for virus isolation and purification, 1 batch of prawns yielded hemolymph fractions dominated by a previously undescribed non-occluded baculovirus rather than YHV. Injection of test shrimp with a semipurified preparation of this virus gave rapid mortality, and examination with the transmission electron microscope revealed a dual infection where cells containing the new virus dominated, but some cells containing YHV could also be seen. The tissues infected by the 2 viruses were similar. However, in contrast to YHV, the new virus was assembled completely in the nucleus and in the absence of occluding protein (polyhedrin). By normal histology, the most characteristic feature of infection was eosinophilic Cowdry A-type inclusions in hypertrophied nuclei with marginated chromatin, especially in epithelial cells of the stomach. These intranuclear inclusions became lightly basophilic in late stages of infection. In the epithelial cells of the gills, ultrastructural pathology included nuclear hypertrophy and cytoplasmic disintegration leading to large voids at lysed cell sites. By negative staining, completely assembled, enveloped virions were ellipsoid to obovate with a distinctive multifibrillar appendage and they measured 276 x 121 nm (excluding the appendage). Enveloped and unenveloped nucleocapsids were significantly different In size, indicating posslble shortening and thickening of the viral core and nucleocapsid during viral assembly. Isolation and punficat~on of the nucleic acid from the new virus yielded double-stranded DNA of approximately 168 lulo base pairs. This DNA did not cross-hybridize with DNA fragments isolated from YHV-infected shrimp or from monodon baculovirus (MBV). The features placed t h~s virus in the family Baculoviridae, subfamily Nudibaculovirinae as PmNOBII, but for convenience we have named it informally as Systemic Ectodermal and Mesodermal Baculovirus (SEMBV).
Subadult Penaeus monodon shrimp were sampled from 4 farms in Queensland, Australia. Histological observations on lymphoid organs showed the formation of abnormal cell foci, which resembled tubules lacking a central hemolymph vessel. These formations contained hypertrophied nuclei with marginated chromatin, vacuolated cells and inclusion bodies which stained positive with Feulgen's reaction. Male and female F! monodon broodstock were sampled from 1 farm and showed the same abnormal histological features within the lymphoid organ as the subadults. Electron microscopy revealed tightly enveloped, cylindrical particles, 163-200 nm X 36-63 nm, packed into paracrystalline arrays within the cytoplasm and infrequently within the nucleus. Arrays of virions were also observed within the cytoplasm of broodstock gill cells. Cross-sections of the particles revealed electron dense nucleic acid cores and envelopes. Nucleocapsids, 83-590 nm X 13-15 nm, were seen free and in vesicles within the cytoplasm. A negatively stained preparation of partially purified lymphoid organ revealed a nucleocapsid with one conical end. Lymphoid organ virus (LOV) resembles yellow-head virus (YHV) from Thailand in its morphology and cytopathology and rhabdovirus of penaeid shrimp (RPS) from the Americas in its cytopathology.
Stably transformed sugarcane plants were produced by the biolistic introduction of DNA into tissue-cultured cells. Constructs containing genes in sense and antisense orientation of polyphenol oxidase and sense orientation of sucrose phosphate synthase were used in the transformations. Regenerated plants were grown in a series of field experiments that incorporated commercial varieties, including Q117, from which the transgenic clones were derived and plants regenerated from tissue culture but not subjected to biolistic bombardment. In all experiments, the mean yield of transgenic sugarcane was lower than commercial varieties and the transgenic clones often exhibited lower sugar content, although individual transgenic clones in some experiments were not significantly different from Q117. Those plants regenerated from tissue culture but not bombarded were intermediate in their yield, and more clones were equivalent to Q117 in agronomic performance. Transformed plants produced by the bombardment of callus performed poorly but the results from the tissue-cultured controls indicated that not all of this could be due to somaclonal variation. Some aspect(s) of the process of transformation itself was deleterious and in most cases more significant than the effects due to tissue culture. Of the transgenic clones grown at Ayr, Queensland, 1.6% were equivalent to Q117 in sugar content and yield, suggesting that large numbers of transgenic clones would have to be generated using the current method in order to allow for selection of clones with acceptable agronomic performance.
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