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Graduation date: 1950

Copies of author's previously published articles inserted. / Bibliography :leaves 65-80 / x, 80, [64] leaves, [24] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Ice nucleation active (INA) bacteria were detected in a pea field in South Australia. They were identified as strains of Pseudomonas syringae and Pseudomonas flourescens biotype 1. Some chemical agents were tested on the two ice nucleating species, as cryoprotectants. / Thesis (Ph.D.)--University of Adelaide, Dept. of Crop Protection, 1991

Thesis (Ph. D.)--University of Adelaide, Dept. of Crop Protection, 1991. / Copies of author's previously published articles inserted. Includes bibliographical references (leaves 65-80).

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.

Sclerotinia sclerotiorum causes white mold and severe yield losses of pea. 484 accessions from the Pisum core collection were screened for resistance using a mini-agar plug technique. 49, 41, and 13 accessions were identified with partial resistance based on lesion expansion inhibition (LEI), nodal transmission inhibition (NTI), and both traits combined, respectively. A genetic linkage map based on F2 DNA from the cross, Lifter/PI240515, was developed with 78 markers on 9 linkage groups (LG) spanning 734 cM. Two quantitative trait loci (QTL) were identified based on phenotypic data from F2:3 and F3:4 families. A single QTL on LGIII explained 34.1% of the phenotypic variation for LEI, while a second QTL on LGII(b) explained 2.5% of the phenotypic variation for NTI. This is the first report of QTL for S. sclerotiorum resistance in pea which will be useful in development of resistant pea varieties.

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by Lam Suet Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 109-121). / Acknowledgements --- p.i / Table of contents --- p.ii / Abstract --- p.vii / List of Abbreviations --- p.ix / List of Tables --- p.x / List of Figures --- p.xi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Ribosome-inactivating proteins (RIPs) --- p.3 / Chapter 1.1.1 --- Types of RIPs --- p.4 / Chapter 1.1.1.1 --- Type I RIPs --- p.5 / Chapter 1.1.1.2 --- Type II RIPs --- p.7 / Chapter 1.1.2 --- Physicochemical properties --- p.7 / Chapter 1.1.3 --- N-glycosidase activity of RIPs --- p.8 / Chapter 1.1.3.1 --- Specificity of N-glycosidase activity --- p.10 / Chapter 1.1.3.2 --- Inhibition of protein synthesis --- p.11 / Chapter 1.1.4 --- Other enzymatic and biological activities of RIPs --- p.11 / Chapter 1.1.4.1 --- Enzymatic activities --- p.11 / Chapter 1.1.4.2 --- Multiple depurination --- p.13 / Chapter 1.1.4.3 --- RNase activity --- p.14 / Chapter 1.1.4.4 --- DNase activity --- p.15 / Chapter 1.1.4.5 --- Biological activities --- p.16 / Chapter 1.1.5 --- Storage of RIPs in plant cells --- p.17 / Chapter 1.1.5.1 --- RIPs targeted to subcellular compartments --- p.18 / Chapter 1.1.5.2 --- Cytoplasmic RIPs --- p.20 / Chapter 1.1.6 --- Physiological roles of RIPs --- p.22 / Chapter 1.1.6.1 --- Defensive role in plants --- p.22 / Chapter 1.1.6.2 --- Metabolic role of RIPs --- p.26 / Chapter 1.1.6.3 --- RIPs as storage proteins --- p.26 / Chapter 1.1.7 --- Application of RIPs --- p.27 / Chapter 1.1.7.1 --- Therapeutic applications --- p.27 / Chapter 1.1.7.2 --- Possible use of RIPs in agriculture --- p.30 / Chapter 1.2 --- Objectives of the present study --- p.31 / Chapter 1.2.1 --- Rationale of the study --- p.31 / Chapter 1.2.2 --- Outline of the thesis --- p.32 / Chapter Chapter 2 --- Screening of hitherto unexplored plant species for RIPs --- p.33 / Chapter 2.1 --- Introduction --- p.34 / Chapter 2.2 --- Materials and methods / Chapter 2.2.1 --- Materials --- p.36 / Chapter 2.2.2 --- Preparation of crude powder --- p.36 / Chapter 2.2.3 --- Protein determination --- p.38 / Chapter 2.2.4 --- Preparation of rabbit reticulocyte lysate --- p.38 / Chapter 2.2.5 --- Protein synthesis inhibition assay --- p.39 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Preparation of crude powder --- p.41 / Chapter 2.3.2 --- Protein synthesis inhibition assay --- p.41 / Chapter 2.4 --- Discussion --- p.43 / Chapter Chapter 3 --- Purification of RIP from garden pea (Pisum sativum) --- p.45 / Chapter 3.1 --- Introduction --- p.46 / Chapter 3.2 --- Materials and methods / Chapter 3.2.1 --- Materials --- p.50 / Chapter 3.2.2 --- Purification of RIP from garden pea --- p.52 / Chapter 3.2.3 --- Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.54 / Chapter 3.2.4 --- Precautions for working with RNA --- p.56 / Chapter 3.2.5 --- N-glycosidase assay --- p.57 / Chapter 3.2.6 --- Quantitation of RNA --- p.60 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Quantitation of RNA --- p.61 / Chapter 3.3.2 --- Affinity chromatography on Affi-gel Blue gel --- p.61 / Chapter 3.3.3 --- Iminodiacetic acid-agarose chromatography --- p.64 / Chapter 3.3.4 --- Cation exchange chromatography on Resource-S --- p.66 / Chapter 3.3.5 --- Gel filtration on Superose 12 HR 10/30 --- p.69 / Chapter 3.3.6 --- "Assessment of purity, yield and activity" --- p.72 / Chapter 3.4 --- Discussion --- p.74 / Chapter Chapter 4 --- Physicochemical and biological properties of garden pea RIP --- p.77 / Chapter 4.1 --- Introduction --- p.79 / Chapter 4.2 --- Materials and methods / Chapter 4.2.1 --- Materials --- p.81 / Chapter 4.2.2 --- Molecular weight determination --- p.82 / Chapter 4.2.3 --- Subunit composition --- p.82 / Chapter 4.2.4 --- Isoelectric focusing (IEF) --- p.83 / Chapter 4.2.5 --- Detection of glycoproteins --- p.84 / Chapter 4.2.6 --- N-terminal amino acid sequence --- p.84 / Chapter 4.2.7 --- Inhibition of cell-free protein synthesis --- p.86 / Chapter 4.2.8 --- N-glycosidase activity --- p.86 / Chapter 4.2.9 --- Deoxyribonuclease activity --- p.87 / Chapter 4.2.10 --- Activity towards tRNA --- p.88 / Chapter 4.3 --- Results / Chapter 4.3.1 --- Molecular weight determination --- p.89 / Chapter 4.3.2 --- Subunit composition --- p.91 / Chapter 4.3.3 --- Isoelectric focusing (IEF) --- p.92 / Chapter 4.3.4 --- Detection of glycoproteins --- p.94 / Chapter 4.3.5 --- N-terminal amino acid sequence --- p.96 / Chapter 4.3.6 --- Inhibition of cell-free protein synthesis --- p.97 / Chapter 4.3.7 --- N-glycosidase activity --- p.99 / Chapter 4.3.8 --- Deoxyribonuclease activity --- p.101 / Chapter 4.3.9 --- Activity towards tRNA --- p.102 / Chapter 4.4 --- Discussion --- p.103 / Chapter Chapter 5 --- General discussion and conclusion --- p.106 / References --- p.109