Investigation into response of wheat genotypes to drought and optimum conditions in the Eastern Cape Province, South Africa
- Authors: Mzileni L S
- Date: 2023
- Subjects: Bonanza farms , Wheat , Drought - tolerant plants
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/27732 , vital:69397
- Description: Wheat is generally one of the dominant crops globally, being mainly used for human food and livestock feed. Due to climate change, drought makes it challenging to produce enough wheat mostly under dryland production regions in South Africa. Drought stress has severely reduced wheat yield by up to 70 percent, and adversely compromised wheat grain quality. The adoption of drought-tolerant cultivars offers a sustainable and low-cost solution for increasing wheat yields and minimise importing the crop to meet national requirements. The main objective of this study was to investigate the response of different wheat genotypes to drought and optimum conditions in the Eastern Cape Province, South Africa. Forty diverse wheat genotypes were evaluated in this study. The specific objectives were: (i) to evaluate the response of wheat genotypes under optimum and drought-stressed field conditions; (ii) to determine the effect of terminal drought stress on wheat grain quality composition; and (iii) to identify appropriate drought tolerance indices that can be used as selection tools under field conditions. This study was conducted in the field using a 5x8 alpha lattice design, replicated twice under two water regimes (drought and optimum) over two consecutive winter seasons of 2020 and 2021 at two different sites namely University of Fort Hare Research Farm in Alice, and Zanyokwe irrigation scheme in Keiskamahoek. Drought stress was imposed from 50% flowering up to physiological maturity. Data on agro-physiological traits such as duration to heading (DTH); flowering (DTF); maturity (DTM); plant height (HT); spike length (SL); number of spikelets per spike (SPS); kernels per spike (KPS); and grain yield (GY (kg/ha)) was subjected to the analysis of variance using Genstat 18th edition. As the study took place over two sites, a combined ANOVA table revealed significant differences (p0.001) among genotypes, and all interactions such as genotype by water regime (GWR); genotype by seasons (GS) for all studied traits. Notably, the extent and severity of drought differed between geographical regions and between seasons. This necessitated the adoption of the additive main effect and multiplicative interaction analysis (AMMI) for the identification of stable genotypes under two different water regimes over two sites. Regarding grain yield, superior and/or stable genotypes included G5 (4334 kg/ha under optimum, and 2871kg/ha under drought), and G22 (4418 kg/ha under optimum, and 2624kg/ha under drought) at the UFH site. G21 (3194 kg/ha under optimum, and 2938 kg/ha under drought), G33 (2552kg/ha under optimum, and 3810 kg/ha under drought), and G35 (2688 kg/ha under optimum, and 3309 kg/ha under drought) at the ZAN site. Stable genotypes across sites included G21 and G33. There were generally weak correlations between agro-physiological traits and grain yield. From the experiment, grain quality traits such as fixed protein (PF); wet gluten (WG); hectolitre mass (HLM); and thousand kernel weight (TKW) were also examined. A combined ANOVA revealed significant differences (p0.001) among the interaction of genotypes by environments (GE) for all traits except PF. This implies that the performance of wheat genotypes across sites was also different, and therefore, necessitated separate analysis of variance for each site. Significant differences (p0.001) among genotypes (G), water regimes (WR), and the interaction of genotypes by water regime (GWR) were observed for all studied quality traits except PF in both sites. GWR showed no significant differences for TKW in the ZAN site. The stability in the performance of genotypes across water regimes was further determined. G38 was stable for wet gluten; G31 and G26 were stable for PF; G36 was stable for HLM; and G11, G15, and G29 were stable for TKW at the UFH site. G6 was stable for both WG and PF; G13 and G15 were stable for HLM; and G35, G21, and G40 were stable for TKW at the ZAN site. These results suggest that the quality of wheat grains was affected under drought stress conditions except PF. Average grain yield data under both stressed (Ys) and optimum (Yp) conditions was used to compute a number of different drought tolerance indices. These include mean productivity (MP); geometric mean productivity (GMP); harmonic mean (HM); Tolerance index (TOL), stress susceptible index (SSI), sensitive drought index (SDI), and stress tolerance index (STI). The aim was to identify appropriate drought tolerance indices that can be used as selection tools under drought stress. MP, GMP, and HM were the more appropriate indices as they had a strong and positive correlation with grain yield under both drought and optimum conditions. However, genotypes G5, G22, G8, and G21 were more tolerant and stable as they showed high mean values. Based on the results, G19, G16, G2, and G20 were more sensitive to drought as they showed low values of MP, GMP, and HM. Overall, genotype: G5, G21, G22, and G33 are recommended for production under drought and optimum conditions, as they showed stable performance across water regimes. Principal component analysis also revealed that MP, GMP, and HM were the only indices that had positive loadings into the first principal component. , Thesis (MSc) -- Faculty of Science and Agriculture, 2023
- Full Text:
- Date Issued: 2023
- Authors: Mzileni L S
- Date: 2023
- Subjects: Bonanza farms , Wheat , Drought - tolerant plants
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/27732 , vital:69397
- Description: Wheat is generally one of the dominant crops globally, being mainly used for human food and livestock feed. Due to climate change, drought makes it challenging to produce enough wheat mostly under dryland production regions in South Africa. Drought stress has severely reduced wheat yield by up to 70 percent, and adversely compromised wheat grain quality. The adoption of drought-tolerant cultivars offers a sustainable and low-cost solution for increasing wheat yields and minimise importing the crop to meet national requirements. The main objective of this study was to investigate the response of different wheat genotypes to drought and optimum conditions in the Eastern Cape Province, South Africa. Forty diverse wheat genotypes were evaluated in this study. The specific objectives were: (i) to evaluate the response of wheat genotypes under optimum and drought-stressed field conditions; (ii) to determine the effect of terminal drought stress on wheat grain quality composition; and (iii) to identify appropriate drought tolerance indices that can be used as selection tools under field conditions. This study was conducted in the field using a 5x8 alpha lattice design, replicated twice under two water regimes (drought and optimum) over two consecutive winter seasons of 2020 and 2021 at two different sites namely University of Fort Hare Research Farm in Alice, and Zanyokwe irrigation scheme in Keiskamahoek. Drought stress was imposed from 50% flowering up to physiological maturity. Data on agro-physiological traits such as duration to heading (DTH); flowering (DTF); maturity (DTM); plant height (HT); spike length (SL); number of spikelets per spike (SPS); kernels per spike (KPS); and grain yield (GY (kg/ha)) was subjected to the analysis of variance using Genstat 18th edition. As the study took place over two sites, a combined ANOVA table revealed significant differences (p0.001) among genotypes, and all interactions such as genotype by water regime (GWR); genotype by seasons (GS) for all studied traits. Notably, the extent and severity of drought differed between geographical regions and between seasons. This necessitated the adoption of the additive main effect and multiplicative interaction analysis (AMMI) for the identification of stable genotypes under two different water regimes over two sites. Regarding grain yield, superior and/or stable genotypes included G5 (4334 kg/ha under optimum, and 2871kg/ha under drought), and G22 (4418 kg/ha under optimum, and 2624kg/ha under drought) at the UFH site. G21 (3194 kg/ha under optimum, and 2938 kg/ha under drought), G33 (2552kg/ha under optimum, and 3810 kg/ha under drought), and G35 (2688 kg/ha under optimum, and 3309 kg/ha under drought) at the ZAN site. Stable genotypes across sites included G21 and G33. There were generally weak correlations between agro-physiological traits and grain yield. From the experiment, grain quality traits such as fixed protein (PF); wet gluten (WG); hectolitre mass (HLM); and thousand kernel weight (TKW) were also examined. A combined ANOVA revealed significant differences (p0.001) among the interaction of genotypes by environments (GE) for all traits except PF. This implies that the performance of wheat genotypes across sites was also different, and therefore, necessitated separate analysis of variance for each site. Significant differences (p0.001) among genotypes (G), water regimes (WR), and the interaction of genotypes by water regime (GWR) were observed for all studied quality traits except PF in both sites. GWR showed no significant differences for TKW in the ZAN site. The stability in the performance of genotypes across water regimes was further determined. G38 was stable for wet gluten; G31 and G26 were stable for PF; G36 was stable for HLM; and G11, G15, and G29 were stable for TKW at the UFH site. G6 was stable for both WG and PF; G13 and G15 were stable for HLM; and G35, G21, and G40 were stable for TKW at the ZAN site. These results suggest that the quality of wheat grains was affected under drought stress conditions except PF. Average grain yield data under both stressed (Ys) and optimum (Yp) conditions was used to compute a number of different drought tolerance indices. These include mean productivity (MP); geometric mean productivity (GMP); harmonic mean (HM); Tolerance index (TOL), stress susceptible index (SSI), sensitive drought index (SDI), and stress tolerance index (STI). The aim was to identify appropriate drought tolerance indices that can be used as selection tools under drought stress. MP, GMP, and HM were the more appropriate indices as they had a strong and positive correlation with grain yield under both drought and optimum conditions. However, genotypes G5, G22, G8, and G21 were more tolerant and stable as they showed high mean values. Based on the results, G19, G16, G2, and G20 were more sensitive to drought as they showed low values of MP, GMP, and HM. Overall, genotype: G5, G21, G22, and G33 are recommended for production under drought and optimum conditions, as they showed stable performance across water regimes. Principal component analysis also revealed that MP, GMP, and HM were the only indices that had positive loadings into the first principal component. , Thesis (MSc) -- Faculty of Science and Agriculture, 2023
- Full Text:
- Date Issued: 2023
Isolation, purification and kinetic characterization of prolyl endopeptidase from Titicum aestivum
- Authors: Abrahams, Adriam Mark
- Date: 2013
- Subjects: Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11269 , http://hdl.handle.net/10353/d1004356 , Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Description: PEP activity has been described in several locations and has mostly been linked to a variety of neurological disorders such as schizophrenia, amnaesia, depression as well as other disease states such as anorexia nervosa, bulimia nervosa and blood pressure regulation. The enzyme has also been previously isolated from a variety of archae, microorganisms and several eukaryotic species but no prolyl endopeptidases have been isolated from plants. Plants have high levels of proline and glutamine rich peptides in seeds. We therefore hypothesize plants must express PEPs during germination. Bioinformatics tools were used to identify known PEPs and putative plant PEPs. A global sequence alignment of putative plant PEPs and other known PEPs indicated that the active site amino acids Ser, His and Asp are conserved in putative plant PEP sequences. Furthermore, putative plant PEPs showed similar secondary structures to known PEPs and when a rice PEP was modelled onto porcine brain PEP structure, a high degree of similarity was found. Germination studies of wheat seed showed an increase of PEP activity over time with maximum PEP activity reached after 4 days of germination, which remained at this level until 9 days of germination, implying a function for PEP in plant seed germination. Wheat PEP was purified using ion exchange and gel filtration chromatography with a final yield of less than 1 percent and a relative purity (only 2 bands detected by SDS-PAGE). The purified wheat PEP had a molecular weight of approximately 55kDa, substrate specificity for chymotrypsin-like substrates (N-Suc-Ala-Ala-Pro-Phe-pNa, Km value of 0.58 mM, Kcat of 29.37 s–1; Kcat /Km 50813.14s–1 M–1); a pH optimum of 7.9; temperature optima of 37oC and a high sensitivity to temperature as indicated by loss of activity at temperatures above 40oC. Inhibition studies using E64, Leupeptin and PMSF confirmed that the wheat PEP is from the Serine protease family and is most likely a trypsin-like protease.
- Full Text:
- Date Issued: 2013
- Authors: Abrahams, Adriam Mark
- Date: 2013
- Subjects: Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11269 , http://hdl.handle.net/10353/d1004356 , Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Description: PEP activity has been described in several locations and has mostly been linked to a variety of neurological disorders such as schizophrenia, amnaesia, depression as well as other disease states such as anorexia nervosa, bulimia nervosa and blood pressure regulation. The enzyme has also been previously isolated from a variety of archae, microorganisms and several eukaryotic species but no prolyl endopeptidases have been isolated from plants. Plants have high levels of proline and glutamine rich peptides in seeds. We therefore hypothesize plants must express PEPs during germination. Bioinformatics tools were used to identify known PEPs and putative plant PEPs. A global sequence alignment of putative plant PEPs and other known PEPs indicated that the active site amino acids Ser, His and Asp are conserved in putative plant PEP sequences. Furthermore, putative plant PEPs showed similar secondary structures to known PEPs and when a rice PEP was modelled onto porcine brain PEP structure, a high degree of similarity was found. Germination studies of wheat seed showed an increase of PEP activity over time with maximum PEP activity reached after 4 days of germination, which remained at this level until 9 days of germination, implying a function for PEP in plant seed germination. Wheat PEP was purified using ion exchange and gel filtration chromatography with a final yield of less than 1 percent and a relative purity (only 2 bands detected by SDS-PAGE). The purified wheat PEP had a molecular weight of approximately 55kDa, substrate specificity for chymotrypsin-like substrates (N-Suc-Ala-Ala-Pro-Phe-pNa, Km value of 0.58 mM, Kcat of 29.37 s–1; Kcat /Km 50813.14s–1 M–1); a pH optimum of 7.9; temperature optima of 37oC and a high sensitivity to temperature as indicated by loss of activity at temperatures above 40oC. Inhibition studies using E64, Leupeptin and PMSF confirmed that the wheat PEP is from the Serine protease family and is most likely a trypsin-like protease.
- Full Text:
- Date Issued: 2013
Evaluation of various proteomic techniques to identify proteins involved in cereal stress responses to aphid infestation
- Authors: Nqumla, Ntombekhaya
- Date: 2012
- Subjects: Aphids , Wheat , Plant proteomics , Rice
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11270 , http://hdl.handle.net/10353/d1004572 , Aphids , Wheat , Plant proteomics , Rice
- Description: All plants are exposed to abiotic and biotic stresses and have developed intricate signalling responses to survive. They respond to cell-structure disruption caused by herbivore probing and feeding by the formation of callose. Callose is a linear homopolymer made up of β-1,3-linked glucose residues with some β-1,6-branches. Plant responses to abiotic or biotic stress share events such as phosphorylation, membrane depolarization, calcium influx and the release of reactive oxygen species such as hydrogen peroxide. These events lead to the up-regulation of several pathways leading to biosynthesis of signalling molecules such as salicylic acid, jasmonate, abscisic acid and ethylene pathways. The aim of this study was to determine the most suitable proteomic approach for identifying proteins and signalling pathways involved in cereal response to aphid infestation. An in silico approach was first evaluated in which the 5ʹ upstream regulatory region of proteins belonging to the family of callose synthases was scanned for cis-regulatory elements in order to identify which callose synthases are possibly expressed in plants during biotic or abiotic stresses. Bioinformatics tools were used in the identification of twelve Arabidopsis and ten rice callose synthase coding regions. Genome sequences for rice and Arabidopsis were scanned for the 2000 bp 5ʹ region upstream of the start codon of each callose synthase coding region. PlantCare, PLACE and Athena software were used to identify putative cis-regulatory elements present in the 2000 bp 5ʹ upstream sequences. The majority of cis-acting elements identified were involved in drought and high temperature responses and only one cis-acting element was involved in wound stress. These results therefore indicated a probable role for plant callose synthases in drought stress responses rather than in biotic stress responses. Genevestigator analysis of Arabidopsis results of micro-array experiments indicated that AtGSL10 is highly up-regulated, with AtGSL1, 3, 5, 6, 7, 8, 11 and 12 showing medium up-regulation and AtGSL2, 4 and 9 no up-regulation during aphid infestation of Arabidopsis plants, implicating a possible role for AtGSL10 in the plant response to aphid infestation. An LC/MS/MS approach was used to identify specific signalling pathways involved in wheat resistance or stress response to aphid infestation. Eight proteins were identified as being up-regulated during aphid feeding in wheat, and 11 proteins were identified as possibly involved in the wheat resistance mechanism against aphid infestation. Several proteins were also identified as constitutively expressed proteins, during normal conditions and aphid infestation. Most pathways identified with proteins up-regulated in the resistance mechanisms of TugelaDN plants, were related to energy metabolism and located in the chloroplast. Evaluation of two dimensional gel electrophoresis to identify phosphoproteins differentially regulated in wheat during aphid infestation, revealed the up-regulation of three proteins namely photosystem II oxygen-evolving complex protein 2, HVUNKNOWN from Hordeum vulgare subsp vulgare and HSKERAT9 NID from Homo sapiens. Additional 57 proteins were partially identified as involved in the stress response but due to low protein levels, the percentage of matching peptides to these proteins was below the required confidence level. Although these protein identifications were below the confidence level, it is interesting to note that several of the proteins are known stress response proteins, and therefore could serve as potential targets for future investigations. In conclusion, the down and up-regulation of wheat proteins after aphid feeding reported in this study suggest that several signalling pathways are involved in the cereal stress response to aphid feeding. In silico approaches require knowledge or identification of potential proteins whereas 2D and LC/MS can identify numerous proteins still unknown to be involved in specific stress responses. The 2D approach is also limited in that the proteins of interest may be in low abundance and therefore not detected in the gels due to the presence of high abundant proteins. Therefore the best approach to identify proteins and signalling pathways involved in the stress response of wheat to aphid infestation, is the LC/MS/MS approach, as this proved to be the most sensitive and robust, identifying the most proteins with a high degree of confidence.
- Full Text:
- Date Issued: 2012
- Authors: Nqumla, Ntombekhaya
- Date: 2012
- Subjects: Aphids , Wheat , Plant proteomics , Rice
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11270 , http://hdl.handle.net/10353/d1004572 , Aphids , Wheat , Plant proteomics , Rice
- Description: All plants are exposed to abiotic and biotic stresses and have developed intricate signalling responses to survive. They respond to cell-structure disruption caused by herbivore probing and feeding by the formation of callose. Callose is a linear homopolymer made up of β-1,3-linked glucose residues with some β-1,6-branches. Plant responses to abiotic or biotic stress share events such as phosphorylation, membrane depolarization, calcium influx and the release of reactive oxygen species such as hydrogen peroxide. These events lead to the up-regulation of several pathways leading to biosynthesis of signalling molecules such as salicylic acid, jasmonate, abscisic acid and ethylene pathways. The aim of this study was to determine the most suitable proteomic approach for identifying proteins and signalling pathways involved in cereal response to aphid infestation. An in silico approach was first evaluated in which the 5ʹ upstream regulatory region of proteins belonging to the family of callose synthases was scanned for cis-regulatory elements in order to identify which callose synthases are possibly expressed in plants during biotic or abiotic stresses. Bioinformatics tools were used in the identification of twelve Arabidopsis and ten rice callose synthase coding regions. Genome sequences for rice and Arabidopsis were scanned for the 2000 bp 5ʹ region upstream of the start codon of each callose synthase coding region. PlantCare, PLACE and Athena software were used to identify putative cis-regulatory elements present in the 2000 bp 5ʹ upstream sequences. The majority of cis-acting elements identified were involved in drought and high temperature responses and only one cis-acting element was involved in wound stress. These results therefore indicated a probable role for plant callose synthases in drought stress responses rather than in biotic stress responses. Genevestigator analysis of Arabidopsis results of micro-array experiments indicated that AtGSL10 is highly up-regulated, with AtGSL1, 3, 5, 6, 7, 8, 11 and 12 showing medium up-regulation and AtGSL2, 4 and 9 no up-regulation during aphid infestation of Arabidopsis plants, implicating a possible role for AtGSL10 in the plant response to aphid infestation. An LC/MS/MS approach was used to identify specific signalling pathways involved in wheat resistance or stress response to aphid infestation. Eight proteins were identified as being up-regulated during aphid feeding in wheat, and 11 proteins were identified as possibly involved in the wheat resistance mechanism against aphid infestation. Several proteins were also identified as constitutively expressed proteins, during normal conditions and aphid infestation. Most pathways identified with proteins up-regulated in the resistance mechanisms of TugelaDN plants, were related to energy metabolism and located in the chloroplast. Evaluation of two dimensional gel electrophoresis to identify phosphoproteins differentially regulated in wheat during aphid infestation, revealed the up-regulation of three proteins namely photosystem II oxygen-evolving complex protein 2, HVUNKNOWN from Hordeum vulgare subsp vulgare and HSKERAT9 NID from Homo sapiens. Additional 57 proteins were partially identified as involved in the stress response but due to low protein levels, the percentage of matching peptides to these proteins was below the required confidence level. Although these protein identifications were below the confidence level, it is interesting to note that several of the proteins are known stress response proteins, and therefore could serve as potential targets for future investigations. In conclusion, the down and up-regulation of wheat proteins after aphid feeding reported in this study suggest that several signalling pathways are involved in the cereal stress response to aphid feeding. In silico approaches require knowledge or identification of potential proteins whereas 2D and LC/MS can identify numerous proteins still unknown to be involved in specific stress responses. The 2D approach is also limited in that the proteins of interest may be in low abundance and therefore not detected in the gels due to the presence of high abundant proteins. Therefore the best approach to identify proteins and signalling pathways involved in the stress response of wheat to aphid infestation, is the LC/MS/MS approach, as this proved to be the most sensitive and robust, identifying the most proteins with a high degree of confidence.
- Full Text:
- Date Issued: 2012
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