The effects of phosphorus (P) deficiency on growth and nitrogen fixation of Virgilia trees from the Cape Floristic Region (CFR)

Magadlela, Anathi (2013-03)

Thesis (MSc)--Stellenbosch University, 2013.

Thesis

ENGLISH ABSTRACT: The aim of this study was to determine how P nutrition affects biological nitrogen fixation (BNF) via effects on the N2-fixing bacteria in the nodules of Virgilia species native to the Cape Floristic Region (CFR), South Africa. This was evaluated in 3 separate studies: The first study aimed to determine how phosphorus deficiency affects N nutrition of two legume tree species from the Mediterranean Fynbos ecosystem. This study showed that during prolonged P deficiency, V. divaricata maintained a constant biomass, while V. oroboides showed a decreased biomass. V. oroboides showed a decrease in nutritional concentrations, which resulted in the decrease of symbiotic nitrogen fixation (SNF). Both plants utilized atmospheric N more efficiently per nodule under P deficiency. Maximum photosynthesis decreased in V. oroboides, while V. divaricata maintained its photosynthesis. Both species also had greater carbon construction costs during P deficiency. V. divaricata showed clear adaptive features during P-deficiency, as it maintained its growth respiration. The two legume species appear to have different adaptations to P deficiency, which may influence their performance and distribution in their naturally low P environment. The second study aimed to evaluate if soil environmental conditions and mineral nutrient concentration play a major role in microbial communities in plant rhizosphere and nodulation during N2 fixation in legumes. Therefore this study firstly aimed to determine the composition of the N2 fixing bacterial population in the rhizosphere and nodules of V. divaricata. Secondly, it aimed to determine the contribution of these bacteria to N2 fixation during conditions of P deficiency in the Fynbos environment. In the study, the effects of phosphate (P) nutrition on N2 fixing bacterial community structures in Virgilia divaricata rhizosphere and nodules were examined in a pot experiment. V. divaricata were germinated in Fynbos soil as natural inoculum, transferred to clean sand cultures and supplied with 500 μM P and 5 μM P. The N2 fixing bacterial communities in the rhizosphere and nodules were examined based on the PCR-DGGE banding patterns of 16S rDNA and sequencing methods. The GenBank blast results revealed that V divaricata was efficiently nodulated by a wide range of root-nodule bacterial strains, including Burkholderia phytofirmans, Burkholderia sp. and Bradyrhizobium sp. during low P supply. The 15N natural abundance data also confirmed that 40-50% of the N nutrition was acquired through symbiotic N2 fixation. This is not only evidence of nodulation, but also an indication of the adaptation of a range of N2 fixing bacterial strains / species to the nutrient poor, sandy, acidic soil of the Mediterranean-type ecosystems of the Fynbos. The third study examined the physiological effects, such as N2 fixation parameters, plant dependence on N2 fixation, N preference, legume plant growth, carbon costs and amino acid biosynthesis during P deficiency and mineral N supply as NH4NO3 in a slow-growing, Fynbos legume tree, Virgilia divaricata. Continued application of NH4NO3 to the legume plant showed a greater increase in plant dry matter compared to plants with two nitrogen sources (mineral N and atmospheric N2) or plants that relied on atmospheric N2 fixation. Carbon construction costs were more pronounced in plants supplied with two N sources and during P deficiency. Maximum photosynthetic rates per leaf area increased during prolonged P deficiency, irrespective of the N sources. Though plants nodulated, plant dependence on N2 fixation decreased with the addition of NH4NO3. Roots and nodules of the P deficient plants showed an increase in asparagine content, most strikingly so in plants treated with a single source of N. These studies reveal that different legume species of the same genus, may employ contrasting adaptations in order to maintain N nutrition under P deficiency.

AFRIKAANSE OPSOMMING: Die doel van hierdie studie was die bepaling van die wyse waarop fosfaat (P) voeding die biologiese stikstof binding (BNF) deur middel van die effek op N2-bindingsbakterië in die wortelknoppies van Virgilia spesies wat inheems tot die Kaap floraryke area (CFR), Suid Afrika is, affekteer. Drie aparte eksperimente is uitgevoer om die doel te evalueer: Die eerste studie het gepoog om te bepaal hoe 'n fosfaat tekort N voeding van twee peulplant spesies van die Mediterreense Fynbos ekosisteem affekteer. Hierdie studie het getoon dat V. divaricata 'n konstante biomassa tydens verlengde P tekort behou, terwyl V. oroboides ‟n verlaagde biomassa getoon het. V. oroboides het 'n verlaging in voedingskonsentrasies getoon, wat tot 'n verlaging in simbiotiese stikstof binding (SNF) gelei het. Beide plante benut atmosferiese N meer doeltreffend per nodule tydens P tekort. Die maksimum fotosintese in V. oroboides het afgeneem, terwyl V. divaricata sy fotosintese gehandhaaf het. Beide spesies het ook 'n groter koolstof konstruksie koste tydens P tekort gehad. V. divaricata toon duidelike aanpassingsmeganismes tydens P-tekort, aangesien hierdie species sy groei respirasie konhandhaaf. Dit wil voorkom asof die twee peulplant spesies verskillend aangepas is vir P tekort, wat hulle producksie en verspreiding in hulle natuurlike lae P omgewing mag beïnvloed. Die doel van die tweede studie was om te bepaal of grond omgewingskondisies en minerale voedingskonsentrasie 'n belangrike rol speel in die mikrobiese gemeenskappe in die plant risofeer en wortelknoppie vorming tydens N2 binding in peulgewasse. Eerstens het die studie dus gepoog om die samestelling van die N2 bindende bakteriële populasie in die risosfeer en die wortelknoppies van V. divaricata te bepaal. Ten tweede, is die bydrae van die bakterië tot N2-binding tydens P tekort kondisies in die Fynbos omgewing bepaal. In die studie is die effek van fosfaat (P) voeding op die N2-bindende bakteriële gemeenskapstrukture in die Virgilia divaricata risofeer en wortelknoppies in 'n pot eksperiment ondersoek. V. divaricata sade is in fynbos grond as 'n natuurlike inokulum ontkiem, waarna dit na skoon sand kulture oorgedra is en van 500 μM P en 5 μM P voorsien is. Die N2-bindende bakteriële gemeenskappe in die risofeer en wortelknoppies is op grond van die PCR-DGGE band patrone van die 16S rDNA en volgorde bepalingsmetodes ondersoek. Die GenBank Blast resultate het getoon dat V. divaricata doeltreffend deur 'n wye reeks wortel-wortelknoppie bakteriële stamme genoduleer is, insluitende insluitende Burkholderia phytofirmans, Burkholderia sp. en Bradyrhizobium sp. tydens lae P toediening. Die natuurlike 15N voorkoms data het ook bevestig dat 40-50% van die N voeding deur simbiotiese N2 binding bekom is. Dit dien nie net as bewys vir wortelknoppie vorming nie, maar ook 'n aanduiding van die aanpassing van 'n reeks N2 bindende bakteriële stamme/ spesies tot die voedingsarme, sanderige, suur grond van die Mediterreanse ekosisteem van die Fynbos. Die derde studie het die fisiologiese effekte soos bv. N2 fikserings faktore, die afhanklikheid van plante op N2 fiksering, N voorkeur, peulgewas groei, koolstof kostes en aminosuur biosintese tydens P tekort en minerale N toediening soos NH4NO3 in 'n stadig-groeiende, Fynbos peulgewasboom, Virgilia divaricata ondersoek. Volgehoue toediening van NH4NO3 aan die peulplant toon 'n groter toename in plant droë weefsel. Tydens P tekort is die koolstof bou koste meer verhoog in plante wat met twee N bronne voorsien is. Tydens verlengde P tekort het die maksimum fotosintese tempo per blaaroppervlakte toegeneem, ongeag die N bron. Alhoewel die plante wortelknoppies gevorm het, het die plant se afhanklikheid van N2 binding tydens die toediening van NH4NO3 afgeneem. Wortels en wortelknoppies van die P tekort plante het 'n toename in asparagien inhoud getoon, veral in die plante wat met 'n enkele bron van N behandel is.

Please refer to this item in SUNScholar by using the following persistent URL: http://hdl.handle.net/10019.1/79992
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