Botany

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Above-ground allometry, biomass and nutrient content of acacia mearnsii across four ages and three sites in the Kwazulu-Natal Midlands.
(2005) Dovey, Steven B.; Pammenter, Norman William.
Acacia mearnsii (black wattle) is one of the few tree crops of which both timber and bark are utilised, with branches sometimes being collected for fuel wood. There is a great potential for nutrient loss from plantations with intense harvesting practices. Allometric relationships were developed to estimate above-ground biomass across four ages and three site qualities of A. meamsii stands. The three sites were based on high, medium and low site quality classes of productivity. Differences in biomass and the distribution of biomass between the stem, bark, live branches, dead branches and foliage components are described in relation to site and age. Relationships between biomass and light interception and plant area index are investigated and show some merit. Nutrient concentrations were used with the above-ground biomass data to estimate quantities of nutrients held in the various biomass components in each of the stands. Nutrient distributions in the above-ground biomass (AGB) were examined and compared to other studies. Foliar phosphorus (P) and potassium (K) concentrations were shown to be sub optimal, agreeing with results and recommendations of South African fertilisation stupies. Some concentration differences were observed between site and age classes for certain nutrients, although these differences may have been due to seasonal effects. Biomass and nutrient quantities were adjusted to yield a wood production of 100 t ha01 and compared with adjusted calculations for similar studies on other plantation crops. Total nutrients contained in the AGB of the adjusted calculations were 540.8 kg ha01 nitrogen (N), 20.4 kg ha-I P, 200.6 kg ha01 K, 241 kg ha-I calcium (Ca), 55.7 kg ha-I magnesium (Mg), with a half to two thirds of the nutrients held in the stem and bark alone. Nutrients losses vary with harvesting intensity as bark and branches may be harvested with the stem wood. Levels of nutrient removal with harvesting intensity are discussed with reference to estimated losses and gains from natural processes and management practices. An incomplete nutrient budget calculation indicated that P, K, Ca and Mg might potentially be removed in quantities greater than replaced by natural processes under stem and bark harvesting. The budget calculations lack processes such as leaching and N-fixation. It is highly probable that these processes, once quantified, may yield more negative budget results, especially for the base cations K, Ca and Mg.
Abscisic acid and other hormonal effects on growth in Spirodela.
(1969) Van Staden, Johannes.; Bornman, Chris H.
The effects of abscisic acid In particular, as well well as gibberellic acid and the cytoklnlns, 6-benzyladenine, kinetin, and 6-dlmethylal lalylamlnopurine, on the growth of Spirodela oligorrhiza were investigated. Abscisic acid effectively arrested growth permanently at concentrations down to 10¯¹ mg/I. Normal growth tended to be resumed at concentrations of 10¯² and 10¯³ mg/l between nine and twelve days after treatment. A concentration of 10[-8] mg/l, however, resulted in a significant increase in dry weight at both eight, nine and twelve days after introduction into the culture medium. It is suggested that the resumption of growth twelve days after treatment at those concentrations which inhibit growth up to nine days, was due to a possible progressive inactivation of abscisic acid resulting in a lowering of its concentration to a level that is promotive. It was furthermore found that the growth response of Spirodela in terms of dry weight production over a period of eight days is proportional to the log[10] concentration of abscisic acid. It is suggested that this curve can be used as a relatively reliable and easily performed bioassay to detect amounts of abscisic acid as low as 10[-5] μg. The assay is more reliable over the range 0.01 to 10,000 μg and appears not to be affected by gibberellin, benzyladenine and kinetin. The inhibitory effect of abscisic acid on growth in Spirodela was shown to be reversed by benzyladenine, kinetin and dimethylallalylaminopurine, although they were not equally effective in doing so. Benzyladenine at 1.0 mg/l was the most effective In overcoming growth inhibition by abscisic acid. Gibberellic acid, however, proved ineffective in reversing the inhibitory effect of abscisic acid on Spirodela oligorrhiza. The apparent Increases in growth obtained in some cases may have resulted more directly from gibberellic acid stimulation than from the Interaction of gibberel lie acid with abscisic acid.
Achene biology and the chemical control of Chromolaena odorata.
(1985) Erasmus, Daniël Jacobus.; Van Staden, Johannes.
Abstract viewable in PDF document.
The actions of, and interactions between, auxins and cytokinins and their effect on in vitro rooting of selected Eucalyptus clones.
(2011) Nakhooda, Muhammad.; Watt, Maria Paula Mousaco Deoliveira.; Mycock, David John.
Clonal propagation of Eucalyptus spp. and its hybrids allows for competitiveness in the commercial forestry industry through the propagation and preservation of superior/elite genotypes. Vegetative propagation through rooted cuttings is the industry‟s standard and the choice of clones selected for plantations are determined by their rooting ability. However, as many potentially valuable genotypes are recalcitrant to adventitious rooting, micropropagation is the only effective means of propagating them. Micropropagation results in high plantlet yields, achieved primarily through the empirical use of the key plant growth regulators (PGRs) cytokinins and auxins, for shoot and root production, respectively. Their selection for use in vitro is driven by their effects on percent rooting rather than root quality. Little is known regarding the quality of the roots of the plantlets ex vitro, but there is some evidence that they are different from those of seedlings and cuttings. It was therefore hypothesized that the properties of exogenous PGRs and their interaction with other exogenous and endogenous PGRs, influenced root development and subsequent root quality. This was tested in vitro using a good-rooting E. grandis (TAG31) and two poor-rooting E. grandis x nitens hybrid clones (GN155 and NH58). In the former, the auxins supplied during the pre-rooting culture stages (multiplication and elongation) were sufficient for 100% rooting in an auxin-free rooting medium. Different combinations of PGRs in the two pre-rooting stages, followed by rooting without auxins, revealed a direct relationship between the stability of the supplied auxin and the rooting ability of TAG31. Gas chromatographymass spectrometry (GC-MS) analyses indicated that endogenous shoot levels of indole- 3-acetic acid (IAA) influenced graviperception. Also, low IAA content was associated with atypical starch grain accumulation or its absence from root tips (53.1 nmol IAA gˉ¹ DW compared with 325.7 nmol IAA g-¹ DW in gravisensing roots). The specific roles of the natural auxins IAA and IBA on root morphogenesis were then investigated using 2,3,5-triiodobenzoic acid (TIBA; inhibits IAA transport), ρ-chlorophenoxyisobutyric acid (PCIB; inhibits auxin signal transduction), and the auxin antagonist kinetin in the rooting medium, following root induction. After 3 weeks, the mean root diameter was significantly reduced from 552.8μm (control) to 129.2μm (with PCIB) and 278.6μm (with kinetin). TIBA increased root diameter to 833.4μm, decreased Δ root length, increased root vasculature and resulted in agravitropism. Hence, whereas rooting could be induced by IBA, IAA was necessary for the maintenance of vascular integrity and graviperception. This critical role of IAA in root development is of importance as IBA, owing to its higher stability, has been traditionally relied upon for root induction in the majority of micropropagation protocols. The potential of incorporating IAA into the media formulations of in vitro protocols for poor-rooters that do not respond well to IAA was then investigated, using GN155 and NH58. While PCIB in the rooting medium of GN155 completely inhibited rooting, the addition of dihydroxyacetophenone (DHAP), an inhibitor of auxin conjugation, to the rooting medium, did not significantly increase % rooting in the presence of 0.1 mg 1ˉ¹ IBA (i.e. 50% rooting with 2mM DHAP and IBA, compared with 45% with IBA alone). The results suggested that the inability of some eucalypts to induce roots easily in vitro was not due to a deficiency in auxin signal transduction or to auxin conjugation. Instead, rooting was inhibited by an accumulation of kinetin within shoots during the pre-rooting culture stages. The endogenous levels of PGRs in shoots of GN155 and NH58 showed a strong relationship (R² = 0.943) between the shoot kinetin:auxin and shoot rootability. Substituting kinetin with the relatively less stable natural cytokinin trans-zeatin in the elongation stage resulted in a significant increase in % rooting in both clones, from 19% to 45% (GN155) and from 31% to 52% (NH58), with 0.1 mg 1ˉ¹ IAA in the rooting medium. However, omitting all cytokinins from the elongation medium, resulted in over 95% and 75% rooting of shoots of GN155 and NH58, respectively, with 0.1 mg 1ˉ¹ IAA. These results suggest that IAA is a requirement for root development and cannot be substituted by its analogues in certain root developmental events. Hence, IAA should be the preferred auxin for eucalypt micropropagation. As fundamental research, the approach taken in this study circumvents the empirical method used in improving micropropagation protocols. The importance of the properties and the interactions between endogenous and exogenous PGRs in regulating root morphogenesis, and the practical implications of these findings is emphasised.
Adventitious rooting in stem cuttings of Eucalyptus grandis Hill ex Maid.
(1988) Wilson, Philip John.; Van Staden, Johannes.
Adventitious rooting in stem cuttings of Eucalyptus grandis Hill ex Maid. was thought to be influenced by a putative inhibitor. In previous studies it has been usual to infer the presence of putative rooting inhibitors and promoters from the mung bean bioassay, but the possibility was raised that treatment responses in this assay could be mediated more by the concentration of the treatment solution than by the chemical identity of the solute. This appeared to be so: several solutes, including hydrochloric acid and common salt, were found to promote the rooting of mung bean cuttings when present in the treatment solution at an apparently injurous concentration. The concept of promoters and inhibitors of adventitious rooting, as constituted at present, was considered to be an unfavourable approach for further studies. Stem cuttings must contain a morphogen, broadly defined, which operates the 'switch' from stem to adventitious root. The leaves and buds of E.grandis stem cuttings did not appear to be sole sources of a morphogen (as is often assumed), but nevertheless the activity of the leaves and buds was good for rooting. This activity was reflected in the pattern of root emergence. A slight preponderance emerged from the leaf trace sectors of the stem, suggesting that the leaves and buds cause a morphogen (of unknown origin) to circulate in the cutting. The existence of a vascular morphogen was confirmed and it proved to be very mobile in the stem, suggesting that it is well distributed circumferentially at the base of the cutting rather than confined to the leaf trace sectors. It appeared to be super-abundant at the base of easy-to-root cuttings, but it was not possible to tell to what extent the morphogen was rendered accessible to the sites where roots initiate. In general, the rate of efflux from the transporting tissues, the rate of attenuation of the morphogen after efflux, and the number of potential sites for root initiation must interact on a small scale to determine rooting ability. The relative prominence of these groups of factors would be expected to vary with circumstances, for example at different locations within a single stem cutting, so the traditional concept of a limiting morphogen ('rhizocaline') is unhelpful in its simplest form. Nevertheless, the rhizocaline concept provides a starting point towards a more comprehensive view of adventitious rooting, which is required in order to predict and improve rooting ability. This view remains a remote objective because many of the factors which could be important have recieved very little attention and will be difficult to elucidate.
Alkaloids of Catha Spp.
(2001) Field, Courtney Robin.; Smith, Michael Trevor.
The levels of the psychoactive alkaloids S-(-)-cathinone and its primary metabolite cathine, consisting of the diastereomers (+) -norpseudoephedrine and (-)- norephedrine were determined in Catha edulis (Vahl) Forssk. ex Endl., Catha transvaalensis Codd and Catha abbottii Van Wyk & Prins. Alkaloid levels were investigated in C. edulis plants collected from three different localities in South Africa, and one from a Nairobi khat market. The efficiency of three different methods for the extraction and isolation of cathinone and cathine were investigated, viz. an aqueous acid extraction, an organic solvent extraction and an aqueous acid extraction using the commercially available Extrelutᴿ procedure. The aqueous acid extraction resulted in the rapid loss of cathinone and yielded variable alkaloid levels in replicate studies. This was also observed when this method was coupled with the Extrelutᴿ procedure. In contrast, the organic solvent extraction did not result in a loss of cathinone and provided consistent results over a number of replicates; it also proved to be a simple and rapid method for extracting and isolating cathinone and cathine. A trifluoroacetic acid (TFA) derivatization procedure which has been suggested to produce characteristic diagnostic fragments for gas chromatography / mass spectrometry (GC/MS) identification, was investigated, but failed to produce consistent TFA derivatives of cathinone and cathine. However, underivatized cathinone and cathine were easily identified by GCMS due to their unambiguous mass spectra. All subsequent studies were undertaken using the organic solvent extraction and isolation method, coupled with GC analysis and GC/MS identification of underivatized cathinone and cathine. Leaves of C. edulis were found to contain cathinone and cathine at levels 100 times higher than those of C. transvaalensis. The alkaloids were undetectable in C. abbottii. Plants grown from cuttings of C. edulis collected from the Durban Botanical Gardens were found to contain cathinone and cathine at levels of 0.410 mg per gram fresh weight and 0.157 mg per gram fresh weight in leaves, respectively, while these levels in plants derived from different localities decreased in the order: Eastern Cape (0.319 mg/g f.w cathinone and 0.029 mg/g f.w cathine), Mpumalanga (0.139 mg/g f.w. cathinone and 0.171 mg/g f.w. cathine) and Nairobi (0.032 mg/g f.w. cathinone and 0.025 mg/g f. w. cathine). In an investigation of the cathinone levels in the different plant parts it was found that the highest levels were found in leaves of the shoot tip (0.243 mg/g f.w.) but decreased with the age of the leaf and developmental stage of the plant in the order: juvenile leaves (0.124 mg/g f.w.), mature leaves (0.035 mg/g f.w.), young stem (0.033 mg/g f.w.) and mature stem (0.004 mg/g f.w.). Concomitantly, cathine levels increased with the age of the leaf: leaves of the shoot tip (0.006 mg/g f.w.), juvenile leaves (0.011 mg/g f.w.), mature leaves (0.019 mg/g f.w.). The cathine level in the young stem material was found to be the highest in the entire plant (0.270 mg/g f.w.) but decreased markedly in the mature stem (0.052 mg/g f.w.). Both cathinone and cathine levels in the mature root were greater than levels in the mature stern, being 0.012 mg cathinone per gram fresh weight, and 0.063 mg cathine per gram fresh weight. Neither cathinone nor cathine were detectable in young root material.
Aluminium-induced gene expression in sugarcane roots.
(2002) Graham, Natalie Jane.; Watt, Derek Alexander.
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.
Analysis of historical flowering data, investigations into aspects of pollen biology and selected biotechniques to complement sugarcane breeding in South Africa.
(2016) Mhlongo, Nonsikelelo Yvonne.; Snyman, Sandra Jane.
Abstract available in PDF file.
Another culture of Solanum genotypes.
(1995) Liebenberg, Denise.; Van Staden, Johannes.
Being the third most cultivated crop in South Africa, potatoes are of great economic importance. As potatoes originated from cooler areas in the world, they do not easily adapt to South African conditions. The main objective of potato breeding is, therefore, to extend the crop's limited genetic base. Progress in crop improvement is slow due to dominance, segregation and other factors caused by the tetraploid character of cultivated potatoes. A new breeding program for rapid progress has been initiated at the Vegetable and Ornamental Plant Institute, Roodeplaat, South Africa, which comprises the combination of conventional and unconventional breeding techniques. The program is based on the reduction of the ploidy level from the tetraploid to the dihaploid level to facilitate crossings with diploid wild species. Anther culture is the preferred technique for the rapid reduction of the ploidy level and has been successfully applied on different members of the Solanaceae. Cultivated potato, Solanum tuberosum is, however, an important exception. In this study various potato genotypes (tetraploid cultivars, dihaploid breeding lines and a diploid wild species) were used in experiments concerning microtechniques, alternative culture methods and medium manipulation. The main objectives were to evaluate and compare the androgenetic ability of the various genotypes used and to try and identify the factors limiting their in vitro response. Regarding microtechnique, the study focussed on the investigation of the frequency of androgenesis - as a function of plant age - and the determination of defined flower bud lenqths representative of the correct microspore developmental stage for optimal androgenetic response. Combined with an extensive histological study on the microspore development within anthers, from the time of flower selection, after a cold-pretreatment and at various time-intervals during the culture period of 42 days, the following conclusions were reached: In vitro androgenetic response proved optimal when flowers of responsive genotypes were selected during the first seven to 21 days of the flowering period. Both microspore derived embryoid- and callus development were visible within responsive anthers after a culture period of only seven days. The flower bud length required for anthers to be in the optimal stage of microspore development, e.g. the uninucleate stage, varied between the different genotypes but could readily be determined with the DAPI (4,6-diamidino-2- phenylindole) technique. It was also concluded that anthers of the tetraploid cultivar Atzimba should be selected later, between the late-uninucleate and the early-binucleate developmental stages. This suggested a limited selection period for Atzimba anthers, as starch depositioning - which prevent embryogenesis - occurs within anthers during the binucleate stage. Histologically, Atzimba showed limited embryoid development with no embryoid release, while the diploid wild species, S. canasense, proved androgenetically unresponsive. Alternative culture methods were applied to study the effect of different culture phases (liquid, double layered and agar solidified) and anther orientations (lateral, dorsal and ventral) on the androgenetic response of the potato genotypes used. Liquid cultures, based on the so-called shed-pollen technique, enhanced the androgenetic response of the tetraploid cultivar Atzimba. Optimal embryogenesis was obtained for responsive breeding line 87.2002/3 with the utilization of agar solidified media, with maximal response when anthers were cultured in the lateral orientation. No response was observed from S. canasense. The effect of medium manipulation on the androgenetic response of the three genotypes was investigated. The utilization of various combinations of different concentrations of indole-3-acetic acid (1M) and benzyladenine (BA), the alteration of the initial time of incubation of anthers on the initiation media and the use of media without growth regulators compared to that containing gibberellic acid (GA[3]), were investigated. BA had to be present in the initiation media and had a major, though not exclusive, effect on embryogenesis compared to 1M. The optimal BA concentration varied between the two trials. IAA also had an increasing effect on anther response, both in the absence of BA and, especially, in addition with relatively high BA concentrations. In this experiment, only breeding line 87.2002/3 responded. The initial culture of anthers, during the first seven to 21 days of the culture period, on media containing growth regulators proved essential for microspore derived embryoid production in the tetraploid cultivar Atzimba. As these growth regulators are metabolized in the culture media, the regular transfer at shorter, two-weekly intervals to media containing metabolically active substances, proved important. GA[3] had no enhancing-effect on embryogenesis in any of the three tetraploid cultivars. The results obtained in this study suggest that the first 21 days is the critical stage in the anther culture period in terms of the optimal time for flower selection, embryoid induction and the increase in embryogenetic response due to growth regulator influence. It is important to pre-determine the developmental stage when most microspores were in the uninucleate stage of development and to correlate this stage with a specific flower bud length. This would assure maximum response of those genotypes amenable to anther culture. It also implies a more practical and economical starting pOint to anther culture experiments. Following the determination of microspore developmental stage and pollen fertility, flowers should be selected from the donor plants only during the first three weeks of the flowering period. The composition of the nutrient media used for potato anther cultures were sufficient with respect to growth regulators. The growth regulators SA, IAA and the amines glutamine and asparagine had to be present in the initiation media, especially during the first three weeks of the culture period. As microspore development within anyone anther was found to be asynchronous, the regular transfer of anthers to fresh media is recommended to assure proper development of all microspores. The use of a slightly higher IAA concentration could be considered, but care should be taken as too-high concentrations would induce callus production. Microspore derived embryoid production is preferred, as the ploidy level of callus derived plantlets normally varies and somaclonal variation can occur. Liquid media should be considered for anther culture of tetraploid genotypes, while embryoid production can be increased by culturing the anthers of responsive genotypes on agar solidified media on the lateral orientation. Finally, the diploid wild species S. canasense seemed androgenetically unresponsive, or the media and culture conditions used did not satisfy the specific requirements of this genotype. Androgenetic amenability should first be transferred by means of interspecific crossings with a responsive dihaploid genotype, such as the breeding line 87.2002/3.
Anti-inflammatory and anti-bacterial activity of South African Erythrina species.
(2000) Pillay, Candice Claudia Natasha.; Van Staden, Johannes.; Jäger, Anna Katharina.
An investigation was undertaken to determine whether Erythrina species indigenous to South Africa contained the same type of compounds as Erythrina species not found in South Africa and to determine whether they displayed any anti-inflammatory and antibacterial activity. Phytochemical analysis was conducted using thin layer chromatography. A great similarity was found in the leaf profiles of the species being studied. The leaf and bark extracts of E. caffra and E. lysistemon appear to have similar profiles when viewed under normal light and ultraviolet light, (254 and 366 nm). These two species have similar banding patterns when stained with fast blue reagent for flavonoids and potassium hydroxide reagent for coumarins. The five species that were tested appear to contain alkaloids, flavonoids, coumarins and triterpenes just like the species not found in South Africa from this genus. Dried bark and leaves from E. caffra, E. humeana, E. latissima, E. lysistemon and E. zeyheri were screened for anti-inflammatory and anti-bacterial activity. Ethanol, ethyl acetate and water extracts were screened for both anti-inflammatory and anti-bacterial activity. The cyclooxygenase bioassay was used to test for anti-inflammatory activity. The ethanol and ethyl acetate extracts generally displayed activity while the water extracts displayed no activity for both the bark and the leaves. The bark generally displayed more cyclooxygenase inhibitory activity than the leaves. The bark of E. caffra and E. lysistemon displayed the highest cyclooxygenase inhibitory activity. The disc diffussion bioassay was used to screen for anti-bacterial activity. Anti-bacterial activity was only detected in the water extracts of the leaves. The water extracts of the bark showed very little or no activity. The bark yielded more anti-bacterial activity than the leaves. Anti-bacterial activity was mainly displayed against Gram positive bacteria. The bark of E. caffra and E. lysistemon displayed the highest anti-bacterial activity. On the basis of the screening results it was decided to use bioasssay guided fractionation in an attempt to isolate putative anti-inflammatory and anti-bacterial compounds. A hexane extract from the bark of E. lysistemon was prepared and purified using a range of chromatographic methods. Vacuum liquid chromatography, separation using a chromatotron, thin layer chromatography and high performance liquid chromatography were used to isolate anti-inflammatory compound(s). The isolation proved to be unsuccessful as the pure compound had no cyclooxygenase inhibitory activity. It was subsequently determined that the compounds were lost during the HPLC procedure. An ethanolic extract of the bark of E. Iysistemon was purified in an attempt to isolate an anti-bacterial compound(s). Vacuum liquid chromatography and separation using the chromatotron was used to purify the crude extract. The more sensitive microtitre bioassay was used to test for anti-bacterial activity against S. aureus. The isoflavone, Wighteone was isolated.
The application of microsatellites to sugarcane parentage determination and varietal identification.
(2002) Hack, Simon Matthew.; Huckett, Barbara Isobel.; Butterfield, Michael.
The use of microsatellite markers has matured and become commonplace for plant genome analyses and is now poised for widespread practical application in sugarcane. Sequence Tagged Microsatellite Site (STMS) amplification is the most prevalent microsatellite-based approach and involves the amplification of a microsatellite by designing primers that flank and hence define the microsatellite site, revealing variation in the length of repeat motifs between individuals. Twenty-six microsatellite primer pairs received from the International Sugarcane Microsatellite Consortium (ISMC) were evaluated and the STMS protocol was optimised to ensure robust and reproducible results. The objectives of this study were to use STMS for sugarcane parentage analysis and fingerprinting. Previously, Restriction Fragment Length Polymorphism (RFLP) marker data had suggested that the parentage of a genetic mapping population, sugarcane cross AA40 (N18 x CP57/614), was incorrect. Based on the assertion that the incorrect parentage was as a result of either mislabelling at planting or at seed collection, microsatellite parentage analysis was carried out on eight potential parent pairs (13 cultivars). A total of 75 markers were scored with non-parental bands (12 on average) being observed for all of the potential parent pairs and none could be identified as the true AA40 parents. It has been suggested in other plant species that PCR artefacts could give rise to non-parental bands and to investigate this the marker data of single parent DNA reactions and pooled parent pair DNA reactions or 'synthetic offspring' were compared. The results suggested that either a certain percentage of non-parental bands, perhaps 10% (maximum value observed), should be tolerated in microsatellite parentage analysis or a marker should only be considered to be discriminating for parentage if it is absent in both the parents and the pooled parent pair amplifications. Fingerprinting of 20 cultivars using 14 microsatellite primer pairs was conducted to evaluate the potential of the STMS approach for sugarcane varietal identification. It was found that only two microsatellite primer pairs were required to discriminate between all 20 cultivars with a theoretical number of non-differentiated pairs of cultivars (XK) of only 0.03. This estimator was used to determine the approximate number of microsatellites necessary for large-scale sugarcane fingerprinting.
The application of the heat pulse velocity technique to the study of transpiration from Eucalyptus grandis.
(1994) Olbrich, Bernard Wolfgang.; Pammenter, Norman William.
This thesis examines the application of the heat pulse velocity technique (HPV) to plantation-grown Eucalyptus grandis in the Eastern Transvaal, South Africa. The work addresses the application of the technique per se and is ultimately focused on improving the prediction of the hydrological impact of afforestation, to assist in the equitable management of South Africa's limited water resources. The verification of the HPV technique on E. grandis against the cut-tree method showed that the technique accurately reflected the water uptake in four three-year-old trees and a sixteen-year-old tree. It was found that accurate measurement of wound size and probe separation was essential for accurate water use estimates. The optimal probe allocation strategy for accurate measurements of transpiration in individual trees and stands of trees was examined. Stratifying the depths of implanted probes resulted in greater precision and repeatability in the HPV-derived estimates of sap flow in E. grandis. Given a limitation in the number of probes available to estimate stand transpiration, the results showed that sampling many individuals with a low sampling intensity (few probes per tree), rather than sampling few individuals intensively, improved the estimate of stand transpiration. An examination of the influence of tree age and season on transpiration rates showed that the transpiration rate per unit leaf area of E. grandis declined with age. Also, transpiration rates were higher in summer than under equivalent conditions of evaporative demand in winter. A seasonal change in the response of transpiration to VPD was implicated as the primary cause of this shift. A number of models were derived to predict transpiration from E. grandis. The variables vapour pressure deficit (VPO) and photosynthetically active radiation (PAR) were found to account for a large proportion of the observed variation in transpiration from the age sequence of trees studied. The models developed are applicable to trees of varying age, but are valid only for conditions where minimal soil water stress is experienced. The derived models were tested against two sets of independent data. This confirmed that a simple linear multiple regression adequately describes the relationship between transpiration and the two driving meteorological variables, PAR and VPO, in E. grandis. The application of a selection of the developed models on a sample data set from Sabie showed that transpiration from a three-year-old stand of E. grandis in summer may be more than double that for a sixteen-year-old stand under the same conditions. Simulated results also showed that transpiration in summer was about 25 to 50% higher than that from the same stand during winter conditions. Simulated transpiration rates from the young E. grandis stands were high, suggesting that further validation of the estimated rates is required before the models are applied. It is concluded that the HPV method is an ideal technique to estimate water use in E. grandis trees. The models developed represent a major advancement on previous models used to predict the hydrological impact of afforestation on mountain catchments.
Aspects of avian thermal physiology and frugivory of indigenous and invasive fruits in South Africa.
(2014) Thabethe, Vuyisile.; Downs, Colleen Thelma.
Indigenous and invasive plants have long been recognised as an important food source for avian frugivores worldwide. It has been suggested that seed germination success can vary amongst avian and plant species and that ingestion by avian frugivores can enhance, reduce or have no effect on seed germination. Few studies have looked at the role of both invasive and indigenous avian species on the germination success of invasive and indigenous plants in South Africa. Therefore the first aim of this study was to determine the effects of invasive rose-ringed parakeets (Psittacula krameri) and indigenous Knysna (Tauraco corythaix) and purple-crested (Gallirex porphyreolophus) turacos on seed germination of invasive alien plant species (Solanum mauritianum, Cinnamomum camphora, Psidium guajava, and Morus alba). The second aim was to determine the effect of invasive rose-ringed parakeets on seed viability and germination of indigenous Ficus species (i.e. F. lutea, F. burkei, F. sur and F. natalensis). Previous studies have shown that many avian frugivores prefer invasive alien fruits suggesting that they may be able to efficiently assimilate energy from these fruits. However few studies have focused on the consumption and digestive efficiency of invasive fleshly fruits by indigenous and invasive birds simultaneously in one study. Consequently this study also investigated the digestive efficiency of invasive and indigenous avian frugivores feeding on invasive fleshy fruits. Finally, this study assessed the seasonal effects on the thermoregulation of invasive rose-ringed parakeets. As invasive bird species are spreading in South Africa, understanding the physiological responses that equip them to tolerate a wide range of climatic conditions would be useful for modelling potential distributions and effectively managing them. Seed ingestion by Knysna and purple-crested turacos resulted in significantly higher germination success than those from whole fruits for all invasive alien plant species in this study. Germination success of manually pulp-removed seeds did not differ from that of turacos ingested seeds, suggesting that seed coat abrasion was not important for germination of these invasive alien plants. Seed passage through the digestive tract of rose-ringed parakeets resulted in significantly reduced germination success and viability of all ingested plant species, suggestion that seed ingestion by this species is disadvantageous to these plant species. These results suggest that Knysna and purple-crested turacos are legitimate seed dispersers of the four fleshy-fruited invasive plants, while rose-ringed parakeets are mainly seed predators. Results obtained from this study also suggest that rose-ringed parakeets are mainly seed predators of Ficus fruit, as none of the ingested seeds managed to germinate. In addition, all three avian species investigated managed to meet their energetic demands by feeding on invasive alien fruit only, suggesting that these plant species may sustain avian frugivores especially during periods of food scarcity. Furthermore, the results obtained also suggest that rose-ringed parakeets show seasonal thermoregulatory responses representing energy conservation, as expected. This suggests that rose-ringed parakeets are physiologically equipped to cope with a range of environmental conditions and this partly explains their global success as an invasive species.
Aspects of seed propagation of commonly utilised medicinal trees of KwaZulu-Natal.
(1996) Netshiluvhi, Thiambi Reuben.; Pammenter, Norman William.; Cooke, John Anthony.
Due to over-exploitation of commonly-used medicinal plants, mainly from KwaZuluNatal, because of ever-increasing human population growth, many of the useful medicinal plants are becoming depleted in their natural habitats. Some species like Warburgia salutaris, which is currently declared very rare in the KwaZulu-Natal province, appear to be on the verge of extinction. In order to counteract this overexploitation, this study sought to provide information that could help resource users to grow these threatened species through ex situ conservation methods. A short list of heavily utilised medicinal tree specles was selected from the approximately 700 tree species indigenous to KwaZulu/Natal. The criteria considered for short listing were; life form, species scarcity, past population status and part used. A total of 23 species were short listed, but a subset of 12 species was selected based on the availability of fruits and seeds. The aim of short-listing was to work on a manageable number of commonly utilised medicinal tree species. The seed physiology and growth of these species were studied. With the exception of Erythrophleum lasianthum and Curtisia dentata, all of them had a moisture content of 2': 20 % (on a dry mass basis), which is indicative of a recalcitrant behaviour. However, it could not be concluded that these seeds were truly recalcitrant because desiccation sensitivity was not directly assessed. Using the triphenyl tetrazolium chloride (TTC) viability test, most of the seeds of the 12 species seemed to be of good quality. Results of the TTC test for seed viability were similar to results obtained v using direct germination for most species. Results of flotation test for seed viability were different from the results obtained using direct germination for most spcies. The pre-treatment which achieved the highest germination percentage in almost all the seed types was cracking the outer coverings. Cracking pre-treatment appeared to be efficient in enhancing the removal of some substances which might inhibit germination of seeds. Hot water and acid pre-treatments frequently reduced germination. Growth of young seedlings was assessed in terms of stem diameter, height, and leaf area under sun and shade. Seedling growth in terms of stem diameter and height of most species did not show any significant difference. One of the few species which showed statistically significant differences in stem diameter growth was Ekebergia capensis. It was found that 3 out of lO of the species showed statistically significant differences in height growth. Two of the statistically significant differences in height occured on seedlings in the sun while one had statistically significant difference in the 40% shadecloth while 7 did not. Significant differences in leaf area occured on 7 out of lO species. Of these, 4 species had higher growth in the shade than in the sun while 3 had higher growth in the sun than in the shade. Generally, it appears that young developing seedlings establish themselves well under shade environment; this could be because most of the species used in this study are forest species.
Aspects of structure, growth and morphogenesis in a new filamentous red alga (Ceramiaceae, Rhodophyta)
(1993) Stirk, Wendy Ann.; Van Staden, Johannes.
Pteroceramium, a descriptive name given to an undescribed winged species closely related to Ceramium, has uniaxial filamentous thallus construction with pseudodichotomous branching. Alternate branches become dominant. This pattern of growth is referred to as cellulosympodial growth. All growth is from an apical cell which cuts off subapical cells. The subapical cells develop into axial cells. Each axial cell cuts off six pericentral cells in a ring around its apical pole. The pericentral cells divide further to form the cortical band. Pc1 always forms on the outer face of the thallus as determined by the preceding pseudodichotomy and gives rise to the larger outer wing which is a lateral expansion of the cortical band. The smaller inner wing forms from Pc6 on the inner face. The other pericentral cells give rise apically to uniseriate spines. The pericentral cells also give rise to rhizoids and adventitious lateral branches. Each axial cell has a large central vacuole with a few peripheral chloroplasts, mitochondria and floridean starch granules. The smaller wing cells have a much denser cytoplasm with fewer small vacuoles, many chloroplasts which are more closely packed together and more floridean starch granules than axial cells. Chloroplasts have a typical Rhodophyta ultrastructure with single, evenly spaced thylakoids with phycobilisomes. Pit connections have a plug core but no plug cap. Pteroceramium has a typical Polysiphonia-type triphasic life history. The carposporophyte is naked and tetraspores are produced in a characteristic decussate cruciate arrangement. The effect of a number of physical and chemical factors on growth and morphogenesis was investigated. Pteroceramium grew best at irradiance levels between 79 μmol m⁻² S¯¹ and 129 μmol m⁻² S¯¹ with growth being limited at 30 μmol m⁻² S-I. The largest axial cells and wings were obtained from the material grown at 79 μmol m⁻² S¯¹ and the smallest measurements for material grown at 129 μmol m⁻² S¯¹. Monochromatic light fields of red, green and blue caused reduced growth rates compared to the control replicates grown in a white light from both incandescent and fluorescent lights. Light quality had no effect on morphogenesis. The critical daylength for maximum rates of cell elongation was 10 hours or longer, although 16 hours light caused a decrease in final axial cell volume. Optimum temperatures for growth of Pteroceramium were between 20°C and 25°C with growth decreasing at 15°C and 30°C. Axial cell volume was reduced and wing size was stunted at these two extreme temperatures tested. Scouring by sand caused axial cells to decrease in volume although the wings were unaffected. Smothering by sand did not prevent growth although axial cells and wings were greatly decreased in size, with the wings consisting of only one or two other cells. Tumbling to disrupt gravity did not affect the angle of each pseudodichotomy. Decreased levels of nitrogen and phosphorus limited growth but had little effect on axial cell volume and wing development. Pteroceramium was stenohaline with maximum growth at 34°/[00] and reduced growth at 300/[00] and 40°/[00]. Pteroceramium grew best at pH 7.5 and pH 8.5 with decreased growth at pH 6.5 and pH 5.5. The various pHs tested had little effect on morphogenesis. The best photosynthetic responses were obtained from material preconditioned at 80 μmol m⁻² S¯¹ compared with that at 30 μmol m⁻² S¯¹ and 150 μmol m⁻² S¯¹. There was a decrease in pigment content with increasing irradiance at which the alga was grown. Phycoerythrin was the dominant pigment. Exposure to a high irradiance (3000 μmol m⁻² S¯¹) for 30 minutes or longer inhibited photosynthesis. Plants did not fully recover even 24 hours later, indicating that this damage was permanent. Pteroceramium was able to acclimatize slowly over a week to temperature changes within the range of 15°C to 25°C. Rapid increases of 5°C within this temperature range increased photosynthetic performance and a rapid drop of 5°C decreased photosynthetic performance. However, a 10°C increase or drop reduced Pteroceramium's photosynthetic performance. Photosynthetic rates were decreased in alkaline conditions and increased in acidic conditions. Pteroceramium has well defined developmental patterns with basal band growth of axial cells and tip growth in the rhizoids. The pericentral cells are formed in a set sequence similar to Ceramium species with Pcl forming on the outer face, Pc2 and Pc3 forming on the lower and upper surface nearest to Pel respectively, Pc4 and PcS forming on the lower and upper surface respectively farthest from Pel, and Pc6 forming on the inner face. This sequence is unaffected by the direction of illumination or gravity. Exogenous application of plant hormones (IAA, GA3 and kinetin) in the concentration range of 10[-9] M to 10[-5] M had no effect on growth and morphogenesis in Pteroceramium. Application of polyamines and their precursors caused a decrease in growth and a reduction in cell size at concentrations higher than 10[-4] M. Polyamine inhibitors caused a reduction in growth and cell size at concentrations higher than 10[-5] M. Arginine increased growth at concentrations 10[-5] M and 10[-6] M. High power liquid chromatography (HPLC) separation of Pteroceramium extracts indicated that spermidine was present in Pteroceramium at approximately 38 μg spermidine g¯¹ fresh weight. The apical tip exerts an apical dominance effect on the subordinate branches, suppressing their elongation. Removal of the dominant apical tip increased adventitious branch formation. This effect was not reversed by application of exogenous IAA at concentrations of 10[-9] M to 10[-4] M.
Aspects of the influence of temperature on the desiccation responses of seeds of Zizania palustris (Wild rice)
(1996) Ntuli, Tobias M.; Berjak, Patricia.; Pammenter, Norman William.
Seeds of wild rice (Zizania palustris var. interior) have been reported to show highest survival when dehydrated at 25 QC. It has also been reported that axis cells sustained least damage at this drying temperature. In the present study, a linear relationship between drying rate and dehydration temperature was established. Whereas highest positive tetrazolium staining and lowest leakage were recorded for seeds that were dehydrated at 25 QC, maximum germination was recorded for seeds dried at 20 QC. A proportion of seeds showed the presence of glasses, irrespective of the dehydration temperatures used. Parameters of the glass to liquid transition, however, correlated with neither water content nor sugar profiles. The ratio of raffinose to sucrose was similar among all the treatments. A hydroperoxide test revealed a linear relationship between peroxide levels and temperature of drying although the levels of fatty acids were not correlated with hydroperoxide levels. Butanal levels and total aldehydes evolved, on the other hand, showed a high negative correlation with peroxide levels. Electron microscopy showed that the variability and relative abundance of peripheral membrane complexes (PMCs) was the highest for cells of embryonic axes dehydrated at 25 QC and the lowest for embryonic axes of seeds dried at 10 QC. Furthermore, intramembrane particles (IMPs) were evenly distributed in cells of axes dried at 25 or 37 QC. In contrast, membranes of cells of axes dehydrated at 10 QC showed large IMP-free areas. The relative abundance of IMPs was the highest for cells of embryonic axes dried at 25 QC, and the lowest for cells of axes dehydrated at 10 QC. From these observations, it is suggested that membrane phase transition, with the concomitant elimination of proteins, accompanies dehydration of Z. paluslris seeds at 10 QC, whereas at 37 QC peroxidation may predominate.
Aspects of the invasion of southern tall grassveld by Aristida junciformis subsp. junciformis Trin. et Rupr.
(1998) Van Zyl, Douglas Dirk.; Granger, James Edmund.; Zacharias, Peter John Kenneth.
Aristida junciformis subsp. junciformis Trin. et Rupr. is an indigenous, densely tufted, evergreen perennial grass associated with the degeneration of grasslands over large areas of South Africa. More than two hundred thousand square kilometres of veld, c. 17% of the total land area of South Africa, contains A. junciformis. The aim of this study was to improve our understanding of the mechanisms by which this species invades and dominates grassland, specifically in this study, Southern Tall Grassveld. Aristida junciformis also has a low nutritional value resulting in a very low grazing value. The unpalatability of the plant is due to the high tensile strength of the leaves, whose hard, fibrous laminas are very difficult for a grazing animal to crop once their length attains 30 cm or more. This species lack of response to conventional grazing practices has often resulted in a grassland in which the carrying capacity has diminished to such an extent as to be virtually useless for grazing in a relatively short time. The extent of encroachment of veld by A. junciformis appears dependant on the frequency of disturbance and rest afforded to the veld. Annual burning and mowing maintained the species composition of A. junciformis at levels <10% whilst protection from fire, burning or grazing allows this species to dominate the herbaceous layer at levels approaching 90%. Aristida junciformis does not become moribund and is unlikely to die if left undefoliated. As the abundance of A. junciformis increases, veld deterioration gradually accelerates through increased selective grazing on remaining palatable species. Competition from adult A. junciformis plants increases the mortality of seedlings and constrains growth of surviving seedlings. Tiller production of Aristida junciformis seedlings declined from an average of 5.2 tillers per seedling in a no competition situation to 2.2 tillers per seedling when subjected to full competition. Tiller production of T. triandra seedlings decreased from 9.6 tillers per seedling free from competition to 3.3 tillers per seedling subject to full competition for resources. Once these seedlings have become adult plants they are avoided by grazing animals and grazing pressure on the remaining palatable species consequently increases. This allows the A. junciformis plants to increase their size and density in the sward. Once this density is sufficiently high, grass seedlings of either A. junciformis or more desirable grass species such as T. triandra are unable to establish, eventually resulting in a monospecific stand of A. junciformis if left undisturbed. Large amounts of caryopses are produced by A. junciformis - up to a 19 000 caryopses from a large mature plant (c. 38 000 caryopses/m²). Of these c. 40% is likely to be infertile but the remaining c. 60% viable caryopses are dispersed in a typically leptokurtic distribution, the number of caryopses dispersed rapidly declining within a 10 metre radius. A large proportion of the caryopses was trapped in surrounding foliage but in open swards caryopses had greater opportunity to be blown further distances than in a closed sward. The density of A. junciformis caryopses on the soil surface was positively correlated with the density of flowering adult plants in the area and varied from 400 caryopses/m² (density of parent plants c. 0.6 plants/m²) in less effected areas to 11000 caryopses/m² (density of adult plants c. 6 plants/m²) in severely encroached areas. The primary function of the three awns appears to be orientating the caryopsis correctly for in its descent from the parent plant to expedite germination. Caryopses orientated vertically with the awns uppermost exhibited the highest and most rapid germination (67%) compared to caryopses lying horizontally (35%) whilst only 1% of inverted caryopses germinated. Caryopses trapped in litter and effectively held off the soil surface failed to germinate. Removal of the glumes from A junciformis seed greatly enhanced the rate and overall germination of the seed except for inverted seed of which <1% germinated. The highest numbers of A. junciformis seedlings (32 seedlings/m²) were found in those areas with the highest density of caryopses on the soil surface. Despite the large amounts of caryopses produced, dispersed and landing in apparently suitable micro-sites for germination, comparatively few A. junciformis seedlings (n=992) were found and overall germination ranged between 1 % and 4% of the initial caryopses density on the soil surface. Seedling survival through winter was low with only 13% surviving to the following spring. The basal areas of A. junciformis increased overall by 66% whilst that of other grass species increased overall by only 3% in the time monitored. These results suggest that the primary method of encroachment of A. junciformis in the grass sward appears to be through vegetative expansion and not seedling recruitment. Frequent defoliation of the sward and avoidance of overgrazing to enhance the competitive abilities of palatable species and provide as high a fuel load as possible appear to be the most economically and logistically feasible ways to remove or at least inhibit A. junciformis veld encroachment at present.
Aspects of the role of cytokinins in adventitious root formation.
(1996) Taylor, Joslyn Leanda Susan.; Van Staden, Johannes.
The initiation and development of adventitious roots in cuttings are highly complex processes, influenced by both endogenous and exogenous factors. These vary from the environmental factors prior to the striking of the cutting, to the anatomical and physiological factors, within the stem. Encompassed are the nutrient status, physiological age and degree of differentiation of the tissues, and the balance of endogenous rooting inhibitors and/or promoters (including hormones). The role of cytokinins in root initiation and development has been perceived as that of an inhibitor. This investigation considered several aspects of the role played by cytokinins in the process of root development. A qualitative/quantitative analysis of the cytokinin-like activity in stem cuttings of several plants, including both easy- and difficult-to-root species was conducted on a comparative basis. There was no clear correlation between the type / level of cytokinins detected in the cuttings and the relative ease of root formation. Both qualitative and quantitative changes in the compounds exhibiting activity in the soybean callus bioassay were observed over the period of root formation in Impatiens stem cuttings. The effects on root formation in cuttings of exogenously applied auxins and cytokinins were investigated. Auxins generally promoted root number and elongation at relatively high concentrations (10[-4] M), but showed less effect on lateral root initiation and development. At high concentrations, cytokinins strongly inhibited root development, but did promote lateral root growth. In suspension culture, the effect of these hormones differed slightly, with IAA and IBA having no significant effect on root development, but NAA strongly stimulating lateral root initiation. Zeatin (10¯¹¹ M) significantly increased root length and the number of lateral roots produced. The effect of treatment of the stem cuttings with potassium permanganate and centrifugation was examined. While both these treatments have been perceived to increase root production in cuttings, no significant improvement in rooting ability following centrifugation (relative to the control) was observed. Impatiens cuttings centrifuged in the presence of distilled water showed a significantly reduced rooting ability relative to those centrifuged in the dry state. Treatment with an 8-hour pulse in 0.05 % potassium permanganate significantly increased the average root length. These treatments had an effect on the cytokinin levels and distribution in the stem cuttings. Slightly higher levels of cytokinins were associated with the increase in root number and length in both experiments.
Aspects of the structure and functioning of the vegetation of the Hlatikulu Vlei.
(1996) Guthrie, Iain Andrew.; Granger, James Edmund.
Hlatikulu Vlei, situated in the foothills of the Natal Drakensberg, is one of the priority wetlands in KwaZulu-Natal, occupying an area of 733 Hectares. The importance of Hlatikulu Vlei lies in its functions to store water, regulate stream flow and attenuate floods, as well as to provide suitable habitat for wildlife and grazing for livestock. Hlatikulu Vlei is a threatened system and has suffered the effects of human mismanagement. Forty-nine percent of the vlei has been classified as disturbed or destroyed, mainly due to the construction of two large dams and past drainage of vlei to facilitate pasture planting. The effects of grazing and fire on the plant communities has been considerably less. Vegetation communities at Hlatikulu Vlei have similarities with those at Ntabamhlope Vlei. The main plant communities present at Hlatikulu Vlei are: vlei grassland, sege-meadows, bulrushes and reedswamp. Species compositions of the mixed sedge and grass sedge-meadow community have a notably higher species diversity than similar communities sampled at Ntabamhlope Vlei and the mires at Highmoor. Soil type and moisture content are shown to be the most significant environmental factors determining the distribution of plant communities and species within the vlei. A wetland re-establishment and rehabilitation programme in the Hlatikulu Crane and Wetland Sanctuary has been effective in allowing many wetland plants to become reestablished. The sanctuary communities bear greater similarity to the sedge and rush sedge-meadow community, than the mixed sedge and grass sedge-meadow communities that were originally present. This is also reflected in the seed bank. All three Southern African crane species (Blue, Wattled and Crowned Crane) and fourteen species of waterfowl have been recorded in the Hlatikulu Crane and Wetland Sanctuary since the wetland rehabilitation programme. The waterfowl play a role in the dispersal of seeds into the sanctuary, particularly those of Schoenoplectus decipiens and Eleocharis dregeana and are in part responsible for the return of certain wetland plants to the sanctuary. The flooding of soils, the fluctuating water level and the soil type related to hummocks and to channels are shown to be responsible for the location of Cyperus denudatus, Arundinella nepalensis and Aristida junciformis in differing positions in the channels and on the hummocks and are also responsible for the maintenance and functioning of the hummocks and channels. Seed banks on the hummocks are similar to seed banks in the channels, however the extant vegetation on the hummocks is distinctly different to that in the channels. Certain species represented in the channel seed bank are being excluded from surviving to maturity.
Aspects related to the germination of Themeda triandra seed.
(1996) Baxter, Brent J. M.; Van Staden, Johannes.
Themeda triandra is a grass species of economic importance in Southern and Eastern Africa, and Australia. The species is being lost from grasslands and savannas in these areas due to poor agricultural practice, rangeland degradation, opencast mining and increased afforestation. Based on the poor re-establishment of the species from seed in sub-climax grasslands, dogma holds that T. triandra can not be re-established from seed. Recent research has, however, highlighted the potential to establish this species from seed, but the use of seed of T. triandra in re-vegetation of disturbed areas is limited by poor understanding of the seed biology of the species and low seed availability. In this Thesis ways to maximise the use of available seed are reported. Areas investigated include optimisation of seed storage conditions, overcoming primary seed dormancy, promoting germination of available seed and pre-treatment of seed to improve germination. The Thesis closes with an investigation of the environmental limits of tolerance of seedlings from the T. triandra ecotypes studied, when grown under field conditions at reciprocal sites. Two altitudinally and geographically distinct populations of T. triandra were studied; a high altitude grassland population at Cathedral Peak (Drakensberg: 1800 m) and a low altitude savanna population from the Umfolozi Game Reserve (Zululand: 90 m). At seed shed T. triandra seed is dormant. The depth and duration of primary seed dormancy varies between populations, but appears to reflect severity of the winter period experienced. More than 95% of T. triandra seed from the Drakensberg population was dormant at seed shed, compared to 55% of seed from the Zululand population. In both populations dormancy is lost during dry after-ripening. At seed shed T. triandra seed displays a high level of seed viability (> 80%). Seed temperature range -15°C to 70°C, was achieved at 25°C (± 2°C), at which temperature seed was held for 40 months. During this period viability decreased from over 80% to 50% and dormancy was lost through dry after-ripening within four (Zululand) to eight (Drakensberg) months. Loss of dormancy can be accelerated at higher temperatures, but is accompanied by rapid loss of seed viability. In contrast, viability can be maintained in storage at sub zero temperatures, but loss of dormancy is retarded. Loss of dormancy coinsides with the onset of spring. Dormant seed is capable 'of germination at a narrow range of constant temperatures (25 ° C to 40 ° C). With after-ripening, the range of temperatures at which germination takes place increases (15 ° C to 40 ° C) and the optimum temperature for germination decreases from 30 ° C in both populations to 25 ° C. After-ripened seed is capable of germination at lower water potentials than dormant seed. Similarly, seed from the low altitude population is capable of germination at lower water potentials (-1.0 MPa dormant: -1.5 MPa after-ripened) than seed from the high altitude population (-0.5 MPa dormant: -1.0 MPa afterripened). Dormancy in seed from the high altitude population is overcome by prolonged stratification (30d). In contrast, seed from the low altitude population responds to short duration stratification (5d) with longer periods proving detrimental to seed germination. Germination of dormant and non-dormant seed of T. triandra does not differ significantly in the light or dark. Neither does photoperiod, or red / far-red light exposure significantly affect germination. Seed response to light and temperature, as characterised under controlled conditions, was verified in a field seed burial experiment undertaken at the high altitude Drakensberg site during winter. Burial in soil does not affect the response of T. triandra seed to light or temperature. Loss of dormancy is accelerated in buried seed. After-ripened seed germinates over a wider range of temperatures than dormant seed. The mechanisms governing T. triandra seed dormancy and germination appear to be universal between ecotypes. Dormancy is enforced, in part, by the seed covering structures (glumes) which impose a mechanical restraint to radicle emergence. Approximately 85% of dormant seed, however, contains a dormant embryo. Embryo dormancy is enforced at seed shed by compounds inhibitory to seed germination. The germination process in T. triandra appears to be governed by endogenous gibberellins. Bioassay results reveal that endogenous gibberellin synthesis commences up to six hours sooner in after-ripened seed than in dormant seed and that the level, or concentration, of gibberellin-like compounds is substantially lower in after-ripened seed than in dormant seed. Similarly, the concentration of applied gibberellic acid required to achieve maximum germination of T. triandra seed decreased from 500 mg.l ¯¹ (8 week old seed) to 50 mg.l ¯¹ (78 week old seed) as dormancy is lost during after-ripening. Contrary to previous reports, boron does not promote T. triandra seed germination. Plant-derived smoke significantly promotes T. triandra seed germination (5% to 43% for dormant seed from the Drakensberg population). The effectiveness of smoke in promoting germination increased with increasing seed imbibition suggesting smoke action at a metabolic level. This suggestion is reinforced by the ability of smoke to bring about the germination of seed which had failed to germinate in water. Moreover when smoke is applied in combination with gibberellic acid the final level of seed germination following combined treatment is significantly greater than the level of germination achieved in the presence of either smoke or gibberellic acid alone. A similar result is achieved with joint application of smoke and kinetin, although the results were not statistically significant. Furthermore, smoke treatment reversed ABA-induced inhibition of germination of non-dormant T. triandra, wheat, radish and sunflower seed to a level equal to or greater than that achieved using GA[3] or kinetin. The possibility that smoke promotes seed germination by mimicking, or promoting the synthesis of endogenous gibberellins was investigated. Bioassay results revealed that smoke had no effect on increasing the level of endogenous gibberellin-like activity in T. triandra caryopses. The mechanism by which smoke acts to promote seed germination remains elusive, however results presented suggest that smoke may act to remove an ABA-induced block to seed germination. Consequently, it is suggested that smoke plays a permissive role in promotion of T. triandra seed germination by removing a block to the seed germination process thereby allowing endogenous gibberellins to act. Treatments which significantly improved the level of T. triandra seed germination were evaluated as seed pre-treatments. Significant improvement in germination was obtained following smoke (aq) and gibberellic acid (100 mg.l ¯¹) pre-treatment of seed. The effects of pre-treatment were evident on germination of seed for up to 21 days after pre-treatment. Seed pre-treatment with smoke had no affect on subsequent seedling growth, but gibberellic acid pre-treated seedlings developed abnormally. In contrast, short duration exposure of dormant seed to high temperature (70 0 C for 7 days) increased germination, seedling height and tiller number. Priming of seed in polyethylene glycol (PEG 8000) for 7 days significantly improves the level of T. triandra seed germination. The use of seed pre-treatment to maximise germination of T. triandra seed is discussed. Reciprocal transplanting of seedlings from both the Drakensberg and Zululand populations confirmed that the T. triandra populations under investigation are distinct ecotypes. Field transplant gardens were established in the Drakensberg, Zululand and at an intermediate altitude in Pietermaritzburg (800m). Less than 10% of planted seedlings died at any site. With increasing altitude of the field site, tiller number increased, but tiller allocation to reproduction decreased. Similarly, for both Zulu land and Drakensberg seedling transplants the time taken to reach anthesis increased with increasing altitude and the proportion of transplants which flowered decreased. These data are consistent with the climate of the field sites where the high altitude site experiences a short growing season and harsh winter while the Zululand site experiences a prolonged growing season and mild winter period. These data indicate that T. triandra ecotypes are tolerant of a wide range of environmental variables. The application of the data presented in this Thesis, in maximising the use of available seed of T. triandra for use in re-vegetation, is discussed.

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