Typescript. Bibliography: leaves [137]-[157]

The use of chronological age for example, using days after sowing (DAS), or days after germination (DAG) as a time variable may result in the inherent variability between plants resulting in differences which can be large enough to obscure subtle developmental trends that become evident among plants sown at the same time. An alternative to DAS or DAG is the plastochron index (PI), first used by Erickson and Michelini (1957) as a morphological time scale and numerical index; which to according to the authors suggested and represented a more accurate reflection of the developmental status of a plant. The research presented in this thesis was therefore aimed specifically at utilizing the index in qualitative and quantitative analyses, to confirm its usefulness in analyzing and predicting plant growth and development. Specifically this research focused on investigating various morphological and physiological events that together, hopefully, would serve as a template for the prediction of the growth, development and reactions of Pisum sativum L. to different growth conditions. In Chapter 3, the use of the average length of the first pair of leaflets on each node as a suitable parameter for calculating PI in P. sativum is suggested. The results presented in Chapter 3 suggest that plant age is best expressed using the plastochron index, as this reflects the time interval between the initiations of successive pairs of leaflets. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2005) A re-evaluation of plastochron index in peas - a case for using leaflet length. South African Journal of Botany 71: 76-80”. The PI formula developed was subsequently used in this research to conduct qualitative and quantitative investigations of plant growth and development in which all data and observations were related directly to the plastochron index. In Chapter 4, the sink to source transition in Pisum sativum L. leaves at different plastochron ages in nodulating plants was investigated using the phloem-mobile fluorescent marker, 5,6-carboxyfluorescein (5,6-CF). The results demonstrated that young leaves remained strong sinks up until LPI 0, after which sink-source transition occurred up to LPI 1.8 and leaflets transitioned to strong source systems by LPI 2.0. A well-developed cross-connected phloem system between paired leaflets in peas, and the petiole and the stem vascular supply was observed. The data presented in the second part of Chapter 4 suggest that the phloem transport between leaflet pairs is independent of the sink/source state of the leaflets, or of movement along the source to sink gradient. The data support the presence of a modular transport system which may ensure re-allocation and balancing between leaflets of the same physiological age and photosynthetic and transport status, thereby load-balancing the local transport system, before exporting to other younger (sink) regions. The investigation of leaf development using the plastochron index (Chapter 5) revealed that the formation of air spaces in the palisade and spongy mesophyll, one of the preparatory events for transition from sink to source state in developing leaves, occurs between LPI 0 and LPI 1 in pea leaflets. Results of the anatomical and ultrastructural study related to PI are presented in Chapter 5. The density of wall ingrowths in transfer cells of minor veins increased with LPI and appeared to be associated with the probable transition to source state and the related potential increase in the production of assimilates for export. The onset of wall ingrowth development in leaflets at LPI 0 provided evidence that sink-to-source transition commences at LPI 0 in P. sativum. Presumably-functional plasmodesmata as well as a few mature sieve elements were evident in class IV veins in the apical region of young and older leaflets at LPI 0. The number of mature sieve elements per vein however, increased with increasing LPI. Most class V veins were still undergoing division at LPI 0 and their sieve elements did not show signs of maturity until LPI 1. The increase in the number of mature metaphloem sieve elements in young, supposedly importing tissue at LPI 0 to older, supposedly exporting tissues at LPI 2 is evidence of the association between phloem maturation and transition from importing to exporting status. In Chapter 6, I report on the effects of elevated CO[subscript 2] on the growth and leaf development of nodulating and non-nodulating Pisum sativum L var. Greenfeast grown under controlled environment of the same nitrogen (6mM) and nitrogen- free nutrient solution conditions. Shortterm exposure to elevated CO[subscript 2] induced rapid plant growth, irrespective of treatment. However, long-term elevated CO[subscript 2] treatment did not affect rate of leaf appearance (RLA) in nodulated plants, irrespective of mineral N supply but enhanced RLA in non- nodulating plants supplied with mineral N. Supplied N resulted in a significant increase in leaflet elongation rate (LfER) under both ambient and elevated CO[subscript 2], but LfER was not significantly affected by nodulation but was increased by high CO[subscript 2]. This suggested that the growth of nodulating P. sativum L may not be significantly affected under CO[subscript 2] levels as high as 1000 μmol mol[superscript -1]. The data suggest that elevated CO[subscript 2] will enhance canopy size, provided adequate soil N is available and more so in non-nodulating plants. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2004) The effects of elevated CO[subscript 2] and nitrogen availability supersedes the need for nodulation in peas grown under controlled environmental conditions. South African Journal of Botany 70: 816 – 823”. This thesis demonstrates that the similarity in the qualitative analyses results obtained from plants from different CO[subscript 2], nitrogen and nodulation treatment conditions, highlights the fact that plants of same PI value are at the same developmental state, irrespective of the growth condition. Furthermore, changes in plant structure and function observed under different growth conditions can be related simply to changes in plastochron index. The work presented in this thesis demonstrate that changes in plant structure and function analyzed are related to changes in PI. An important finding of this thesis is that with the use of PI, results can be compiled as a template for predicting the structure- function state of pea plants at any plastochron age, under any growth conditions, before using small representative sample populations.

Freshly isolated protoplasts from pea leaves were used to investigate the responses of antioxidant enzymes to oxidative stress. Two cultivars, Progress (tolerant) and Nugget (sensitive), that have differing resistance with respect to oxidative stress at the whole plant level were used. Sulfite and the superoxide generating herbicide, paraquat, were used as the oxidants. Final sulfite concentrations during photosynthetic incubations ranged from 1.5 mM to 30.0 mM. During the polarographic estimation of photosynthesis, CO₂-dependent O₂ evolution did not decrease. At sulfite concentrations of 3.0 mM or less, light-dependent O₂ evolution increased and was probably due to a concomitant SO₂-dependent O₂ evolution. Photosynthesis determined as ¹⁴CO₂ fixation was not increased at these low concentrations of sulfite. Concentrations greater than 7 mM = sulfite inhibited photosynthetic ¹⁴CO₂ fixation. No difference in these responses was found between the two cultivars. At 0.1 µM paraquat, the relative resistance to oxidative stress was reversed compared to previous studies at the whole plant level. With the tolerant cultivar, activity of the plastid antioxidant enzyme, glutathione reductase, increased after a three-hour exposure. Changes in the steady state level of glutathione reductase protein, as judged by immunoblots, did not correlate with the observed changes in enzyme activity. No change in the de novo synthesis of glutathione reductase occurred over the same period as a consequence of paraquat application. A mechanism, unrelated to oxygen free radical scavenging, may contribute to the relative tolerance to low concentrations of paraquat. On the other hand, after an eight-hour exposure to 0.1 mM PQ in the presence of Gamborg’s basal salts, superoxide dismutase activity of Progress protoplasts was enhanced 288% above the preexposure levels while glutathione reductase activity decreased 70% and ascorbate peroxidase activity decreased 90%. The relationship of these changes to oxidative damage to the photosynthetic machinery remains to be assessed. / Ph. D.

Typescript / Bibliography: leaves [137]-[157] / xx, 136 leaves : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Agronomy, Waite Agricultural Research Institute, 1987