Varieties, plant breeding and release

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Construction of synthetic Fiji virus resistance genes for use in sugarcane : SRDC final report BS86S
(1998) Smith, GR; Handley, JA; Dale, JL; Harding, RM
Fiji disease of sugarcane, caused by Fiji disease fijivirus (FDV), is one of the most important diseases affecting sugarcane in Australia. FDV is a member of the reovirus family and has a multipartite genome consisting of ten segments of double stranded (ds)RNA ranging in size from 1.8 to 4.4 kilobasepairs (KBP). The total genome size of FDV is approximately 30 kbp. Approximately 80% of the fdv genome has now been clones and sequenced. The majority of the FDV segments characterised to date encode a single protein product, indicated by the presence of a single open reading frame (ORF). Two of the segments, 7 and 9, were found to contain two ORFs each and hence encode two proteins. The predicted functions of segments 1, 3, 5, 7, 8, 9 and 10 have been assigned based on homology to equivalent segments in related reoviruses, and\or protein expression studies. A construct containing ORF 1 from segment 9 has been prepared and used to transform the sugarcane cultivars, Q117 and Q124. Transgenic plants have been produced for each of these cultivars and are currently being prepared for challenge experiments with FDV.
Meristem transformation for sugarcane genetic engineering : SRDC final report BSS209
(1999) Smith, GR; Lakshmanan, P; Elliott, AR; Grof, CPL
Maintaining access to foreign germplasm by developing methods to detect unidentified viruses in sugarcane in quarantine : SRDC final report BS116S
(1998) Braithwaite, KS; Smith, GR
Four types of gneeric tests for detecting unidentified viruses in plants were trialed for their suitability for sugarcane. The tests were; double stranded RNA (dsRNA) analysis, sap inoculation of indicator plants, sap examination by electron microscopy and polymerisation chain reaction (PCR) amplification using 'group-specific' primers. The aim of this project was to develop and implement general diagnostic tests for unidentified viruses in sugarcane germplasm held in quarantine. The availability of these general tests has provided an opportunity to significantly improve the security of sugarcane quarantine.
Production and evaluation of SCMV resistant transgenic sugarcane pests derived from transformed callus : SRDC final report BS94S
(1997) Smith, GR; Joyce, PA
We have successfully demonstrated that sugarcane can be genetically engineered for resistance to sugarcane mosaic virus, using the coat protein gene of the virus to mediate resistance. At present the frequency of plants showing moderate to high transgene-mediated resistance to viral infection is low, but the second generation of constructs with stronger promoters should address this issue. Transgenic sugarcane plants of cultivars Q95, Q153 and Q155 that contained a sugarcane mosaic coat protein gene construct under control of the Emu promoter were produced. These plants were regenerated from embryogenic callus which had been co-transformed by microprojectile transformation with the coat protein gene construct and the neomycin phosphotransferase II (NPT II) selection gene. The plants were established in the PC2 glasshouse at the Pathology Farm, Eight Mile Plains and were then mechanically inoculated with an inoculum of sugarcane mosaic virus prepared from infected sugarcane grown at the Pathology Farm. One transgenic line of Q155 did not develop mosaic symptoms over the time course of the trial, while another 6 lines of cultivars Q95, Q153 and Q155 showed a delay in the development of symptoms. Molecular analysis of these lines by PCR indicated the presence of both the NPT II and CP genes. Further analysis of selected lines by Southern blot hybridisation indicated the presence of one to three copies of the CP gene.
Selection of a non-sugarcane gene for control of canegrubs : SRDC final report BS95S
(1996) Allsopp, PG; McGhie, TK; Shepherd, K; Smith, GR
Four types of non-sugarcane genes were investigation for potential toxictiy to canegrubs. They were: toxins from the bacterium Bacillus thuringiensis (Bt); plant proteinase inhibitors; plant lectins; and avidin. Each of these was tested in an artificial diet based on the wheatgerm and casein and developed within the project.A Bt toxin with toxicity to canegrubs was not identified. The Bt isolates tested, which had activity against New Zealand grassgrub, bound to the gut lining of larvae of Antitrogus consanguineus. This binding proved to be not sufficient for toxicity.
The production of transgenic sugarcane plants : SRDC final report BS44S
(1994) Smith, GR
Transgenic sugarcane plants, which expressed the coat protein of SCMV at a very low level, were produced by micoprojectile bombardment of sugarcane meristems. Transgenic plants expressing the luciferase (Luc) and B-glucuronidase (GUS) reporter genes were also produced. A paper describing regeneration of plantlets expressing the reporter gene GUS (Plant Cell Reports 12:343-346) was the first report of the use of sugar meristem tissues as a transformation target, and the second report of transgenic sugarcane plants. The level of expression of the coat protein gene in the regenerated plantlets was very low, possibly due to chimaerism, ie. mixtures of transformed and non-transformed cells in the same tissues. We have established that sugarcane meristems are a useful target for microprojectile transformation of sugarcane, although more research is necessary before this target can be routinely used for sugarcane genetic engineering.

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