This thesis gives the results of an investigation of the strengths and particularly the crust forming potential of arable soils from Botswana and the relationship to seedling emergence of sorghum, the most important crop in Botswana and one that because of its small seed weight (about 2 0 mg) can fail to emerge through a hard soil crust. A review of the literature suggested that the soil factors which had to be considered were dispersibility of clay and factors which affect this, and the bulk density of the soils. Soils which are found in many tropical and sub-tropical regions, with low organic matter and inactive clays can set hard after a simple wetting and drying cycle. In these soils, the strength is very strongly dependent on the water content, showing a hyperbolic or exponential relationship-, and the strengths when dry may be very greatly increased by remoulding the wet soil. The crust strength required to prevent seedling emergence varies with the size of the seedling and for cotton (seed weight about 80 mg), 1-3 MPa penetration resistance measured with a penetrometer is sufficient to prevent emergence. Seedlings exert a total force proportional to their number. In the experimental programme, seedlings' forces were measured, seedling emergence observed in a field experiment under crusting conditions, and the strength characteristics of a group of soils, representative of arable soils in Botswana, studied. A sorghum seedling was found to exert a maximum force of about 1 N or dividing by the area of the plumule, a pressure of about 0,5 MPa. The field experiment showed that much better emergence was obtained from planting 15 seeds together than from planting 4 seeds together when a crust formed after planting. A study of 32 soils, most of them sand to sandy loam in texture but with a few clays and hydroirorphic soils, from arable areas in Botswana showed the sandy to sandy loam soils to have high bulk densities ( 1,45-1 ,75 Mg/m3) and extremely low organic carbon contents (0,12-0,85 g/100g). The bulk densities of all the soils were inversely related to the organic carbon content and this was itself related to the clay content of the soils. The bulk densities of the sands were dependent on the grading of the sand fraction. Many of the soils were sensitive to remoulding in the Emerson test and the sands to loany sands had 0,4-1,0 g/100 g water dispersible clay. Measurements of tensile strength on air-dry samples showed that all the soils, except for one sand, set hard after a wetting and drying cycle, giving for vacuum wet samples indirect tensile strengths 1,0-14,4 kPa. For the sands to sandy loams this strength was related to the water dispersible clay content. Samples wet at atmospheric pressure were weaker than the vacuum wet samples, the reduction in strength was related to the air porosity of the non-vacuum wet soils prior to drying. Remoulding the soils prior to drying them increased the strength by a factor of up to 50 times, giving strengths from 4 kPa to 600 kPa. The strength after remoulding was dependent on the Emerson index. Compacting the soils increased their strength greatly and to an extent that agreed with the hypothesis that the strength obtained was proportional to the area of contact between the particles. Experiments on penetration resistance at a range of water contents were performed on a few soils. A hyperbolic relationship between water content and penetration resistance of the surface soil was found for sand to sandy loam soils, with the maximum resistance of dry soils above 2 MPa. The penetration resistance of the sandy loam soil was Increased three times by disturbing it when wet. Sprinkler wetting the sieved soils was not found to affect the penetration resistance by a large amount compared and other methods of welting. Penetration resistance was measured on air-dry samples of most of the main group of soils following varying degrees of wetting with a rainfall simulator. The clays and hydromorphic soils gave very low values of penetration resistance under these conditions, showing that at organic carbon contents of about 1% and clay contents from 20 to 30%, the decreased bulk density and tendency to form aggregates' on drying overcame the tendency to set hard. The mean values for the sands to sandy loams were from 1 to 6 MPa so all these soils could offer significant resistance to a sorghum seedling. The penetration resistance of the sands and loam/ sands depended on their bulk densities and water dispersible clay contents, while the penetration resistance of the sandy loams depended only on the water dispersible clay content.

In this work the effects of mother plant nutrition and flower position on the plant in relation to different harvest stages on French beans (Phaseolus vulgaris L. cv. Cascade); seed yield, quality and progeny performance have been studied. In three pot experiments under glass and one in the field, different levels of nitrogen, phosphorus, potassium and molybdenum were applied to the mother plant and it was found that higher seed yields were obtained with the higher nutrient levels tested, but these high seed yields were not necessarily accompanied by high quality. Seed yield and quality increased with the increase of nitrogen and potassium. The phosphorus effect however, was different, in that the seed yield increased in relation to the levels but the seed quality decreased. Molybdenum was found to be necessary in moderate amounts, especially for the seed quality. The interactions between nitrogen and phosphorus levels (NP) and between nitrogen, phosphorus and potassium (NPK) were found to be very important for bean seed quality, because their significant effect was similar and constantly present in most of the seed vigour components examined. It seems that the combinations of high nitrogen levels with moderate amounts of phosphorus applied to the mother plant resulted in seed of good quality. In one pot experiment under glass the progeny performance was examined, using seed from the 1st mother plant nutrition experiment and it was found that the mother plant nutrition affected the progeny in it's early stages. This effect disappeared later and no difference in progeny seed yield was found. In two pot experiments under glass the bean pods were harvested at different stages and the pods from the main axis were separated from the pods on the secondary branches grown mainly from the axils of the primary leaves. It was found that the quality of seed from the secondary branches was lower than the quality of seed from the main axis. However, this difference in seed quality became less with the later harvests. The following tests were used to assess seed quality: the official germination test, seedling evaluation test, cold test, and electrical conductivity test. In addition, the seed size was determined and the seeds were analysed for total nitrogen, phosphorus and potassium content.

Digitized by Kansas Correctional Industries

Seed vigor, although not well understood, is a key critical component for yield and is in part due to a well establishment and vigorous stand of canola (<i>Brassica napus</i>) seedling under less than ideal conditions in Western Canada. My objective was to determine what constitutes vigor by studying the response of a black seed line and a yellow seed line imbibed at 8 ºC in either water, saline or osmotic solutions, abscisic acid (ABA), ABA biosynthesis inhibitor, gibberellin (GA4+7), inhibitor of GA biosynthesis and a germination promoter, fusicoccin. Also tested was the effect of seed coat (testa) on seed germination rate and percent germination. Previous studies have established that seed vigor is in part hormonal controlled and genetically controlled. In our study, gene expression was investigated by using transcriptome analysis and hormonal analysis was used to quantitate the changes in hormones and their metabolites during germination. <p> Both the black and the yellow canola seed lines were very sensitive to increasing concentrations of saline and osmotic solutions; however, at the same osmotic potential, osmotic solutions were more inhibitory. The yellow seed line was more sensitive to these conditions than the black seed line. As expected, ABA delayed seed germination, whereas GA4+7 enhanced seed germination and GA4+7 partially overcame the inhibitory effect of ABA. The seed coat was a major factor affecting the germination rate of the yellow seed line; however, GA4+7 overcame the inhibitory effect of the seed coat, whereas ABA exacerbated it. Fusicoccin was more stimulatory to germination than GA4+7; however, unlike GA4+7, it was unable to overcome the inhibitory effect of paclobutrazol, a GA biosynthesis inhibitor. Fluridone, an ABA biosynthesis inhibitor, was unable to overcome the inhibitory effects of a saline solution suggesting that the inhibitory effect was not due to elevated ABA levels. Ethylene, a stimulator of germination, did not appear to be involved in the germination of these two lines. Controlled deterioration at 35 ºC, 85% RH was either partially or completely overcome by exogenous GA4+7. This study demonstrates that the role of hormones, salinity and seed coat on the germination of canola seed under low temperature environmental conditions. <p>During germination, ABA declined while GA4 increased. Higher ABA was found in un-germinated seeds compared to germinated seeds. GA4+7 was lower in seeds imbibed in the saline solution compared to seeds imbibed in water. Un-germinated seeds imbibed in ABA had lower GA4+7 compared to un-germinated seeds imbibed in water; however, the contents of GA4+7 were similar for germinated seeds imbibed in either water or ABA. Phaseic acid (PA) and dihydrophaseic acid (DPA) increased in seeds imbibed in either water, the saline solution or ABA, while they decreased in seeds imbibed in GA4+7. In addition, we found that ABA inhibited GA4 biosynthesis, whereas, GA had no effect on ABA biosynthesis, but altered the ABA catabolic pathway. <p> Gene expression profiles revealed that there are significant differences between un-germinated and germinated seeds. Seeds imbibed in water, GA4+7, a saline solution or ABA had different gene profiles. LEA genes, hormone-related genes, hydrolase-related genes and specific seed germination-related genes were identified and their expression profiles were finely associated with seed germination performance.

Seed vigor, although not well understood, is a key critical component for yield and is in part due to a well establishment and vigorous stand of canola (<i>Brassica napus</i>) seedling under less than ideal conditions in Western Canada. My objective was to determine what constitutes vigor by studying the response of a black seed line and a yellow seed line imbibed at 8 ºC in either water, saline or osmotic solutions, abscisic acid (ABA), ABA biosynthesis inhibitor, gibberellin (GA4+7), inhibitor of GA biosynthesis and a germination promoter, fusicoccin. Also tested was the effect of seed coat (testa) on seed germination rate and percent germination. Previous studies have established that seed vigor is in part hormonal controlled and genetically controlled. In our study, gene expression was investigated by using transcriptome analysis and hormonal analysis was used to quantitate the changes in hormones and their metabolites during germination. <p> Both the black and the yellow canola seed lines were very sensitive to increasing concentrations of saline and osmotic solutions; however, at the same osmotic potential, osmotic solutions were more inhibitory. The yellow seed line was more sensitive to these conditions than the black seed line. As expected, ABA delayed seed germination, whereas GA4+7 enhanced seed germination and GA4+7 partially overcame the inhibitory effect of ABA. The seed coat was a major factor affecting the germination rate of the yellow seed line; however, GA4+7 overcame the inhibitory effect of the seed coat, whereas ABA exacerbated it. Fusicoccin was more stimulatory to germination than GA4+7; however, unlike GA4+7, it was unable to overcome the inhibitory effect of paclobutrazol, a GA biosynthesis inhibitor. Fluridone, an ABA biosynthesis inhibitor, was unable to overcome the inhibitory effects of a saline solution suggesting that the inhibitory effect was not due to elevated ABA levels. Ethylene, a stimulator of germination, did not appear to be involved in the germination of these two lines. Controlled deterioration at 35 ºC, 85% RH was either partially or completely overcome by exogenous GA4+7. This study demonstrates that the role of hormones, salinity and seed coat on the germination of canola seed under low temperature environmental conditions. <p>During germination, ABA declined while GA4 increased. Higher ABA was found in un-germinated seeds compared to germinated seeds. GA4+7 was lower in seeds imbibed in the saline solution compared to seeds imbibed in water. Un-germinated seeds imbibed in ABA had lower GA4+7 compared to un-germinated seeds imbibed in water; however, the contents of GA4+7 were similar for germinated seeds imbibed in either water or ABA. Phaseic acid (PA) and dihydrophaseic acid (DPA) increased in seeds imbibed in either water, the saline solution or ABA, while they decreased in seeds imbibed in GA4+7. In addition, we found that ABA inhibited GA4 biosynthesis, whereas, GA had no effect on ABA biosynthesis, but altered the ABA catabolic pathway. <p> Gene expression profiles revealed that there are significant differences between un-germinated and germinated seeds. Seeds imbibed in water, GA4+7, a saline solution or ABA had different gene profiles. LEA genes, hormone-related genes, hydrolase-related genes and specific seed germination-related genes were identified and their expression profiles were finely associated with seed germination performance.

Thesis--Auburn University, 2010. / Abstract. Includes bibliographic references.

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. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1996.

A protocol is presented for the rapid (28 days) induction of microtubers on micropropagated layered potato plantlets of 'Kennebec', 'Russet Burbank' and 'Superior' in medium devoid of growth regulators. With this method the addition of coumarin, 6-(2-chloroethyl)-trimethylammonium chloride and 6-benzylamino-purine to the microtuberization medium either had no effect or significantly reduced microtuber weight per plantlet. Increasing the incubation period from 28 to 56 days significantly increased the weight of microtubers per plantlet and the proportion of microtubers heavier than 1 gram. Increasing the volume of microtuberization medium from 50 to 100 ml significantly increased the number of microtubers per plantlet. Microtuber dormancy periods were cultivar-specific and microtubers $ le$250 mg had longer dormancy periods as compared to microtubers $>$250 mg. A positive correlation was established between endogenous abscisic acid levels and microtuber dormancy periods. Microtubers $ le$250 mg had lower specific gravity, fewer eyes and produce fewer sprouts than microtubers $>$250 mg. Microtuber-derived plants were generally single-stemmed. Severe physiological ageing treatment ($>$2500 degree-days) had no effect on microtuber sprout development, stem number, tuber number and only minimally influenced tuber weight of microtuber-derived plants. Decreasing field in-row planting density from 30 to 10 cm reduced tuber weights and numbers per plant but increased them on a per hectare basis. Economic analysis indicated that optimum planting density varied depending on plantlet cost. The optimum planting density was 10 cm if the cost of the plantlet was $0.10 or less, 20 if plantlet cost were from $0.10 and $0.20 and 30 cm for plantlet cost greater than $0.20. A potato seed tuber certification program adapted to the needs and constraints of Egypt is presented.

This study has demonstrated and investigated the expression of a cDNA, coding for the pea seed storage protein vicilin, in the yeast, Saccharomyces cerevisiae. The cDNA was contained in the plasmid pLG1.63 and has been characterised and sequenced. The sequence showed that the cDNA coded for a 47KDa type of vicilin with a putative 24 amino-acid signal peptide, a proteolytic cleavage site and one glycosylation signal. The cDNA was cloned into two yeast expression vectors. The first utilised the GALIO promoter rendering expression of the cDNA inducible galactose, the construct was called pDUB2300. The second construct, pDUB2302, placed the cDNA under the control of the PGK promoter, rendering the cDNA constitutively expressed. When transformed into yeast, both constructs produced an immunoreactive vicilin species of M(_r) =49KDa. In the case of pDUB2302 the protein was produced at up to 5.5% of total cell protein. The protein was shown to be associated with a particulate fraction and displayed altered precipitation characteristics when compared with pea vicilin. By using tunicanydn and N-glycosidase, the protein was shown to be unglycosylated. Partial purification and (^35)S-methionine labelling demonstrated that the signal pep tide remained uncleaved. Cell fractionation studies indicated that vicilin was enriched in the yeast microsomal fraction, suggesting that vicilin was located in the EH. This was confirmed electron microscopy of immuno-gold labelled yeast which showed vicilin associated with the ER. The electron micrographs also suggested that a small proportion of the protein might be reaching thecolgi apparatus and the vacuole membrane. The presence of specific cleavage products on some western blots suggested that vicilin possessed a cleavage site for a yeast protease, though whether this was the same site as the pea proteolytic cleavage site was not determined. The pattern and nature of the expression of vicilin from this cDNA was discussed in the context of heterologous protein expression in yeast in general and plant storage protein expression in yeast in particular.

Many of the world's tropical forests are under threat, with anthropogenic deforestation and degradation occurring at an alarming rate. Seed dispersal in an important process in forest restoration and regeneration, however seed rain is often low in degraded habitats, hindering reforestation efforts. Up to 90% of tropical fruit are dispersed by vertebrates, animal seed dispersers are incredibly important in maintaining forest health. Additionally, frugivores that disperse seeds into degraded areas may be of great importance in aiding natural reforestation. I therefore, investigated the potential role of the frugivorous monkey, Chlorocebus tantalus tantalus, in forest regeneration via seed dispersal. I assessed its patterns of habitat use, the quality and quantity of seed dispersal it provides, the effectiveness of current conservation management actions and the density of C. t. tantalus at Ngel Nyaki Forest Reserve. I found C. t. tantalus utilised forest, edge and grassland habitats, and dispersed seeds of 28 pioneer and forest edge species into these habitats. Moreover, the number of seeds dispersed per faeces was significantly higher in the grassland than the forest with means of 16.4 +/- 6.1 and 3.4 +/- 0.97 seeds >2 mm in these habitats respectively. Germination of C. t. tantalus dispersed seeds was highest in grazed grassland and lowest in grassland protected from grazing and fire, suggesting the current practice of fencing off grassland to protect from cattle grazing may not be sufficient on its own, due to seed-seedling conflict in habitat suitability. These findings combined with an estimated density of 28 +/- 10.8 C. t. tantalus individuals km⁻² suggests C. t. tantalus may benefit forest regeneration via its role as a seed disperser, provided appropriate management actions are implemented. This and other frugivorous species may play similar roles in other locations but such roles need to be investigated in order to implement management actions that ensure their seed dispersal benefits are maximised for forest restoration and regeneration.