Diuraphis noxia biotypes ascribed through genotypic characterization and utilization of their endosymbiont, buchnera aphidicola

Date
2022-11
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Stellenbosch : Stellenbosch University
Abstract
ENGLISH ABSTRACT: The aphid Diuraphis noxia (more commonly known as the Russian wheat aphid), is a pest of wheat and other small grains which has successfully spread to all global wheat producing regions. Feeding damage of the aphid results in characteristic longitudinal streaking, leaf rolling, and head trapping which all contribute to a lowered crop yield. Although wheat cultivars resistant to the aphid have been successful in curbing yield losses, the aphid has overcome most of these resistant lines through the process of biotypification. As reproduction of the aphid is limited to cyclical parthenogenesis, and thus unable to generate genetic diversity through chromosomal crossover, the exact means of how biotypification occurs remains elusive. With the availability of the only known genealogically linked D. noxia biotypes SA1 and SAM, this study attempted to ascertain their genomic backgrounds to better understand how they could occupy the extremes of the virulence scale. Sequencing, assembly, and comparisons of the genomes of these biotypes revealed an unexpected number of genetic polymorphisms and an extremely AT rich genome. The roughly 400 Mb genome, containing just over 32,000 protein coding genes, appears to have many polymorphisms in genes responsible for protein expression and turnover, which supports previous results indicating that biotypes differ in their ability to regulate their transcription. The unexpected number of polymorphisms would indicate that there is a yet unknown mechanism that enables these aphids to induce genomic variation in the absence of sexual reproduction. Further analysis of the salivary protein genes shared between the biotypes indicates that copy number and polymorphisms in genes associated with defence against plant volatiles also play a role in how these biotypes interact with their wheat host. Through comparative transcriptomics with other aphid transcriptomes, this study was also able to identify genes preferentially expressed in aphid head and salivary tissue through a proof-of-concept comparison, thereby enabling the future discovery of D. noxia salivary effectors. Lastly, this study was able to assemble and compare the genome of the sole endosymbiont of D. noxia, Buchnera aphidicola, with that of nine other genomes. From this the overall importance of several genic features such as gene length and percentage GC content was found to be critical for the maintenance of Buchnera genes when compared to their closest free-living relative, Escherichia coli. It was also revealed that D. noxia contains multiple Buchnera isotypes and that their genes are polyadenylated which may indicate a mechanism for their regulation. With the availability of these datasets, the as of yet unanswered question of the process of biotypification within D. noxia is becoming clearer.
AFRIKAANSE OPSOMMING:Die plantluis Diuraphis noxia (meer algemeen bekend as die Russiese koringluis), is 'n plaag van koring en ander klein grane wat suksesvol na alle koringproduserende streke wêreldwyd versprei het. Voedingskade van die plantluis lei tot kenmerkende longitudinale strepe, blaarrol en kopvanging wat alles bydra tot 'n verlaagde oesopbrengs. Alhoewel koringkultivars wat teen die plantluis bestand is suksesvol was in die verlede om opbrengsverliese te bekamp, het die plantluis die meeste van hierdie weerstandbiedende lyne deur die proses van biotipifisering oorkom. Aangesien voortplanting van die plantluis beperk is tot sikliese partenogenese, en dus nie in staat is om genetiese diversiteit deur chromosomale oorkruising te genereer nie, bly die presiese manier van hoe biotipifisering plaasvind ontwykend. Met die beskikbaarheid van die enigste bekende genealogies gekoppelde D. noxia biotipes SA1 en SAM, het hierdie studie gepoog om hul genomiese agtergronde beter te verstaan en hoe hulle die uiterstes van die virulensieskaal kon beset. Volgordebepaling, samestelling en vergelykings van die genome van hierdie biotipes het 'n onverwagte aantal genetiese polimorfismes en 'n uiters AT-ryke genoom aan die lig gebring. Die ongeveer 400 Mb-genoom, wat net meer as 32 000 proteïenkoderende gene bevat, blyk om baie polimorfismes te hê in gene wat verantwoordelik is vir proteïenuitdrukking en -omset, wat vorige resultate ondersteun wat aandui dat biotipes verskil in hul vermoë om hul transkripsie te reguleer. Die onverwagte aantal polimorfismes sou aandui dat daar 'n nog onbekende meganisme is wat hierdie plantluise in staat stel om genomiese variasie in die afwesigheid van seksuele voortplanting te veroorsaak. Verdere ontleding van die speekselproteïengene wat tussen die biotipes gedeel word, dui daarop dat kopiegetal en polimorfismes in gene wat verband hou met verdediging teen plantvlugtige stowwe ook 'n rol speel in hoe hierdie biotipes met hul koringgasheer in wisselwerking tree. Deur vergelykende transkriptomika met ander plantluis-transkriptome, was hierdie studie ook in staat om gene te identifiseer wat verkieslik in plantluiskop en speekselweefsel uitgedruk word deur 'n bewys-van-konsep-vergelyking, waardeur die toekomstige ontdekking van D. noxia speekseleffektore moontlik gemaak word. Laastens kon hierdie studie die genoom van die enigste endosymbiont van D. noxia, Buchnera aphidicola, saamstel en vergelyk met dié van nege ander genome. Hieruit is gevind dat die algehele belangrikheid van verskeie genekenmerke soos geenlengte en persentasie GC-inhoud krities is vir die instandhouding van Buchnera-gene in vergelyking met hul naaste vrylewende familielid, Escherichia coli. Dit is ook aan die lig gebring dat D. noxia veelvuldige Buchnera isotipes bevat en dat hul gene gepoliadenileerd is wat 'n moontlike meganisme vir hul regulering kan aandui. Met die beskikbaarheid van hierdie datastelle word die nog onbeantwoorde vraag van die proses van biotipifisering binne D. noxia duideliker.
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Thesis (PhD)--Stellenbosch University, 2022.
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