A preliminary study on the effects of elevated CO2 on aphid resistance of Tugela Dn and the population dynamics of the Russian wheat aphid (Homoptera: Aphididae), Diuraphis noxia
- Authors: Mundondo, Daphine
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11402 , http://hdl.handle.net/10353/d1020244
- Description: Food security is of major importance due to the increasing world population with 8.9 billion people expected by 2050 (Cohen, 2003). Diuraphis noxia (RWA), have caused aggravating, massive losses to wheat farmers in many areas of the world. If unchecked, RWA are able to destroy plants resulting in major economic impacts (Botha, 2013). Due to ineffective use of other control methods, the Small Grains Institute in Bethlehem, South Africa, have therefore developed resistant cultivars to the known RWA subtypes over the past decades through intensive breeding programmes (Tolmay et al., 2006). Climate change has however become a major factor threatening food security especially with the observed increase in CO2 from less than 300 ppm in pre-industrial period to the current 385 ppm and is predicted to reach 550 ppm by 2050 (IPCC, 2007; Meehl et al., 2007). Elevated CO2 concentration may affect individual species of a community hence the need to understand the wheat-aphid interactions. In this study, population growth rates and virulence of RWA SA1 at ambient (385 ppm) and elevated (450 ppm) CO2 concentration were evaluated on two wheat cultivars: Tugela Dn (resistant) and Scheepers (susceptible). Fluorescence microscopy techniques using aniline blue were used to investigate feeding related damage caused by RWA SA1 through an examination of callose deposition at the two CO2 concentration. A two-dimensional gel electrophoresis method was developed in order to determine the effect of RWA SA1 on the wheat cultivars proteome at the two CO2 concentration. Differentially expressed proteins that were up or down regulated more than two fold were identified using PDQuestTM Basic 2D Gel analysis software. Populations of RWA SA1 increased significantly on the two wheat cultivars at both CO2 concentration. Although the population growth rate for RWA SA1 on both cultivars was generally exponential at all treatments, growth at elevated CO2 concentration was noticeably faster with populations increasing 3 fold in 14 days as compared to the 2 times at ambient CO2 concentration. Hence, both cultivars provided a better quality host for RWA SA1 at 450 ppm than 385 ppm. There was no significant difference between RWA SA1 population on Tugela Dn and on Scheepers at elevated CO2 concentration on day 14 after infestation which means there was a change in the resistance mechanism in Tugela Dn at this condition. Approximately 70% of the total leaf showed chlorosis by 21 days of aphid infestation for both cultivars although the susceptible cultivar was more vulnerable. There was low callose deposition in the controls (uninfested plants) but heavy callose in infested plants due to aphid feeding. A proteomics approach was used as a pilot study to investigate whether it would be possible to identify the changes in the resistance mechanism during aphid infestation under elevated CO2 levels. The major changes in the proteome of the control group (uninfested Tugela Dn at ambient versus elevated CO2 concentration) occurred in the early events (day 1-7) in the molecular weight range of approximately 25 kDa to 55 kDa are mainly within the basic to neutral pH range. This was suggested to be a result of mechanisms to adjust to the CO2 concentration. Elevated CO2 results in instant higher photosynthetic rates and C:N ratios as well as changes in expression levels of SA-dependant defense genes (Lindroth 1995; Hughes and Bazzaz, 2001; Sun et al., 2013). Because most of these changes are directly regulated by proteins, it is expected that the most differential protein expression will occur immediately after the atmospheric changes (early events) as was shown in the study. Infested plants under elevated and ambient conditions showed that the stress conditions gave rise to differentially regulated proteins within the wheat proteome. Most changes occurred elevated CO2 levels. It can be suggested that the changes were a result of differentially regulated plant defence proteins which fall in this range (25 kDa - 80 kDa) such as peroxidases, chitinases and β-1.3-glucanases as well as protein kinases, heat-shock proteins and photosynthetic proteins. These results indicate that there has been changes in the resistance due to elevated CO2 because of the evident changes in the proteome. If so, then the results will be similar to those documented by Louw (2007) where up-regulation was due to putative storage proteins, proteins involved in photosynthesis, heat shock proteins and defense proteins. Of course, the pI value and molecular mass of the proteins and the identification of the proteins in these spots, must be determined in future work to specifically identify whether these suggestions are authentic. However, Louw (2007) also reports that the susceptible Betta wheat cultivar, displayed a defence response similar to the HR although it was unable to up-regulate specific defensive proteins against RWA infestation but proteins for broad resistance. Although the changes in the proteins in infested Tugela Dn under elevated CO2 concentration were not accurately identified, the defense mechanism is similar to that portrayed by the susceptible Betta wheat cultivar which shows that the resistance mechanism had been overcome. Because this was a pilot study and preliminary results were obtained due to limited funding and time constraints, suggestions were made on how to further develop the method to obtain statistically significant results.
- Full Text:
- Authors: Mundondo, Daphine
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11402 , http://hdl.handle.net/10353/d1020244
- Description: Food security is of major importance due to the increasing world population with 8.9 billion people expected by 2050 (Cohen, 2003). Diuraphis noxia (RWA), have caused aggravating, massive losses to wheat farmers in many areas of the world. If unchecked, RWA are able to destroy plants resulting in major economic impacts (Botha, 2013). Due to ineffective use of other control methods, the Small Grains Institute in Bethlehem, South Africa, have therefore developed resistant cultivars to the known RWA subtypes over the past decades through intensive breeding programmes (Tolmay et al., 2006). Climate change has however become a major factor threatening food security especially with the observed increase in CO2 from less than 300 ppm in pre-industrial period to the current 385 ppm and is predicted to reach 550 ppm by 2050 (IPCC, 2007; Meehl et al., 2007). Elevated CO2 concentration may affect individual species of a community hence the need to understand the wheat-aphid interactions. In this study, population growth rates and virulence of RWA SA1 at ambient (385 ppm) and elevated (450 ppm) CO2 concentration were evaluated on two wheat cultivars: Tugela Dn (resistant) and Scheepers (susceptible). Fluorescence microscopy techniques using aniline blue were used to investigate feeding related damage caused by RWA SA1 through an examination of callose deposition at the two CO2 concentration. A two-dimensional gel electrophoresis method was developed in order to determine the effect of RWA SA1 on the wheat cultivars proteome at the two CO2 concentration. Differentially expressed proteins that were up or down regulated more than two fold were identified using PDQuestTM Basic 2D Gel analysis software. Populations of RWA SA1 increased significantly on the two wheat cultivars at both CO2 concentration. Although the population growth rate for RWA SA1 on both cultivars was generally exponential at all treatments, growth at elevated CO2 concentration was noticeably faster with populations increasing 3 fold in 14 days as compared to the 2 times at ambient CO2 concentration. Hence, both cultivars provided a better quality host for RWA SA1 at 450 ppm than 385 ppm. There was no significant difference between RWA SA1 population on Tugela Dn and on Scheepers at elevated CO2 concentration on day 14 after infestation which means there was a change in the resistance mechanism in Tugela Dn at this condition. Approximately 70% of the total leaf showed chlorosis by 21 days of aphid infestation for both cultivars although the susceptible cultivar was more vulnerable. There was low callose deposition in the controls (uninfested plants) but heavy callose in infested plants due to aphid feeding. A proteomics approach was used as a pilot study to investigate whether it would be possible to identify the changes in the resistance mechanism during aphid infestation under elevated CO2 levels. The major changes in the proteome of the control group (uninfested Tugela Dn at ambient versus elevated CO2 concentration) occurred in the early events (day 1-7) in the molecular weight range of approximately 25 kDa to 55 kDa are mainly within the basic to neutral pH range. This was suggested to be a result of mechanisms to adjust to the CO2 concentration. Elevated CO2 results in instant higher photosynthetic rates and C:N ratios as well as changes in expression levels of SA-dependant defense genes (Lindroth 1995; Hughes and Bazzaz, 2001; Sun et al., 2013). Because most of these changes are directly regulated by proteins, it is expected that the most differential protein expression will occur immediately after the atmospheric changes (early events) as was shown in the study. Infested plants under elevated and ambient conditions showed that the stress conditions gave rise to differentially regulated proteins within the wheat proteome. Most changes occurred elevated CO2 levels. It can be suggested that the changes were a result of differentially regulated plant defence proteins which fall in this range (25 kDa - 80 kDa) such as peroxidases, chitinases and β-1.3-glucanases as well as protein kinases, heat-shock proteins and photosynthetic proteins. These results indicate that there has been changes in the resistance due to elevated CO2 because of the evident changes in the proteome. If so, then the results will be similar to those documented by Louw (2007) where up-regulation was due to putative storage proteins, proteins involved in photosynthesis, heat shock proteins and defense proteins. Of course, the pI value and molecular mass of the proteins and the identification of the proteins in these spots, must be determined in future work to specifically identify whether these suggestions are authentic. However, Louw (2007) also reports that the susceptible Betta wheat cultivar, displayed a defence response similar to the HR although it was unable to up-regulate specific defensive proteins against RWA infestation but proteins for broad resistance. Although the changes in the proteins in infested Tugela Dn under elevated CO2 concentration were not accurately identified, the defense mechanism is similar to that portrayed by the susceptible Betta wheat cultivar which shows that the resistance mechanism had been overcome. Because this was a pilot study and preliminary results were obtained due to limited funding and time constraints, suggestions were made on how to further develop the method to obtain statistically significant results.
- Full Text:
Identification of agricultural and industrial pollutants in the Kat River, Eastern Cape and their effect on agricultural products found along the river banks
- Authors: Mutingwende, Nhamo
- Date: 2015
- Subjects: Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11291 , http://hdl.handle.net/10353/d1020242 , Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Description: There is growing concern that commonly used Pharmaceuticals and Personal Care Products (PPCPs) and pesticides are entering and contaminating drinking water supplies. The use of targeted quantitation of PPCP has been well established but there is an emerging trend to also screen for and identify unexpected environmental pollutants. Chemicals like pesticides hormones and antibiotics are especially of interest because of proven endocrine disrupting effects and a possible development of bacterial resistance. Powerful screening methods are required to detect and quantify the presence of these compounds in our environment. PPCP encompass a wide range of pollutants, including Endocrine Disrupting Compounds (EDC), pesticides, hormones, antibiotics, drugs of abuse, x-ray contrast agents and drinking water disinfection by-products to name a few. In order to properly assess the effects of these compounds on our environment, it is necessary to accurately monitor their presence. The diversity of chemical properties of these compounds makes method development challenging. LC/MS/MS is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range. The new AB SCIEX TripleTOF™5600 LC/MS/MS was used to profile environmental samples for unexpected pollutants, to identify and characterise the chemical composition and structure of the pollutants, and to quantify (based on intensity) the concentration in collected water samples. Liquid Chromatography coupled to tandem Mass Spectrometry (LCMS/ MS) is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range, such as pesticides, antibiotics, drugs of abuse, x-ray contrast agents, drinking water disinfection by-products etc. More recently there is a growing interest from environmental researchers to also screen for and identify non-targeted compounds in environmental samples, including metabolites and degradates, but also completely unexpected pollutants. The new AB SCIEX TripleTOF™5600 LC/MS/MS system is capable of performing highly sensitive and fast MS scanning experiments to search for unknown molecular ions while also performing selective and characteristic MS/MS scanning for further compound identification and, therefore, is the instrument of choice for this challenging task. General unknown screening workflows do not use a target analyte list and compound detection is not based on any prior knowledge, including retention times and information on possible molecular and fragment ions. Therefore, acquired chromatograms are very rich in information and can easily contain thousands of ions from both any compounds present in the sample as well as from the sample matrix itself. Thus, powerful software tools are needed to explore such data to identify the unexpected compound. Water samples were collected both upstream and downstream of two WWTPs (Seymour and Fort Beaufort) and were directly injected on the AB SCIEX TripleTOF™5600 LC/MS/MS after being filtered. 15 sample points along the Kat River, ranging from a point as close to the source as possible to a point just before it joins the Great Fish River were used. The samples collected from the source were used as the control in each of the experiments, the assumption being the closer you get to the source, the less contaminated the water would be for the analysis of pesticides. Points were selected where the Kat River crosses the R67 or on farms where the river was accessible using farm roads. Samples were collected from October 2013 to November 2014.The Peak view software and Analyst software were used in the analysis of PPCPs. The XIC Manager allows you to manage large lists of compounds and perform automatic extracted ion chromatogram (XIC) calculations and review results operations. The results were displayed in the chromatogram pane and the XIC table (see results). The results reported here in this thesis indicate that there is contamination in the Kat River water due to both pesticides and PPCPs. The results also indicate that the food products are also contaminated and hence both the Kat River agricultural produce and its water need to be closely monitored for both pesticide and PPCPs contaminants. Further studies to investigate the quantitative levels of pesticides and PPCPs in the Kat river water to determine if the concentration levels of the detected pesticides are below the reported Maximum Residues Limits will be explored in the future.
- Full Text:
- Authors: Mutingwende, Nhamo
- Date: 2015
- Subjects: Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11291 , http://hdl.handle.net/10353/d1020242 , Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Description: There is growing concern that commonly used Pharmaceuticals and Personal Care Products (PPCPs) and pesticides are entering and contaminating drinking water supplies. The use of targeted quantitation of PPCP has been well established but there is an emerging trend to also screen for and identify unexpected environmental pollutants. Chemicals like pesticides hormones and antibiotics are especially of interest because of proven endocrine disrupting effects and a possible development of bacterial resistance. Powerful screening methods are required to detect and quantify the presence of these compounds in our environment. PPCP encompass a wide range of pollutants, including Endocrine Disrupting Compounds (EDC), pesticides, hormones, antibiotics, drugs of abuse, x-ray contrast agents and drinking water disinfection by-products to name a few. In order to properly assess the effects of these compounds on our environment, it is necessary to accurately monitor their presence. The diversity of chemical properties of these compounds makes method development challenging. LC/MS/MS is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range. The new AB SCIEX TripleTOF™5600 LC/MS/MS was used to profile environmental samples for unexpected pollutants, to identify and characterise the chemical composition and structure of the pollutants, and to quantify (based on intensity) the concentration in collected water samples. Liquid Chromatography coupled to tandem Mass Spectrometry (LCMS/ MS) is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range, such as pesticides, antibiotics, drugs of abuse, x-ray contrast agents, drinking water disinfection by-products etc. More recently there is a growing interest from environmental researchers to also screen for and identify non-targeted compounds in environmental samples, including metabolites and degradates, but also completely unexpected pollutants. The new AB SCIEX TripleTOF™5600 LC/MS/MS system is capable of performing highly sensitive and fast MS scanning experiments to search for unknown molecular ions while also performing selective and characteristic MS/MS scanning for further compound identification and, therefore, is the instrument of choice for this challenging task. General unknown screening workflows do not use a target analyte list and compound detection is not based on any prior knowledge, including retention times and information on possible molecular and fragment ions. Therefore, acquired chromatograms are very rich in information and can easily contain thousands of ions from both any compounds present in the sample as well as from the sample matrix itself. Thus, powerful software tools are needed to explore such data to identify the unexpected compound. Water samples were collected both upstream and downstream of two WWTPs (Seymour and Fort Beaufort) and were directly injected on the AB SCIEX TripleTOF™5600 LC/MS/MS after being filtered. 15 sample points along the Kat River, ranging from a point as close to the source as possible to a point just before it joins the Great Fish River were used. The samples collected from the source were used as the control in each of the experiments, the assumption being the closer you get to the source, the less contaminated the water would be for the analysis of pesticides. Points were selected where the Kat River crosses the R67 or on farms where the river was accessible using farm roads. Samples were collected from October 2013 to November 2014.The Peak view software and Analyst software were used in the analysis of PPCPs. The XIC Manager allows you to manage large lists of compounds and perform automatic extracted ion chromatogram (XIC) calculations and review results operations. The results were displayed in the chromatogram pane and the XIC table (see results). The results reported here in this thesis indicate that there is contamination in the Kat River water due to both pesticides and PPCPs. The results also indicate that the food products are also contaminated and hence both the Kat River agricultural produce and its water need to be closely monitored for both pesticide and PPCPs contaminants. Further studies to investigate the quantitative levels of pesticides and PPCPs in the Kat river water to determine if the concentration levels of the detected pesticides are below the reported Maximum Residues Limits will be explored in the future.
- Full Text:
Parent characterization of quality protein maize (Zea mays L.) and combining ability for tolerance to drought stress
- Authors: Pfunde, Cleopatra Nyaradzo
- Date: 2012
- Subjects: Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Language: English
- Type: Thesis , Masters , MSc Agric (Crop Science)
- Identifier: vital:11869 , http://hdl.handle.net/10353/d1007536 , Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Description: Quality protein maize (QPM) has enhanced levels of two essential amino acids, lysine and tryptophan compared to normal maize. This makes QPM an important cereal crop in communities where maize is a staple crop. The main abiotic factor to QPM production is drought stress. Little information is available on the effect of drought stress on QPM. Therefore, the objectives of this study were to: (i) conduct diversity analysis of QPM inbred lines using morpho-agronomic and simple sequence repeat markers, (ii) screen available QPM inbred lines and F1 progeny for tolerance to seedling drought stress, (iii) determine the combining ability and type of gene action of QPM inbred lines for tolerance to seedling drought stress, grain yield and endosperm modification. The study was conducted in South Africa, at the University of Fort Hare. Morphological characterisation of 21 inbred lines was done using quantitative and qualitative traits. A randomised complete block design with three replicates was used for characterizing the inbred lines in the field. Genstat statistical software, version 12 (Genstat ®, 2009) was used for analysis of variance (ANOVA) and descriptive statistics. Analysis of variance was performed on all quantitative data for morphological traits. Data for qualitative traits was tabulated in their nominal classes. Traits that contributed most to the variation were days to anthesis, days to silking, anthesis-silking interval, plant height, number of kernel rows, ear length and grain yield. Cluster analysis grouped the inbred lines into three main clusters. The first cluster was characterised by tall and average yielding lines, while the second cluster showed the least anthesis-silking interval, and had the highest yield. Cluster three consisted of lines that were early maturing, but were the least yielding. Genetic distances between maize inbred lines were quantified by using 27 simple sequence repeat markers. The genetic distances between genotypes was computed using Roger’s (1972) genetic distances. Cluster analysis was then carried out using the neighbour-joining tree method using Power Marker software version 3.25. A dendrogram generated from the genetic study of the inbred lines revealed three groups that concurred with expectations based upon pedigree data. These groups were not identical to the groups generated using morpho-agronomic characterisation. Twenty one QPM inbred lines were crossed using a North Carolina design II mating scheme. These were divided into seven sets, each with three inbred lines. The three inbred lines in one set were used as females and crossed with three inbred lines in another set consisting of males. Each inbred line was used as a female in one set, and as a male in a second set. Sixty three hybrids (7 sets x 9 hybrids) were formed and evaluated in October 2011, using a 6x8 alpha-lattice incomplete block design with three replicates under glasshouse and optimum field conditions. A randomised complete block design with three replicates was used for the 21 parental inbred lines. Traits recorded for the glasshouse study were, canopy temperature, chlorophyll content, leaf roll, stem diameter, plant height, leaf number, leaf area, fresh and dry root and shoot weights. Data for the various traits for each environment, 25 percent (stress treatment) and 75 percent (non-stress) of field capacity, were subjected to analysis of variance using the unbalanced treatment design in Genstat statistical package Edition 12. Where varietal differences were found, means were separated using Tukey’s test. Genetic analyses for grain yield and agronomic traits were performed using a fixed effects model in JMP 10 following Residual Maximum Likelihood procedure (REML). From the results, inbred lines that were not previously classified into heterotic groups and drought tolerance categories were classified based on their total dry weight performance and drought susceptibility index. Inbred lines L18, L9, L8, L6 and L3, in order of their drought tolerance index were the best performers under greenhouse conditions and could be recommended for breeding new varieties that are tolerant to seedling drought stress. Evaluation of maize seedlings tolerant to drought stress under glasshouse conditions revealed that cross combination L18 x L11 was drought tolerant, while cross L20 x L7 was susceptible. Total dry weight was used as the major criteria for classifying F1 maize seedlings as being resistant or susceptible. General combining ability effects accounted for 67.43 percent of the genetic variation for total dry weight, while specific combining ability effects contributed 37.57 percent. This indicated that additive gene effects were more important than non-additive gene action in controlling this trait. In the field study (non-drought), the experimental design was a 6x8 alpha lattice incomplete block design with three replicates. On an adjacent field a randomised complete block design with three replicates was used to evaluate the parental inbred lines. The following variables were recorded: plant height, ear height, ears per plant, endosperm modification, days to silking and days to anthesis, anthesis-silking interval, number of kernels per row, number of rows per ear and grain yield. General analyses for the incomplete lattice block design and randomised complete block design for hybrid and inbred data respectively were performed using JMP 10 statistical software. Means were separated using the Tukey's test. Genetic analyses of data for grain yield and agronomic traits were conducted using a fixed effects model using REML in JMP 10. The importance of both GCA (51 percent) and SCA (49 percent) was observed for grain yield. A preponderance of GCA existed for ear height, days to anthesis, anthesis-silking interval, ears per plant and number of kernels per row, indicating that predominantly, additive gene effects controlled hybrid performance under optimum field conditions. The highest heritability was observed for days to silking (48.27 percent) suggesting that yield could be improved through selection for this trait. Under field conditions, variation in time to maturity was observed. This implies that these inbred lines can be recommended for utilisation in different agro-ecologies. Early maturing lines such as L18 can be used to introduce earliness in local cultivars, while early maturing single crosses such as L18 x L2, L5 x L9, L3 x L4 and L2 x L21 could be recommended for maize growers in drought prone areas such as the former Ciskei. Single crosses L18xL11, L16xL18, L8xL21 and L9xL6 had good tolerance to seedling drought stress. On the other hand, single crosses L18xL11 and L11xL13 had high grain yield and good endosperm modification. All these single crosses could be recommended for commercial production after evaluation across locations in the Eastern Cape Province. Alternatively they can be crossed with other superior inbreds to generate three or four way hybrids, which could then be evaluated for potential use by farmers in the Eastern Cape.
- Full Text:
- Authors: Pfunde, Cleopatra Nyaradzo
- Date: 2012
- Subjects: Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Language: English
- Type: Thesis , Masters , MSc Agric (Crop Science)
- Identifier: vital:11869 , http://hdl.handle.net/10353/d1007536 , Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Description: Quality protein maize (QPM) has enhanced levels of two essential amino acids, lysine and tryptophan compared to normal maize. This makes QPM an important cereal crop in communities where maize is a staple crop. The main abiotic factor to QPM production is drought stress. Little information is available on the effect of drought stress on QPM. Therefore, the objectives of this study were to: (i) conduct diversity analysis of QPM inbred lines using morpho-agronomic and simple sequence repeat markers, (ii) screen available QPM inbred lines and F1 progeny for tolerance to seedling drought stress, (iii) determine the combining ability and type of gene action of QPM inbred lines for tolerance to seedling drought stress, grain yield and endosperm modification. The study was conducted in South Africa, at the University of Fort Hare. Morphological characterisation of 21 inbred lines was done using quantitative and qualitative traits. A randomised complete block design with three replicates was used for characterizing the inbred lines in the field. Genstat statistical software, version 12 (Genstat ®, 2009) was used for analysis of variance (ANOVA) and descriptive statistics. Analysis of variance was performed on all quantitative data for morphological traits. Data for qualitative traits was tabulated in their nominal classes. Traits that contributed most to the variation were days to anthesis, days to silking, anthesis-silking interval, plant height, number of kernel rows, ear length and grain yield. Cluster analysis grouped the inbred lines into three main clusters. The first cluster was characterised by tall and average yielding lines, while the second cluster showed the least anthesis-silking interval, and had the highest yield. Cluster three consisted of lines that were early maturing, but were the least yielding. Genetic distances between maize inbred lines were quantified by using 27 simple sequence repeat markers. The genetic distances between genotypes was computed using Roger’s (1972) genetic distances. Cluster analysis was then carried out using the neighbour-joining tree method using Power Marker software version 3.25. A dendrogram generated from the genetic study of the inbred lines revealed three groups that concurred with expectations based upon pedigree data. These groups were not identical to the groups generated using morpho-agronomic characterisation. Twenty one QPM inbred lines were crossed using a North Carolina design II mating scheme. These were divided into seven sets, each with three inbred lines. The three inbred lines in one set were used as females and crossed with three inbred lines in another set consisting of males. Each inbred line was used as a female in one set, and as a male in a second set. Sixty three hybrids (7 sets x 9 hybrids) were formed and evaluated in October 2011, using a 6x8 alpha-lattice incomplete block design with three replicates under glasshouse and optimum field conditions. A randomised complete block design with three replicates was used for the 21 parental inbred lines. Traits recorded for the glasshouse study were, canopy temperature, chlorophyll content, leaf roll, stem diameter, plant height, leaf number, leaf area, fresh and dry root and shoot weights. Data for the various traits for each environment, 25 percent (stress treatment) and 75 percent (non-stress) of field capacity, were subjected to analysis of variance using the unbalanced treatment design in Genstat statistical package Edition 12. Where varietal differences were found, means were separated using Tukey’s test. Genetic analyses for grain yield and agronomic traits were performed using a fixed effects model in JMP 10 following Residual Maximum Likelihood procedure (REML). From the results, inbred lines that were not previously classified into heterotic groups and drought tolerance categories were classified based on their total dry weight performance and drought susceptibility index. Inbred lines L18, L9, L8, L6 and L3, in order of their drought tolerance index were the best performers under greenhouse conditions and could be recommended for breeding new varieties that are tolerant to seedling drought stress. Evaluation of maize seedlings tolerant to drought stress under glasshouse conditions revealed that cross combination L18 x L11 was drought tolerant, while cross L20 x L7 was susceptible. Total dry weight was used as the major criteria for classifying F1 maize seedlings as being resistant or susceptible. General combining ability effects accounted for 67.43 percent of the genetic variation for total dry weight, while specific combining ability effects contributed 37.57 percent. This indicated that additive gene effects were more important than non-additive gene action in controlling this trait. In the field study (non-drought), the experimental design was a 6x8 alpha lattice incomplete block design with three replicates. On an adjacent field a randomised complete block design with three replicates was used to evaluate the parental inbred lines. The following variables were recorded: plant height, ear height, ears per plant, endosperm modification, days to silking and days to anthesis, anthesis-silking interval, number of kernels per row, number of rows per ear and grain yield. General analyses for the incomplete lattice block design and randomised complete block design for hybrid and inbred data respectively were performed using JMP 10 statistical software. Means were separated using the Tukey's test. Genetic analyses of data for grain yield and agronomic traits were conducted using a fixed effects model using REML in JMP 10. The importance of both GCA (51 percent) and SCA (49 percent) was observed for grain yield. A preponderance of GCA existed for ear height, days to anthesis, anthesis-silking interval, ears per plant and number of kernels per row, indicating that predominantly, additive gene effects controlled hybrid performance under optimum field conditions. The highest heritability was observed for days to silking (48.27 percent) suggesting that yield could be improved through selection for this trait. Under field conditions, variation in time to maturity was observed. This implies that these inbred lines can be recommended for utilisation in different agro-ecologies. Early maturing lines such as L18 can be used to introduce earliness in local cultivars, while early maturing single crosses such as L18 x L2, L5 x L9, L3 x L4 and L2 x L21 could be recommended for maize growers in drought prone areas such as the former Ciskei. Single crosses L18xL11, L16xL18, L8xL21 and L9xL6 had good tolerance to seedling drought stress. On the other hand, single crosses L18xL11 and L11xL13 had high grain yield and good endosperm modification. All these single crosses could be recommended for commercial production after evaluation across locations in the Eastern Cape Province. Alternatively they can be crossed with other superior inbreds to generate three or four way hybrids, which could then be evaluated for potential use by farmers in the Eastern Cape.
- Full Text:
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