School of Agriculture, Food and Ecosystem Sciences - Theses
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ItemRegeneration and genetic transformation of Australian rice (Oryza sativa L.) varieties( 2001)The rice industry is a major contributor to Australia's agricultural production, which also puts Australia as one of the highest rice yield producing countries in the world. One of the priorities in Australian rice improvement program includes the generation of plants with useful qualitative and quantitative traits that affect agronomic performance and consumer preferences, which can be facilitated by genetic transformation techniques. An efficient tissue culture and regeneration system for four commercial varieties of Australian rice namely; Amaroo, Millin, Langi and Pelde are described. In this study, efficient plant regeneration via organogenesis was achieved in a short time, by optimising explant source as well as the composition of culture medium. MS medium containing BAP (2-4 mg/L) + NAA (1 mg/L) was found to be ideal for shoot initiation from mature embryo derived callus. Of the four varieties tested, Millin showed the best regeneration frequency followed by Amaroo, Pelde and Langi. The development of protocols for Agrobacterium-mediated transformation of Australian rice varieties was discussed. Using a binary vector, pIG121Hm, several parameters affecting Agrobacterium infection were optimised, including the choice of embryogenic calli; density of Agrobacterium; co-cultivation conditions including composition of medium, temperature and light, the presence of acetosyringone and the concentration of hygromycin in the medium. With the optimised protocol, transgenic rice plants were obtained in 3 to 4 months, with an average transformation efficiency of 11.6% and 2.0% for Amaroo and Millin, respectively. The plants grew normally and set seeds under glasshouse conditions. The integration of the transgenes was confirmed by Southern blot analysis. The optimised Agrobacterium-mediated transformation protocol was used to study expression of a pollen- and a generative cell-specific gene, Ory s 1 and LGC1, repectively in transgenic rice plants. Analysis of transgenic plants carrying Ory s 1- uidA (GUS) confirmed spatial and temporal expression of Ory s 1 in mature pollen. Deletion analysis (-405 bp and -812 bp) of promoter region of Ory s 1 gene (-1524 bp, full-length) showed that the 5' regulatory region contains enhancer/quantitative and pollen-specificity elements upstream of -405 bp and within the -405 bp regions, respectively. Further experiments using antisense construct resulted in reduction of Ory s 1 protein in two types of transgenic plants. Analysis of plants carrying LGC1- uidA (GUS) confirmed the spatial LGC] expression in generative cell of pollen. Whilst analysis of transgenic plants carrying each of the cytotoxic genes, barnase (Hartley, 1989) and diptheria toxin A (DT-A) (Greenfield et al., 1983) driven by LGC1 promoter showed generative cell-specific ablation caused by expression of barnase, resulting in arrested pollen development at late binucleate stage, reduced pollen size and lesser starch production, and 50-75% pollen sterility in transgenic plants. These studies showed that it is possible to transform Australian commercial varieties of rice. And the optimised transformation conditions can be used to introduce foreign genes for desired manipulation of important traits.
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ItemThe persistence and productivity of subterranean clover in southern Australia with special reference to rate of development in different cultivars( 1971)Subterranean clover, Trifolium subterraneum L. (commonly referred to as sub clover) has long been recognised as the key to improvement of annual pastures in southern Australia. Although the precise acreage of sub clover is difficult to determine, Donald (1970) has suggested that it may have been sown on as much as 80% of the present estimated area of 50 million acres (20 million ha) 0f sown pasture in this region (see Fig.1). This area (especially the western part of it) has a Mediterranean-type climate with mild wet winters and hot, dry summers. The total annual rainfall varies from about 8 inches (200 mm) to 40 inches (1000 mm) and the length of the growing season (the period during which rainfall relative to evaporation is sufficient to support plant growth) ranges from less than 4 months in the low rainfall areas of Western Australia to 10-11 months in some parts of south-eastern Australia. The species of which the pastures in this area are composed, are predominantly annuals. They become re-established each year following the autumn break, and flower and set seed during the spring prior to over-summering in the seed phase. The autumn break is on average later and the onset of the dry summer period earlier as the annual rainfall becomes smaller, but, in any one location, the year to year variation in the length of the growing period is considerable. In isolated areas reasonable constancy in length of growing period is achieved by autumn and spring irrigation. Pastures may be permanent, becoming re-established each year over an indefinite period, or temporary, in which case a pasture phase of one to several years alternates with a cropping phase, mainly cereals, of 1-3 years. Sub clover has proved to be well adapted to the conditions prevailing in southern Australia and it has played a key role in the pastures because of its capacity to fix atmosphere nitrogen. This together with liberal dressings of superphosphate which have been the rule has greatly improved the fertility status of the soils, thereby increasing the productivity of the pastures themselves and the crops grown in sequence with them. That sub clover had agricultural potential was first realised by Mr A. W. Howard in 1889 in the Mt. Barker district of South Australia, and his efforts to publicise it in the face of public indifference, have been well documented (Hill 1936; Davies 1951; Morley 1961; Symon 1961). There was a period of little progress. Then, through the efforts of many research workers, came an advance in knowledge of the plant and how it could best be used, and, with this, a tremendous increase in its use. There still exists opportunities for its greater use in new areas in southern Australia and for better use in some existing areas. This will require additional research to provide new cultivars and a better understanding of the interaction between genotype and environment. Matching of the genotype with the environment and the importance of this in determining the persistence and productivity of the species will be the main theme of this thesis. The thesis begins with a literature review in which variation within the species and how this is implicated in its widespread use in agriculture are the underlying considerations. This is followed by a report of work concerning the effects of various factors of the environment on the developmental physiology of the plant.
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ItemGrowth and yield of rice cultivars of differing growth duration in New South Wales( 1993)The growing season for rice in New South Wales is limited by low temperatures in Spring and Autumn. With current cultivars, almost all of the possible growing season is used in producing a crop. Cultivars with short growth duration have been sought by rice growers in southern New South Wales for reasons of decreased water use, increased flexibility in designing rotations and added time for rice field preparation in the event of a wet spring. This study examines the growth and yield of a short-duration cultivar compared with commercial long-duration cultivars, when subjected to a range of fertiliser applications and sowing times. Biomass accumulation of the short-duration cultivar was smaller than that of the long-duration cultivars, however yield potential was similar. This was achieved by the production of similar number of florets/m2 despite smaller biomass at flowering, and greater harvest index. Early sowing resulted in smaller yield due to restricted biomass accumulation and floret production of the short-duration cultivar. The short-duration cultivar had a greater proportion of filled grains and thus yielded more than the other cultivars when sown late. While yield of all cultivars was reduced with late sowing, the short-duration cultivar was affected least. Cultivars of this type are therefore recommended when late-sowing is unavoidable. Variation in yield was not due entirely to low-temperature damage at the critical growth stages of pollen microspore development and anthesis. Yield was also associated with growth after anthesis. A summary model of post-anthesis growth, which included the effect of low temperature on growth, predicted growth with an accuracy similar to that of the growth measurements. Thus, in determining yield, the importance of environmental conditions during grain filling was highlighted. It was concluded that the yield of short-duration cultivars may be less stable when stress occurs during grain filling, because there is less reserve available to fill the grain. Continued work on short-duration cultivars is advocated, with emphasis on faster growth to provide greater biomass production in the shorter vegetative stage. Incorporation of tolerance to low temperatures during the grain-filling stage is also suggested, to give greater yield stability.