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Typecript Includes bibliographical references

Nod factors, lipo-chitooligosaccharides (LCOs), are rhizobial signal molecules important in the establishment of nodule formation, leading to atmospheric dinitrogen fixation in legume-rhizobium symbioses. Recently, LCOs were also found to regulate other plant processes. We demonstrated that, at 10 -6 M, four LCOs produced by Bradyrhizobium japonicum enhanced soybean seed germination. Evaluation of G-protein inhibitors showed that U-73122, a phospholipase C inhibitor, also increased soybean seed germination, similar to the increase by LCO NodBj-V(C18:1 MeFuc), indicating different mechanisms for the plant perception to LCOs for nodule initiation and seed germination. This was confirmed as LCOs were not able to break dormancy of skotodormant lettuce seeds. Soybean early seedling growth was also increased by the application of LCOs. Pulse 14Ca2+ experimentation showed that the increase might also be related to an increase in Ca 2+ uptake by shoots. We confirmed this with both genistein-induced and non-induced B. japonicum 532C; however, strain 168 (a mutant unable to produce LCO) and non-host rhizobia (Rhizobium leguminosarum, Sinorhizobium meliloti), did not increase Ca2+ uptake. Addition of 1.6 g L-1 casein hydrolysate in yeast extract mannitol broth drastically increased bacterial growth and increased volume-basis LCO production, but decreased LCO production per cell. Best conditions for sterilizing and storing LCOs were determined. LCO should be sterilized by using polyestersulfone filter or autoclaving for up to 30 minutes. LCO was degraded faster when stored at room temperatures (23 +/- 2°C) than low temperature (4 +/- 1°C) and can be stored more than one year.

In Africa, the grain sorghum (Sorghum bicolor (L.) Moench), is malted to provide the most important ingredient in brewing, malt, which is used primarily for the production of traditional (opaque) sorghum beer. Malting is the germination of cereal grain in moist air under controlled conditions, the primary objective being to promote the development of hydrolytic enzymes which are not present in the ungerminated grain. The malting process can be physically split into three distinct unit operations (viz. steeping, germination and drying). To date, little attention has been given to optimising the conditions of steeping for sorghum. The effects of different steeping variables (time, temperature and aeration) on the quality (in terms of diastatic power (amylase activity), free amino nitrogen and hot water extract) of sorghum malt for brewing were investigated. Malt quality was found to increase with steeping time, over the range 16-40 hours and the optimum steeping temperature was found to be in the range 25 to 30°C. Aeration during steeping appeared to be necessary to maximise the malt quality, particularly when steeping was conducted for long periods at high temperatures. Of particular significance was the observation that final sorghum malt quality was highly significantly correlated (p<0.01) with grain moisture content at steep-out (the end of the imbibition period). When steeping conditions based on these findings were used, a germination temperature of 25-30°C was found to be optimal for sorghum malt quality. As with steep-out moisture, green malt (grain after the specified germination time) moisture content was correlated Significantly (p<0.01) with final sorghum malt quality. The finding that sorghum malt quality is related to steepout moisture content was given further substance when it was shown that the stimulatory effect on sorghum malt quality of steeping sorghum in a dilute solution of alkali, actually increases the amount of water taken up during steeping probably because the alkali disrupted the pericarp cell wall structure of the grain. Barley malting practices have taken advantage of the knowledge that the exogenous application of gibberellic acid can enhance the synthesis of the critically important malt hydrolytic enzyme, a-amylase. To date, literature on the effect of exogenous application of gibberellic acid on sorghum malt quality has been inconclusive; with reports both of no effects, and of positive effects, on amylase activity. To elucidate the possible control mechanisms involved in sorghum germination, a combined HPLC-radioimmunoassay technique was used to determine the levels of selected plant growth regulators from the groups auxin, cytokinins, gibberellins and abscisic acid in sorghum at various stages of germination. Levels of gibberellic acid were low throughout germination. During germination the levels of the other plant growth regulators declined, but a peak in cytokinins followed the first visible signs of root protrusion. The high level of the germination inhibitor and gibberellic acid antagonist, abscisic acid, in the germ (embryo inclusive of scutellum) portion of the mature non-germinated grains was noteworthy. Based on these findings, it was determined that sorghum malt quality could in fact be improved significantly by the application of exogenous gibberellic acid. However, this was effective only if it was administered during the end of steeping or at the beginning of the germination step. By optimising the conditions of steeping and germination and by steeping in dilute NaOH or in gibberellic acid not only should it be possible to enhance the quality of sorghum malt, it should be possible to reduce the time required to obtain the specific quality, thereby offering a saving to the sorghum maltster in terms of operation costs and enhancing the total throughput possible from the malting plant. / Thesis (Ph.D.)-University of Natal, Durban, 1997.

Thesis (MScAgric (Agronomy)--University of Stellenbosch, 2007. / Soil salinity is a serious constrain to crop production in many countries. In the Western Cape canola production area, large variability in soil properties within the same field is a very common feature. Some of this variability may be ascribed to differences in salinity. These saline patches affected the germination and survival of cereal crops like wheat and may therefore also be a reason for the high variability and generally low canola yields obtained in this area. It will therefore be important to quantify the effect of such saline conditions on the germination and seedling establishment of canola in an effort to improve yields. In this study several experiments were conducted under controlled conditions in either Petri dishes in an incubator or pots in a temperature controlled glasshouse to determine the salinity response of canola. Results of the experiments done in Petri dishes, showed that the germination response of canola (Brassica napus L., cv. Tornado) to increasing levels of salinity is very similar to that of wheat (Triticum aestivum L., cv SST 88). Although canola germinated more rapidly than wheat at a temperature of 20 ºC, the final percentage germination for both crops decreased severely when NaCl induced salinity exceeded levels of an EC of 8 dS m-1, but less severe responses were obtained with NaCl/CaCl2 induced salinity. Under soil conditions an interaction between soil type and salinity level showed that the responses in sandy soil to an increase in salinity from 0 to 8 iv dS m-1 were less severe than in loamy soil and especially when salinity stress was induced by NaCl. Although cultivar Spektrum showed more tolerance to saline conditions during the germination and seedling stages in pot experiments when compared to other cultivars tested, these results will have to be confirmed in field trails where grain yield and quality can be measured.

The biochemical changes taking place during soybean seed development and gennination, and some aspects of desiccation tolerance were assessed with reference to lipid peroxidation and antioxidant systems. During nonnal seed development, fresh weight and dry weight increased between 20 and 50 days after flowering (DAF), concomitant with the accumulation of triacylglycerols and sugar reserves, after which dry weight remained almost unchanged, and fresh weight decreased. Seed moisture content decreased rapidly during the last stages of development. High levels of lipid peroxidation were evident between 20 and 45 DAF, and decreased thereafter. An examination of antioxidant systems revealed that whereas total glutathione levels accumulated continuously throughout the 80 days of seed development, both dehydroascorbic acid (DHA) reductase and ascorbate free radical (AFR) reductase increased concurrently with the increase in total ascorbate content, and the overall levels did not decrease markedly during maturation drying. Ascorbate peroxidase (ASC POD) activity was high during the period ofgreatest ascorbate accumulation. Both catalase (CAn and superoxide dismutase (SOD) activities increased progressively during early seed development (20-40 DAF), but showed variable patterns of change during maturational drying, in marked contrast to ASC POD which declined from 40 DAF to undetectable levels at 70 DAF. An assessment of the relationship between the antioxidant systems and lipid peroxidation was made during imbibition and gennination, as it has been suggested that controlling free radicals was a critical event in early imbibition. Unexpectedly, lipid peroxidation increased progressively in both seeds and isolated axes, and were eight-fold higher at 48 hours of imbibition compared to dry tissues. A progressive, and co-ordinated, increase in CAT, total glutathione, total ascorbate pool, guaiacol POD, ASC POD, and SOD appeared to parallel the rise in lipid peroxidation in both whole seeds and axes. Variable responses were evident between seeds and axes for the enzymes AFR reductase and DHA reductase In order to gain a further insight into the dynamics of desiccation-tolerance and desiccationsensitivity, imbibing seeds were subjected to an unscheduled dehydration treatment, and then rehydrated for up to 24 hours. During these hydration-dehydration-rehydration (H-D-R) treatments, changes in lipid peroxidation and antioxidant systems were measured. Concurrent with the loss of viability in the axes of seeds dehydrated after 24 and 36 hours of imbibition, there were increases in both lipid peroxidation and solute leakage. Unscheduled drying was seen to be a critical stage, as intolerant axes showed four- to eightfold increases in lipid peroxidation, which were only partially reduced on subsequent rehydration. Tolerant axes, on the other hand, were able to maintain low, basal levels of lipid hydroperoxides on drying. The relationship between these observations and the antioxidant systems showed that the antioxidant enzymes CAT, ASC POD, AFR reductase, DHA reductase, guaiacol POD and SOD declined markedly during the unscheduled drying, whereas GSH and ASC declined only slightly. On rehydration, most of the enzymes, total glutathione, and total ascorbate pool increased, the only exception being the loss of ASC POD activity. ORA reductase, which was seen to decrease as a part of nonnal gennination, declined progressively also in H-D-R treatments. These results suggested that loss of viability was not attributable to a decline of the antioxidant systems but rather to the combined deleterious effects of increased lipid peroxidation, and a generalized and moderately compromised antioxidant system. These studies have indicated that the occurrence of lipid peroxidation can be seen as a nonnal part of seed development and gennination. The H-D-R studies, on the other hand, supported the concept that the balance between peroxidation reactions and the protective systems was critical to the development of desiccation tolerance. / Thesis (Ph.D.)-University of Natal, Durban, 2003.

Master of Science / Department of Agronomy / P. V. Vara Prasad / In many countries producers have been growing varieties of hard white winter (HWW) wheat since decades. The cause of concern is most varieties of HWW wheat are susceptible to pre-harvest sprouting (PHS) which affects grain quality. Environmental conditions like high humidity, precipitation, heavy dew and hormonal activity at physiological maturity stimulate PHS in HWW. To alleviate these conditions research was carried out at KSU. KS01HW163-4, a sprouting tolerance line was crossed with Heyne, a sprout susceptible cultivar. A total of 224 doubled haploid (DH) lines thus produced were phenotyped in the present study through experiments conducted in controlled environments. The objectives of this research were to (i) characterize and phenotype the doubled haploid lines for PHS in controlled environments; (ii) understand the impact of growth environment (high temperature and/or drought) and; (iii) impact of exogenous application of growth hormones on tolerance to pre-harvest sprouting in the parental lines of the doubled haploid population. The phenotypic data collected from this research will be ultimately combined with the genotypic data to identify DNA markers related to PHS tolerance and provide DNA markers for marker assisted selection. Based on my results of the germination percentages from the 224 DH lines, the population was distributed as susceptible, and tolerant to PHS showing a bimodal distribution and X[superscript]2 analysis indicating a complimentary gene action. From the study of the influence of environmental factors on PHS, my results confirmed a definite influence of stress on sprouting. Under optimum temperature (OT), KS01HS163-4 was tolerant to PHS, but at HT and/or drought it became susceptible to PHS. Growth under stressed conditions changed the tolerance levels to PHS. Seed dry-weight, and harvest index were also influenced negatively due to stress. Therefore multi-location tests must be conducted with variable environments to test the stability of a variety to PHS. From the study of the influence of phytohormones on PHS, the results suggest that tolerance to sprouting was seen in seeds from plants sprayed with abscisic acid (ABA) and paclobutrazol (GA-inhibitor) treatments where as those from gibberellic acid (GA) treatment showed susceptibility to sprouting.

Thesis (MScAgric)--University of Stellenbosch, 1994 / 127 Leaves printed single pages, preliminary pages i-viii and numberd pages 1-118.Includes bibliography,tables and figures. / Date on t.p.: Dec. 1994. / ENGLISH ABSTRACT: Effects of pH,sucrose, boric acid and temperature on in vitro germination of pollen of Protea repens (L.) L. cv. 'Embers' were investigated in hanging-drop culture to establish optimum conditions for germination. Optimum values were found within ranges pH: 5 - 8, sucroseconcentration:0.4 - 0.7 M, boric acid concentration:50 - 500 mg.e-1 , and incubation temperature: 5 - 30°C. Storage temperature and humidity on pollen viability was studied in four Pro tea clones. Pollen was stored at a range of temperatures and relative humidities for up to one year and tested for ability to germinate in vitro. Pollen of P. repens cv. 'Sneyd', P. eximia cv. 'Fiery Duchess' andP. magnifica clone 'T 84 07 OS', stored in liquid nitrogen (-196°C) and in a freezer (-14° to -18°C), retained a germination percentage as high as that of fresh pollen regardless of humidity. The study showed that long-term storage of protea pollen is not feasible at temperatures above O°C. The correlations between the fluorochromatic reaction (FCR) and germinability were found to be low and nonsignificant. Fifteen month old cryopreserved 'Sneyd' pollen was shown to retain its ability to fertilize and set seed equal to that of fresh pollen. 'Sneyd', 'Fiery Duchess' and 'T 84 07 OS' pollen could be repeatedly thawed and frozen in liquid nitrogen before its germinability in vitro decreased. The morphology and size of Protea pollen was studied, using light and scanning electron microscopy. Polymorphic grains were observed in two interspecific hybrids. Very small differences in pollen grain size were recorded between clones/species. The male fertility of 25 interspecific Pro tea hybrids, based on in vitro pollen germinability, was investigated. The majority of hybrids were found to be sufficiently fertile to be used in a breeding programme. Pistil structure and pollen tube pathways were investigated in 'Sneyd' using light and scanning electron microscopy. The pistil had four distinct regions, consisting of the stigma, the vertebra-shaped upper style, the heart-shaped lower style, and the ovary. The pistil had a stylar canal along its entire length and this canal was also the route by which pollen tubes grew to the ovary. Very low numbers of pollen tubes reached the ovary. The breeding system of 'Sneyd' and 'Fiery Duchess' were determined from pollen tube and seed set data, after controlled hand-pollinations. Both clones were found to be fully selfcompatible. Very low percentages autogamous seed set were recorded. Interspecific crosses had a low success rate. An incompatibility reaction probably occurred on the stigma and/or in the upper style regions.The attainment of maximum stigma receptivity of two Protea cultivars was investigated by means of seed set experiments, pollen tube growth observations and measurement of the degree of opening and closing of the stigmatic groove. Both cultivars were found to be protandrous. The maximum stigmatic groove width of both cultivars never exceeded the pollen grain diameter. It was concluded that Protea spp. must be hand-pollinated two to six days after anthesis in order to obtain maximum seed set; while for the observations of pollen tubes in the ovary, inflorescences must not be harvested before seven days after pollination. / AFRIKAANSE OPSOMMING: Ten einde 'n optimale medium vir die in vitro-ontkieming van Protea-stuifmeel te ontwikkel, is die effek van pH, sukrose, boorsuur, en temperatuur op die in vitro-ontkieming van Protea repens (L.) L. cv. 'Embers'-stuifmeel deur middel van die hangdruppel-metode ondersoek. Die volgende reekse van veranderlikes wat getoets is, is as optimaal gevind; pH: 5 - 8, sukrosekonsentrasie: 0.4 - 0.7 M, boorsuurkonsentrasie: 50 - 500 mg.e-1 en inkubasietemperatuur: 5 - 30°C. Die invloed van bergingstemperatuur en humiditeit op stuifmeel-Iewenskragtigheid is in vier Protea-klone ondersoek. Stuifmeel is gestoor by 'n reeks temperature en relatiewe humiditeite vir tot eenjaar, en vir in vitro-ontkiemingsvermoe getoets.' Stuifmeel van P. repens <?v. 'Sneyd', P. eximia cv. 'Fiery Duchess', en P. magnifica kloon'T 84 07 OS', in vloeibare stikstof (-196°C) en in 'n vrieskas (-14° tot - 18°C) geberg, het 'n ontkiemingspersentasie gelykstaande aan die van vars stuifmeel gehandhaaf, ongeag van die humiditeit. Hierdie studie het verder aangetoon dat langtermynberging van Protea-stuifmeel bokant O°C me die moeite werd is me. Die korrelasie tussen die fluorochromatiese reaksie (FCR) en ontkieming was laag en me betekemsvol me. 'Sneyd' -stuifmeel wat vir 15 maande in vloeibare stikstof gestoor is, het die bevrugtings- en saadsetvermoe gelykstaande aan vars stuifmeel behou. 'Sneyd', 'Fiery Duchess' en 'T 84 07 OS'-stuifmeel kon herhaaldelik in vloeibare stikstof gevries en ontdooi word voordat hul ontkiemingsvermoe afgeneem het. Die morfologie en grootte van Proteastuifmeel is deur middel van lig- en skandeerelektronmikroskopie bestudeer. Polimorfiese stuifmeelkorrels is in twee interspesie-hibriede waargeneem. Baie klein verskille in stuifmeelkorrelgroottes het tussen klone/spesies voorgekom. Die manlike vrugbaarheid van 25 Protea-interspesiehibriede, gebaseer op die in vitro-ontkiembaarheid, is ondersoek. Dit is gevind dat die meerderheid hibriede 'n voldoende graad van vrugbaarheid het om in 'n teelprogram te gebruik. Die stamperstruktuur en stuifmeelbuiswee in P. repens is deur middel van lig- en skandeer-elektronmikroskopie ondersoek. Die stamper bestaan uit vier kenmerkende gebiede, naamlik die stempel, die werwelvormige bo-styl, die hartvormige onderstyl, en die vrugbeginsel. Die stamper het 'n stylkanaal regdeur die totale lengte van die stamper, en hierdie kanaal is ook die weg waarvolgens stuifmeelbuise na die vrugbeginsel gegroei het. Min stuifmeelbuise het die vrugbeginsel bereik. Die teelsisteem van 'Sneyd' en 'Fiery Duchess' is deur middel van stuifmeelbuis- en saadsetdata na gekontroleerde handbestuiwings ondersoek. Beide kIone was ten volle selfverenigbaar. Die persentasie outogame saadset was baie laag. Interspesiekruisings het 'n baie lae sukses gehad. Dit is voorgestel dat die onverenigbaarheidsreaksie in die stempel en/of in die bopunt van die styl plaasvind. Die bereiking van maksimum stempelontvanklikheid van twee Protea-cultivars is deur middel van saadseteksperimente, stuifmeelbuisdata en waarnemings van die oop- en toemaak van die stempelgroef ondersoek. Beide cultivars was protandries. Die maksimum stempelgroefwydte het nooit die stuifmeelkorreldeursnee oorskry nie. Dit is afgelei dat Protea-spesies twee tot ses dae na antese handbestuif moet word vir optimale saadset. Vir die waarneming van stuifmeelbuise in die vrugbeginsel, moet bloeiwyses nie voor sewe dae na bestuiwing geoes word nie.

Woody plant encroachment is one of the major problems worldwide because it affects negatively the herbaceous layer, which provide forage for livestock production. However, the role of ruminants particularly browsers in the dispersal of woody plant seeds still remains a concern for farmers interested in grass production. Seedpods of various woody plant species constitute a crucial part of the diet of herbivores during the dry season because of their high nutritional quality compared to herbaceous material. The interaction of associated diet quality, seed characteristics and animal species among other factors play a pivotal part in the success of livestock faecal seeds dispersion. Furthermore, dispersed seeds that successfully grow into mature woody plants become an important source of protein for herbivores. The use of woody plants as a source forage is known to be limited by plant secondary metabolites (PSMs) such as condensed tannins. The objectives of this study were to determine 1) the effects of condensed tannins and crude protein of Vachellia tortilis and Dichrostachys cinerea pods in seed recovery and germination fed to goats, and 2) the effects of diet mixing on the feed intake of plant species by goats. In the first experiment, a total of 12 female indigenous goats and 12 female Pedi sheep were utilised in this study, with the average body weights of 29.50 kg ± 1.60 (S.E) and 28.70 kg ± 1.60, respectively. Twelve goats were grouped into two groups of six goats per group, one group was fed D. cinerea pods and the other group was fed V. tortilis pods. The group of 12 sheep were divided similarly, the one group was fed D. cinerea pods and the other group was fed V. tortilis pods. Each animal was given V. tortilis and D. cinerea pods at 2.50% of their body weight. All animals were allowed to consume D. cinerea or V. tortilis pods within 24 h, after which the remaining pods were collected and weighed. Faecal collection commenced immediately after the 24 h pods feeding and was carried on until no seeds were discovered in faeces. All faeces extracted from sheep and goats were collected daily in the morning from the faecal bags. In the second experiment, a total of 24 indigenous goats with average body weight of 26.6 kg ± 0.51 were utilised. Goats were arbitrarily selected and grouped into four groups of six goats per group (goats were placed individually in 2 m2 pens). Each group was fed one of the following diets: diet one - Searsia lancea, diet two - S. pyroides, diet three - Euclea crispa and diet four - was a combination of the three plant species (Searsia lancea, S. pyroides and Euclea crispa). Searsia lancea, S. pyroides and E. crispa branches were collected every morning prior to feeding, and were weighed before offering the animals. Refusals were gathered and weighed, and intake was calculated as distinction between weight in and refusals. Plant species foliage were analysed for crude protein, condensed tannin, acid detergent lignin, acid detergent fibre and neutral detergent fibre. During the first experiment, the cumulative percentage seed recovery of V. tortilis from goats (46.00 % ± 1.90) and sheep (52.00 % ± 2.93) was significantly higher than D. cinerea from goats (13 % ± 1.47) and sheep (24.00 % ± 1.16). Germination percentage of D. cinerea seeds that passed through the gastro-intestinal tract of goats (33.12 % ± 2.94) and sheep (36.00 % ± 2.68) was significantly higher than V. tortilis seeds that passed through the gastro-intestinal tract of goats (28.98 % ± 2.68) and sheep (23.04 % ± 2.81). Average D. cinerea (34.56 % ± 1.99) and V. tortilis (26.02 %± 2.10) seeds that went through the gastro-intestinal of goats and sheep had a significantly higher germination rate than the control (i.e. no passage through the gut; D. cinerea = 2.31 % ± 1.55, V. tortilis = 5.07 % ± 2.68). The high mean cumulative percentage seed recovery of V. tortilis (18.80 %) may be attributed to the relatively higher crude protein than D. cinerea (12.20 %). This may encourage animal seed dispersal and germination of woody plant species with relatively high crude protein content. In the second experiment, Searsia lancea contained 8.50 % CP, 21.46 % acid detergent fibre (ADF), 12.50 % ADL and 39.37 % NDF. Searsia pyroides had 9.03 % CP, 27.07 % ADF, 10.89 % ADL and 40.30 % NDF. Euclea crispa had 6.19 % CP, 26.20 % ADF, 16.63 % ADL and 30.02 % NDF. Mixed diet (combination of the three plant species) had 8.96 % CP, 23.72 % ADF, 11.13 % ADL and 38.28 % NDF. Searsia lancea had 2.70 % of CTs while S. pyroides had 5.20 % CT, E. crispa had 6.44 % CT and mixed diet had 7.20 % CT. The mean dry matter intake varied significantly among dietary groups (P < 0.001). Similarly, goats offered a mixed diet consumed more CTs (P < 0.01) than those offered individual forage species. The high mean cumulative percentage seed recovery of V. tortilis may be attributed to the higher crude protein of V. tortilis (18.80 %) than D. cinerea (12.20 %). Higher passage rate may encourage animal seed dispersal and germination of plant species. The results from experiment two support the postulation that animals foraging in mixed diet systems consume more PSMs and achieve higher dry matter intake than animals confined to monocultures or single species feeding systems. Given that woody plant encroachment is already reducing farm-grazing capacities in African savannas and this problem is predicted to double by 2050, strategies that improve herbivore ability to consume woody plants will increase forage availability and inform bush control programmes and policies. Moreover, the concomitant increase in CTs by goats exposed to diets with diverse species also has positive implications for animal / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)