The total protein, carbohydrate, lipid and ash compositions, and fatty acid contents of two species of marine microalgae, the eustigmatophyte Nannochloropsis oculata (formerly 'Chlorella sp., Japan') and the chrysophyte Isochrysis sp. (Tahitian) used in tropical Australian mariculture, were studied. The microalgae were grown under a range of culture conditions (4 1 and 60 1 laboratory culture, 300 1 bag culture, and 8000 1 outdoor culture) and four light regimes (100 to 107 ktE m -2 -, 240 to 390 E m-2 s -', 340 to 620 iE m-2 s -1, and 1100 to 1200 ,tE m-2 s-1 respectively) to determine the effect of light intensity on the chemical composition of large scale outdoor cultures. Laboratory and bag cultures were axenic and cultured in Walne medium while outdoor cultures were grown in a commercial medium designed for optimum nutrition in tropical outdoor aquaculture operations. Change in growth medium and photon flux density produced only small changes in the proximate biochemical composition of both algae.
Phytoplasmas were found in 33 plant species that were not described as host
plants in an earlier Australian survey. Plants displayed characteristic
symptoms of little leaf, proliferation, and floral abnormalities. Restriction
fragment length polymorphism analysis revealed 13 different restriction
patterns. The majority of phytoplasmas showed a restriction pattern identical
to that of either the tomato big bud (TBB) or sweet potato little leaf V4
(SPLL-V4) phytoplasma. Phytoplasmas from 6 plant species showed a restriction
pattern similar to that of the pigeonpea little leaf (PLL) phytoplasma. One
phytoplasma from garden bean displayed a restriction pattern identical to that
found in papaya dieback and Australian grapevine yellows (AGY) phytoplasmas.
Seven new restriction fragment patterns have been detected and sequence
analysis of the 16S/23S spacer region revealed that 3 of these
phytoplasmas are related to the faba bean phyllody (FBP) group. The spacer
region of a graminaceous phytoplasma was most similar to phytoplasmas from the
sugarcane white leaf group. Another graminaceous phytoplasma was identical to
a phytoplasma from Indonesia. The spacer region of a phytoplasma from
poinsettia (PoiBI) was identical to the western X-disease phytoplasma from
North America and Europe. The spacer region of a phytoplasma in stylosanthes
contained no tRNAIle. Full-length 16S rRNA gene
sequences from selected new phytoplasmas were determined to corroborate
results obtained from the spacer region analyses. Three of these phytoplasmas
(galactia little leaf, vigna little leaf, and stylosanthes little leaf) are,
along with the PoiBI phytoplasma and the graminaceous phytoplasmas, members of
phytoplasma groups that have not been reported before in Australia.
A diagnostic test using the polymerase chain reaction is described for the detection of phytoplasma DNA in grapevines collected from South Australia and Victoria. Grapevines with Australian grapevine yellows disease tested positively for a phytoplasma but those with ‘restricted spring growth syndrome’ (formerly called ‘grapevine decline’) tested negatively. Restriction fragment length polymorphism analyses were done to determine the relationships between phytoplasmas of the Australian grapevine yellows and of representatives from both the aster yellows group (which includes phytoplasmas of grapevine yellows from Italy) and the elm yellows group (which includes phytoplasmas of flavescence dorée). Results showed that Australian grapevine yellows is associated with a unique phytoplasma that is more closely related to the phytoplasmas of the aster yellows group than to those of the elm yellows group.
To further understand the genomic diversity and genetic architecture of phytoplasmas, a physical and genetic map of the sweet potato little leaf (SPLL) strain V4 phytoplasma chromosome was determined. PFGE was used to determine the size of the SPLL-V4 genome, which was estimated to be 622 kb. A physical map was prepared by two-dimensional reciprocal digestions using the restriction endonucleases BssHII, SmaI, EagI and I-CeuI. Sixteen cleavage sites were located on the map. Southern hybridizations of digested SPLL-V4 chromosomal DNA were done using random clones and PCR-amplified genes as probes. This confirmed fragment positions and located the two rRNA operons and the linked fus/tuf genes encoding elongation factors G and Tu, respectively, on the physical map. An inversion of one of the rRNA operons was observed from hybridization data. Sequence analysis of one of the random clones identified a gid gene encoding a glucose-inhibited division protein. Digestions of the tomato big bud (TBB) phytoplasma chromosome with the same four enzymes revealed genome heterogeneity when compared to the closely related SPLL-V4, and a preliminary chromosome size for the TBB phytoplasma of 662 kb was estimated. This mapping information has revealed that significant genome diversity exists within the phytoplasmas.
Using molecular tools, the spread of phytoplasma diseases in a papaya plantation was investigated for 3 years to identify phytoplasma strains affecting papaya, insect vectors and alternative plant hosts. Five phytoplasma strains (SPLL‐V4, TBB, CaWB, StLL and WaLLvar) were associated with papaya yellow crinkle disease and one phytoplasma strain (PDB) was associated with papaya dieback disease. The most prevalent strains were TBB and SPLL‐V4 which occurred in 94% of infected papaya. There was a significant correlation between phyllody and TBB, and virescence and SPLL‐V4, although other phytoplasma types could also be associated with either phyllody or virescence. No mixed infections were detected in diseased papaya. Disease progress curves for TBB and SPLL‐V4 showed a sigmoid response reaching a maximum disease incidence of 16% after 24 months. The rate of disease spread was best described by a logistic model which showed that TBB spread at a slightly higher rate than SPLL‐V4. Ten phytoplasma strains were detected in 14 alternative plant species; however, TBB and SPLL‐V4 were present in only a few individual plants of some of these species, so these alternative hosts would probably not have provided a significant infection source to papaya. Very few phytoplasmas were detected in leafhoppers collected over 3 years with TBB and SPLL‐V4 only detected in Orosius spp.
Chemical analysis and sensory assessment of river waters from the Murrumbidgee and Murray Rivers showed geosmin is an important odor compound. Experiments in culture using Anabaena circinalis isolated from the Murrumbidgee River, New South Wales, Australia showed that changes in chlorophyll per unit dry weight resulting from alterations in light intensity were paralleled by similar changes in geosmin/dry weight such that geosmin concentration was correlated with chlorophyll a over an extreme range of light conditions. A.circinalis was a prolific producer of geosmin, but a substantial proportion of the total geosmin was often retained intra-cellularly and released on sonication or treatment with copper. Therefore water treatment processes should avoid cell lysis.
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