Four sampling techniques were evaluated to estimate clover aphid, Nearctaphis bakeri (Cowen), abundance in red clover fields in the Willamette Valley. The Berlese funnel method detected one to four times more aphids than other sampling techniques. Visual assessment often underestimated the high aphid densities during the flowering and seeding of clover. Sweep net and Schuh shaker techniques had limitations and underrated the numbers of aphids present in the clover fields. Strong correlations (r² = 0. 74 - 0.87) were found between the Berlese funnel technique and visual assessment technique in a 2nd year established field. Numerical differences in the spatial distribution of N. bakeri on clover stems were shown during various stages of plant development over the season. During development of buds and axils, aphid numbers on the lower half of the plant averaged 1.5 times greater than those on the upper half. As the season progressed into the flowering and seeding stage, aphid numbers on the upper stem halves were approximately five times greater than those on the lower half. Winged clover aphids were too few to show definite peaks of flight using water traps, averaging 2.19 aphids per yellow bucket during the June-August period. No significant differences in aphid attractiveness to the different colors were found among water pan traps. Yellow buckets captured approximately five times more alate aphids than did yellow, red, or green pan traps. Traps placed on the south side of the field contained higher numbers of aphids than in other locations in the red clover fields. / Graduation date: 1991

Some mounted ill. Bibliography: leaves 156-186.

Some mounted ill. / Bibliography: leaves 156-186 / ix, 186, 43 leaves, 35 leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Pathology, Waite Agricultural Research Institute, 1984

To make use of the information on infestation estimates of forage insect pests obtained by the conventional methods of sampling, it is important to estimate the actual losses taking place in association with definite numbers of the insect. Six month old red clover, alfalfa, and birdsfoot trefoil plants were exposed for a month to spittlebug infestations of 0, 50, 100, and 150 adults. Although the spittlebug mortality was lowest when reared on alfalfa and highest on birdsfoot trefoil, yellowing appeared on alfalfa and birdsfoot trefoil. On red clover there was a consistent increase in protein and nitrogen free extract and a reduction in crude fiber and ash content indicating slight reduction in growth. None of these differences were significant. On alfalfa the loss of xylem sap due to feeding caused a consistent general reduction of proteins, fat and ash content and an increase in dry matter and nitrogen free extract. Of these constituents only the increase in dry matter was significant with an infestation of 150 spittlebugs per cage. No differences in dry matter, proteins and fat were significant. Another test on 4 month old alfalfa with infestation levels of 0, 100, 200 and 300 showed similar consistent patterns but the difference in dry matter was significant only when the infestation was 200 spittlebugs or more. On birdsfoot trefoil there was a consistent increase of dry matter, proteins, and ash and reduction of fat and crude fiber. Only the difference of dry matter and fats was consistent showing that an infestation of 100 or more spittlebugs per sq ft will affect the general growth of the plants. The survival of 1st instar nymphs of the pea aphid (a phloem feeder) was very poor on birdsfoot trefoil. There was a fair amount of survival on red clover and those on alfalfa survived fully. There was no visible symptoms on birdsfoot trefoil and red clover but the alfalfa showed retarded growth. On red clover there was a consistent increase of dry matter, fat, crude fiber and nitrogen free extract but a reduction of proteins. The difference in crude fiber was significant when the infestation was 1000 pea aphid nymphs or more. A consistent reduction of proteins and crude fiber was also recorded on alfalfa but these differences were not significant. On birdsfoot trefoil, there was a decrease in dry matter and an increase in fat and ash content. The difference in ash content was significant when the infestation of pea aphid nymphs was 1000 or more. Potato leafhopper (phloem feeder) injury was very characteristic on birdsfoot trefoil. The terminal 1 to 2 inches of the sterns became wilted and bent down. On alfalfa there was gradual yellowing and the retarded growth was evident. The red clover did not show any characteristic symptoms. Infestations of 20 or more caused a consistent increase in protein, fat, crude fiber and a reduction of nitrogen free extract. On alfalfa there was consistent decrease in dry matter. On birdsfoot trefoil, there was consistent increase in dry matter, fat, and nitrogen free extract but a reduction of crude fiber showing that feeding on birdsfoot trefoil probably retards growth and condensation of carbohydrates to form crude fibers. The increase in fat was significant when the infestation was 40 potato leafhoppers or more. Alfalfa weevil adult feeding caused characteristic stripping of leaves, girdling of stem and drying of the entire plant. Feeding of 10 or more weevils for 18 days on a two month growth of alfalfa after the 1st cutting, cause a consistent reduction of proteins, fat and crude fiber and an increase of dry matter, ash, and nitrogen free extract. Increase of percent dry matter was significant when the infestation of alfalfa weevil adults was 20 or more. In another test, the dry matter, fat and ash gave similar consistent results, when 40, 60, and 80 adults were fed on 4 month old alfalfa for 18 days. Reduction of percent fat was significant when the infestation was 40 or more alfalfa weevil adults. Alfalfa weevil larval feeding showed stripping of the leaves and buds. Infestation of 30 or less, 2nd to 4th instar larvae for 12 days caused a consistent reduction of fat and ash and an increase of dry matter, proteins and crude fiber. The infestation was not high enough to show a significant change in plant constituents. / Ph. D.

Small broomrape, a holoparasitic weed, is a relatively new weed introduction in the Pacific Northwest that has contaminated a limited number of red clover fields in Oregon. Greenhouse and field studies were conducted to evaluate small broomrape response to common crop and weed species in the Pacific Northwest. Host species in the greenhouse or field study included alfalfa, arrowleaf clover, carrot, celery, common vetch, crimson clover, lettuce, prickly lettuce, red clover, spotted catsear, subterranean clover, white clover, and wild carrot. False-host species included barley, birdsfoot trefoil, creeping bentgrass, cucumber, field corn, fine fescue, flax, Italian ryegrass, nasturtium, oat, orchardgrass, perennial ryegrass, snap bean, sugar pea, sunflower, sweet corn, tall fescue, tomato, and wheat. Non-host species included sugar beet and curly dock. The greenhouse polyethylene bag system provided a rapid and inexpensive screening for plant species host status to small broomrape. Germination and attachment to host roots are initiated by chemical exudates, that may change concentration in response to nutrient availability and microorganisms. Red clover was grown in varying concentrations of ammonium sulfate fertilizer with and without Rhizobium inoculation, and with small broomrape seeds. Neither Rhizobium inoculation nor ammonium concentration influenced the number of small broomrape attachments to red clover roots. A survey was conducted of red clover seed growers with small broomrape-contaminated fields in the Pacific Northwest. Red clover seed from six respondents were cleaned at the same cleaning facility, and the same respondents purchased their seed stock from this cleaning facility. Small broomrape was not identified in red clover fields prior to or during the first clover seed harvest of fall planted red clover in small broomrape-contaminated sites. / Graduation date: 2003

Trifolium L. (clover) are annual or perennial species established in pasturelands to improve forage productivity and quality. In the southeastern United States, Trifolium repens L. (white clover) and Trifolium pratense L. (red clover) are important species, susceptible to virus infection. Objectives of this research were to isolate bean yellow mosaic (BYMV), clover yellow vein (CYVV) and peanut stunt (PSV) viruses from naturally infected white and red clovers from different locations in Virginia; and, to compare Indirect Enzyme-Linked ImmunoSorbent Assay (i-ELISA) and tissue immunoblot assay (TIBA) as methods for virus detection. A total of five white clover samples from Augusta, Richmond and Washington Counties were positive against CYVV antiserum and four white clover samples from Augusta County were positive against PSV antiserum. Single red clover samples from Frederick and Montgomery Counties were positive against BYMV antiserum. There were notable differences in host range with samples that tested positive for CYVV and BYMV, indicating they may be different strains. PSV was evenly distributed in the plant, whereas CYVV was higher in older plant parts. Viruses were successfully detected by blotting leaf samples directly onto membranes, thereby simplifying the sample preparation step. A number of membranes, such as nitrocellulose, nylon, chromatography paper, filter paper and writing pad could be used to detect viruses. In terms of specificity, immunoblots were equal or superior to i-ELISA. The TIBA should be useful in support of breeding and plant pathology studies as it is simple and rapid, and is less laborious and less expensive than i-ELISA. / Master of Science

Experiments were conducted to investigate effects of endophyte (Acremonium coenophialum Morgan-Jones and Gams) infection of tall fescue (Festuca arundinacea Schreb.) on grazing cattle and subsequent feedlot performance, serum minerals and prolactin, intake, digestibility, bite size, and biting rate. Grazing animal performance was measured April to October, 1989 and 1990, at two locations in VA. Effects of grazing low (<5%) and high (>70%) endophyte infected tall fescue, with and without white clover (Trifolium repens L.), were studied with yearling steers. At each location, two replicates were used with a stocking rate of .3 ha/steer (116 steers/yr). At the end of grazing, all animals were finished on a high corn silage diet and slaughtered. Daily gains of steers grazing low endophyte-infected pastures were higher (P<.01) than for those grazing high endophyte-infected pastures. Cattle grazing high endophyte-infected tall fescue with clover at Glade Spring gained at the same rate as those grazing the low endophyte-infected tall fescue without clover during Summer, 1989. During 1990, cattle at Glade Spring had higher (P<.05) daily gains than did those at Blackstone. Rectal temperatures were .5 to 1 °C higher (P<.05) for steers grazing infected pastures by June of each year at both locations. Prolactin concentrations were lower (P<.01) in cattle grazing endophyte-infected fescue within 28 d of initiation of grazing at both locations. Visual evaluation indicated that steers grazing high-endophyte pastures had rough hair coats compared to those grazing low endophyte pastures (P<.01), showing evidence of tall fescue toxicosis. Serum mineral analyses indicated no consistent influence of the endophyte. During feedlot finishing, there were no detrimental effects from previously grazing endophyte-infected tall fescue on animal performance. Forage DM yield, and CP in 1990, and IVDMD in 1989 and 1990 from Glade Spring were higher (P<.05) compared to Blackstone. Forage from Blackstone was typically higher in NDF, ADF, and lignin compared to Glade Spring. Forage from Glade Spring was higher in Ca, Mg, P, and Cu, compared to Blackstone. Diet selection, digestibility, biting size, and biting rate were measured with esophageally-fistulated steers grazing low (<5%) and high (>70%) endophyte-infected tall fescue, with and without white clover. Bite size was smaller (P<.05) in July compared to May, corresponding to decreased quantity of forage available. Bite size and biting rate were not affected by endophyte infection or inclusion of clover. In vitro DM digestibility was higher (P<.05) for high endophyte-infected tall fescue in July. Animals consumed a higher percentage of clover than was available in the sward. Intake and digestibility were estimated using 48 experimental animals and four esophageally-fistulated steers. Chromic oxide sustained-release boluses and forage IVDMD were used to estimate intake. The chromic oxide boluses were determined to be reliable for predicting intake of steers during a 21 d trial involving total fecal collection. Intake was higher (P<.01) in cattle grazing low endophyte-infected tall fescue compared to animals grazing high endophyte-infected fescue. Twenty-one Angus steers (256 kg) were used to compare the effect of 0 and 40% endophyte-infected tall fescue and orchardgrass hay on DM intake, daily gain, body temperature, serum minerals, and prolactin concentrations during an 8 wk study. Quality of the diets was low, as indicated by low CP (8.1 to 10.3%), and IVDMD (41.3 to 47.1%). Daily DM intake and daily gain were higher (P<.05) for animals consuming the noninfected tall fescue compared to animals fed the other hays. Body temperature and serum prolactin were not affected by diet. These data suggest that other factors, besides endophyte infection, are important in the etiology of tall fescue toxicosis. Some of the decreased performance of steers grazing endophyte-infected tall fescue was related to lower intake. Additional research is needed to evaluate higher levels of clover to possibly ameliorate some of the problems of tall fescue toxicosis. / Ph. D.

No description available.

Characterization of the most frequently isolated virus found during a survey of eastern Ontario red clover fields, which was designated as the Ottawa isolate, was undertaken. Determination of the biological and physico-chemical properties of the virus showed that the virus studied differed in some respects from other WCMV isolates. An ultrastructural examination of virus localization of cytopathological changes in infected red clover and pea plants revealed various types of viral inclusions, including a membrane-bound mass of tubules, not usually associated with infection by potexviruses, in red clover leaves, and severe organelle disorganization within pea leaf tissues. WCMV infection significantly reduced several processes relevant to symbiotic nitrogen fixation, including plant growth, nodulation, nitrogenase activity, leghemoglobin content, and Rhizobium population, but nitrate reductase and acid phosphatase specific activities were increased. The correlation between leghemoglobin content and nitrogenase activity shown at ten-leaf and pre-blooming stages was absent during the flowering and senescence stages. Infectivity assays, immunosorbent electron microscopy, and an ultrastructural examination showed the presence of the virus in nodular tissues; the cytopathological changes seen in bacteroids suggested that virus infection accelerated nodule senescence. Application of a cytochemical technique using diaminobenzidine showed that the leghemoglobin was located in the peribacteroidal space.