Pea root plastids were isolated by differential centrifugation and resulting crude plastid fraction was purified by centrifugation through 10%(v/v) Percoll. Marker enzymes indicated that greater than 50% of the plastids were recovered essentially free from mitochondrial and endoplasmic reticulum contamination. The optimum in vitro conditions for glycerolipid biosynthesis from (U-$ sp{14}$C) glycerol-3-phosphate have been determined. Total glycerolipid biosynthesis was approximately 15 nmole/hr/mg protein in the presence of 200 $ mu$M glycerol-3-phosphate, 0.5 mM each of NADH and NADPH, 15 mM KH$ sb2$CO$ sb3$, 0.05 mM CoA, and 2 mM each of ATP and MgCl$ sb2$, 100 mM Bis Tris Propane (pH 7.5) and incubated at the standard temperature of 25$ sp circ$C. ATP, Coenzyme A and a divalent cation are absolutely required for glycerolipid biosynthesis, whereas reduced nucleotides and bicarbonate improve the synthesis to varying degrees. Dihydroxyacetone phosphate had little effect, while dithiothreitol, detergent and Mn$ sp{2+}$ inhibited activity. Under the optimum conditions, isolated pea root plastids mainly synthesized approximately 15% phosphatidic acid, 16% phosphatidylcholine, 13% phosphatidylglycerol, 32% triacylglycerol. Galactolipid synthesis occurred only when UDP-galactose was supplied. Different concentrations of some cofactors resulted in alterations of glycerolipid distribution. Phospholipase A$ sb2$ and Rhizopus lipase digestions of phospholipids and neutral lipids revealed that radioactive fatty acids were preferentially esterified to position sn 2 of each glycerolipid with generally 2-4 times as much radioactivity as position sn 1. Pea root plastids are composed of approximately 62% phospholipid, 24% neutral lipid and 14% glycolipid. Within these classes PG, TAG, and the galactolipids are the major components representing 24, 12, and 12% of the total plastid lipids.

Studies of corn root morphology, canopy description, light and nutrient relationships, have focused on conventional corn hybrids. We are now extending these studies to other corn types with contrasting canopy and root architectures. Field and greenhouse experiments were carried out in order to characterize root morphology, N status in the plant and its relationship with yield and yield components, canopy architecture and light interception of these genotypes. The indoor experiments investigated root morphology and how N affects it. Root fractal geometry and its relationship with standard measured root variables were investigated. The field research, at two sites and over two growing seasons, examined (i) maize canopy architecture with regard to light interception and (ii) nitrogen effects on grain yield of different maize genotypes. Four genotypic types were included: (i) Leafy reduced-stature, Lfy1rd1 (LRS), (ii) non Leafy-reduced stature, lfyrd1 (NLRS), (iii) Leafy normal stature, Lfy1Rd1 (LNS), and (iv) conventional commercial hybrids, lfy1Rd1. Pioneer 3905 served as the check hybrid for late maturity, and Pioneer 3979, the check for early maturity. The work allowed development of following methods: (i) root sampling for measurement of large root systems, (ii) staining to enhance root contrast for measurement with a scanner-based software system, (iii) sample size determination for SPAD meter readings, and (iv) the design and construction of a mobile and multi-strata device for measurement of light interception. Data were collected for mathematical characterization of canopies (i.e. leaf angle, co-ordinates of the maximum height of the leaf, co-ordinates of the leaf tip), plant N status (SPAD meter readings), light interception, yield and grain yield components. Conventional hybrids generally showed greater root length and surface area than their leafy genotypic counterparts at early developmental stages (i.e. up to 15 days from emergence). However, Leafy geno

Extracts of gelling agents have been shown to stimulate rooting and this study was initiated to investigate the presence of root stimulating substances in gelling agents. After screening a number of gelling agents, four were selected, namely; Agar Bacteriological, Agar Commercial Gel, Difco Bacto Agar and Gelrite were selected and examined for the presence of root-stimulating substances using mungbean bioassay. Water extracts of Agar Bacteriological, Agar Commercial Gel and Difco Bactol Agar stimulated rooting of mungbean cuttings. Addition of Charcoal neither reduced nor increased rooting produced by the water extract of the first two agars but when added in conjunction with Difco Bacto Agar rooting was reduced. Autoclaving, however reduced rooting in extracts of the gelling agents. The possibility that root-stimulating substances may not be the same in all the gelling agents can not be excluded. Extraction of Gelrite with water was problematical and was therefore excluded. IBA solution and water extracts of the gelling agents separately promoted good rooting in mungbeans cuttings. Rooting in extracts of autoclaved frozen-thawed gelling agents was poor, however, IBA + gelling agents gave high rooting at the 100% concentration and this could possibly be due to an additive effect of the IBA. Addition of charcoal reduced rooting significantly in extracts of IBA + gelling agents. Using 80% acidic methanol, reasonable levels of rooting substances were obtained from the residue extract of this complex (IBA + gelling agent+charcoal) of all the gelling agents except Gelrite indicating that root-promoting substances were adsorbed by charcoal. The low rooting in the presence of the Gelrite extract was attributed to the matrix of the polymer of the Gelrite. Ethyl acetate fractionated extracts (EA-pH 8.0; EA-pH 3.0; and Aqueous) obtained from the four gelling agents stimulated rooting indicating the presence of numerous root promoting substances. Gelrite gave good rooting with both the 50 and 100% concentrations of all the fractions. Purified water and ethanol extracts of the gelling agents exhibited auxin-like activity when separated by paper chromatography and compared with IBA and IAA standards. Using HPLC, IAA was quantified in all the gelling agents with Difco Bacto Agar and Agar Commercial Gel having the highest IAA concentration and Gelrite the lowest IAA concentration. IAA concentration in Agar Bacteriological was a third of the level detected in Difco Bacto Agar. The information from this work may enable researchers to consider gelling agents as sources of auxin-like compounds and other plant hormones as well as support media for use in tissue culture procedures and also increase the enthuse for further research into the nutrient types and levels in gelling agents. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2003.

Due to the increasing prevalence and severity of Al phytoxicity in certain regions of the South African sugar industry, a research programme has been initiated at SASEX to elucidate the molecular mechanisms by which sugarcane detects and responds to the metal. As part of this larger investigation, the current study aimed to assess the response of a reportedly Al tolerant cultivar, Saccharum spp. hybrid cv. N12, to phytotoxic levels of Al. Hydroponically-grown plants of this commercial genotype were used in Al inhibition studies, the results of which indicated that exposure of plants to 250µM Al for 24 hours resulted in maximum reduction of root elongation. Under these conditions, root growth was inhibited by approximately 36%, compared with only 4% for the 50µM Al treatment. Subsequently, this exposure regime was used to gather the terminal 5 to 10mm of root tips, the site of the primary Al lesion, of challenged and control, unchallenged plants for molecular analysis. Total RNA was extracted from the Al challenged and control root tips, from which mRNA was subsequently isolated, reverse transcribed and converted to double-stranded cDNA. The two populations of cDNA were reciprocally subtracted from each other and used to construct subtractive cDNA libraries in Lambda ZAP®II phages. Randomly selected clones, 576 representatives from each of the libraries, were screened using membrane-based array technology. Results indicated that only 33% (190) of the Al-treatment specific library cDNAs were found to be more highly expressed under conditions of Al stress than under control conditions. Of these potentially Al response-related cDNAs, 25 were sequenced and submitted to sequence databases for the assignment of putative identities. No genic sequences known to be directly associated with the Al stress response were identified, however, several were found to be related to pathogenesis or general stress pathways. Although further Northern hybridisation work is required to validate these results, they suggest that the induction of general stress response pathways may be involved in the aluminium stress response of this sugarcane cultivar. Such Al stress-related sequences could have applications in marker-assisted breeding programmes and as candidate genes for the genetic engineering of tolerant genotypes. / Thesis (M.Sc.)-University of Natal, Durban, 2002.

Denna uppsats är en litteraturstudie av Roots: The Saga of an American Family. Studien syftar till att undersöka argumentet att rasism har en långvarig effekt på de människor som utsätts för rasism. I detta fall, karaktärerna i romanen som utsätts för rasismens olika hemskheter så som slaveri. Genom användningen av Robert Jensen, WEB Du Bois och Edward Saids teorier (dvs. Privileged/unprivileged, The Veil, Double Consciousness och Others) har romanen Roots analyserats. Slutsatsen visar på hur slaveriet påverkade huvudpersonerna och hur effekten av slaveri lyckats beröva de svarta människorna i romanen deras ursprungliga identitet. Slaveriet ersattes istället av en kollektiv identitet av mindre värde än vita människors identitet vilket lämnat långvariga psykiska ärr för karaktärerna i romanen. Det pedagogiska fokus som används i uppsatsen är den så kallad cooperative learning metoden.

The root system is a vital and dynamic part of a plant throughout its lifetime. Its spatial distribution is the consequence of multiple interactions with the surrounding soil medium. In particular, the presence of pathogens in soil may influence the development of the plant, especially the below-ground part, in both its physiology and its structure. Studies of diseased plant roots may take different approaches and investigate disease effects at different levels. In this study, two groups of four potato (Solanum tuberosum L.) plants were grown in middle-sized plastic pots in a greenhouse, and their root systems, together with the soil medium (i.e., sieved and autoclaved homogeneous sand), were submitted to computed tomography (CT) scanning every two weeks until 10 weeks after planting. For the "diseased" group, sand was inoculated with Streptomyces scabies EF-35, the causal agent of potato common scab, at the time of planting. Disease effects on tissue density of roots and below-ground organs, space occupancy and complexity were assessed by analyzing the CT scanning data in the spatio-temporal approach. Fluctuations of tissue density over time were different, on average, between the two groups. They were characterized by an increase of density in Week 2 for the diseased group, reflecting a hardening of tissue, and a delayed decrease relative to the "healthy" group, suggesting a slower transfer of energy from the seed potato to growing roots. Space occupancy, which was studied via volumetric growth rates evaluated from CT scan data, and complexity, which was quantified by the fractal dimension estimated from skeletonized 3-D images constructed from CT scan data, also showed differences in the first part of the experiment. Original analytical procedures based on data transformation and curve fitting in histogram analysis of CT numbers were developed to obtain those results. In conclusion, the new approach presented here, which is based on the advanced processing of CT scanning data collected over time on developing plant root systems and below-ground organs, can be recommended for future phytopathological applications.

Soybean (Glycine max L. Merr.) and Bradyrhizobium japonicum can form a nitrogen fixing symbiosis. This symbiosis is important for most sustainable agriculture systems. This thesis examines two ways to enhance nodulation and nitrogen fixation by this symbiosis: coinoculation of plant growth promoting bacteria (PGPB) with B. japonicum, and addition of RNA to a bradyrhizobial culture medium. The optimal coinoculation dose of Serratia proteamaculans 1--102 and S. liquefaciens 2--68 was determined as 108 cells per plant under both optimal and suboptimal root zone temperatures (RZTs). Nodulation dynamics studies indicated that coinoculation of these two PGPB caused earlier nodule initiation and a higher nodulation rate, contributing to the higher nodule number and nodule weight. The coinoculation also increased nitrogen fixation efficiency under both optimal and suboptimal RZTs. A novel inducible activator only produced by the bacteria after addition of flavonoids to the culture system was prepared and evaluated in greenhouse and field experiments. Fourteen non-bradyrhizobial endophytic bacteria (NEB) were isolated from the surface sterilized root nodules, and three of these, designated NEB4, NEW and NEB17, showed soybean plant growth promotion under both greenhouse (with controlled RZTs) and field conditions. Alone, they were neither nodule inducers nor nitrogen fixers. Biolog tests and partial 16S rRNA gene sequence analyses placed the three strains in genus Bacillus: NEB4 and NEB5 are B. subtilis and NEB17 B. thuringiensis. Bradyrhizobium species grow slowly, making the culture process long and the cost of inoculant production higher. Addition of commercial yeast RNA to the bacterial culture medium accelerated the bacterial growth rate, shortened the culture time and increased the lipo-chitooligosaccharide (LCO) yield in flask cultures. Inoculation experiments in the greenhouse also showed that bradyrhizobial inoculant produced in the presence of RNA had better symb

xv, 217 leaves : ill., tables, graphs, photos ; 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, 1977

In strongly acidic soils (pH <4.5) aluminium (Al) becomes soluble in quantities that can lead to Al phytotoxicity. It is estimated that approximately 30 % of the worlds’ potentially arable lands are acidic, with Al toxicity the most limiting factor for plant growth on acid soils. With increasing use of marginal land in cropping systems, this area could reach 70 %. Cell wall pectin provides up to 70 % of the root cation exchange capacity. Pectin is suggested to control a number of physiological properties of the plant cell wall such as porosity, charge density, microfibril spacing and pH. The ability of pectin to bind cations is not only important for the uptake of nutrients but is implicated in metal toxicity, in particular Al. Despite over a century of research, the mechanisms of Al toxicity are yet to be fully elucidated or agreed upon. Gluconacetobacter xylinus is a gram-negative, soil dwelling bacterium which produces extracellular cellulose. It is an established archetype for the study of cellulose biogenesis. In the presence of pectin in the growth medium, the bacterium can form cellulose-pectin composites. Recently, the bacterium has been used to form composites as model cell walls to understand plant cell wall deposition. Additionally, bacterial cellulose composites in their natural hydrated state mimic the hydration state of primary plant cell walls. The aim of this project was to attempt to incorporate this novel cell wall analog into laboratory investigations into metal interactions with plant cell walls. Preliminary work was undertaken to optimise the bacterial culture medium, growth conditions, analysis of the composites and developing an overall general methodology. The medium buffering system was altered, growth under non-optimal pH conditions was evaluated and Al was successfully incorporated into the composites. Appropriate sample preparation for scanning electron microscopy (SEM) of the composites was determined. This work resulted in the successful production of bacterial cellulose-pectin composites with 30 % w/w pectin incorporation. The effect of Al on the tensile properties of the composites was examined. Aluminium had no effect on the stress/strain profiles, confirming the hypothesis that pectin is not the main load bearing component of the cell wall. The composites were used to investigate the effects of Al and other trace metals (copper (Cu), gadolinium (Gd), lanthanum (La), ruthenium (Ru) and scandium (Sc)) on the hydraulic conductivity of the composites. Hydraulic conductivity was reduced to ≈ 30 % of the initial flow rate by 39 μM Al and 0.6 Cu μM, ≈ 40 % by 4.6 μM La, 3 μM Sc and 4.4 μM Ru, and ≈ 55 % by 3.4 μM Gd. These metal concentrations were selected based on the concentrations causing a 50 % reduction in root elongation in cowpea (Vigna unguiculata L.). This study demonstrated that all the trace metals caused a similar decrease of hydraulic conductivity, despite the different concentrations of the metals used. Scanning electron microscopy showed changes in pectin porosity with metal binding which may account for the decreases in hydraulic conductivity observed. As the composites could not be used as a model material in all investigations, pectin-only systems were employed in a rheological study to investigate the effect of increasing concentrations of Al, Ca, Cu or La at pH 4 on pectin (degree of esterification 30 %, 1 % w/v) gel physical strength. Comparing similar saturation levels, La formed the weakest gel, followed by Ca, which was similar to Al, while the strength of Cu gels was almost an order of magnitude stronger than the other cations. This study was the first to investigate Al and La pectate gel strength. The swelling of the gels also varied, with Ca gels being the most swollen. Pectin was also used to determine the exchange selectivity of Al, Cu, Gd, La, Ru and Sc toward Ca pectate. The order of selectivity was found to be Sc>Gd>La>Cu>Ru>Al. There were some parallels between this sequence and the rhizotoxicity data of the metals, suggesting that the strength with which metals bind to pectin is an indication of their rhizotoxicity. Through the use of synthetic pectate gel systems new information was discovered about the strength of pectin gels and the selectivity of trace metals towards pectin. These findings were in keeping with those of a number of related studies, as well as with studies of plant root tissue. Overall, the novel bacterial cellulose-pectin cell wall analog was successfully integrated into research into Al and other metal toxicity in plants, and offers a useful system that can overcome some of the difficulties encountered when using plant cell wall tissue. Further research may be warranted on manipulating the growth system to produce composites in the presence of the metal (ie. metal added to the growth medium), as opposed to post-formation treatments. Moreover, the production of a three way composite of cellulose, hemicellulose and pectin would likely be another useful analog for plant cell wall material.

[Truncated abstract] The hydraulic properties of the roots of three crop species important to Western Australia were examined: wheat (Triticum aestivum), narrow-leafed lupin (Lupinus angustifolius) and yellow lupin (L. luteus). Generally, the hydraulic conductivity (Lpr) of root systems differs between species and can change in response to adverse conditions. To determine the significance of root anatomy and aquaporin activity on the pathway of water flow through roots, water flow was measured across cell membranes, individual roots and whole root systems. The combination of measurements identified that wheat and lupin roots have contrasting hydraulic properties. Wheat roots absorb water preferentially in the apical region, whereas lupin roots appear to absorb water more evenly along the entire root length. Lupin roots have a greater axial hydraulic conductance than wheat, due to more abundant xylem vessels and axial conductance increases with root length, in conjunction with xylem vessel development. However, water flow through the radial pathway is the limiting factor in whole root hydraulic conductance, in all species. Modelling and the inhibition of aquaporin activity with mercuric chloride demonstrated that radial water flow in wheat roots occurs by a combination of the cell-to-cell and apoplastic pathways, but in lupins, water flow appears to be predominantly apoplastic. Despite the presence of aquaporins in root cell membranes of all species, their role in regulating bulk water flow across roots is not clear in lupins, because of the significance of the apoplastic pathway ... After draining the chambers, the root systems of yellow lupin resumed growth, but there was no subsequent recovery in narrow-leafed lupin root systems. The growth and survival strategies of wheat and lupin root systems are disparate. Wheat root systems are comprised of numerous fine, highly branched, individual roots that extract water near the root tips and have the ability to regulate flow. These attributes may be advantageous in non-uniform or variable environments. Moreover, the ability of wheat roots to regulate flow may not only support survival during waterlogging, but also enhance recovery. In comparison, lupin root systems are designed like conduits, for the rapid uptake and transport of water when conditions are favourable. However, their thick taproots and lack of regulation of water transport or anatomical changes make them unsuitable for very wet soils.