Nabis kinbergii is a native polyphagous predator in Australia. It has been found in all states and territories of Australia. N. kinbergii has been regarded as an efficient predator of many insect pests in lucerne, cotton and particularly brassica crops. The circadian rhythms, the effects of different prey on development and reproduction, and prey preferences of N. kinbergii have not been studied in South Australia. These are the subjects of this thesis. N. kinbergii is more active at night than in the day. They seemed to be still more frequently at dawn and more active at dusk under natural environmental conditions. Yet, their behaviour was possibly different when they were held in a controlled environment with artificial light. Under both natural and artificial lighting conditions, they would spend more time moving on plants at night. They spent less time moving than other activities. Furthermore, they were more active during the second day of the observations, probably because of the hunger. A mixed diet including Plutella xylostella, Myzus persicae and Brevicoryne brassicae brought the most significant positive influences to the development, survival and longevity of N. kinbergii. It had a shorter preoviposition period and greater egg production when fed on P. xylostella than when fed on M. persicae. Among the three prey, B. brassica was the poorest food because the survival rate between egg hatch and adult eclosion was only 7.5 %, compared to 85 %, 92.5 % and 97.5 % when N. kinbergii fed on M. persicae, P. xylostella and a mixed diet, respectively. Evidence of prey preference was exhibited by N. kinbergii. B. brassica seemed to be the least preferred food. N. kinbergii possibly attacked less mobile prey and delayed eating prey with poor nutrition or that were toxic. In wind tunnel experiments, N. kinbergii may have been attracted by plant volatiles rather than prey odour. These findings may help to refine the timing of experiments and improve the understanding of the role of this predator in integrated pest management. / Thesis (M.Sc.) -- University of Adelaide, School of Agriculture, Food and Wine, 2008

In the late 1990s, a series of landmark publications described RNA interference (RNAi) and related RNA silencing phenomena in nematodes, plants and fungi. By manipulating RNA silencing, biologists have been able to create tools for specifically inactivating genes. In organisms from trypanosomes to insects, RNA silencing is now indispensible for studying gene function. RNA silencing has been used in a project aimed at systematically knocking out all genes in the model plant Arabidopsis thaliana. RNA silencing has a natural role in defending eukaryotic cells against virus replication. By assembling virus DNA sequences in a form that triggers RNA silencing, biologists have created plants resistant to specific viruses. In this study, we set out to test if a similar approach would protect plants against infection by the agriculturally important Brassica pathogen, Plasmodiophora brassicae. P. brassicae is an obligate intracellular biotroph, from the little studied eukaryotic supergroup, the Rhizaria. To identify the gene sequences that would be starting material for P. brassicae RNA silencing, new P. brassicae genes were gathered by cDNA cloning or genomic PCR-walking. Using suppression subtractive hybridisation (SSH) and oligo-capping cloning of full-length cDNAs, 76 new gene sequences were identified. A large proportion of the cDNAs were predicted to contain signal peptides for ER translocation. In addition to the new cDNA identified here, partial sequences for the P. brassicae actin and TPS genes were published by other researchers close to the beginning of this study. Using PCR-walking, full-length genomic DNA sequences from both genes were obtained. Later, genomic DNA sequences spanning or flanking a total of 24 P. brassicae genes were obtained. The P. brassicae genes were rich in typical eukaryotic spliceosomal introns. Transcription of P. brassicae genes also appears likely to begin from initiator elements rather than TATA-box-containing promoters. A segment of the P. brassicae actin gene was assembled in hairpin format and transformed into Arabidopsis thaliana. Observation of simultaneous knockdown of the GUS marker gene as well as detection of siRNAs indicated that the hpRNA sequences induced RNA silencing. However, inoculation of these plants with P. brassicae resulted in heavy club root infection. We were unable to detect decreases in actin gene expression in the infecting P. brassicae, at either early or late stages of infection. We conclude that, within the limits of the techniques used here, there is no evidence for induction of RNA silencing in P. brassicae by in planta produced siRNAs.

In this thesis, I combined observational studies with field and greenhouse experiments to examine selection on life history traits and variation in tolerance and resistance against herbivores in natural populations of the annual herb Arabidopsis thaliana in its native range. I investigated (1) phenotypic selection on flowering time and plant size, (2) the effects of timing of germination on plant fitness, (3) the effect of leaf damage on seed production, and (4) correlations between resistance against a specialist and a generalist insect herbivore. In all three study populations, flowering time was negatively related to plant fitness, but in only one of the populations, significant selection on flowering time was detected when controlling for size prior to the flowering season. The results show that correlations between flowering time and plant fecundity may be confounded by variation in plant size prior to the reproductive season. A field experiment detected conflicting selection on germination time: Early germination was associated with low seedling survival, but also with large leaf rosette before winter and high survival and fecundity among established plants. The results suggest that low survival among early germinating seeds is the main force opposing the evolution of earlier germination, and that the optimal timing of germination should vary in space and time as a function of the relative strength of selection acting during different life-history stages. Experimental leaf damage demonstrated that tolerance to damage was lowest among vegetative plants early in the season, and highest among flowering plants later in the season. Given similar damage levels, leaf herbivores feeding on plants before flowering should thus exert stronger selection on defence traits than those feeding on plants during flowering. Resistance against larval feeding by the specialist Plutella xylostella was negatively correlated with resistance against larval feeding by the generalist Mamestra brassicae and with resistance against oviposition by P. xylostella when variation in resistance was examined within and among two Swedish and two Italian A. thaliana populations. The results suggest that negative correlations between resistance against different herbivores and different life-history stages of herbivores may contribute to the maintenance of genetic variation in resistance.

In the late 1990s, a series of landmark publications described RNA interference (RNAi) and related RNA silencing phenomena in nematodes, plants and fungi. By manipulating RNA silencing, biologists have been able to create tools for specifically inactivating genes. In organisms from trypanosomes to insects, RNA silencing is now indispensible for studying gene function. RNA silencing has been used in a project aimed at systematically knocking out all genes in the model plant Arabidopsis thaliana. RNA silencing has a natural role in defending eukaryotic cells against virus replication. By assembling virus DNA sequences in a form that triggers RNA silencing, biologists have created plants resistant to specific viruses. In this study, we set out to test if a similar approach would protect plants against infection by the agriculturally important Brassica pathogen, Plasmodiophora brassicae. P. brassicae is an obligate intracellular biotroph, from the little studied eukaryotic supergroup, the Rhizaria. To identify the gene sequences that would be starting material for P. brassicae RNA silencing, new P. brassicae genes were gathered by cDNA cloning or genomic PCR-walking. Using suppression subtractive hybridisation (SSH) and oligo-capping cloning of full-length cDNAs, 76 new gene sequences were identified. A large proportion of the cDNAs were predicted to contain signal peptides for ER translocation. In addition to the new cDNA identified here, partial sequences for the P. brassicae actin and TPS genes were published by other researchers close to the beginning of this study. Using PCR-walking, full-length genomic DNA sequences from both genes were obtained. Later, genomic DNA sequences spanning or flanking a total of 24 P. brassicae genes were obtained. The P. brassicae genes were rich in typical eukaryotic spliceosomal introns. Transcription of P. brassicae genes also appears likely to begin from initiator elements rather than TATA-box-containing promoters. A segment of the P. brassicae actin gene was assembled in hairpin format and transformed into Arabidopsis thaliana. Observation of simultaneous knockdown of the GUS marker gene as well as detection of siRNAs indicated that the hpRNA sequences induced RNA silencing. However, inoculation of these plants with P. brassicae resulted in heavy club root infection. We were unable to detect decreases in actin gene expression in the infecting P. brassicae, at either early or late stages of infection. We conclude that, within the limits of the techniques used here, there is no evidence for induction of RNA silencing in P. brassicae by in planta produced siRNAs.

In the late 1990s, a series of landmark publications described RNA interference (RNAi) and related RNA silencing phenomena in nematodes, plants and fungi. By manipulating RNA silencing, biologists have been able to create tools for specifically inactivating genes. In organisms from trypanosomes to insects, RNA silencing is now indispensible for studying gene function. RNA silencing has been used in a project aimed at systematically knocking out all genes in the model plant Arabidopsis thaliana. RNA silencing has a natural role in defending eukaryotic cells against virus replication. By assembling virus DNA sequences in a form that triggers RNA silencing, biologists have created plants resistant to specific viruses. In this study, we set out to test if a similar approach would protect plants against infection by the agriculturally important Brassica pathogen, Plasmodiophora brassicae. P. brassicae is an obligate intracellular biotroph, from the little studied eukaryotic supergroup, the Rhizaria. To identify the gene sequences that would be starting material for P. brassicae RNA silencing, new P. brassicae genes were gathered by cDNA cloning or genomic PCR-walking. Using suppression subtractive hybridisation (SSH) and oligo-capping cloning of full-length cDNAs, 76 new gene sequences were identified. A large proportion of the cDNAs were predicted to contain signal peptides for ER translocation. In addition to the new cDNA identified here, partial sequences for the P. brassicae actin and TPS genes were published by other researchers close to the beginning of this study. Using PCR-walking, full-length genomic DNA sequences from both genes were obtained. Later, genomic DNA sequences spanning or flanking a total of 24 P. brassicae genes were obtained. The P. brassicae genes were rich in typical eukaryotic spliceosomal introns. Transcription of P. brassicae genes also appears likely to begin from initiator elements rather than TATA-box-containing promoters. A segment of the P. brassicae actin gene was assembled in hairpin format and transformed into Arabidopsis thaliana. Observation of simultaneous knockdown of the GUS marker gene as well as detection of siRNAs indicated that the hpRNA sequences induced RNA silencing. However, inoculation of these plants with P. brassicae resulted in heavy club root infection. We were unable to detect decreases in actin gene expression in the infecting P. brassicae, at either early or late stages of infection. We conclude that, within the limits of the techniques used here, there is no evidence for induction of RNA silencing in P. brassicae by in planta produced siRNAs.

PhD (Agric) (Plant Production) / Department of Plant Production / Brassicas are important vegetable crops grown for home consumption and market gardening in eastern and southern Africa. However, productivity is affected by aphids, through both direct feeding and disease transmission. Botanical insecticides have potential to control the aphids, but so far few plants have been evaluated for use on brassicas. This study was conducted to evaluate the effectiveness of Solanum panduriforme to control aphids on brassicas. Botanical extracts from three parts of S. panduriforme were assessed for their insecticidal effects on the cabbage aphid, Brevicoryne brassicae. The extracts from leaf powder (LP), ripe berry powder (BP), fresh ripe berries (RB) and fresh unripe berries (UB) were extracted with four solvents; water, ethanol, hexane and diethyl ether, using homogenisation, maceration and solvent-assisted / sequential extraction methods. The effectiveness of the extracts was determined by laboratory bioassays as well as by plant assays in the screen house and under field conditions. The experiments were replicated three or four times depending on the assays and the design used was completely randomized design (CRD). The immature (LP and UB) plant parts were generally more effective than the mature (BP and RB) plant parts, with mortalities ranging from 100 % down to 40 % respectively depending on assays. Ethanol extracts were more effective than aqueous extracts (LP 96% and 63%; BP 96% and 64%; RB 100% and 64%; UB 100% and 90%). The dried crude extracts from hexane were more effective than di-ethyl ether extracts. The group chemical analysis indicated presence of alkaloids in the berries (BP, RB and UB), which were absent in the leaves (LP). Phenolic compounds and flavonoids were present in all the extracts (LP, BP, RB, and UB). Saponins were present in the fresh parts (RB and UB). The results show how the locally available S. panduriforme plants can be used as an aphicide to control aphids on brassicas. Farmers can directly prepare an easy and cheap botanical / NRF

Abiotic and biotic stresses lead to crop yield losses and ultimately negatively affect agriculture production. Elucidation of the mechanisms of plant stress responses and their regulation could help to understand plant defence and improve stress tolerance. Phytohormones stand behind both plant growth and developmental changes as well as stress signalling. This thesis summarises the results published in two articles focused on phytohormone dynamics in response to abiotic and biotic stresses, namely phosphate shortage and Plasmodiophora brassicae infection. A review article focuses in depth on strigolactones as master regulators of phosphate deficiency responses. The main emphasis is put on organ-specific reactions and exogenous phytohormone treatment with the potential to convey stress tolerance. In the case of phosphate shortage, the universal reaction in all organs was a decrease in active cytokinin trans-zeatin and gibberellin GA4 with a concurrent elevation of abscisic acid. Also, the high- affinity phosphate transporters (PHT1;4 and PHT1;7) exhibited increased gene expression within the whole plant. Shoot apical meristems showed numerous changes in gene expression and were the most affected organ during the lack of phosphate. Only in roots, we observed a substantial elevation of low active...

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