Involvement of lipoxygenase in resistance of wheat against the Russian wheat aphid

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Date
1999
Authors
Swart, Elsabe
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Publisher
University of the Free State
Abstract
English: The upstream role of lipoxygenase (linoleate: oxygen reductase, E.C. 1.13.11.12) in the induced defence response against the Russian wheat aphid (Diuraphis noxia) was studied in resistant wheat (Triticum aestivum L.) cultivars Tugela-DN (Dn1), Tugela (Dn2) and Tugela (Dn5) and the near isogenie susceptible cultivar, Tugela. The RWA induced LOX was partially characterised. The pH for optimum LOX activity was 5.6 and it had no requirement for cofactors such as K+, Mg2+ or Ca2- The enzyme also had a high affinity for linoleic acid, with a Km-value of -5.8 x 10-4M. LOX activity was selectively induced in infested resistant Tugela cultivars containing the (Dn1), (Dn2) and, to a lesser extent, (Dn5) resistant genes. The differential induction of LOX activity corresponded with the induction of the hypersensitive response during the resistance response of Tugela-DN against the RWA. The incorporated resistant gene affected the level of induced LOX activity, which roughly corresponded with the cultivars' levels of resistance. Localised RW A infestation led to a systemic increase in LOX activity. All indications were that the induced LOX activity was involved in the resistance response in wheat against the RWA. Inhibitory studies were performed with LOX inhibitors, piroxicam, salicylhydroxamic acid and n-propyl gallate, to shed light on the possible upstream role of LOX. Piroxicam was the most suitable inhibitor due to the fact that it only inhibited LOX activity in vitro. Inhibition of LOX activity in vivo by piroxicam, led to the inhibition of 'downstream' defence responses such as peroxidase and β-l ,3-glucanase activities. The selective increase in LOX activity did not correlate with a selective increase in jasmonic acid concentrations. The effect of intercellular injected JA on LOX, peroxidase and β-1 ,3- glucanase activities, indicated that jasmonic acid was not the primary product of the RWA induced LOX activity and that it was not the signal molecule in the RWA /wheat resistance response. The products of the RWA induced LOXs appeared to be derived from the 9-hydroperoxide derivative pathway, rather than the 13- hydroperoxide derivative (jasrnonic acid precursor) pathway. Neither was abscicic acid a product of the selective increased LOX activity, nor playing a role in the RWA resistance response in wheat. Other studies in our laboratory showed that salicylic acid is probably the signal molecule, activating defence genes after elicitation. It is evident that the RWA resistance response corresponded with that of pathogenesis, and that each defence response follows its own distinctive defence cascade.
Afrikaans: Die 'stroom-op' rol van lipoksigenase (linoleaat: suurstofreduktase, E.C. 1.13.11.12) in die geïnduseerde weerstandsreaksie teen die Russiese koringluis (Diuraphis noxia) was in weerstandbiedende koring (Triticum aestivum L.) kultivars, Tugela-DN (Dn1), Tugela (Dn2) en Tugela (Dn5) en die naby-genetiese vatbare kultivar, Tugela ondersoek. Gedeeltelike karakterisering van die Russiese koringluis (RKL) geïnduseerde lipoksigenase het getoon dat die optimum pH vir maksimale lipoksigenase-aktiwiteit, 5.6 was. Die geïnduseerde lipoksigenase het geen behoefte aan ko-faktore soos Kt., Mg2+ en Ca2+ gehad nie en dit het 'n hoë affiniteit vir linoleïensuur getoon, met 'n Km-waarde van -5.8 x 10-4M. Lipoksigenase-aktiwiteit was selektief geïnduseer in geïnfesteerde weerstandbiedende Tugela kultivars bevattende weerstandsgene (Dn1), (Dn2) en, tot 'n mindere mate, (Dn5). Die differensiële induksie van lipoksigenase-aktiwiteit het met die induksie van die hipersensitiewe-reaksie in weerstandbiedende koring Tugela-DN tydens RKL infestering gekorrelleer. Die geïnkorporeerde weerstandsgene het die vlak van geïnduseerde lipoksigenase-aktiwiteit beïnvloed. Die lipoksigenase-aktiwiteitsvlak het ook rofweg met die weerstandbiedendheid van die kultivars ooreengestem. Gelokaliseerde RKL infestering het tot die lokale en sistematiese induksie van lipoksigenase-aktiwiteit gelei. Alles dui daarop dat die geïnduseerde lipoksigenaseaktiwiteit in die weerstandsrespons teen die RKL in koring betrokke is. Inhibisieondersoeke was met behulp van lipoksigenase-remstowwe, piroksikam, salisielhidroksaamsuur en n-propielgallaat gedoen, om lipoksigenase se 'stroom-op' rol tydens die RKL-weerstandsreaksie te bepaal. Piroksikam was die geskikste remstof, aangesien dit alleenlik lipoksigenase-aktiwiteit in vitro gerem het. Die in vivo remming van lipoksigenase-aktiwiteit, het gelei tot die 'stroom-af remming van peroksidase- en β-I,3-glukanase-aktiwiteite. Die selektiewe toename In lipoksigenase-aktiwiteit het nie met die selektiewe toename in jasmoonsuurkonsentrasies in vivo gepaard gegaan nie. Die aktiverende invloed van jasmoonsuur, intersellulêr toegedien, op lipoksigenase-, peroksidase- en β P-I,3-glukanase aktiwiteite, bevestig dat jasmoonsuur me die produk van die RKL geïnduseerde lipoksigenase-aktiwiteit was nie, en dat dit nie opgetree het as die seinmolekuul tydens die RKL-weerstandsreaksie in die koring nie. Die RKL geïnduseerde lipoksigenase-aktiwiteit se produkte blyk vanaf die 9-hidroperoksied-derivaat gesintetiseer te wees, en nie vanaf die 13-hidroperoksiedderivaat (jasmoonsuur voorloper) nie. Soortgelyk was absisiensuur (ABA) ook nie 'n produk van die selektief geïnduseerde lipoksigenase-aktiwiteit nie, en het dit ook nie 'n rol in die RKL-weerstandsreaksie in koring gespeel nie. Ander studies in ons laboratorium het getoon dat salisielsuur moontlik die seinmolekuul is wat verdedigingsgene aktive~r na elisitering. Dit is duidelik dat die RKL-weerstandsrespons ooreenstem met die van patogenese, en dat elke verdedigingsreaksie sy eie unieke verdedigings-sein-kaskade meganisme volg.
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Keywords
Wheat (Triticum aestivum L.), Russian wheat aphid (Diuraphis noxia), Lipoxygenase, Resistance, Resistance genes, Systemic, Inhibitors, Signalling, Jasmonic acid, Abscisic acid, Salicylic acid, Wheat -- Disease and pest resistance, Dissertation (M.Sc. (Botany and Genetics))--University of the Free State, 1999
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