on Colorado potato beetle performance. Results are discussed in the context of host-plant resistance and insect-plant interactions.     

Ovariole number of Italian geographic populations of Colorado Potato Beetle (Leptinotarsa decemlineata Say)

Populations of Colorado Potato Beetle (CPB) have been collected in seven Italian locations. The number of ovarioles per ovary amounted to an average of 32.72±3.71, with a range of 21 to 41. These figures are consistant with the reports for CPB from North Eastern North America. The populations studied did not differ significantly among themselves suggesting that the character “ovariole number” is not sufficient to discriminate among CPB populations collected in different Italian areas.     

The number of ovarioles of the Colorado potato beetle,Leptinotarsa decemlineata (Say), in four North Eastern North American localities

The number of ovarioles of the Colorado potato beetle averages 32.04 ± 0.27 per ovary in North Eastern North America, seven more than previously reported for that area. It differs between the left and right ovary in 84% of beetles. The lack of significant differences among beetle populations from Maine, New Brunswick, New York and Ontario suggests that there is no geographical variation within the area studied. There could, however, be variation over a larger area: the number observed is similar to the one for Belgium and Poland but higher than the one for Russia.     

Use of reciprocal grafts to elucidate mode of resistance to Colorado potato beetle (Leptinotarsa decemlineata (Say)) and potato aphid (Macrosiphum euphorbiae (Thomas)) in six wildSolanum species

Reciprocal grafts were done between potato (Solanum tuberosum) and six wildSolanum species known to be resistant to the Colorado potato beetle (Leptinotarsa decemlineata (Say)). Potato or wildSolanum foliage from each reciprocal graft was fed to first instar beetles or adult potato aphids (Macrosiphum euphorbiae (Thomas)). The proportion of second instar larvae after 48 h and survival and fecundity (number of nymphs per female) of the aphids were used to i estimate the resistance level of the foliage. Survival and fecundity of potato aphid were reduced on wildSolanum foliage (except withS. circaeifolium ssp.capsicibaccatum andS. okadae) used as scion or rootstock, but were not different from the control (potato-potato graft) on potato foliage. Development of Colorado potato beetle onSolanum foliage was reduced. In some cases, potato foliage grafted to wildSolanum became resistant to the beetle. Results suggest that a chemical factor or factors providing resistance to Colorado potato beetle are translocated from foliage of some wildSolanum species to potato foliage. They also indicate that mode of resistance to the two insects differs quantitatively or qualitatively.     

Feeding behavior and survival of Colorado potato beetle,Leptinotarsa decemlineata (Say), larvae onSolanum berthaultii Hawkes and an F6 S. tuberosum L. XS. berthaultii hybrid

Newly hatchedLeptinotarsa decemlineata larvae were less likely to initiate feeding onSolanum berthaultii than onS. tuberosum. Among larvae that fed there was no difference in the time to initiation of feeding; however, the duration of the initial feeding bout was less onS. berthaultii than onS. tuberosum. Larvae spent less time feeding and more time walking onS. berthaultii than onS. tuberosum. All larvae caged onS. tuberosum fed within 24 hours, but larvae were less likely to feed on a F₆ S. tuberosum XS. berthaultii hybrid and least likely to feed onS. berthaultii. Lack of feeding during the first 24 h was highly correlated with mortality at 72 h. Whether or not larvae feed within the first five minutes of being placed on a leaflet is a reliable predictor of larval mortality at 72 h and is a useful test for determining the susceptibility of plants to feeding byL. decemlineata. The amount of trichome exudate accumulated on the tarsi over a 72 h period is correlated with resistance. Resistance ofS. berthaultii to neonateL. decemlineata is characterized by a refusal to initiate feeding and a reduced feeding rate by larvae that do feed.     

A Comparison of Resistance to Imidacloprid in Colorado Potato Beetle (<Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> Say) Populations Collected in the Northwest and Midwest U.S.

The Colorado potato beetle, Leptinotarsa decemlineata Say, is a serious pest of potato, Solanum tuberosum L., worldwide. Leptinotarsa decemlineata has a history of repeated adaptation to insecticides, and exhibits a geographic pattern of decreasing insecticide resistance from east to west in the USA. Imidacloprid is one of the most widely used insecticide in western states. In this study, we measured imidacloprid resistance among larval and adult L. decemlineata from ten locations in the Columbia Basin (southeastern Washington and northeastern Oregon) using topical LD₅₀ bioassays, and compared them to estimates from ten locations in Minnesota and Wisconsin. Larval and adult imidacloprid LD₅₀’s and mean percent mortality were generally lowest in Washington and Oregon, but some sites exhibited reductions in mortality comparable to those observed at some Wisconsin sites. Adult LD₅₀’s suggest L. decemlineata in the Columbia Basin may be evolving in response to selection by neonicotinoid insecticides, but larval data suggest high susceptibility to imidacloprid remains in most populations. Future work should expand resistance monitoring efforts to include more regions in the West and other insecticide modes of action.     

Evaluation of tuber-bearingSolanum species for symptomology,as diagnostic hosts,and sources of immunityto potato virus Y necrotic strain (PVYN)

A total of 258 Plant Introductions (PI) belonging to 69Solarium species were evaluated in the greenhouse for their reaction to the tobacco veinal necrosis strain of potato virus Y (PVY N). One hundred and thirty-one (50.7%) of the PI accessions produced mosaic symptoms ranging from mild to severe. Local lesion and veinal necrosis symptoms were observed in 19 PI accessions (7.3%) and a variety of other symptoms were observed in another 11 PI accessions (4.2%). Only 97 PI accessions (37.5%) were symptomless carriers of PVY^N. PI accession 473505 ofS. sparsipilum and PI accession 498021 ofS. brachycarpum developed local lesions and veinal necrosis with PVY^N, but necrotic spots and mosaic with PVY^o. Common mechanically-transmitted potato viruses A, S, M, and X did not interfere with PVY symptom development inS. sparsipilum andS. brachycarpum. Thus, PI 473505 and PI 498021 can be used as indicator plants for specific identification of PVY^N. PI accession 472819 ofS. chacoense developed local lesions with systemic spread in PVY^o, but without systemic spread in PVY^N. Thus, this can be used as a differential host plant for PVY strains. Two PI accessions ofS. stoloniferum, PI 160372 and 161171 were immune to PVY^N.     

Identification of late blight, Colorado potato beetle, and blackleg resistance in three Mexican and two South American wild 2x (1EBN)Solanum species

Wild potatoes are important sources of genes for resistance to disease and insect pests. A collection of wild Mexican and South AmericanSolarium species from the US potato Genebank was evaluated under laboratory and/or field conditions for their reaction to late blight (Phytophthora infestans), Colorado potato beetle (CPB,Leptinotarsa decemlineata Say), and blackleg (Erwinia carotovora subsp.atroseptica (van Hall) Dye) in order to identify individual genotypes with multiple resistance genes. Late blight inoculations using aggressive isolates (US-8/A2 and US-11/A1 mating types) of P.infestans revealed a wide range of variation for resistance between and within the accessions of the wild species tested. For late blight, susceptible as well as moderately to highly resistant genotypes were observed in all the species tested. However, at least one accession from the three Mexican and one South American wild diploid species tested showed a relatively uniform high level of resistance toP. infestans. These includedS. bulbocastanum, S. pinnatisectum, S. cardiophyllum, andS. circaeifolium. Two accessions from South American speciesS. commersonii were highly susceptible to late blight. For the Colorado potato beetle test, only one species,S. pinnatisectum appeared uniformly resistant to CPB under field conditions. Results of screening for blackleg resistance showed that there were major differences between genotypes in the wild species. Accessions ofS. circaeifolium PI 498119 andS. bulbocastanum PI 243504 were identified as having significantly higher blackleg resistance than cultivated potato and the other wild species tested. However, genotypes from these two accessions were more susceptible to late blight and CPB. Characterization of theP. infestans isolate P1801C.16 used for late blight evaluation and multi-locus isolate tests using US-8/A2 and US-11/A1 races revealed that the resistance inS. pinnatisectum genotypes tested corresponded to a race-non-specific genetic system, which was different from any existing R genes.Solanum pin-natisectum genotypes with both high levels of late blight and CPB resistance as well as blackleg resistance genotypes identified in the present study represent a diverse gene pool that may be useful for development of new potato cultivars with multiple disease and insect resistance. The potential utilization of these valuable sources for improvement of cultivated potato is discussed.     

Comparison of three methods for assessing varietal differences in resistance to common scab disease (Streptomyces scabies) of potato

Summary Consistent varietal differences in scab severity were obtained when plants were grown in an open field or under a polythene tunnel in soil naturally infested withStreptomyces scabies, or in a glasshouse in pots filled with artificially inoculated sand. It was concluded that the glasshouse pot-inoculation technique would be the most suitable method to use in a breeding programme in which there are many clones to be tested, because a large number of plants can be grown in a small area under closely controled conditions.     

Wild potato (Solanum sect.Petota) germplasm collecting expedition to Argentina in 1990, and status of Argentinian potato germplasm resources

Summary A joint Argentina/United States expedition collected wild potato (Solanum sect.Petota) germplasm in Argentina between February 4–28 and March 28–April 18, 1990. This was followed by a short expedition between February 27 and March 3, 1991. Collections were made in the Andes Mountains from southern Tucumán Province (27°20′S), south to southern Rio Negro Province (40°52′S) and comprised 88 lots of germplasm of ten species, 76 as true-seed and 12 as tubers. The most important new collections included germplasm ofS. brevidens, S. maglia, S. x rechei andS. venturii. By summarizing details of these collections with those of existing collections, we have been able to suggest future collecting in Argentina.     

The use of additives,temperature, and plant position to increase efficacy of dinoseb for potato (Solanum tuberosum) vine desiccation

Chemical vine desiccation is an important management practice for late season potatoes (Solanum tuberosum L.) in Michigan. A number of desiccants are registered; however, most Michigan growers apply dinoseb (2-sec- butyl-4,6-dinitrophenol). Greenhouse experiments were initiated to evaluate the potential of ammonium salts and ethephon (2-chloroethyl)phosphonic acid) to increase the efficacy of the chemical harvest aids dinoseb and endothall (7-oxabicyclo[2,2,1]heptane-2,3-dicarboxylic acid) on potato vines. Temperature and potato vine position were evaluated to determine their influence on vine desiccation. All treatments with endothall applied at 0.6 kg/ha resulted in unsatisfactory potato vine desiccation. Ethephon and ammonium sulfate increased efficacy of dinoseb. High temperature both before and after treatment contributed to greater dinoseb efficacy. The horizontal placing of potato vines resulted in the desired desiccation rating of 9.0 or greater using the combination of dinoseb at 2.8 kg/ha, plus ammonium sulfate at 9 kg/ha plus 2.4 L/ha X-77 surfactant.³     

Effect of thermal treatments on the chemical and biological value of irradiated and non-irradiated cowpea bean (Vigna unguiculata L. Walp) flour

Chemical and nutritional characteristics of irradiated and nonirradiated cowpea bean at 0.2 kGy submitted to several thermal treatments were studied. The cowpea bean flours irradiated at 0.2 kGy were superior to those non-irradiated concerning digestibility and the flours submitted to microwave oven were superior to those cooked under low pressure, autoclaved or even raw from the nutritional point of view. As to the presence of anti-nutritional factors, the results showed that the variety of cowpea bean under research presented neither trypsin inhibitor nor haemagglutinin activity, however, regarding tannin, a 0.006% content was found and considered negligible.     

Chemical,sensory and rheological properties of porridges from processed sorghum (Sorghum bicolor,bambara groundnut (Vigna subterranea L. Verdc) and sweet potato (Ipomoea batatas) flours

The chemical, sensory and rheological properties of porridges made from blends of sprouted sorghum, bambara groundnuts and fermented sweet potatoes were examined. Sorghum and bambara groundnuts were sprouted for 48 h while sweet potatoes were fermented for the same period. Blends were formulated from the processed ingredients in the ratio of 60:40:0, 57:42:1, 55:44:1 and 52:46:2 (protein basis) of sorghum, bambara groundnuts and sweet potatoes. Porridges were prepared from the composite flours and the traditional sorghum complementary food. Standard assay methods were used to evaluate the flours for nutrient composition. The porridges were also tested for sensory properties and viscosity. Processing increased the levels of most of the nutrients evaluated. Relative to the sorghum traditional complementary food, thecomposite flours had higher levels of lipids, protein, ash, crude fiber and minerals (p<0.05). The porridges from the composite flours were generally liked slightly by the panelists and were about seven times less viscous than the porridge from the traditional sorghum complementary food. Use of the composite flours, particularly the 52:46:2 blend, as a traditional complementary food should be encouraged in Nigeria especially with the increasing cost of commercial complementary foods.     

A simple and rapid method for the detection of potato spindle tuber viroid (PSTVd) by RT-PCR

Summary A test procedure for PSTVd is described based on immobilisation of plant sap on filter paper, by dotting or tissue printing followed by RT-PCR. Tests were carried out using primarily and secondarily infected potato plants, primarily infected in vitro plants, and potato tubers. Print PCR was shown to be suitable for testing large samples of potato plants whereas dot PCR is recommended for in vitro plantlets and tuber tissue. Bulking one infected plant to 4 or 9 healthy plants gave reliable results with secondarily infected potato plants, but sometimes the test failed to detect PSTVd in primarily infected in vitro plants. Dotted and printed paper squares could be stored at 4°C for at least 2 weeks in Triton X-100 solution or under dry conditions. Storing at room temperature can lead to unreliable results.     

Simultaneous detection of potato viruses Y,leafroll, X and S by DAS-ELISA technique with artificial polyvalent antibodies (APAs)

Summary A modification of the double antibody sandwich-enzyme linked immunosorbent assay (DASELISA) was used to try to detect simultaneously PVY, PLRV, PVX and PVS in both leaf and sprout samples, using artificial polyvalent antibodies (APAs). Each virus, singly or in a mixture, was detected with similar sensitivity using either specific antibodies or APAs. The APA for all four viruses is suitable for routine leaf testing, but only the APA for PVY+PVX+PVS is suitable for routine sprout testing.     

Evaluation of Allorhogas pyralophagus Marsh (Hymenoptera: Braconidae) for the biological control of Diatraea spp. (Lepidoptera: Pyralidae) in sugar-cane in Guyana

In 1989, a mass production and field release programme of Allorhogas pyralophagus Marsh (Hymenoptera: Braconidae) for the control of the Diatraea spp., particularly D. centrella (Möschler) (Lepidoptera: Pyralidae) in sugar-cane, was initiated by the Guyana Sugar Corporation. The programme was terminated in 1995 after the release of approximately two million adult parasites resulted in a mean field parasitism in Diatraea spp. larvae of only 0.22%. In contrast, 62% of Diatraea spp. larvae were parasitised in vitro as were 32% of larvae in young cane shoots exposed to A. pyralophagus in the laboratory. Neither Diatraea larvae in young cane plants caged in the field nor those recovered from Echinochloa polystachya (Kunth) Hitchc. and Paspalum virgatum L. around release fields showed any parasitism by A. pyralophagus. The reasons for the low rate of host acquisition outside the laboratory are not known, but it is clear that A. pyralophagus offers little prospect for significant control of Diatraea spp., particularly D. centrella in sugar-cane in Guyana.     

Resistance to gangrene (Phoma exigua var.foveata) and dry rot (Fusarium solani var.coeruleum) in potato tubers. 2. Evaluation of genotypic differences in pectin-bound magnesium and calcium for screening purposes

Summary The levels of incorporation of Mg and Ca into tuber cell walls and middle lamellae were compared with the intensity of attack byPhoma exigua var.foveata (P.e.f.) (gangrene) andFusarium solani var.coeruleum (F.s.c.) (dry rot) in six genotypes varying in their levels of resistance. The Mg:Ca ratio was higher in those genotypes more resistant toP.e.f., while the cation effect on attack byF.s.c was variable. Although genotype resistance may be improved by extra Mg bridges in the pectin structure the genotypic differences in susceptibilty to gangrene could not be explained by differences in Mg content assessed from alcohol-insoluble solids. Analyses of breeders' selections showed no general relationship between Mg or Ca contents and disease incidence. Therefore, genotypic differences in incorporation of these cations could not be used for screening for improved resistance.     

Economic implications of herbicide resistance and high labour costs for management of annual barnyardgrass (Echinochloa crus-galli) in Philippine rice farming systems

Implications of increasing labour costs and the development of herbicide resistance for profitable weed management in Philippine rice farming systems are investigated. The study employs RIMPhil (Resistance and Integrated Management in the Philippines), a bioeconomic simulation model developed to provide a comprehensive assessment of integrated weed management programmes for the control of annual barnyardgrass (Echinochloa crus-galli) in rice crops. Results indicate that herbicide application will become increasingly economically attractive, relative to manual weeding, as labour cost increases. This is important since urban migration in the Philippines continues to increase the scarcity of rural labour. Results also show that the onset of herbicide resistance results in substantial losses in farm profit. It is worthwhile for farmers to take management actions to prevent or delay the onset of herbicide resistance, provided that these changes are effective and not too costly. The study highlights the complexity of decision making about integrated weed management on rice farms in the Philippines.