Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1990. / Vita. Abstract. Includes bibliographical references (leaves 102-104). Also available via the Internet.

The impact of nutrient addition on growth and nutrient accumulation in insectivorous plants was studied in field populations and greenhouse plantings. Drosera rotundifolia was studied in the field, and D. binata var. multifida and D. capensis were studied in long-established plantings in the greenhouse. In each case, experiments were performed by enclosing insects and/or adding phosphorus and/or nitrogen to the soil. None of the species significantly benefitted from insect capture nutritionally or energetically in nutrient-poor or rich soils. Added nutrients to the soil or by foliar insect feeding decreased phosphorus retention in hibernacula by 50% (D. rotundifolia). Nutrient additions reduced D. rotundifolia vegetative growth in both N and P addition treatments. In addition, reproductive output (inflorescences) decreased flowering by 98% when N was added to the soil. Nutrient addition to soil increased nutrient concentration significantly in D. rotundifolia (N and P), and to a greater extent in D. capensis (N and P) and D. binata (P), and increased growth in D. capensis (N and P). In natural settings, insectivory was not found to be a significant source of nutrients for the species of Drosera studied. Larger subtropical species such as D. capensis and D. binata var. multifida is found in relatively richer (nutrient) soil than cool temperate species (D. rotundifolia) and are better able to utilize available nutrients in a substrate by high absorption rates and luxury consumption. / Master of Science

Le travail présenté porte sur le développement d’une nouvelle technologie innovante, nommée PAT Friday®, visant à produire des protéines recombinantes au sein des sécrétions extracellulaires de plantes carnivores génétiquement modifiées. Deux objectifs ont été fixés : Réaliser la preuve de concept de la technologie sur le modèle expérimental Drosera rotundifolia, en transformant la plante avec des gènes marqueurs et humains afin de mettre en évidence la présence des protéines recombinantes dans la glu ; et développer, après évaluation, la technologie sur un modèle potentiellement industrialisable, Nepenthes alata. Les résultats ont indiqué la présence des deux protéines marqueurs GFP et GUS dans les tissus et dans la glu de Drosera rotundifolia transformées. Les plantes ont également été transformées génétiquement avec les gènes humains de l’interféron gamma et du facteur intrinsèque. Les protéines recombinantes humaines ont été mises en évidence au sein des tissus végétaux. Le potentiel industriel du modèle Nepenthes alata a ensuite été étudié : 10 à 15 kg de protéines totales par hectare et par an peuvent être produits, grâce notamment à des récoltes successives non destructrices, et la possibilité de contrôler l’activité des protéases digestives naturelles. L’élaboration d’un protocole de régénération de la plante a été entreprise par embryogénèse somatique et organogénèse indirecte, en vue de sa transformation génétique. La technologie PAT Friday®, avec des étapes simplifiées d’extraction et de purification des protéines d’intérêt produites dans le liquide digestif, offre de nouvelles perspectives dans le domaine des protéines thérapeutiques produites à partir de plantes / The present work focuses on the development of a new innovating technology, called PAT Friday®, aiming at producing recombinant proteins into the extra-foliar fluid of modified carnivorous plants. Two objectives were assigned to this work : 1- to realize a proof of concept of the technology on the experimental model Drosera rotundifolia, transformed with marker and human genes, to confirm the occurence of the recombinant proteins into glu ; and 2 - to evaluate and develop, the technology on the model Nepenthes alata, more adapted to industrial scaling-up. The results indicate the presence of two marker proteins GUS and GFP inside the tissues and into the glu of modified Drosera rotundifolia plants. The same plant species has also been transformed with human gamma interferon and intrinsic factor genes. The corresponding human recombinant proteins have been detected into the plant tissues. Potential industrial scaling-up has been studied with the species Nepenthes alata. The results show a potential productivity of 10 to 15 kg of total proteins per hectare per year, thanks to non-destructive repeated harvests, and possibility to efficiently control the natural proteinase activity. The elaboration of a regeneration protocol has been undertaken through indirect organogenesis and somatic embryogenesis, with a view to transform genetically this plant. PAT Friday® technology, with simplified extraction and purification methods of the proteins of interest targeted into the liquid secretions, opens new perspectives in the field of therapeutical proteins produced in plants

Nitrogen (N) is an essential element to plants for growth, maintenance and reproduction, however most N does not exist in a form that is biologically available to plants. In order to maximise the acquisition and retention of N, plants have evolved a variety of morphological and physiological adaptations and life history strategies, as well as the ability to respond plastically to changes in resource availability in ecological time. Determining the ecophysiological responses of plants to changes in root N availability is crucial to further understanding of the mechanisms underlying competitive interactions between plants, and between plants and other organisms, that ultimately contribute to community structure and ecosystem functioning. Carnivorous plants are ideal systems for investigating ecophysiological responses to N availability as:- (i) they share a unique adaptation for obtaining supplemental N from captured prey, therefore ecological stoichiometry and energetic cost/benefit models may be explored; (ii) the trait of botanical carnivory is widely considered to have independently co-evolved as a response to N-deficient, sunny and wet environments, therefore resource allocation trade-offs between plant investment in N and carbon (C) acquisition may be observed, and (iii) they are extremely sensitive to changes in root N availability in ecological time. In this research, the carnivorous plant Drosera rotundifolia (round-leaved sundew) was used to address several unanswered ecophysiological and evolutionary questions relating to patterns and processes of prey capture and the N nutrition of carnivorous plants. Furthermore, the potential for reducing uncertainty in the calculation of plant reliance on carnivory using a δ15N natural abundance multi-level linear mixing model was explored. A combined approach of in-situ and ex-situ studies was employed, using co-occurring non-carnivorous plants or carnivorous plant species with differing evolutionary lineages or prey capture mechanisms respectively to provide context. Results show that the adaptations of carnivory, high reproductive investment and a relatively short life span enable Drosera rotundifolia to survive and thrive in an extreme, N deficient environment. Phenotypically plastic responses by the plant to light and root N availability provide evidence of resource allocation trade-offs between investment in carnivory for N acquisition and in photosynthesis for C acquisition. Plants invested less heavily in prey capture (measured as the stickiness of leaf mucilage) as N availability increased or light availability decreased. These results show that the energetic costs associated with carnivory are avoided by the plant when less costly sources of N are available for uptake and that the production of carbon-rich mucilage is only made under nutrient-limited and well-lit conditions. Results obtained from the comparison of captured insect prey with background invertebrates of potential prey indicate that Drosera rotundifolia is a dietary generalist, where the quantity of prey captured per plant is positively correlated with leaf stickiness and total leaf area. Plant reliance on prey-derived N decreased with increasing root N availability, providing evidence that carnivory is only of net benefit to the plant in N-deficient and well-lit environments, as the photosynthetic costs of investment in the trait are not exceeded by the energetic gain from prey N uptake in shady or dry habitats. A more accurate and precise method for calculating plant reliance on botanical carnivory is presented which incorporates the insect diet of the plant. This method has wider significance for reducing uncertainty in the calculation of relative source contributions to a mixture for most natural abundance applications using a multi-level linear mixing model. To conclude, results from this research further understanding of the ecophysiological mechanisms underlying plant responses to changes in resource availability and the selective pressures driving the evolution of plant adaptations. These results therefore assist with predicting how plants and plant communities may respond to sustained N deposition inputs and future environmental scenarios.