Crop models have great potential as research tools, for crop system management and policy analysis. One of the most promising future uses of crop models is in crop improvement. The limitation in the use of models for crop improvement has been the inability of crop models to predict variety differences. Currently, the CANEGRO model, a sugarcane crop model developed the South African Sugar Association Experiment Station (Inman-Bamber, 1991a) can only model the performance of the NC0376 variety. Experiments were undertaken in the South East Lowveld of Zimbabwe, which is a hot and dry environment where sugarcane is grown under irrigation, to examine the canopy growth and development of four commercial varieties, ZN6, ZN7, N14 and NC0376. The study aimed at determining variety differences in canopy (tillers and leaves) development, develop parameters that can be used to model variety differences and test the improved CANEGRO canopy model for its ability to predict variety differences in canopy growth and development. For the late season, the numbers of leaves and tillers produced by each variety were counted every fortnight throughout the crop cycle. The total leaf area of the varieties and the individual leaf area on a stalk were determined using a Delta-T leaf area meter every fortnight. The date of emergence of successive leaves on a stalk was recorded daily. The leaf angles of each variety were measured every fortnight. The amount of photosynthetically active radiation (PAR) intercepted by the varieties was measured using a SunScan Ceptometer. Tillering and tiller senescence rates, phyllochron intervals, extinction coefficients and base temperatures were determined for the growth and development processes of varieties ZN6, ZN7, N14 and NC0376. Tiller and leaf population development was varietal. Tillering and leaf emergence were highly correlated to thermal time while tiller and leaf senescence were less correlated to thermal time. The poor correlation of the senescence phases to thermal time could mean that tiller and leaf senescence was driven by other factors other than thermal time. PAR interception could be one of these factors. The data showed that PAR interception could be a trigger of tiller senescence. The study showed that the tiller and leaf population development could be approximated by two linear equations. Tillering will be the first linear phase and tiller senescence the second linear phase. The first linear phase is driven by thermal time. While the second linear phase is triggered by PAR interception, the major driving factors need to be determined. This study proposed the use of two linear equations to model tiller and leaf population development as opposed to the polynomial equations used in the current CANEGRO model. Polynomial equations assume the factors driving tillering and tiller senescence are the same. The green leaf numbers per plant showed that all varieties experienced a decline in green leaf numbers with crop age. Varieties NC0376 and ZN7 had the greatest decline in green leaf numbers per plant while varieties N14 and ZN6 had the least decline. Variety ZN7 had the highest number of green leaves per plant while NC0376 had the least. The tiller growth and development was divided into three phases: the exponential phase during the initiation of stalks, the first linear phase during a period of rapid stalk elongation and the second linear phase during sucrose accumulation and maturation. The first two phases of development were driven by thermal time while the sucrose accumulation was not. There were variety differences in tiller growth and development. There were variety differences in base temperature for the development of various components of the canopy. Internode formation occurred at lower air temperatures than stalk elongation and tillering while canopy heights were correlated with higher air temperatures. This implies that internode formation could occur under conditions unsuitable for stalk elongation and may explain the short internodes frequently observed in stalks exposed to winter during rapid stalk elongation. The basic requirements for physiological parameters are that they should be stable across different environments, have significant differences between varieties and have physiological meaning. The parameters studied were thermal time requirement for shoot emergence, leaf appearance, to reach peak tiller population and to start of stalk elongation; surface area of the youngest biggest leaf, leaf number of the youngest biggest leaf, PAR transmission at the start of tiller senescence, extinction coefficients, and peak and mature tiller population. The difference between varieties in thermal time to shoot emergence was least using a base temperature of 16 QC compared to using 10 QC and therefore 16 QC could be a more appropriate base temperature for shoot emergence. The accumulated soil temperatures were less variable than accumulated air temperature and could therefore be a more reliable driver of shoot emergence. However, the limitations in the use of soil temperature are that it is not a readily available measurement and that it is not easy to measure. The gradual increase in phyllochron intervals appeared to be a better method of predicting leaf appearance compared to using a broken stick model. The phyllochron gradient was proposed, as it is likely to be a more robust way of modelling leaf appearance. The varieties had different phyllochron gradients. Variety ZN7 had highest rate of leaf appearance and produced the highest number of leaves per stalk while NC0376 had the lowest rate and produced the least number of leaves. There were statistically significant differences between varieties (P = 0,05) in PAR transmission at the start of tiller senescence and a base temperature of 16 QC was best at determining accumulated thermal time to the start of tiller senescence. Varieties with higher peak tiller population had higher final tiller population, lower thermal time per tiller and a higher ratio of final to peak tiller population. There were differences between varieties in the youngest leaf number attaining maximum leaf area and the leaf area of the youngest biggest leaf. Variety Nl4 had the biggest leaves and NC0376 had the smallest. Variety Nl4 had the highest leaf area index (LAI) while ZN7 had the lowest. There were significant differences (P = 0,01) in PAR intercepted by the varieties but there were no significant differences in extinction coefficients. Extinction coefficients increased with crop age. The varieties had significantly different (P = 0,01) leaf angles and ellipsoidal leaf angle distribution parameters. The measurement of LAI using SunScan ceptometer provided a better estimate of extinction coefficients than LAI measured using Delta-T leaf area meter. Model evaluation showed that CANEGRO canopy model version 2 was improved compared to than version 1. The model (version 2) was accurate in predicting tiller heights and dead leaf numbers per stalk. It was fairly accurate in predicting green leaf numbers per plant, stalk population and intercepted PAR but was poor in predicting LA!. Version 2 has proved to be a substantial improvement over version 1 in predicting stalk population. Generally, the version 2 model overestimated tiller heights early and underestimated later, overestimated the tiller population and LAI after peak, underestimated green leaf numbers per stalk for varieties ZN6, ZN7 and N14 and overestimated dead leaf numbers per stalk and intercepted PAR. The version 2 model predicted a constant green leaf numbers per plant and LAI from peak to harvest while observed data showed that green leaf numbers per stalk and LAI decreased towards harvest. Version 2 model predicted the tiller population of NC0376 closely but underestimated tiller senescence in N14 and also underestimated final tiller population in varieties ZN6 and ZN7. Future model refinements may need to focus on the prediction of the sigmoid pattern of tiller heights. The model may need to be calibrated to predict the green leaf numbers per stalk accurately, which should possibly improve the prediction of LAI that in turn could improve the prediction of intercepted PAR. The improvement in the timing and rate of tiller senescence should improve the prediction of tiller population particularly in varieties ZN6, ZN7 and N14. The study showed that the broken stick method IS superior in explaining leaf and tiller population development compared to using polynomial equations. The development of variety parameters helped improve the prediction of variety differences in canopy growth and development. A major weakness of most crop models is modelling variety differences in canopy growth and development. The inability of crop models to predict variety differences has limited their use in plant breeding. This study has resulted in an improved version of CANEGRO version 1 that is an initial attempt at modelling variety differences of sugarcane. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

The insight that the fossil fuel reserves are limited, together with concerns over security of supply (i.e. the oil crises), initiated the first raise of interest in biomass and all other renewable energy forms. However, the concern grew that global warming and the resulting climate change were enhanced by carbon dioxide (CO2) emissions resulting from fossil fuel consumption. Meanwhile, biomass energy is thought to account for 14–15% of total world energy consumption. Hence, the exploitation and utilization of biomass energy are effective and necessary for relieving the pressures caused by environmental pollution and fossil fuel shortage (Lv et al., 2010). Recently, extensive research aimed at converting biomass to useful energy have been carried out, especially pyrolysis and gasification, which are particularly suitable for the effective and efficient utilization of biomass. Sugarcane bagasse is the main byproduct of sugarcane mill, ready available at the production site so that it may be a suitable raw material for the production of biofuels, chemicals, and electricity. Among the methods that have been previously studied, is this particular study we explore thermochemical process, gasification. The properties of sugarcane bagasse relevant to gasification are briefly reviewed. The compositions of the bagasse as a fuel are variable, especially with respect to inorganic constituents important to the critical problems of sintering, fouling and slagging. Alkali and alkaline earth metals, in combination with other fuel elements such as silica and sulfur, are responsible for many undesirable reactions in a gasifier system.

The use of genetic markers that are linked to specific traits in sugarcane has the potential to increase the efficiency of the selection of improved varieties. Conventionally, markers are identified by analysing the segregation of potential markers and traits in the progeny of single crosses. However, this approach is not practical for sugarcane breeding programmes where replicated, well characterized progenies do not exist. The objective of this project was to investigate the potential of using commercial varieties for identifying markers associated with some of the important traits in sugarcane. This approach would be far more effective than dealing with single progenies since the traits of commercial varieties have already been characterized. The DNA of fifty commercial varieties of sugarcane was amplified by RAPD PCR using forty-one arbitrary decamer primers. Analysis of the resulting banding profiles, obtained by agarose gel electrophoresis, yielded fifty-four reliable polymorphic fragments. Two approaches were used to identify putative markers linked to the traits of resistance to eldana, sugarcane mosaic virus, and smut: (1) a correlation approach which attempted to identify whether the presence of any polymorphisms could be used to imply the existence of a particular phenotypic state, and (2) multiple regression analysis, in order to determine whether polymorphisms could be used to predict the performance of the varieties for each of the traits. Both approaches appeared to identify associations between polymorphisms and the traits, although multiple regression analysis yielded the most informative results and was able to assign statistical values to the associations. Using multiple regression, the best predictive model was obtained for sugarcane mosaic virus resistance. This model consisted of four polymorphisms and had an r² of 0.40l. By dividing the resistance ratings into three groups (resistant, intermediate and susceptible), 52% of the varieties were correctly classified and only 2% of the varieties were predicted in opposite groups (i .e. predicted susceptible when actually resistant, and vice versa). The predictive model for eldana resistance consisted offour polymorphisms and had an r² of 0.347. This model classified 30% of the varieties in the correct group of three while none of the varieties were predicted in opposite groups. The predictive model for smut resistance consisted of three polymorphisms and had an r² of 0.316. This model classified 30% of the varieties in the correct group of three while 2% of the varieties were predicted in opposite groups. Further analysis of sugarcane varieties using additional polyrnorphisrns has the potential to identify markers linked to important traits. These markers could be used for marker-assisted selection to increase the efficiency of selecting for improved sugarcane genotypes for commercial release. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.

Inheritance studies have previously been undertaken at the South African Sugar Association Experiment Station (SASEX) under irrigated conditions. Since most sugarcane is grown in South Africa under dryland (raingrown) conditions, heritability estimates were calculated under these conditions in this study and compared to those previously obtained under irrigated conditions. A sugarcane population consisting of 12 crosses, 32 offspring in each cross, and their parents were planted in the first two selection stages of the SASEX selection programme to ascertain which stage provided the most useful information when selecting parent cultivars. Data collected from Stage 2 was more reliable than data collected from Stage 1. Variance components, narrow and broad sense heritabilities, correlations among traits, and clonal repeatabilities between seasons were determined for 11 sugarcane traits at Stages 1 and 2. These traits studied included: stalk population; stalk diameter; stalk height; cane mass; dry matter % cane; fibre % cane; brix % cane; brix % dry matter; purity; pol % cane; and ers % cane. Narrow sense heritabilities of the sugarcane traits were estimated by mid-parent offspring regression . Alternative heritability estimates were obtained through restricted maximum likelihood (REML) analysis of the unbalanced North Carolina design II at Stage 2. Although narrow sense heritabilities determined by mid-parent-offspring regression were comparable with those previously determined at SASEX and by other workers, REML was more efficient than regression. Use of REML enabled additive and non-additive genetic variance components to be estimated by allocating degrees of freedom to treatments and the interactions between the different treatments. Heritability estimates varied for different traits and compared favourably with those obtained under irrigated conditions and by other workers. Additive genetic variance was more important than non-additive genetic variance for some characters, but not for stalk population, cane mass, and dry matter % cane, for which both variances were important. Selection of parent cultivars for all sucrose-related traits, fibre % cane, and stalk diameter should be as successful under raingrown as under irrigated conditions, provided that the environmental variation is determined efficiently under raingrown conditions. Environmental correlations were observed between some traits, particularly between the yield related traits, and may have influenced heritability estimates for those traits determined by mid-parent offspring regression. Stalk diameter, fibre % cane, and brix % dry matter were the most repeatable traits between seasons. Cane mass was the least repeatable trait between Stages 1 and 2 but was highly repeatable between plant (-P) and ratoon (-R) crops of Stage 2. Stalk diameter was positively correlated with brix % dry matter (0.457-P and 0.623-R) and strongly negatively correlated with stalk population (-0.790-P and -0.711-R) and fibre % cane (-0.628-P and -0.651-R). Cane mass was strongly positively correlated with brix % dry matter (0.638-P and 0.679-R). By selecting for brix % dry matter and stalk diameter, indirect selection for cane mass would be possible. Brix % dry matter was determined as the most reliable trait on which to base parental and commercial cultivar selection because it was highly heritable, highly repeatable and highly positively correlated with stalk diameter and cane mass. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1995.

Made available in DSpace on 2014-06-11T19:26:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-16Bitstream added on 2014-06-13T18:29:33Z : No. of bitstreams: 1 melloni_mlg_me_jabo.pdf: 351261 bytes, checksum: d4725b9a138adce6f019c7948559b267 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Este trabalho teve por finalidade o estudo da fisiologia e a viabilidade do pólen para fins de melhoramento de cana-de-açúcar. Para isto três trabalhos foram realizados. No primeiro foi estimada a taxa de autofecundação de famílias obtidas por policruzamento envolvendo quatro genótipos (IACSP95-5000, IAC94-1099, RB867515, SP89-1115) além do impacto da taxa de autofecundação nas taxas de sobrevivência e seleção das progênies obtidas. O policruzamento foi realizado na cidade de Ribeirão Preto-SP. Após o teste de germinação das sementes obtidas, as que apresentaram viabilidade (sementes da IACSP95-5000 e SP89-1115) foram semeadas em campo onde permaneceram por 10 meses para avaliação da taxa de sobrevivência (4, 6 e 10 meses) e seleção (aos 10 meses). A taxa de autofecundação foi determinada por meio de marcadores microssatélites. Para as progênies da IACSP95-5000 e SP89-1115 as taxas de autofecundação foram de 98,5% e 0% respectivamente. A taxa da sobrevivência foi maior para as progênies da SP89-1115 nas duas últimas avaliações bem como a taxa de seleção no valor de 78,26% contra 28,18% das progênies da IACSP95-5000. O segundo trabalho teve como objetivo comparar dois corantes (iodo 0,1N e azul de lactofenol) utilizados para avaliar viabilidade do pólen de inflorescências de cana-de-açúcar com a germinação em meio de cultura. A leitura da viabilidade do pólen foi realizada em microscópio fotônico (250x) para cada corante, dentro de cada horário, para quatro genótipos de cana-de-açúcar na estação de hibridação do IAC em Uruçuca-Ba. O meio de cultura para germinação foi composto de água de chuva, sacarose (300g/L) e ágar-ágar (10g/L). O corante azul de lactofenol mostrou queda da viabilidade do pólen com o passar das horas de avaliação, o corante de iodo... / The present work aimed to investigate the flowering and the physiology of pollen viability for sugarcane breeding. For these purpose three studies were conduce. In the first one, the selfing rate of polycross derived families involving 4 parents (IACSP95-5000, IAC94-1099, RB867515, SP89-1115) was estimate as also as its impact on the survival and selection rate of the progenies. The polycross was done at Ribeirão Preto-SP. After the seed germination test, those that were viable (IACSP95-5000 and SP89-1115 seeds) were planted in the field where stayed for 10 months for the survival (4, 6 and 10 months) and selection rate (at 10 months) evaluation. The selfing rate was estimated through microsatellite markers. For the progenies derived from IACSP95-5000 and SP89-1115 the selfing rates were 98.5% and 0% respectively. The survival rate was higher for SP89-1115 derived progenies at the last two evaluations as well as the selection rate with 78.26% against 28.18% observed for IACSP95-5000 progenies. In the second study two dyes (iodine 0.1N and lactophenol blue) used to evaluate pollen viability from sugarcane tassels were compared with the pollen germination at culture media composed by rain water, sucrose (300g/l) and agar-agar. The read of pollen viability was done with a photonic microscope (250x) for each dye, in each time for 4 sugarcane genotypes at the IAC cross station (Uruçuca-BA). The lactophenol dye showed a degree of pollen viability with the evaluation time while the iodine dye did not. Compared with the germination in culture media at 6 hours, both dyes did not differ, but, over estimated the viability obtained through germination. At 8 and 9 hours, the iodine dye over estimate the pollen germination while the lactophenol blue under estimate. In the third study the behavior of 17 sugarcane... (Complete abstract click electronic access below)

Orientador: Luciana Rossini Pinto / Coorientador: Maximiliano Salles Scarpari / Banca: Rinaldo César de Paula / Banca: Jose Antonio Bressiani / Resumo: Este trabalho teve por finalidade o estudo da fisiologia e a viabilidade do pólen para fins de melhoramento de cana-de-açúcar. Para isto três trabalhos foram realizados. No primeiro foi estimada a taxa de autofecundação de famílias obtidas por policruzamento envolvendo quatro genótipos (IACSP95-5000, IAC94-1099, RB867515, SP89-1115) além do impacto da taxa de autofecundação nas taxas de sobrevivência e seleção das progênies obtidas. O policruzamento foi realizado na cidade de Ribeirão Preto-SP. Após o teste de germinação das sementes obtidas, as que apresentaram viabilidade (sementes da IACSP95-5000 e SP89-1115) foram semeadas em campo onde permaneceram por 10 meses para avaliação da taxa de sobrevivência (4, 6 e 10 meses) e seleção (aos 10 meses). A taxa de autofecundação foi determinada por meio de marcadores microssatélites. Para as progênies da IACSP95-5000 e SP89-1115 as taxas de autofecundação foram de 98,5% e 0% respectivamente. A taxa da sobrevivência foi maior para as progênies da SP89-1115 nas duas últimas avaliações bem como a taxa de seleção no valor de 78,26% contra 28,18% das progênies da IACSP95-5000. O segundo trabalho teve como objetivo comparar dois corantes (iodo 0,1N e azul de lactofenol) utilizados para avaliar viabilidade do pólen de inflorescências de cana-de-açúcar com a germinação em meio de cultura. A leitura da viabilidade do pólen foi realizada em microscópio fotônico (250x) para cada corante, dentro de cada horário, para quatro genótipos de cana-de-açúcar na estação de hibridação do IAC em Uruçuca-Ba. O meio de cultura para germinação foi composto de água de chuva, sacarose (300g/L) e ágar-ágar (10g/L). O corante azul de lactofenol mostrou queda da viabilidade do pólen com o passar das horas de avaliação, o corante de iodo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The present work aimed to investigate the flowering and the physiology of pollen viability for sugarcane breeding. For these purpose three studies were conduce. In the first one, the selfing rate of polycross derived families involving 4 parents (IACSP95-5000, IAC94-1099, RB867515, SP89-1115) was estimate as also as its impact on the survival and selection rate of the progenies. The polycross was done at Ribeirão Preto-SP. After the seed germination test, those that were viable (IACSP95-5000 and SP89-1115 seeds) were planted in the field where stayed for 10 months for the survival (4, 6 and 10 months) and selection rate (at 10 months) evaluation. The selfing rate was estimated through microsatellite markers. For the progenies derived from IACSP95-5000 and SP89-1115 the selfing rates were 98.5% and 0% respectively. The survival rate was higher for SP89-1115 derived progenies at the last two evaluations as well as the selection rate with 78.26% against 28.18% observed for IACSP95-5000 progenies. In the second study two dyes (iodine 0.1N and lactophenol blue) used to evaluate pollen viability from sugarcane tassels were compared with the pollen germination at culture media composed by rain water, sucrose (300g/l) and agar-agar. The read of pollen viability was done with a photonic microscope (250x) for each dye, in each time for 4 sugarcane genotypes at the IAC cross station (Uruçuca-BA). The lactophenol dye showed a degree of pollen viability with the evaluation time while the iodine dye did not. Compared with the germination in culture media at 6 hours, both dyes did not differ, but, over estimated the viability obtained through germination. At 8 and 9 hours, the iodine dye over estimate the pollen germination while the lactophenol blue under estimate. In the third study the behavior of 17 sugarcane... (Complete abstract click electronic access below) / Mestre

Thesis (PhD (Genetics))—University of Stellenbosch, 2007. / Association analysis was used to improve the efficiency of breeding sugarcane varieties for the negatively correlated traits of resistance to sugarcane smut and the eldana stalk borer. 275 RFLP and 1056 AFLP markers were scored across a population of 77 genotypes representing the genetic variation present within the SASRI breeding programme. Genetic diversity analysis did not detect significant structure within the population. Regression analysis identified 64 markers significantly associated with smut rating and 115 markers associated with eldana rating at r2 > 6.25%. Individual markers with the largest effects explained 15.9% of the phenotypic variation in smut rating and 20.2% of the variation in eldana. Five markers were significantly associated with both smut and eldana. In each case the marker effect was negatively correlated between the two traits, suggesting that they are genetically as well as phenotypically negatively correlated.

The effects of increasing periods under sugarcane monoculture (managed by preharvest burning) on soil organic matter content and related soil properties were investigated in the 0 to 10 cm layer of a sandy coastal Ochric Cambisol (Glenrosa soils) and a red Rhodic Ferralsol (Hutton soil) from the sugar belt of KwaZulu-Natal. The organic C content at both sites under undisturbed vegetation is about 48 g C kg-I. This declined exponentially with increasing years under sugarcane. For the Glenrosa site, organic C reached a new equilibrium level of about 20 g kg-I while at the Hutton soil the equivalent value was 41 g kg-I. The higher organic matter content maintained at the Hutton site was attributed mainly to clay protection of organic matter since the clay content of the Hutton soil was 61 % compared to the 18 % for the Glenrosa soil. The loss of soil organic matter under sugarcane resulted in a concomitant decline in soil microbial biomass C, microbial quotient, basal respiration, aggregate stability, arylsulphatase and acid phosphatase activity. The activities of arylsulphatase and acid phosphatase like those for concentrations of microbial biomass and organic C, were higher for the Hutton than Glenrosa soils. At the Glenrosa site, the natural OBC abundance in soils was used to calculate the loss offorest-derived, native soil C and the concomitant input of sugarcane-derived C. Sugarcane-derived C increased over time until it accounted for about 61 % of organic C in the surface 10 cm in soils that had been under sugarcane for greater than 50 years. The effects of agricultural land use (including burnt sugarcane) on organic matter content and related soil properties were compared with those under undisturbed native grassland in KwaZulu-Natal. Two separate farms situated on Oxisols were used and both contained fields with continuous long-term (>20 years) cropping histories. At site 1, soil organic C content in the surface 10 cm followed the order permanent kikuyu pasture> annual ryegrass pasture> native grassland> preharvest burnt sugarcane > maize under conventional tillage (CT). At site 2, organic C in the surface 20 cm decreased in the general order kikuyu pasture> native grassland > annual ryegrass pasture> maize under zero tillage (ZT) ~ maize (CT). Organic C, microbial biomass C, microbial quotient, basal respiration and aggregate stability were substantially greater in the surface 5 cm under maize ZT than maize CT. In the undisturbed sites (eg native grassland and kikuyu pasture) the metabolic quotient increased with depth. By contrast under maize CT and sugarcane there was no significant stratification of organic C, yet there was a sharp decrease in metabolic quotient with depth. Aggregate stability was high under both native grassland and kikuyu pasture and it remained high to 40 cm depth under the deep-rooted kikuyu pasture. Although soil organic C was similar under maize CT and sugarcane, values for microbial biomass C, microbial quotient, basal respiration and aggregate stability were lower, and those for metabolic quotient and bulk density were higher, under sugarcane. This was attributed to the fallow nature of the soil in the interrows of sugarcane fields. It was concluded that the loss of soil organic matter, microbial activity and aggregate stability is potentially problematic under maize CT and sugarcane and measures to improve organic matter status should be considered. For sugarcane, this could include green cane harvesting and the use of green manure crops in rotation. / Thesis (M.Sc.)- University of Natal, Pietermaritzburg, 2002.

Increasing the sucrose yield and the disease resistance of plants are two major objectives of the transgenic sugarcane plant programme in South Africa. The sugarcane culm has thus been identified as one of the main target areas for transgene expression. A shortage of reliable promoter elements as well as patent limitations have necessitated the isolation of promoters that are preferentially expressed in the sugarcane culm. In the present study two different approaches were followed to isolate such promoters, and the bacterial promoter, rolC, was evaluated for tissue-specific expression in sugarcane. Differential display is a non-directed technique that was used to identify genes that are differentially expressed in the mature sugarcane culm. The original method was modified, and four putative culm-preferential fragments were isolated. Sequence and hybridisation analyses revealed that these fragments were false positives, and could therefore not be used to obtain a culm-specific promoter. Activity of the Agrobacterium rolC promoter was evaluated by analysing expression patterns of two reporter genes in the mature culm of transgenic sugarcane plants. Nucleic acid analyses indicated that the foreign DNA was incorporated into the sugarcane genome, and that mRNA transcripts were produced. Histochemical analysis was done to visualise rolC-driven GUS and GFP expression in the mature sugarcane culm. In both cases the reporter gene expression was restricted to the vascular bundles and specifically to the phloem. A directed approach was followed to isolate the gene and subsequently the promoter of the β-subunit of pyrophosphate-dependent phosphofructokinase (PFP-β). An incomplete cDNA clone was obtained from a mature culm cDNA library, and was used for the screening of a sugarcane genomic library. Two clones containing different parts of the PFP-β gene were isolated. A Deletion Factory™ system was used to analyse the clone containing the 5' end of the gene. The first five exons and 1747 bp of the 5' flanking region of the gene were sequenced. Preliminary activity analysis of the promoter region was done by constructing two expression vectors, and analysing transient GUS expression in sugarcane callus. Results indicated that the promoter is capable of driving foreign gene expression in callus. Transient expression levels were lower than that of the maize Ubi-1 promoter. Further analysis of the 5' flanking region will be done to establish whether cis-acting elements outside the analysed area have an influence on the activity of the promoter. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1997.

The eldana stalk borer (Eldana saccharina Walker) is the most serious pest of the Southern African sugarcane industry, and it is imperative that effective control measures are available to minimize economic damage. Because conventional control methods have had limited success, cultivar resistance is seen as the most viable method of controlling infestation. However, due to the space- and time-consuming nature of the present screening methods, only small numbers of cultivars can be tested relatively late in the Plant Breeding selection programme. Increased resistance in breeding and selection populations is therefore slow. Buds are a preferred entry point of eldana larvae as they are softer than the rind that is present on the rest of the stalk surface. Preliminary results by other workers suggested that near infrared spectroscopy (NIRS) could provide a rapid screening method for the chemical profile in bud scales, the outer coating of buds and therefore the first contact point of an invading larva. If feasible, analysis of samples using this method could be done in the South African Sugar Experiment Station's (SASEX) stage two selection trials, providing an early indication of eldana resistance on large numbers of cultivars, without the necessity of separate trials. However, knowledge of how environments, position of bud scales on the stalk and age affect NIRS is required in order to determine the feasibility of the method. Planting of a trial with an identical set of genotypes across a range of environments, sampled at a number of ages, would provide the necessary information on environmental effects, whilst simultaneously providing the necessary range of samples to develop a calibration between bud scale chemical profiles and eldana resistance ratings. Inheritance patterns of the characteristics being measured is also required if they are to be used in a breeding programme. The original work by Rutherford (1993) was carried out on only five calibration sets (a set of standard clones with relatively well-known eldana resistance ratings), and different sets were not comparable due to what was assumed to be environmental differences between calibration sets. One aspect of the current experiment was to examine more closely the effect of genotype x environment interaction (G x E) on the performance of the NIRS technique under a range of conditions. Two sites were chosen to represent the conditions encountered in trials carried out by SASEX. The crops were sampled at three ages, representing the range of ages at which sugarcane is harvested in South Africa. Two locations on the stalk were also examined, top and bottom, for removal of bud scales, based on the assumption that aging of bud scales may affect chemical composition. A new NIRSystems 6500 instrument was acquired during the course of this study. Data from the new instrument indicated that there were no longer differences between the different calibration sets, and therefore no longer differences between environments. Spectra for different samples were very close, the differences being of the same scale as those recorded with repeated measures of the same samples, or between the readings for the standard solvent solution. This led to the conclusion that the differences observed on the original NIRSystems 5000 instrument were due to instrument error, not environmental differences. More importantly, the different calibration sets were not comparable despite being similar to each other. Prediction from one calibration set to another was low. These observations led to the conclusion that NIRS was not a suitable method for determining chemical compounds associated with tolerance of sugarcane genotypes to eldana borer. The original NIRS instrument was subject to error, and the small number of calibration sets included in the study led to the erroneous conclusion that NIRS was suitable for the prediction of varietal tolerance to eldana. With the acquisition of the new instrument, the errors generated by the old instrument became apparent. With the increase in number of calibration sets included in the study, it also became apparent that a global calibration covering all environments was not possible. An analysis of the heritability of the chemical compounds associated with eldana resistance was also included in this study. A biparental progeny design of 24 crosses with 33 unselected offspring per cross was used. This trial would have been analysed once the calibration had been developed using the environmental trial, and it would have provided knowledge of the breeding behaviour of the chemical compounds associated with tolerance to eldana. Because the NIRS technique proved to be unsuitable for detection of chemical compounds associated with eldana resistance, the heritability of these chemical compounds could not be studied. As the NIRS study did not produce data, the G x E interaction analysis and determination of heritability was applied to the bud scale mass data set. This study showed a relatively low positive correlation between bud scale mass and resistance to eldana. The broad sense heritability estimate for bud scale mass from the G x E interaction analysis was 0.45, and the narrow sense heritability estimate from parent-offspring regression analysis was approximately 0.27, suggesting a low degree of genetic determination in bud scale mass. The G x E interaction analyses gave varying results depending on the method used. The ANOVA analysis suggested that ages, sites and years had an effect on bud scale mass, while deviation from maximum plot showed no significance for G x E interactions. The number and choice of genotypes selected as unstable also varied with the method used to determine the stability of individual genotypes. Regression analysis and rank order analysis revealed a number of unstable genotypes, whilst stability variance and ecovalence, which produced similar results, detected only two unstable genotypes. In the rank order analysis correction of data to remove genotype effect, reduced the number of unstable genotypes, suggesting that the G x E interaction effect was partially confounded with the bud scale mass of the genotypes. This was a more reliable method than the uncorrected rank order analysis, and would be the preferred analysis type of all those tried. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.