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Elevated CO2 determines cell damage and nitrogen allocation in barley subjected to aphid herbivory

  • Authors: Gallagher, Sean
  • Date: 2018
  • Language: English
  • Type: text , Thesis , Doctoral , PhD
  • Identifier: http://hdl.handle.net/10962/64337 , vital:28535
  • Description: Expected release date-May 2019
  • Full Text:
  • Date Issued: 2018

An investigation on the effect of Russian wheat aphid (Diuraphis noxia Kurdjumov) population growth and feeding damage on selected barley (Hordeum vulgare L.) cultivars under ambient and elevated CO2

  • Authors: Sacranie, Sattar Farouk
  • Date: 2016
  • Subjects: Russian wheat aphid -- Research -- South Africa Barley -- Disease and pest resistance -- South Africa Callose Aphids
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: http://hdl.handle.net/10962/50311 , vital:25975
  • Description: The Russian wheat aphid (RWA) (Diuraphis noxia Kurdjumov) is a major pest of cultivated small grains. It is particularly devastating because of is high reproductive rate which results in the growth of large populations which become damaging to its host plants. Development of resistant barley (Hordeum vulgare L.) cultivars is complicated as resistance is polygenic. As a result, the industry remains at risk now that the RWA has spread throughout South Africa. It has, as recently as, 2013, been identified in the SW Cape, which was previously geographically isolated. This is South Africa‟s principle barley growing region. Now a potentially huge problem exists. Therefore, it is imperative that an alternative to pesticide use is found. Testing potential innate resistance in barley cultivars is thus, critical. In this thesis, I present data on four barely cultivars where I have examined their resistance/ lack of resistance to three known RWA biotypes, RWASA1, RWASA2 and RWASA3. The barley varieties used were two economically important South African malt barley cultivars (S5 and SSG 564) along with two potentially RWA resistant Afghan accessions (CIho 4125 and CIho 4159). The RWA biotype population growth rates on each of the plants were determined over a 14 day period. The aim was to establish baseline data of the effects of RWA population growth on the host plants under ambient CO2 (380 – 400 ppm) conditions. The extent of RWA feeding damage was investigated at the cell level by examining saliva deposition and cell disruption using Transmission Electron Microscopy; at the tissue/vascular level using fluorescence microscopy, to determine the extent of callose formation; at a whole leaf level by recording percent chlorosis and leaf roll; and finally, at a whole plant level by measuring biomass loss.The experiments were repeated under elevated CO2 (450 ppm) to model any changes in RWA/plant interaction with respect to future climate change. The effects of an elevated CO2 environment and RWA feeding on host plant foliar N and C:N ratio were compared to ambient CO2 conditions, to provide a clearer picture of the potential nutrient drain that a feeding RWA colony exacts on its host. Of the varieties tested, the CIho accessions performed better than the two SA barley cultivars as the CIho accessions appeared to express a mild antibiosis resistance response as RWA populations, particularly those of RWASA1, were smaller than those observed on either S5 or SSG 564. In addition, less damage was evident in the two CIho accessions due to RWA feeding. II RWASA2 was the most virulent of the three RWA biotypes tested, followed by RWASA3 while RWASA1 was the least virulent. Under elevated CO2 conditions, RWA feeding damage was exacerbated but the trend of biotype virulence remained the same. Higher aphid population sizes were recorded under elevated CO2, meant that even the more resistant CIho accessions were overcome by the increased demand made by the larger aphid colonies on the host plants. The % foliar N data showed that under elevated CO2 aphid-free control plants had increased N levels in their leaves. Increased “food” supply (as shown by the increased N levels) therefore allowed significantly larger aphid populations to develop on the plants exposed to elevated CO2, due to improved nutrient status of the phloem sap taken up by RWA. The knock-on effect of a higher aphid population was increased cell disruption as a result of extensive probing, extensive formations of wound callose, with the result that phloem damage impeded nutrient flow through the vascular tissues which contributed to chlorosis and (eventually plant) death. The major conclusion from this study is that even a mild CO2 elevation resulted in an increase aphid population which may pose a severe and very real threat to a barley crop. Therefore, without effort to identify and deploy resistant barley cultivars, it could well be possible that future barley cultivation in South Africa may no longer be viable.
  • Full Text:
  • Date Issued: 2016

Differential expression and regulation of sucrose transporters in rice (Orzya sativa L, cv Nipponbare) during environmental stress conditions

  • Authors: Ibraheem, Omodele
  • Date: 2011
  • Subjects: Crops -- Effect of stress on , Plant molecular genetics , Gene expression , Sucrose , Rice
  • Language: English
  • Type: Thesis , Doctoral , PhD (Biochemistry)
  • Identifier: vital:11249 , http://hdl.handle.net/10353/330 , Crops -- Effect of stress on , Plant molecular genetics , Gene expression , Sucrose , Rice
  • Description: Plant productivity is greatly affected by environmental stresses such as drought, salinity and insect herbivory. Plants respond and adapt to these stresses by exhibiting physiological as well as biochemical changes at the cellular and molecular levels in order to survive. Expression of a variety of genes which encode numerous membrane transporters have been demonstrated to be induced by these stresses in a variety of plants. The nutritional status of plants is controlled by these transporters, which are regulated by the transcription of the corresponding genes. In spite of these adverse stress effects on agricultural yield, only a few studies have focused on gene transcriptional and translational regulation of membrane transporters during environmental stress situations. Rice, like other plants, contains a number of sucrose transporters encoded by a family of genes. However, detailed knowledge of their roles, localization and regulation during environmental stress conditions is lacking. Bioinformatic tools were used to identify putative cis-acting regulatory elements that may be involved in the regulation of rice and Arabidopsis thaliana sucrose transporters. The possible cis-acting regulatory elements were predicted by scanning genomic sequences 1.5 kbp upstream of the sucrose transporter genes translational start sites, using Plant CARE, PLACE and Genomatix Matinspector professional data bases. Several cis-acting regulatory elements that are associated with plant development, plant hormonal regulation and stress response were identified, and were present in varying frequencies within the 1.5 kbp of 5′ regulatory region. The putative cis-acting regulatory elements that possibly are involved in the expression and regulation of sucrose transporter gene families in rice and Arabidopsis thaliana during cellular development or environmental stress conditions were identified as: A-box, RY, CAT, Pyrimidine-box, Sucrose-box, ABRE, ARF, ERE, GARE, Me-JA, ARE, DRE, GA-motif, GATA, GT-1, MYC, MYB, W-box, and I-box. Expression analysis was used to elucidate the role of rice (Oryza sativa L. cv Nipponbare) sucrose transporter (OsSUT) genes during drought and salinity treatments of three week old rice plants ( at four leaf stage) over a 10 days. Among the five rice OsSUT genes identified, only OsSUT2 was observed to be progressively up-regulated during drought and salinity treatments, while OsSUT1, OsSUT4 and OsSUT5 were expressed at low levels, and OsSUT3 showed no detectable transcript expression. Sucrose transport will be essential to meet the cellular energy demands and also for osmoprotectant activities during drought and salinity stresses. It therefore indicates that OsSUT2 which facilitates transport of sucrose from photosynthetic cells will be III essential for rice plants to cope with drought and salinity stresses, and cultivars with a higher OsSUT2 expression should be able to tolerate these environmental stresses better. The role of OsSUT in assimilate transport during rusty plum aphids (Hysteroneura setariae; Thomas) infestation on the leaves of three week old rice (Orzya sativa L. cv Nipponbare) cultivar plants, over a time-course of 1 to 10 days of treatments, was also examined by combination of gene expression and β-glucuronidase (GUS) reporter gene analysis. Real Time PCR analysis of the five OsSUT genes revealed that the expression of OsSUT1 was progressively up-regulated during the course of aphid infestation. OsSUT2 and OsSUT4 expression were comparatively low in both the control and treated plants. OsSUT5 showed no clear difference in transcript expression in both control and treated plants, while no detectable transcript expression of OsSUT3 could be found. The up-regulation of OsSUT1 gene was verified at protein level by western blot analysis in both the control and treated plants. OsSUT1 protein expression was found to increase with time during aphid infestation. A similar trend was noticeable in the control plants, however at a lower expression level. These demonstrate that the cellular expression of OsSUT1is regulated by both developmental and environmental factors. OsSUT1-promoter:::GUS reporter gene expression was observed within the vascular parenchyma and/or companion cells associated with phloem sieve elements of the large and small bundles in the phloem tissues of the flag leaf blade regions where feeding aphids were confined, which progressively increased with time of infestation. It is suggested that OsSUT1 may primarily play an essential role in phloem transport of assimilate to wounded tissues from adjacent health tissues or may be involved in the retrieval of assimilate back into the phloem to minimize loss caused by the infestation. Some OsSUT1-promoter:::GUS expression was also found in the metaxylem at 10 days after infestation, which could signify a recovery system in which sucrose lost into the xylem as a result of aphids feeding are retrieved back into the phloem through the vascular parenchyma. This was supported by the exposure of cut ends of matured OsSUT1-promoter:::GUS rice plant leaf to 2% sucrose solution. OsSUT1-promoter:::GUS expression was observed within the protoxylem, xylem and phloem parenchyma tissues. This indicates that sucrose translocating within the xylem tissues are retrieved into the phloem via the OsSUT1 localized within the parenchyma tissues. In conclusion, the differential expression and regulation of rice (Orzya sativa L. cv Nipponbare) sucrose transporters as reported here suggest that OsSUT2 and OsSUT1 were constitutively expressed compared to other rice sucrose transporters during drought and salinity, and rusty plum aphids (Hysteroneura setariae; Thomas) infestation stresses respectively. Thus, the expression and regulation of the sucrose transporters could be related to the physiological and nutritional requirements of the cells during plant developmental or environmental stress state that allows their differential expression.
  • Full Text:
  • Date Issued: 2011

Plant aphid interactions : effects of diuraphis noxia and rhopalosiphum padi on the structure and function of the transport systems of leaves of wheat and barley

  • Authors: Saheed, Sefiu Adekilekun
  • Date: 2008
  • Subjects: Russian wheat aphid Rhopalosiphum padi Aphids -- Host plants Wheat -- Diseases and pests
  • Language: English
  • Type: Thesis , Doctoral , PhD
  • Identifier: vital:4225 , http://hdl.handle.net/10962/d1003794
  • Description: The infestation of the cultivated grain crops by phloem feeding aphids has generated a great deal of interest over the years, due to the serious damage they cause to the crops and yield losses that result. The mechanism of the interaction between aphids and host plants remains largely unknown in spite of efforts to understand the basis of aphid feeding on grain crops. Greater efforts are required to explain the mechanism(s) of this interaction in order to achieve sustainable agriculture. This thesis focused on an investigation of the mechanism of feeding by the Russian wheat aphid, Diuraphis noxia Mordvilko (RWA) and the bird cherry-oat aphid, Rhopalosiphum padi L. (BCA) on barley and wheat cultivars. These two aphids co-occur naturally, but they inflict very different feeding effects on host plants. Structural and functional approaches were employed to investigate their feeding habits and these were then related to the observed differences in their host plants. Transmission electron microscopy (TEM) techniques were used to study the ultrastructural damage, while fluorescence microscopy techniques – using aniline blue fluorochrome (a specific stain for callose) and 5, 6-CFDA (a phloem-mobile fluorophore) – were employed to investigate the functional response to damage via wound callose formation and phloem transport capacity respectively. RT-PCR and quantitative real-time RT-PCR techniques were used to investigate the regulation of the genes involved in callose synthesis and degradation at the transcriptional level. Morphological observation of the damage caused by the aphids show that infestation by RWA results in extensive leaf chlorosis, necrosis and rolling, while infestation by BCA does not lead to any observable symptoms within the same period. Interestingly, the population study shows that BCA breeds faster than RWA within the two-week experimental period. The ultrastructural study of feeding damage caused by the two aphids on the vascular bundles of susceptible barley cv Clipper, shows a different patterns of damage. Probing the vascular bundles results in the puncturing of vascular parenchyma by both aphids, but severe damage occurs in sieve tubes-companion cell complex during sustained feeding by RWA. In contrast, less damage occurs when BCA feeds on the phloem. Drinking from the xylem by RWA results in deposition of a large quantity of electron-dense watery saliva, which apparently seals the xylem vessels completely, by blocking all the pit membrane fields between the xylem vessels and associated parenchyma cells. In contrast, drinking from xylem by BCA results in deposition of a dense, granular saliva into the xylem vessels only, which does not appear to totally occlude the pit membrane fields. This is the first known report in which ultrastructural evidence of aphids’ drinking in xylem is provided. The comparative effects of RWA feeding on a susceptible Betta and resistant Betta-Dn1 wheat cultivars showed that after two weeks, the Betta cultivar expressed damage symptoms such as chlorosis, necrosis and leaf roll, while few chlorotic patches and necrotic spots occur in resistant Betta-Dn1 cultivars. An ultrastructural investigation of the feeding damage caused to all leaf tissues revealed, for the first time, that RWA is capable of both intra- and inter-cellular probing within mesophyll cells. Probing in the mesophyll cells induces a more severe damage in susceptible Betta than in the resistant Betta-Dn1 counterpart. Similar differences in damage occurred during feeding in the thin-walled sieve tubes of the phloem, with the sieve tubes of the Betta showing more damage than that of the resistant Betta-Dn1. However, drinking from xylem resulted in the characteristic occlusion of metaxylem vessels by copious deposition of saliva by RWA in both Betta and Betta-Dn1 cultivars. In all cases of probing, feeding, and drinking by RWA in both cultivars, all probed cells with evidence of salivary material deposit and those cells adjacent to salivary material deposit, exhibit significant damage in susceptible Betta cultivar, whereas similar cells in Betta-Dn1 cultivars do not show as damage as severe. Investigation of the functional response of the plants to feeding by aphids through the deposition of wound-induced callose shows that formation and deposition of wound callose occurs in both longitudinal and cross veins within 24h of feeding by RWA. This deposition increases through short-term feeding (72h) and prolonged feeding (14d). This is in sharp contrast to the observations with BCA feeding,where little or no callose formation occurs within the same time frame. Callose formation and deposition occurs only when a higher population of BCA feeds on barley leaves. This is the first report of aphid-induced wound callose by BCA. In all cases of callose deposition, aphid stylet tracks were associated with callose and the deposition of callose appears to be a permanent feature, because wound callose remained in the leaf tissues even after 120h of the aphids’ removal. Wound callose signals (defence and anti-defence) are discovered to be transported in the phloem tissues and are dependent on the direction of assimilate flow. Examination of the possible regulation of wound callose genes at the transcriptional level shows that the two expressed glucan synthase gene sequences (GSL – genes involved in callose formation) analysed did not show any significant increase or regulation upon aphid infestation. Contrary to expectation, all three aphid-induced β-1, 3-glucanases (genes which are thought to be involved in callose degradation) showed higher expression in RWA-infested tissue than in BCA-infested tissue. The results of the feeding damage on the transport capacity of the phloem shows that BCA infestation does not lead to a significant reduction in the phloem transport capacity during short-term feeding (72h), while RWA-infested leaves showed considerable reduction in the transport capacity of the phloem within the same period. However, prolonged feeding (14d) by BCA induces a considerable reduction on the transport capacity of the phloem on the infested tissues. In contrast, a marked reduction in the transport capacity of the phloem occurs in RWA-infested leaves and in most cases, complete cessation of transport ensues. In conclusion, these data collectively suggest that RWA is a serious and most destructive phloem feeder in comparison to the BCA. RWA causes severe damage to all cellular tissues of the host plants, which result in apoplasmic and symplasmic isolation of xylem and phloem tissues, while BCA infestation does not result in such isolation within the same time and population levels. Resistance genes appear to function by conferring resistance to cell damage on the resistant cultivars during aphid feeding. Responses by plants to aphid infestation via wound callose deposition are again shown to be species-specific. A quick response results when RWA feeds, even at a very low population level, while a response occurs only at a higher infestation level by BCA, and this response was shown as not regulated at the transcriptional level. Differences in the damage to leaf tissues and wound callose deposition eventually lead to varying degrees of damage to the transport capacity of the phloem. These differences in the damage signatures are hereby suggested to be the cause of the diversity in the observed damage symptoms and the yield losses upon infestation by the two aphid species.
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  • Date Issued: 2008

Wheat stress responses during Russian wheat aphid and Bird Cherry Oat aphid infestation: an analysis of differential protein regulation during plant biotic stress responses

  • Authors: Louw, Cassandra Alexandrovna
  • Date: 2007
  • Subjects: Russian wheat aphid , Plants, Effect of stress on , Wheat -- Diseases and pests , Rhopalosiphum , Plant proteins
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:3995 , http://hdl.handle.net/10962/d1004055 , Russian wheat aphid , Plants, Effect of stress on , Wheat -- Diseases and pests , Rhopalosiphum , Plant proteins
  • Description: Plants possess a complex and poorly understood network of defence mechanisms that enable them to counteract the effects of abiotic and biotic stress. Aphid phloem feeding is source of biotic stress in plants. Russian wheat aphid and Bird Cherry-Oat aphid feeding cause significant losses in the annual wheat crop, and control by conventional methods such as pesticide application, has proved to be ineffective. Infestation by the Russian wheat aphid has a particularly devastating effect in South Africa. Aphid-resistant wheat cultivars have been identified but an incomplete understanding of the mechanism of the plant’s resistance thwarts the development of improved cultivars. A two-dimensional gel electrophoresis method was developed, partially optimised and validated in order to determine the effect of Russian wheat aphid and Bird Cherry-Oat aphid phloem feeding on the Betta and Betta DN wheat proteome. Differentially expressed proteins that were up or down regulated more than two fold were identified using PDQuest™ Basic software and matched to known wheat proteins stored in the SwissProt protein database on the basis of their molecular mass and isolectric point. Initial analysis of the differential protein expression of Betta and Betta DN wheat in response to Russian wheat aphid and Bird Cherry-Oat aphid phloem feeding at different growth stages revealed that younger plants display higher levels of resistance than older plants. Feeding by the Bird-Cherry Oat aphid does not result in the upregulation of proteins implicated in a defence response, which indicates that the damage incurred by the plant due to feeding by this aphid is not enough to trigger a classic defence response. Feeding by the more damaging Russian wheat aphid resulted in a stress response in susceptible wheat cultivar Betta, and a defence response in resistant wheat cultivar Betta DN. The infestation of Betta DN resulted in the upregulation of putative thaumatins and amylase trypsin inhibitors, indicating that the Betta DN resistance response could be due to the combined effect of protease inhibitors that discourage aphid phloem feeding and the activation of the salicylic acid and jasmonic acid plant defence pathways.
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  • Date Issued: 2007

Plastochron index - an indicator of plant structure and function a case study using Pisum sativum L

  • Authors: Ade-Ademilua, Omobolanle Elizabeth
  • Date: 2006
  • Subjects: Plant anatomy Plant physiology Peas -- Anatomy Peas -- Physiology
  • Language: English
  • Type: Thesis , Doctoral , PhD
  • Identifier: vital:4183 , http://hdl.handle.net/10962/d1003751
  • Description: The use of chronological age for example, using days after sowing (DAS), or days after germination (DAG) as a time variable may result in the inherent variability between plants resulting in differences which can be large enough to obscure subtle developmental trends that become evident among plants sown at the same time. An alternative to DAS or DAG is the plastochron index (PI), first used by Erickson and Michelini (1957) as a morphological time scale and numerical index; which to according to the authors suggested and represented a more accurate reflection of the developmental status of a plant. The research presented in this thesis was therefore aimed specifically at utilizing the index in qualitative and quantitative analyses, to confirm its usefulness in analyzing and predicting plant growth and development. Specifically this research focused on investigating various morphological and physiological events that together, hopefully, would serve as a template for the prediction of the growth, development and reactions of Pisum sativum L. to different growth conditions. In Chapter 3, the use of the average length of the first pair of leaflets on each node as a suitable parameter for calculating PI in P. sativum is suggested. The results presented in Chapter 3 suggest that plant age is best expressed using the plastochron index, as this reflects the time interval between the initiations of successive pairs of leaflets. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2005) A re-evaluation of plastochron index in peas - a case for using leaflet length. South African Journal of Botany 71: 76-80”. The PI formula developed was subsequently used in this research to conduct qualitative and quantitative investigations of plant growth and development in which all data and observations were related directly to the plastochron index. In Chapter 4, the sink to source transition in Pisum sativum L. leaves at different plastochron ages in nodulating plants was investigated using the phloem-mobile fluorescent marker, 5,6-carboxyfluorescein (5,6-CF). The results demonstrated that young leaves remained strong sinks up until LPI 0, after which sink-source transition occurred up to LPI 1.8 and leaflets transitioned to strong source systems by LPI 2.0. A well-developed cross-connected phloem system between paired leaflets in peas, and the petiole and the stem vascular supply was observed. The data presented in the second part of Chapter 4 suggest that the phloem transport between leaflet pairs is independent of the sink/source state of the leaflets, or of movement along the source to sink gradient. The data support the presence of a modular transport system which may ensure re-allocation and balancing between leaflets of the same physiological age and photosynthetic and transport status, thereby load-balancing the local transport system, before exporting to other younger (sink) regions. The investigation of leaf development using the plastochron index (Chapter 5) revealed that the formation of air spaces in the palisade and spongy mesophyll, one of the preparatory events for transition from sink to source state in developing leaves, occurs between LPI 0 and LPI 1 in pea leaflets. Results of the anatomical and ultrastructural study related to PI are presented in Chapter 5. The density of wall ingrowths in transfer cells of minor veins increased with LPI and appeared to be associated with the probable transition to source state and the related potential increase in the production of assimilates for export. The onset of wall ingrowth development in leaflets at LPI 0 provided evidence that sink-to-source transition commences at LPI 0 in P. sativum. Presumably-functional plasmodesmata as well as a few mature sieve elements were evident in class IV veins in the apical region of young and older leaflets at LPI 0. The number of mature sieve elements per vein however, increased with increasing LPI. Most class V veins were still undergoing division at LPI 0 and their sieve elements did not show signs of maturity until LPI 1. The increase in the number of mature metaphloem sieve elements in young, supposedly importing tissue at LPI 0 to older, supposedly exporting tissues at LPI 2 is evidence of the association between phloem maturation and transition from importing to exporting status. In Chapter 6, I report on the effects of elevated CO[subscript 2] on the growth and leaf development of nodulating and non-nodulating Pisum sativum L var. Greenfeast grown under controlled environment of the same nitrogen (6mM) and nitrogen- free nutrient solution conditions. Shortterm exposure to elevated CO[subscript 2] induced rapid plant growth, irrespective of treatment. However, long-term elevated CO[subscript 2] treatment did not affect rate of leaf appearance (RLA) in nodulated plants, irrespective of mineral N supply but enhanced RLA in non- nodulating plants supplied with mineral N. Supplied N resulted in a significant increase in leaflet elongation rate (LfER) under both ambient and elevated CO[subscript 2], but LfER was not significantly affected by nodulation but was increased by high CO[subscript 2]. This suggested that the growth of nodulating P. sativum L may not be significantly affected under CO[subscript 2] levels as high as 1000 μmol mol[superscript -1]. The data suggest that elevated CO[subscript 2] will enhance canopy size, provided adequate soil N is available and more so in non-nodulating plants. This section of the research has been published as “Ade-Ademilua OE, Botha CEJ (2004) The effects of elevated CO[subscript 2] and nitrogen availability supersedes the need for nodulation in peas grown under controlled environmental conditions. South African Journal of Botany 70: 816 – 823”. This thesis demonstrates that the similarity in the qualitative analyses results obtained from plants from different CO[subscript 2], nitrogen and nodulation treatment conditions, highlights the fact that plants of same PI value are at the same developmental state, irrespective of the growth condition. Furthermore, changes in plant structure and function observed under different growth conditions can be related simply to changes in plastochron index. The work presented in this thesis demonstrate that changes in plant structure and function analyzed are related to changes in PI. An important finding of this thesis is that with the use of PI, results can be compiled as a template for predicting the structure- function state of pea plants at any plastochron age, under any growth conditions, before using small representative sample populations.
  • Full Text:
  • Date Issued: 2006

The anatomy and distribution of the cyperaceae in the Eastern Cape region of South Africa

  • Authors: Sonnenberg, Bernd Jürgen
  • Date: 2005
  • Subjects: Cyperaceae Cyperaceae -- Anatomy Cyperaceae -- South Africa -- Eastern Cape
  • Language: English
  • Type: Thesis , Doctoral , PhD
  • Identifier: vital:4228 , http://hdl.handle.net/10962/d1003797
  • Description: The principal objective of this investigation, was to collect the family Cyperaceae and to study their leaf, bract and culm anatomy. The second was to examine the collection for unique structures or forms, whilst a third was to classify the Cyperaceae according to their photosynthetic structures and types. Distribution of the Cyperaceae within the broad region defined as the Eastern Cape would be influenced by rainfall pattern. It was expected that C₃ species would predominate in more mesic environments and habitats, whilst the C₄ species would be found in drier less favourable habitats. Collection within the region (November 1993 to late January 1997), yielded 106 species, totalling some 600 specimens. Both sub-families of the Cyperaceae (Caricoideae and Cyperoideae), eight tribes (Abildgaardieae, Cariceae, Cypereae, Hypotvtreae, Rhynchosporeae, Shoeneae, Sirpeae and Slerieae) and twenty five genera were found to be present. Sixty percent of the species were C₃ and forty percent were C₄. Sixteen new species, which had not been collected within the boundaries of the region previously were also found. Over 43 percent of the species collected had unique anatomical characteristics that appeared to be influenced by habitat and or climate influenced. These are the characters influenced by hydromorphic, mesomorphic and xeromorphic environments. These anatomical characters: Thickness of leaves and bracts; thickness of the adaxial and abaxial epidermis of the leaves and bracts; flush, sunken and raised stomata; presence or absence of bulliform cells; presence and distribution of sclerenchymatous structures; presence or absence of a hypodermis; presence or absence of cavities in the leaves, bracts and culms; mesophyll or ground tissue structure, and the presence or absence of secretary structures. A few noteworthy anatomical characters that are influenced by climate are present in Cladium mariscus subsp. jamaicense (Schoeneae), the Cariceae, the Cypereae and the Sclerieae. Within Cladium mariscus subsp. jamaicense the pseudo-dorsiventral leaves and bracts, as well as the large lamina cavities, containing trans-lamina girders are unique. Papillate epidermal cells are limited to the tribe Cariceae. In the Cypereae many of the species lack bulliform cells and hypodermal layers. In Pycreus cooperi (Cypereae) the vascular bundles of the leaves and bracts, appear to be stacked in rows, that are inter-spaced with lamina cavities. In the Sclerieae the mesophyll structure is specific to the species level. Unique anatomical characteristics were also present in the leaves, bracts and culms of the genus Carpha. In this genus distinctive lateral vascular bundles were present abutting the large bundles and/or midrib bundle. The most distinctive anatomical characteristics that could be used to separate the members of the Cyperaceae were the structures and associated structures related to the photosynthetic pathway. The Eastern Cape Cyperaceae could be divided into three distinct groups based on photosynthetic structure, namely one C₃, a C₄ and a potential C₃-C₄ intermediate group. The C₃ group was found to have non-radiate mesophyll and an outer parenchymatous sheath with small chloroplasts (Cariceae, Cyperaceae [in part], Hypolytreae (Chrysithrix capensis], Rhynchosporeae [R. brownii], Schoeneae, Scirpeae and Sclerieae). The C₄ group has radiate mesophyll and an inner parenchymatous sheath with enlarged chloroplasts (Kranz or PCR sheath). The C₄ species are present in tribes Abildgaardieae, Cypereae (in part) and Rhynchosporeae (R. barrosiana). In the Eastern Cape, a few species with C₃ anatomy have anatomical characteristics that are similar to the species with C₄ anatomy (Cyathocoma hexandra [bracts], Cyperus tennellus var. tennellus [leaves and bracts], Ficinia bulbosa [leaves], F. dura [leaves and bracts], F. lateralis coastal [leaves and bracts], F. oligantha [bracts], F. pingiour [bracts], F. stolonifera [leaves and bracts], F. tribracteata [leaves and bracts], F. zeyheri [leaves and bracts], Isolepis cernua [leaves and bracts], I. costata var. macra [bracts], Schoenus nigricans [leaves], Scirpus nodosus [bracts] and Tetraria cuspidata [leaves and bracts)). The vascular bundles within this intermediate group, fall within the Hattersleyand Watson (1975) minimal cell lateral count and maximal cell distal count criteria for C₄ grass species. However, no biochemical data exists to see whether they are C₃-C₄ intermediates or whether the Hattersley and Watson (1975) C₄ criteria for grasses applies to smaller, or scutiform Cyperaceae or not. Based on the results presented here, five distinct structural forms/types were found to be present in the C₃, C₄ and C₃-C₄ intermediate groups. The C₃ and the potential C₃-C₄ intermediate species may be divided into two types, based on the number of vascular sheaths present. In the first or A-type, vascular bundles are surrounded by two sheaths and in the more dominant B-type, by three. The A-type was found in the Cypereae (Cyperus denudatus and C. textilis) and most of the Scirpeae. B-type anatomy occurred in the Cariceae, Cypereae (c. difformis, C. pulcher, C. sphaerospermus, C. tennellus var. tennellus and P. mundii), Hypolytreae, Rhynchosporeae (R. brownii), Schoeneae, Scirpeae (Bolboschoenus maritimus, Ficinia cinnamomea, F. fascicularis, F. lateralis both, F. pingiour, the genus Fuirena, I. diabolica, I. fluitans, I. prolifera and Schoenoplectus paludicola) and Sclerieae tribes. Based on the vascular sheath structure, the C₄ species could be divided into three groups, namely bulbostyloid, chlorocyperoid and fimbristyloid, where the bulbostyloid structure occurred in Bulbostylis schoenoides. Cyperus (in part), Kyllinga, Mariscus and Pycreus (except P. mundii) had a chlorocyperoid structure. Genera with fimbristyloid structure were recorded in the genera Abildgaardia, Bulbostylis and Fimbristylis. The bulbostyloid type represents a potential a fifth C₄ anatomical type within the C₄ Cyperaceae. As a result of this observation, it is possible that the C₄ syndrome may have evolved five times in the Cyperaceae and not four as previously suggested by Bruhl and Perry (1995) and by Soros and Dengler (2001). The C₃ Cyperaceae species within the Eastern Cape are more dominant in higher elevation habitats the C₄ species, similar to the C₃ grasses. The only C₄ species that occur at high elevations are those with three sheaths. The C₃ and C₄ species within the region occur in similar low rainfall habitat ranges, where the C₄'s are more dominant in xeric habitats on drier soils than the C₃ species, similar to the grasses. Where more C₃ species occur in higher rainfall habitats than the C₄ species. With the exception of the Afromontane Bulbostylis schoenoides and R. barrosiana, the C₄ species similar to the grasses are dominant in high light and temperature habitats with low rainfall, unlike the C₄ Cyperaceae of Japan and America. Only five species occur in the desert like conditions of the Karoo-Namib biome (Cyperus laevigatus, C. rupestris var. rupestris, I. cernua, M. capensis and M. uitenhagensis), which have less than 250mm of rainfall per annum. Only three species are habitat-specific or may be endemic to a specific area within the Eastern Cape, namely A. capensis, Chrysithrix capensis and R. barrosiana. A. capensis in marshes on the Amatole mountains near Alice and Hogsback. C. capensis to the Tstsikamma mountains of the Wite Els Bosch forests. R. barrosiana to the marshlands of the Cape Morgan coastal Nature reserve at Kei Mouth. The anatomical types of the C₃ and more especially C₄ Cyperaceae are not specifically found in a particular rainfall regime or habitat type, which is contrary to the thesis hypothesis. However, the C₃ species are mostly correlated with hydrophytic to mesic habitats, with the exception of Ficinia and the two sheathed species. Ficinia is dominant in mesic grasslands and halophytic habitats. The two sheathed C₃ species are mostly present in halophytic habitats. The C₄ species are also more dominant in mesic to xerophytic grasslands, as expected in the hypothesis. Where only a few species occur in habitats correlated with increasing rainfall and temperature similar to the C₄ Cyperaceae of Japan and America. It may thus be that the development and evolution of the different C₄ anatomical forms (or phylogenetic forms) within the Cyperaceae may have enabled these species to establish themselves in habitats that were alien to their origins. It may be that the ability to regulate photoassimilate and water transport within the Cyperaceae enables their success in a dynamic and unpredictable climate, such as the Eastern Cape. Many of the anatomical characteristics reported in this thesis and its appendices are unique to the tribes, genera and/or species of the Eastern Cape Cyperaceae and thus may be valuable to future taxonomic classifications of the family. The research presented here should provide a good working platform for future, more detailed research on this often forgotten component of the vegetation.
  • Full Text:
  • Date Issued: 2005

Confirmation of a slow symplasmic loading and unloading pathway in barley (Hordeum Vulgare L.) source and sink leaves

  • Authors: Buwa, Lisa Valencia
  • Date: 2003
  • Subjects: Phloem , Plant translocation , Barley -- Metabolism
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4253 , http://hdl.handle.net/10962/d1007804
  • Description: Visualization of the transport pathway in barley (Hordeum vulgare) leaves was carried out using a combination of aniline blue and a symplasmically transported fluorochrome, 5,6 carboxyfluorescein (5,6-CF). When applied to a source leaf, basipetal movement of 5,6-CF was observed after 3 h and the fluorochrome front was observed about 3-4cm away from the point of application. The fluorochrome was taken up into the symplasm of the mesophyll and was loaded into the bundle sheath cells and then subsequently the vascular parenchyma and finally into the sieve tubes. In sink leaves, acropetal movement was observed after 3 h and the fluorochrome had moved approximately 3 cm away from the point of application. Unloading of 5,6-CF occurred from all classes of longitudinal veins. Studies on solute retrieval showed that 5,6 CF-diacetate was transferred to xylem parenchyma where it was metabolized. 5,6-CF was then transferred from the xylem parenchyma to the vascular parenchyma cells, and it would appear that thick-walled sieve tubes were the first to show 5,6-CF labeling. Counterstaining with aniline blue demonstrates the presence of plasmodesmata and this suggests a potential symplasmic pathway from the mesophyll to the sieve tubes. Application of 5,6 CF-diacetate revealed a slow symplasmic pathway, which involved transfer of 5,6-CF, which was effected via plasmodesma.
  • Full Text:
  • Date Issued: 2003

Symplasmic pathway in phloem loading and unloading in source and sink leaves of Zea mays L. as evidenced under normal and elevated CO₂ conditions

  • Authors: Nogemane, Noluyolo
  • Date: 2003
  • Subjects: Phloem , Plant translocation , Plant cells and tissues , Corn -- Metabolsim
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4254 , http://hdl.handle.net/10962/d1007813
  • Description: Zea mays plants kept at ambient (ca 375ppm) and elevated CO₂ (ca 650 to 700ppm) were used to examine the possibility of a symplasmic loading, unloading and transport pathway in dark-adapted and illuminated (200μmolm⁻²sec⁻¹ ) sink and source leaves. 5,6-carboxyfluorescein diacetate was introduced into the mesophyll cells and symplasmic transfer observed 3h after application. In sink and source leaves exposed to ambient CO₂ and illuminated at 200 molm-2sec-1, the fluorescence front was observed approximately 3cm from the point of application, while in dark-adapted plants, the fluorescence front was observed approximately 1cm from the point of application. Under elevated CO₂ conditions the fluorescence front in illuminated plants appeared to transport faster moving approximately 5cm from the point of application, and in dark-adapted plants, only 3cm from the point of application. Based on the increase in 5,6-CF accumulation under elevated CO₂ conditions, the present study suggests that there was an increase in capacity for assimilate loading and transport under elevated CO₂ conditions. In source leaves, 5,6-CFDA was taken up into the mesophyll cells, loaded symplasmically and transported basipetally. In sink leaves 5,6- CFDA was taken up from basal mesophyll and after symplasmic loading, was transported acropetally where it was offloaded into the younger immature sink region. Transport in the sieve tubes was confirmed by using aniline blue, which was applied 3h after 5,6-CF transport. Aniline blue coupled with 5,6-CF transport studies showed that the sieve tubes of both cross and longitudinal veins are involved in symplasmic unloading, loading and transport processes in sink and source leaves. Apoplasmic uptake of 5,6-CFDA by cut leaves showed that after apoplasmic transport via the transpiration stream, 5,6-CFDA was offioaded to the xylem parenchyma where it was metabolically cleaved , releasing fluorescent 5,6-CF into the xylem parenchyma. Transverse sections cut after 3h of uptake were observed after 120 and 180 min suggesting that a retrieval of solutes occurs from the xylem to the xylem parenchyma, bundle sheath, phloem parenchyma and to the th in-walled sieve tubes. It was not possible to determine if the thick-walled sieve tubes were involved or if they took up 5,6-CF. Given the available data on loading and offioading of assimilates in sink and source leaves respectively, this study demonstrated that a slow symplasmic pathway exists from the mesophyll to the phloem, and that offloading from the phloem in sink leaves can occur via a symplasmic route.
  • Full Text:
  • Date Issued: 2003

The phloem unloading and sucrose-sequestration pathway in the internodal stem tissue of the Saccharum hybrid var. NCo376

  • Authors: Gerber, Jacqués
  • Date: 2001
  • Subjects: Saccharum , Sugarcane , Sugar -- Synthesis , Sugar growing
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4194 , http://hdl.handle.net/10962/d1003763
  • Description: Internodes 5-8, 10, 13 and 15 of Saccharum sp. var. NCo376 were examined for evidence of symplasmic phloem unloading of sucrose from the phloem, via the bundle sheath to the storage parenchyma. The vascular bundle possesses wellisolated phloem comprised of large diameter sieve elements and small diameter companion cells. A layer of phloem parenchyma surrounds the phloem, except where the phloem abuts the crushed protophloem. Outside this is a sclerenchymatous sheath, directly endarch to a parenchymatous bundle sheath, which is surrounded by storage parenchyma. The bundle sheath is interrupted at the centrifugal pole of the vascular bundle by a phloem fibre cap. Scanning Electron Microscopy revealed plasmodesmal fields throughout the bundle sheath and pith tissue. Transmission Electron Microscopy studies provided evidence of plasmodesmal occlusion, but not in all tissues. Aniline blue reactions under UV light indicate the presence of occluded plasmodesmal fields at the phloem parenchyma / sclerenchymatous sheath interface, and in localised regions of cells which are smaller than the surrounding storage parenchyma cells. This suggests a symplasmic transport pathway at these locations, and, based on these positive aniline blue reactions, with regulation via callose-mediated transplasmodesmal transport. Osmotic stress experiments, which included the addition of Ca2+, did not reveal further callose occlusion in the parenchyma, suggesting that the plasmodesmata in these regions may be closed via a noncallosic mechanism. Dye-coupling studies, using Lucifer Yellow (LYCH), which was iontophoretically injected following turgor-pressure equalisation, showed only rare, limited symplastic transport, usually only between the injected cell and one adjacent cell. Most injections did not result in transport of LYCH, suggesting either a lack of plasmodesmal connectivity, occlusion, or gating of any plasmodesmata present. This limited symplasmic transport, combined with the presence of occluded plasmodesmata at the phloem parenchyma / sclerenchymatous sheath interface suggests the presence of a two-domain phloem-unloading pathway. While symplastic transport may occur from the phloem to the sclerenchymatous sheath, further sucrose transport to the storage parenchyma appears to proceed apoplasmically from the sclerenchymatous sheath / bundle sheath interface, and into storage parenchyma cells across the cell wall and cell membrane via specialised sucrose transporters.
  • Full Text:
  • Date Issued: 2001

Vein structure in relation to phloem loading in selected Ranunculaceae, Apocynaceae and Asclepiadaceae of the Eastern Cape

  • Authors: Buswell, Alison Mary
  • Date: 2001
  • Subjects: Ranunculaceae Apocynaceae Asclepiadaceae Phloem Plants -- South Africa -- Eastern Cape
  • Language: English
  • Type: Thesis , Doctoral , PhD
  • Identifier: vital:4243 , http://hdl.handle.net/10962/d1007277
  • Description: The relationship between leaf architecture, vein anatomy and phloem ultrastructure, and that of possible routes from mesophyll cells to phloem and potential phloem loading method was investigated using species adapted to the southern African climate. The research was based on the hypothesis of Gamaiei and Van Bel, using nothern hemisphere species only (Gamalei 1985a, b, 1989, 1991, VanBeletal. 1988, Van Bel 1992, 1994, 1996, Van Bel & Gamalei 1991, 1992, Gamalei et al. 1992, 1994, 1996, Van Bel 1992a-c, 1993a, b, 1996). The thesis commenced with a survey ofleafarchitecture of the Ranunculaceae, Apocynaceae and Asclepiadaceae. Anatomical and ultrastructural studies followed. Leaf architecture was described according to Hickey (1973). Within the Ranuncuiaceae, leaf architecture was found to be marginally actinodromous. Venation pattern consisted of a widely spaced reticulum of delicate veins, especially in Ranunculus. Leaf architecture of the Apocynaceae was described as pinnate, camptodromous and brochidodromous. The Asclepiadaceae showed less uniformity in terms of leaf architecture, being pinnate and camptodromous, with mostly brochidodromous and, unexpectedly, eucamptodromous patterns of secondary venation. A predominantly common leaf architecture supported the move to amalgamate the two families. As the less advanced eucamptodromous arrangement could represent a more primitive branch of this huge family, the phylogenetic classification of the new amalgamated family is eagerly awaited for discussion. Allocation of vein order allowed comparisons between species and families to be drawn. Reticulum density and vein order anatomy was used to indicate potential routes from mesophyll to phloem. A definite contrast was obvious between the loose arrangement of mesophyll and veins in the mesic Ranunculus, and the close mesophyll and dense venation of the xeric apocynate and asclepiad species, and was related to habitat. Ultrastructural characteristics of companion cells, together with plasmodesmatal abundance, were considered especially important for the determination of minor vein configuration. Descriptions of plasmodesmatal distribution did not consider functional status. In this thesis, vein structure and ultrastructure were considered in relation to phloem loading, not as a demonstration thereof. All three families were designated minor vein configuration type 2a. Two interesting examples that did not adhere to the familial norm, viz. few plasmodesmata and normal companion cells, occurred in the Asclepiadaceae. Secamone alpinii had abundant aggregated plasmodesmata, forming a potential symplasmic continuum from mesophyll to companion cells. The question of plasmodesmatal functionality remained open. Ceropegia carnosa showed folding of the companion cell membrane, but no accompanying wall ingrowths. The folds were suggested to increase surface area for apoplasmic phloem loading in the noted absence of plasmodesmata. Loading routes and methods suggested were based on anatomical and ultrastructural evidence only. Whilst these results were supported by published data for other species of these families, the prediction of the Gamalei and Van Bel hypothesis did not hold true. The relatively primitive Ranunculaceae were expected to have the least advanced type 1 minor vein configuration, with abundance plasmodesmata providing a symplasmic phloem loading pathway. The relatively advanced Apocynaceae and Asc1epiadaceae were predicted to have the most progressive minor vein configuration, type 2b, with specialised transfer cells to maximise apoplasrnic uptake. As families with type 2a minor vein configurations, the Ranunculaceae were more advanced than expected and the Apocynaceae and Asc1epiadaceae less so.
  • Full Text:
  • Date Issued: 2001

Observations of selective feeding of the aphid, Sitobion yakini (eastop) on leaf blades of barley (Hordeum vulgare L)

  • Authors: Matsiliza, Babalwa
  • Date: 2000
  • Subjects: Aphids , Phloem , Barley
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4212 , http://hdl.handle.net/10962/d1003781 , Aphids , Phloem , Barley
  • Description: Penetration of leaves of barley (Hordeum vulgare L.) plants grown under normal conditions and those exposed to physiological stress, by the aphid, Sitobion yakini was investigated using light and electron microscope techniques. This was carried out to determine if there was evidence of preferential feeding on either thin- or thick-walled sieve tubes in the barley leaf. Under both stress and non-stress conditions, preliminary results of an electron microscope investigation showed that penetration of the epidermis and mesophyll was largely intercellular, becoming partly intercellular and in part intracellular inside the vascular bundle. A total of 317 stylets and stylet tracks were encountered during the examination of 2000 serial sections. In non-stressed plant material, 293 (92%) terminated in thin-walled sieve tubes and only 24 (8%) in thick-walled sieve tubes. Investigation of 1000 serial sections using stressed plant material showed, that 84 of 89 (94%) stylets and stylet tracks encountered terminated in thin-walled sieve tubes. Furthermore, 90 of 94 (96%) stylets and stylet tracks encountered in 1000 serial sections from the second experiment of control non-stressed plant material (control) terminated in thin-walled sieve tubes. The thin-walled sieve tubes were significantly more visited (probed) by the aphid than the thick-walled sieve tubes. Under stress conditions, 50 of 89 (56%) stylets and stylet tracks which terminated in thin-walled sieve tubes were associated with the small longitudinal bundles. Under normal conditions, 65 of 94 (69%) stylets and stylet tracks which terminated in thin-walled sieve tubes were associated with the small longitudinal bundles. There were no significant differences on the number of probes of sieve elements between the two treatments. These data suggest that the aphid S. yakini feeds preferentially on the thin-walled sieve tubes of the small longitudinal vascular bundles in plants grown under normal conditions and those exposed to physiological stress, such as water stress. This further suggests that the thin-walled sieve tubes in barley leaf blades are more attractive to the aphid and are probably most functional in terms of phloem loading and transport.
  • Full Text:
  • Date Issued: 2000

Effects of sustained elevated CO₂ concentration on two cultivars of barley (Hordeum vulgare L.)

  • Authors: Siphugu, Mashudu Victor
  • Date: 1997
  • Subjects: Barley Barley -- Growth Photosynthesis -- Research Plants -- Effect of carbon dioxide on
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4226 , http://hdl.handle.net/10962/d1003795
  • Description: The enormous burning of fossil fuel and deforestation have caused an increase in the atmospheric CO₂ concentration ([CO₂]) during the last century. This will invariably have profound direct and indirect effects on plant carbon metabolism. The majority of research on the effects of CO₂ enrichment on plants are short-term and are done on other crops, but very little have been done on barley. This project aimed to determine the effects of long-term CO₂ enrichment on photosynthesis, growth and grain yield on barley. Hordeum vulgare L. cvs Stirling and Schooner plants were grown from seeds in controlled environment chambers at ambient (350) and elevated (600) μmol molˉ¹ [CO₂]. Measurements of net assimilation rate (NAR), photosynthetic pigments content and growth parameters were started 7 days after germination (DAG) and continued until senescence. The anatomy of matured fully developed leaves was also monitored. Elevated [CO₂l resulted in an increase in NAR in the two cultivars from days 7 until 14, after which the stimulation of NAR of CO₂-enriched plants started to decrease. At the onset of senescence, NAR was almost equal in plants grown under both ambient and elevated [CO₂]. The response of assimilation as a function of internal [CO₂l (C₁) at the end of the experimental period showed a significant decrease in both the initial slope of the A/C₁ curves and the CO₂-saturated photosynthetic rates in the two cultivars. Stirling showed no significant changes in the content of chlorophyll α,chlorophyll б or in total carotenoids. However, Schooner showed a stimulation in chlorophyll α content at day 7, but decreased at day 28. Chlorophyll б and total carotenoids content were not affected by CO₂ enrichment. While total above-ground biomass was not affected by elevated [CO₂] in the two cultivars, total plant height decreased significantly after 14 days in Stirling whereas no significant change occurred in Schooner throughout the experimental period. Leaf area was not significantly affected by CO₂ enrichment in the two cultivars although the leaves in CO₂ enriched plants were slightly shorter. Anatomical studies reveal that leaf thickness was significantly increased by CO₂ enrichment in Stirling, but the increase was not significant in Schooner. Both cultivars did not show any significant effect on chloroplast morphology and ultrastructure as a consequence of elevated CO₂ exposure. No signs of starch accumulation were evident in variety Schooner, but Stirling showed some form of starch accumulation, under increased atmospheric [CO₂]. Elevated CO₂ resulted in a significant reduction by more than 50 % in the number of grain yield per plant in both Stirling and Schooner. Results from this study therefore indicate that CO₂ enrichment will not be beneficial in terms of growth and yield in this important crop.
  • Full Text:
  • Date Issued: 1997

The effect of elevated CO₂ on Phaseolus vulgaris L. cv Contender

  • Authors: Mjwara, Jabulani Michael
  • Date: 1997
  • Subjects: Plants -- Effect of carbon dioxide on Kidney bean
  • Language: English
  • Type: Thesis , Doctoral , PhD
  • Identifier: vital:4215 , http://hdl.handle.net/10962/d1003784
  • Description: The response of Phaseolus vulgaris L. cv. Contender grown in controlled environmental conditions, at either ambient or elevated (360 and 700 μmol mol ̄¹, respectively) CO₂ concentrations ([CO₂]), was monitored from 10 days after germination (DAG) until the onset of senescence. Elevated CO₂ had a pronounced effect on total plant height (TPH), leaf area (LA), dry weight (DW) accumulation and specific leaf area (SLA). All of these were significantly increased by elevated [CO₂] with the exception of SLA, which was significantly reduced. Except for higher initial relative growth rates (RGR) in CO₂-enriched plants, RGR did not differ significantly between the two CO₂ treatments throughout the remainder of growth period. While growth parameters clearly differed between CO₂ treatments, the effects of CO₂ on many physiological processes including net assimilation rate (NAR), Rubisco activity, and some foliar nutrient concentrations were largely transient. For example, CO₂ enrichment significantly increased NAR, but from 20 DAG onward, NAR declined to levels measured on plants grown under ambient CO₂. Similarly, the decline in both foliar N concentration and Rubisco activity in CO₂-enriched plants after 20 DAG was significantly greater than the decline observed for ambient CO₂ plants. Soluble leaf protein and total chlorophylls (a+b) were also significantly reduced in plants grown under elevated CO₂. Chlorophyll (a/b) ratios increased with time underelevated CO₂, indicating that the rate of decline of chlorophyll b was higher than that of chorophyll α. No significant changes in total carotenoid (x+c) levels were observed in either CO₂ treatment. Under enhanced CO₂, the foliar concentrations of K and Mn were increased significantly, while P, Ca, Fe and Zn were reduced significantly. However, changes in Mg and Cu concentrations were not significant. High CO₂-grown plants also exhibited pronounced leaf discoloration or chlorosis, coupled with a significant reduction in leaf longevity. The levels of non-structural carbohydrates (sucrose, glucose, fructose and starch) and nitrogenous compounds (nitrogen, total soluble proteins and free amino acids) were determined for leaves and developing seeds of P. vulgaris. Leaf tissue of elevated CO₂-grown plants accumulated significantly higher levels of both soluble sugars and starch. Leaf ultrastructure revealed considerable erilargement of starch grain sizes with surface areas more than five times larger compared to those of control plants. No apparent differences in structure and membrane integrity of chloroplasts in both CO₂ treatments were noted. Although ambient CO₂-grown plants had comparatively low levels of non-structural carbohydrates (NSC), they accumulated significantly higher levels of nitrogenous compounds. The levels of NSC were consistently higher in seeds of plants grown under elevated CO₂. In comparison to plants grown at elevated [CO₂], pods and seeds of ambient CO₂-grown plants had significantly larger pools of free amino compounds and N. Stomatal conductance (gs) declined significantly, as expected for plants grown under elevated CO₂. This was accompanied by a decline in transpiration rates (E). Reduced gs and E led to high AlE ratio, which meant improved water use efficiency (WUE) values for CO₂-enriched bean plants. Leaf carbon isotope discrimination (∆) against the heavier isotope of carbon (¹³C), has been used to select for high WUE in C₃ plants. In plants grown at elevated CO₂ concentration, ,1 was significantly reduced. Although ∆ was negatively correlated with WUE in both CO₂ treatments, the correlation was steeper and highly negative for CO₂-enriched plants. These results indicate underlying differences in gas-exchange physiology, including stomatal responses between ambient and elevated CO₂-grown plants. Photosynthetic acclimation was investigated using the response of assimilation to internal carbon dioxide concentration (A/C₁ curves). At early stages of growth, the initial slope of the A/C₁ response curve did not differ with CO₂ treatment. In contrast, CO₂-saturated photosynthetic rate (Amax) was significantly higher in plants grown under elevated versus ambient CO₂ at 15 DAG. However, at subsequent stages of growth both the initial slope and Amax declined in bean plants grown in elevated CO₂. Apparent carboxylation efficiency (ACE, estimated from the initial slope of A/C₁ response) values followed a similar trend and were significantly reduced in CO₂-enriched plants. These results indicate that acclimation or negative adjustment of photosynthesis may have been caused by a combination of both stomatal and biochemical limitations. Bean plants grown under conditions of elevated atmospheric CO₂ flowered 3 to 4 days earlier, and produced significantly more flowers and pods than plants grown at ambient conditions. Plants grown at elevated CO₂ aborted 22 and 20% more flowers and pods, respectively, than plants grown at ambient CO₂. Elevated CO₂ also significantly increased the number of tillers or lateral branches produced by plants, which contributed to a significant increase in pod number and seed yield in these plants. Although plants grown at elevated CO₂ produced on average 8 seeds per pod, while plants grown under ambient CO2 conditions produced 5 seeds per pod, the greater number of seeds was offset by lower seed weights in plants grown under _ elevated CO₂. Thus, despite high seed yield in beans grown under elevated CO₂, the harvest index (HI) did not change significantly between CO₂ treatments.
  • Full Text:
  • Date Issued: 1997

Photosynthetic gas exchange responses to light, temperature, carbon dioxide and water stress, and changes in photosynthetic pigments to light and water stress in two cultivars of Hordeum vulgare L

  • Authors: Logie, Malcolme Ronald Ruxton
  • Date: 1992
  • Subjects: Plants -- Photorespiration , Plants -- Transpiration , Botanical chemistry
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4210 , http://hdl.handle.net/10962/d1003779
  • Description: The gas exchange responses of two cultivars of Hordeum vulgare L., to light, temperature, CO₂ and water stress were investigated in the laboratory. The optimum temperature for net CO₂ assimilation was found to be 25°C and 22.5°C for cv. Clipper and cv. Dayan respectively. Net CO₂ assimilation was reduced at 30°C in cv. Dayan. At low light intensity the highest quantum yield efficiency was 0.051 mol.mol⁻¹ at 30°C for cv. Clipper, and 0.066 mol.mol⁻¹ at 20°C for cv. Dayan. At the same temperature, cv. Clipper had a higher water use efficiency than cv. Dayan, but stomatal conductance for cv. Dayan was higher than cv. Clipper. Stomatal limitation to CO₂ was lowest at the optimum temperature for CO₂ assimilation in both cultivars. Stomata limited CO₂ assimilation in cv. Clipper to a larger degree than in cv. Dayan. Relative stomatal limitation for cv. Clipper at 25°C was 0.280 ± 0.010, and for cv. Dayan at 22.5°C was 0.028 ± 0.011. Short-term exposure to elevated CO₂ concentrations increased CO₂ assimilation in both cultivars, but more so for cv. Clipper. Transpiration rate at elevated CO₂ partial pressures were higher in cv. Dayan than in cv. Clipper. At very high CO₂ (860 μmol.m⁻²s⁻¹) partial pressure water use efficiency in cv. Clipper was higher than cv. Dayan, but at low CO₂ partial pressures water use efficiency in cv. Dayan was higher than cv. Clipper. Water stress reduced the relative leaf water content and net CO₂ assimilation in both cultivars. Cultivar Dayan was more tolerant to water stress, and CO₂ assimilation in this cultivar was less affected by water stress. In both cultivars water stress increased the concentration of chlorophyll a, chlorophyll b, and chlorophyll a+b. The chlorophyll a:b ratio remained relatively constant throughout the stress period. No correlation between relative leaf water content and total carotenoid concentration was observed.
  • Full Text:
  • Date Issued: 1992

The interactive effects of light, temperature and CO₂/O₂ ratios in photosynthesis of Coix lachryma-jobi L

  • Authors: Mjwara, Jabulani Michael
  • Date: 1992
  • Subjects: Coix , Plants -- Effect of light on , Plants -- Effect of oxygen on , Plants -- Effect of carbon dioxide on , Photosynthesis , Plants -- Effect of temperature on
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:4216 , http://hdl.handle.net/10962/d1003785
  • Description: A portable infra red gas analyzer was used to investigate the interactive effects of light, temperature, and CO,jO, ratios under controlled environmental conditions in an attempt to model gas exchange characteristics of Coix Iachryma-jobi L. Plotting light response curves as a function of temperature (20, 25 30 and 35°C) revealed no sign of light saturation even at a photosynthetic photon flux density (PPFD) close to 2000 !Lmol m-' sol. High net assimilation rates (A) of approximately 24 !Lmol CO, m"s'! were realized at 30-35°C. Assimilation (A) versus internal CO, partial pressure (C,) curves showed a steep rise with increase in C, but saturated at approximately 150 (JLII-!) and all the results, either in the absence or presence of 0" showed a similar response under all temperature regimes. C. Iachryma-jobi exhibited low CO, compensation points cr ) between 0 and 10 JLlI-! under similar experimental temperatures and either at 0 or 21%0,. The slopes of double reciprocal plots of llA versus llCi, were nearly identical and crossed the yintercept at almost identical points under all 0, concentrations. These data indicate first; that there was no apparent 0, inhibition and second; indicated that the apparent inhIbitor constant (K,) for 0, at the site of carboxylation did not change with increase in [OJ from 0 to 21% oxygen. These observations were further confirmed by results obtained from the analysis of apparent carboxylation efficiency (CE, as defined as the slope of response of A to increasing CO,), as no inhibition of A with increase of [OJ occurred. These characteristics were consistent with typical features of C,photosynthesis. The absence of 0, inhibition and low r values indicated that an efficient CO, concentrating mechanism which eliminates photorespiration exists in C. Iachryma-jobi. At the light microscope level, leaf anatomy exhibited typical C, structure viz. bundle sheath with large chloroplasts and this sheath is further surrounded by a radiate Kranz mesophyU cells. Furthermore the anatomical features suggested that C. wchryma-jobi was an NADP-ME species. Stomatal conductance (g,) to assimilation (g,/A) indicated an increase in A with decrease in g" an essential feature of improving water use efficiency (WUE), but one which drastically reduces CO, diffusion rate. The physical lintitation (stomatal lintitation, t) to CO, diffusion under various [0,] and temperatures, but constant PPFD, did not exhibit statistically significant change in t values at either 0 or 21% a, within each temperature regime, however there was a marked decrease in t as the plant approached its optimum photosynthetic temperature.
  • Full Text:
  • Date Issued: 1992
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