School of Agriculture, Food and Ecosystem Sciences - Theses
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ItemThe activity of glyphosate and other herbicides in soil( 1989)The effects of herbicides on a legume Rhizobium symbiosis were studied in laboratory experiments. Root applications of all herbicides examined reduced nodulation of legumes grown in aqueous nutrient media. The growth of Rhizobium trifolii TA1 was reduced by 2 - 20 mg ai 1-1 of diquat, 2 mg ai 1-1 of paraquat, 10 mg ai 14 of glyphosate and 2 mg ail-1 of chlorsulfuron. No other herbicide affected rhizobia growth when applied at 2 - 20 mg ai l-1 of nutrient medium. Inoculation with TA1 pre-treated with amitrole, atrazine or glyphosate decreased nodulation of sub-clover plants indicating that these herbicides may affect the nodulation potential of certain strains of Rhizobium. Residues of 2,4-D, amitrole, diquat, trifluralin and glyphosate persisted in a Walpeup sandy loam in sufficient concentration for four months after application to soil to affect growth and symbiotic activity of sub-clover. The behaviour of glyphosate in soil, under various conditions was studied in the laboratory. Adsorption of glyphosate as depicted by Freundlich K constant was greater in an acid soil than in three alkaline soils and values for this constant ranged from 8 - 67.8 at 23.5C and 4.3 - 57.8 at 10C. Rate of decomposition of 14C-glyphosate at 25C decreased slowly over the experimental period in all soils. Two compartments of adsorbed glyphosate in soil were identified as labile glyphosate and non-labile glyphosate and these reflected the strength of adsorption of the chemical. The amount of glyphosate in the labile firm for the soils ranged from 24 - 34.5% of the total and half-life ranged from 6 - 9 days. The amount of glyphosate in the non-labile form for soils ranged from 67 . 75% of the total and half-life ranged from 222 to 835 days. At 10C, the two compartments of glyphosate adsorption were identified for the Walpeup and Rutherglen soils but only one compartment could be identified in the Wimmera and Culgoa soils. Methodology was developed to permit extraction and analysis of glyphosate and AMPA in soil. Recovery of glyphosate from soils where time between fortification and extraction was only 30 sec. was 84.6 - 104%. However where extraction was delayed 13 hours, recoveries were 47.6 - 66.8%. The extractant (0.1 M triethylamine) was shown to be unable to desorb adsorbed glyphosate. Studies revealed that at 25C, the pool of extractable glyphosate was rapidly depleted by decomposition. At this temperature, the pool of extractable glyphosate was supplemented by slow desorption of non-labile glyphosate for each soil. At 10C, depletion of the pool of extractable glyphosate was considerably slower. For the Walpeup and Rutherglen soils, the rate of desorption of glyphosate from the non-labile pool was less than the rate of loss by decomposition of the herbicide. Rate of desorption of non-labile glyphosate in the Wimmera soil was shown to be the same as the rate of loss by decomposition of the herbicide. Loss of extractable glyphosate in the Culgoa soil occurred by decomposition and by slow adsorption of extractable herbicide from the labile to the non-labile form. The effects of residues of glyphosate in the field following an autumn and a summer application were investigated at selected field sites. Following the autumn application, phytotoxic activity of glyphosate was observed in sub-clover plants growing at the Walpeup and Culgoa sites but not at the Wimmera site. Growth and nodulation of plants sown up to 4 weeks after herbicide treatment were reduced at the Walpeup site. Only nodulation of plants sown up to 4 weeks after treatment was reduced at the Culgoa site. Results suggest that residues of glyphosate are only likely to significantly affect the growth of susceptible plants during winter on sandy soils. Following summer application of glyphosate, no phytotoxic activity of the herbicide was observed for sub-clover plants grown in the Walpeup sandy loam. Results suggest that in a hot summer, it is unlikely that residues of glyphosate in any soil would cause significant damage to plant growth.
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ItemStudies of barley yellow dwarf virus infection in ryegrass (Lolium spp.)( 1989)Measurements were made of the effect of barley yellow dwarf virus (BYDV) infection on the early growth of four commercial cultivars of ryegrass (Lolium spp.) under two different temperatures (24C and 16C). At 24C, BYDV infection was associated with reduced root dry weight (30-40%) in all cultivars; the effect of infection on shoot dry weight and leaf area was variable. At 16C, the effect of BYDV infection was variable, being associated with increases in both leaf area and root and shoot dry weight in one cultivar (Grasslands Arild) and decreases in another (Victorian). In two other cultivars, both leaf area and root and shoot dry weight were not significantly affected (p > 0.05) by infection with BYDV. There was no evidence of visible symptoms associated with BYDV infection. At 240C, reductions in root dry weight associated with BYDV infection were not reflected in reductions in the relative growth rates of the roots. Up to at least 28 days after inoculation (46-50 days after germination) reductions in root dry weight were associated with both aphid-feeding damage and virus infection. Experiments with the cultivar Victorian showed that shoot dry weight was not significantly affected (p > 0.05) by feeding with either viruliferous (BYDV) or non-viruliferous aphids (Rhopalosiphum padi L.). At 16C, changes in root and shoot dry weight were associated with changes in the respective relative growth rates. Both visual scoring and changes in plant dry weight were found to be unreliable criteria for assessing disease severity. Two assay systems, an enzyme-linked immunosorbent assay (ELISA) and a nucleic acid, or complementary DNA (cDNA) probe, were developed for the detection of BYDV in ryegrass. The conditions for detecting BYDV with the ELISA (the "double sandwich" variant) were optimized. Exposure of the virus to extracts of ryegrass leaf tissue for 30 min or more resulted in reductions in absorbance, with the amount of the reduction dependent on the antigen (virus) incubation temperature. The optimization of conditions for the immunoglobulin (IgG) used for coating microtitre plates (coat IgG), the IgG - alkaline phosphatase conjugate (conjugate IgG) and the antigen (virus), in conjunction with the use of polyethylene glycol - 6 000 in the preparation of the conjugate buffer, gave a rapid and reliable ELISA procedure for the consistent detection of BYDV at levels of 2 ng/ml. Both ELISA and cDNA probe techniques were used to study the incidence of BYDV isolates in perennial ryegrass at three sites (Balmoral, Mininera and Hamilton) in south-western Victoria. BYDV was detected in perennial ryegrass cultivars from the three sites, with different levels of incidence of BYDV isolates between cultivars. While the PAV-related isolate was prevalent at the three sites, the incidence of the MAV- and RPVrelated isolates varied with the site and the cultivar. One difficulty with the ELISA was that absorbance values did not fall in to distinct infected and uninfected groups within the cultivars, and at Mininera, the incidence of BYDV varied significantly between the various positive - negative ELISA thresholds that were used. At Balmoral, the incidence of BYDV determined with each threshold was compared with that determined by cDNA probing using a Spearman correlation test. A threshold of double the mean negative (uninfected ryegrass) control showed the highest correlation with the cDNA probe for a number of replicate plots at the site. The incidence of the PAV-related isolate was compared for the two cultivars Ellett and Victorian, and significant (p < 0.01) reductions in its incidence in cv. Ellett compared with cv. Victorian at the three sites, suggested that cv. Ellett is resistant to the PAVrelated isolate of BYDV. The comparative resistance of cv. Ellett to at least this isolate of BYDV indicated the feasibility of breeding and selecting for resistance to other isolates of BYDV in perennial ryegrass. In attempts to quantify resistance in perennial ryegrass, the ELISA was further developed to allow measurement of BYDV titre in ryegrass shoot tissue. In twenty selections of cv. Victorian, the performance, as measured by changes in both root and shoot dry weights, of individual selections to infection with BYDV, did not significantly (p < 0.05) correlate with virus titre measurements. Experiments with twenty selections of cv. Ellett indicated that its resistance to BYDV that was e' ident at the three field sites was not associated with resistance to aphid (Rhopalosiphum path) feeding, and consequently, transmission of the virus. Attempts to inoculate cv. Ellett with either the PAV- or RPVrelated isolates were of limited success, and it was speculated that its resistance to the virus may be due to reductions in the level of virus replication.
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ItemStudies on the ecology and control of some important plant parasitic nematodes in Victoria( 1986)Since 1965 I have been employed as a nematologist by the Victorian Department of Agriculture and Rural Affairs, based at the Plant Research Institute, Burnley. My major responsibilities have included: (1) From 1965-1975, funded exclusively by the Wheat Industry, to conduct research on the biology and ecology of the cereal cyst nematode (Heterodera avenae Woll.), and to advise on methods for controlling the disease it causes in wheat and other cereals. (2) From 1975, in addition to research on H. avenae, to conduct research on nematode diseases of other agricultural and horticultural crops in Victoria. My research has always been directed at achieving methods of control which are both practical, and economic to apply. Prior to 1965, cereal cyst nematode was recognised as a major cause of disease in cereals in the southern wheatbelt of Australia; its distribution was known to be related to well structured soils; knowledge of its biology and ecology under Australian conditions was limited; and, crop rotation was the only recommended method of control. There were no sources of resistance suitable for use in breeding programs; the existence of pathotypes was unknown; and, the extent and magnitude of yield losses had not been determined. My research has culminated in the adoption by cereal growers, of several new control strategies, and the results have lead to the establishment of a new pesticide market (valued at millions of dollars per year), and economic benefits from the use of nematicides are already being obtained by rural communities in Victoria and South Australia. The results of this, and other research on various aspects of the biology and control of nematodes causing disease in grapevines, citrus, pastures, vegetable crops, ornamentals, etc. are presented in the papers which follow.