Population structure of phytophthora cinnamomi in South Africa

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Date
1999-01
Authors
Linde, Celeste
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Publisher
University of the Free State
Abstract
English: Research presented in this dissertation describes the population structure of Phytophthora cinnamomi in South Africa. Special emphasis is given to the practical implications of population diversity issues for breeding and selection programs against P. cinnamomi. Levels of gene and genotypic diversity of P. cinnamomi populations, their origin, stability over time, variation in levels of pathogenicity and occurrence of sexual reproduction in vivo and in vitro, are considered. The first chapter represents a literature review on the population biology of P. cinnamomi. Special reference is given to the life-cycle of P. cinnamomi, mechanisms of pathogenicity, and control measures. Available genetic markers useful for population genetic studies of fungal pathogens are described. The origin and maintenance of genetic diversity in fungi, with particular emphasis on P. cinnamomi, is discussed. Research concerning the genetic structure of P. cinnamomi populations is summarised, before the overall and specific aims of the research described in this dissertation are presented. This literature review also emphasises the lack of, or limited number of studies pertaining to the population structure of P. cinnamomi. Chapter two considers levels of genetic diversity as determined in populations of P. cinnamomi in South Africa, using isozymes. Differentiation in regional P. cinnamomi populations, stability over time, and occurrence of sexual reproduction in the South African P. cinnamomi population is discussed. Evidence is presented suggesting that P. cinnamomi is introduced into South Africa and has low levels of gene and genotypic diversity. There is no genetic differentiation between regional P. cinnamomi populations and the populations are stable over time. It is also shown that P. cinnamomi rarely reproduces sexually, if at all. The success of breeding and selection programs against P. cinnamomi is dependant on the levels of variation in pathogenicity of the pathogen. In chapter three, significant differences in levels of pathogenicity to E. smithii in the field, were found. Various factors are shown to affect levels of pathogenicity. These include growth rate in vitro of P. cinnamomi, culture age, season of inoculation, and multi locus isozyme genotype characteristics. On the other hand, mating type and geographic origin of P. cinnamomi isolates did not affect the level of pathogenicity. Significantly more variation was found among A2 than Al mating type isolates, demonstrating the clonality of Al mating type isolates. Results of this study provide valuable information on selection of P. cinnamomi isolates for future resistance/tolerance screening assays, of Eucalyptus germplasm in South Africa. Chapter four deals with levels of gene and genotypic diversity in South African and Australian P. cinnamomi populations, using RAPD and RFLP markers. The high degree of similarity between the South African and Australian P. cinnamomi populations is demonstrated. A low level of genetic differentiation (Dm = 0.003) between the populations provides the most striking evidence of similarity in these two populations. A few isolates from Papua New Guinea, possessed unique alleles not found in the South African or Australian P. cinnamomi populations. The similarity of the South African and Australian P. cinnamomi populations provide opportunities to breed and select for resistance/tolerance in Eucalyptus, using either P. cinnamomi population. Evidence for the occurrence of sexual recombination of P. cinnamomi in vitro and in vivo, is limited. The occurrence of sexual recombination in vitro is investigated in chapter five and the first unambiguous evidence for sexual recombination in P. cinnamomi is provided. This has important consequences for breeding and selection programs, as it could change the genetic structure of the population. Levels of pathogenicity of the F 1 progeny was on average lower than that of parental P. cinnamomi isolates. Other factors concerning fitness of the progeny should be investigated to explain the apparent lack of sexual reproduction in vivo in P. cinnamomi. All chapters of this dissertation deal with some aspects of the population structure and genetic diversity within P. cinnamomi populations from South Africa. This is the first comprehensive study on the genetic diversity of P. cinnamomi populations. It sheds light on the origin, occurrence of sexual reproduction, variation in levels of pathogenicity amongst P. cinnamomi isolates, similarity with the Australian population, and the occurrence of genetic recombination in vitro. It also provides important information for the South African forestry industry, particularly in their efforts to breed and select cold tolerant Eucalyptus spp. that are resistant/tolerant to P. cinnamomi.
Afrikaans: Navorsing in hierdie proefskrif beskryf die populasie struktuur van Phytophthora cinnamomi in Suid Afrika. Spesiale vermelding word gemaak van die praktiese implikasies wat populasie diversiteits aspekte het vir die teling en seleksie programme teen P. cinnamomi. Populasie aspekte wat ondersoek word sluit in, vlakke van geen en genotipiese diversiteit in P. cinnamomi populasies, huloorsprong, stabiliteit, variasie in patogenisiteit, en die voorkoms van geslagtelike voortplanting in vivo en in vitro. Die eerste hoofstuk is 'n literatuur oorsig wat die populasie biologie van P. cinnamomi beskryf. Vermelding word gemaak van die lewenssiklus van P. cinnamomi, patogenisiteits meganismes, and beheer praktyke. Genetiese merkers wat gebruik kan word vir populasie genetika studies van swam patogene word ook beskryf. Verder word die oorsprong en instandhouding van genetiese diversiteit in swamme, en spesifiek in P. cinnamomi, bespreek. 'n Belangrike aspek wat uitgelug word is die klein hoeveelheid studies wat tot dusver bygedra het tot die populasie struktuur van P. cinnamomi. Ten slotte word die navorsing wat betrekking het op die genetiese struktuur van P. cinnamomi populasies saamgevat, en die spesifieke doelwitte van die proefskrif opgesom. Navorsing gedoen in hoofstuk twee beskryf die vlakke van genetiese variasie in Suid Afrikaanse P. cinnamomi populasies soos bepaal met isoensieme. Uit hierdie data blyk dit dat P. cinnamomi nie sy oorsprong in Suid Afrika het nie. Die vlakke van geen en genotipiese variasie is ook baie laag. Daar is nie genetiese differensiasie tussen P. cinnamomi populasies met verskillende geografiese oorspronge nie, en die populasies is stabieloor tyd. Dit word ook gewys dat voortplanting in P. cinnamomi hoofsaaklik ongeslagtig is. Verskille in patogenisiteits vlakke van P. cinnamomi bepaal deels die sukses van teel en seleksie programme daarteen. Resultate van hoofstuk drie wys dat isolate van P. cinnamomi wel betekenisvol verskil in patogenisiteits vlakke op Eucalyptus smithii. Verskeie faktore het die vlakke van patogenisiteit beïnvloed. Dit sluit in groei tempo in vitro van P. cinnamomi, ouderdom van kulture, seisoen van inokulasie, en multigeniese isoensiem eienskappe. Paringstipe en geografiese oorsprong van P. cinnamomi isolate het geen invloed op patogenisiteits vlakke nie. Daar was betekenisvol meer variasie binne A2 as binne Al paringstipe isolate. Dit dui ook op die klonaliteit van Al paringstipe isolate. Hierdie resultate verskaf belangrike inligting ten opsigte van die seleksie van P. cinnamomi isolate vir toekomstige weerstands/toleransie toetse van Eucalyptus in Suid Afrika. Hoofstuk vier handeloor die vlakke van geen en genotipiese diversiteit in die Suid Afrikaanse, vergeleke met 'n Australiaanse P. cinnamomi populasie, soos bepaal met RAPD en RFLP merkers. Beide populasies het lae vlakke van genotipiese diversiteit, plant hoofsaaklik ongeslagtelik voort, het 'n hoë klonale fraksie, en het ooreenstemmende allele. Lae vlakke van genetiese differensiasie (Dm = 0.003) tussen die twee populasies dui ook sterk op die ooreenkoms tussen die twee populasies. Inteenstelling hiermee, het die isolate vanaf Papua New Guinea unieke allele wat nie in die ander populasies voorkom nie. Hoofstuk vyf bied die eerste bewyse vir geslagtelike rekombinasie in vitro in P. cinnamomi. Nege-en-veertig FI hibriede en een self van die Al paringstipe ouer is geïdentifiseer met RAPD's. FI hibriede het gesegregeer in 'n I: I verhouding vir paringstipe. Patogenisiteit van F I hibriede is gemiddeld laer as dié van hulouers. Dit dui daarop dat faktore buiten die verlies in patogenisiteit, ondersoek moet word om die afwesigheid van geslagtelike voortplanting in vitro te verklaar. Geslagtelike rekombinasie het belangrike gevolge vir teel en seleksie programme aangesien dit die genetiese struktuur van die populasie kan verander. Al die hoofstukke in hierdie proefskrif beskryf sekere aspekte van die populasie struktuur van P. cinnamomi in Suid Afrika. Meer inligting is nou beskikbaar aangaande die oorsprong van P. cinnamomi, geslagtelike voortplanting, variasie in vlakke van patogenisiteit tussen P. cinnamomi isolate, ooreenkoms met die Australiaanse populasie, en die moontlikheid van geslagtelike rekombinasie. Dit bied ook belangrike inligting vir die Suid Afrikaanse bosbou industrie met betrekking tot die teel en seleksie van koue tolerante Eucalyptus spp. wat weerstandbiedend is teen P. cinnamomi.
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Soilborne plant pathogens -- South Africa, Fungal populations -- South Africa, Thesis (Ph.D (Microbiology and Biochemistry))--University of the Free State, 1999
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